Nuts and Seeds in Health and Disease Prevention [2 ed.] 0128185538, 9780128185537

Table of contents :
Cover
Nuts and Seeds in Health and Disease Prevention
Copyright
Contributors
Preface
Acknowledgments
Sec 1
1 - Rambutan (Nephelium lappaceum L.) Seed and Its Fat
Introduction
Botanical Descriptions
Historical Cultivation and Usage
Present-Day Cultivation and Usage
Applications in Health Promotion and Disease Prevention
Nephelium lappaceum Seed Fat (RSF) Characteristics, Potential Food, and Therapeutic Uses
RSF Physical and Chemical Properties
RSF Phase Behavior
RSF and Its Potential Food Uses for Health Promotion
Adverse Effects and Reactions, Allergies, and Toxicity
Summary Points
References
2 - Soursop Seed: Soursop (Annona muricata L.) Seed, Therapeutic, and Possible Food Potential
Introduction
Botanical Descriptions
Historical Cultivation and Usage
Present-day Cultivation and Usage
Applications in Health Promotion and Disease Prevention
Annona muricata L. Seed and Its Therapeutic Potential
Annona muricata Seed and Its Possible Potential for Food Use
Adverse Effects and Reactions, Allergies, and Toxicity
Summary Points
References
3 - Red Horse-Chestnut Seeds of Aesculus × Carnea: A New Way for Health and Food Design?
Introduction
Botanical Descriptions
Historical Cultivation and Usage
Present-Day Cultivation and Usage
Red Horse-Chestnuts: Chemical Composition and Characterization
Proximate Analysis
Lipids Analysis
Glucides Analysis
Scanning Electron Microscopy
Applications to Health Promotion and Disease Prevention
Adverse Effects and Reactions (Allergies and Toxicity)
Summary Points
Acknowledgments
References
4 - The African Breadfruit (Treculia africana) Decne Plant Seed: A Potential Source of Essential Food and Medicinal Phytoconsti ...
Introduction
Botanical Description, Cultivation, and Usage
Fruit Processing and Seed Production
Nutritional Value and Food Uses of Treculia africana Seed
Antinutritional Compositions of African Breadfruit Seed
Applications to Health Promotion and Disease Prevention
Future Perspectives
Acknowledgments
References
5 - Perennial Horse Gram (Macrotyloma axillare) Seeds: Biotechnology Applications of Its Peptide and Protein Content – Bowman–B ...
Introduction
Botanical Description
Historical Cultivation and Usage
Present-Day Cultivation and Usage
Applications to Health Promotion and Disease Prevention – Biotechnology Applications of the Protein Seed Content
Macrotyloma axillare Seed Lectin (MaL)
The Bowman–Birk Inhibitors from M. axillare – Biochemistry and Particularities of BBI
Main Biological Activities Related to Bowman–Birk Inhibitors
Bowman–Birk Inhibitors from Macrotyloma axillare and Its Anticarcinogenic Effect on Colorectal Cancer
Macrotyloma axillare Seed Germination and Bowman–Birk Inhibitors
Adverse Effects and Reactions – Toxicity of Macrotyloma axillare
Summary Points
References
6 - Biological Properties of a Partially Purified Component of Neem Oil: An Updated and Revised Work
Introduction
State of the Art in Our Laboratory at the First Edition of the Book
Neem Products: an Update
References
7 - Bioactive Compounds of Oregano Seeds
Introduction
Botanical Description
Historical Cultivation and Use
Current Cultivation and Use
Applications for Health Promotion and Disease Prevention
Adverse Effects and Reactions, Allergies, and Toxicity
Summary Points
References
8 - Mango Seed: Mango (Mangifera indica L.) Seed and Its Fats
Introduction
Botanical Description
Historical Cultivation and Usage
Present-Day Cultivation and Usage
Applications to Health Promotion and Disease Prevention
The Chemical Composition and Lipids of the Mango Seed Kernel
Mango Kernel Fat Composition
Mango Kernel Fat Thermal Behavior and Polymorphism
Current Situation and Direction on Mango Kernel Fat Researches
Adverse Effects and Reactions, Allergies, and Toxicity
Summary Points
References
Sec 2
9 - Biological Functions of Soyasaponins: The Potential Use to Improve Zinc Nutrition
Introduction
Zinc Nutrition and Health
Zinc Absorption in the Small Intestine and Zinc Transporter, ZIP4
Soybean Soyasaponin Bb Increases ZIP4 Abundance at the Apical Membrane
Conclusion
Acknowledgment
References
10 - Purple Wheat (Triticum sp.) Seeds: Phenolic Composition and Antioxidant Properties
Introduction
Botanical Description
Historical Cultivation of Purple Wheat
Phenolic Compounds in Purple Wheat Seeds
Chemical Nature of Phenolic Compounds Identified in Purple Wheat
Extraction of Phenolic Compounds
Methods of Analyses for Phenolic Contents and Phenolic Composition of Purple Wheat Seeds
Analysis of Total Phenolic Content
Analysis of Total Anthocyanins Content
Analysis of Total Flavonoids Content
Analysis of Total Condensed Tannins Content
Analysis of Constituent Phenolic Compounds
Total Phenolic Content and Phenolic Acid Composition
Total Phenolic Content
Phenolic Acid Composition
Total Anthocyanin Content and Anthocyanin Composition
Total Flavonoid Content and Proanthocyanidin Composition
Processing and Utilization of Purple Wheat
Applications to Health Promotion and Disease Prevention
Brief Overview of Oxidative Stress and the Role of Exogenous Sources of Antioxidants
Antioxidant Properties of Purple Wheat Extracts
Adverse Effects and Reactions (Allergies and Toxicity)
Summary Points and Future Perspectives
References
11 - Protective Role of Nigella sativa and Thymoquinone in Oxidative Stress: A Review
Introduction
Free Radicals and Antioxidant Defense
Chemical Constituents
Traditional Medicine
Pharmacological Properties
In vitro Antioxidant Activity of Nigella sativa
In vivo Antioxidant Activity of Nigella sativa
Acetaminophen-Induced Hepatotoxicity
Aging
Arthritis
Carcinogenes
Cardiotoxicity
Chemotherapeutic Agent
Diabetes Mellitus
Encephalomyelitis
Ethanol Toxicity
Fatty Liver
Haematotoxicity
HCV-Related Fibrosis
Hypercholesterolemia
Ischemic/Reperfusion Injury
Morphine-Induced Oxidative Stress
Nephrotoxicity
Neurotoxicity
Plasmodium yoelii Infection
Radiotherapy
Schistosomiasis
Seizure
Subarachnoid Hemorrhage
Ulcerative Colitis
Conclusion
References
12 - Black Soybean Seed: Black Soybean Seed Antioxidant Capacity
Introduction
History, cultivation, and use
Anthocyanin
Antioxidant activity of black soybeans and black soybean–based food products
Health-promoting and disease-preventing effects of black soybean seed
Atherosclerosis and coronary heart disease
Obesity and diabetes
Inflammation and cancer
Summary Points
References
13. - Fenugreek (Trigonella foenum) Seeds in Health and Nutrition
Introduction
Scientific Classification
Morphology of Seed
Earlier Cultivation of Fenugreek Seed
Current Cultivation
Phytochemical Constituents
Therapeutic Potential of Fenugreekenugreek Seed
Treatment of Diabetes
In Cancer Therapy
Fenugreek As Antioxidant
Fenugreek Results in Cholesterol
Anthelmintic
Fenugreek in Bactericide Activity
Fenugreek in Obesity
Fenugreek in Gastroprotection
Fenugreek’s Influence on Digestion
Fenugreek in Inflammation
Fenugreek in Cardiovascular Disease
Adverse Effects
Conclusion
References
14 - Tamarind (Tamarindus indica) Seeds in Health and Nutrition
Introduction
Botanical Description
Vernacular Names
Taxonomical Classification
Historical Cultivation and Usage
Present-Day Cultivation and Usage
Nutritional Characterization of Tamarind Seeds
Phytochemicals Composition of Tamarind Seed
Biological Activities
Tamarind Seed Polysaccharide: a Promising Natural Excipient for Pharmaceuticals
Possible Adverse Effects and Reaction(s)
Summary of Key Point(s)
References
15 - Sesame Seed in Controlling Human Health and Nutrition
Introduction
Plant Profile
Vernacular Names
Geographical Distribution
Morphology
Chemical Composition
Lipids Content
Fatty Acid Composition
Endogenous Antioxidants
Proteins
Carbohydrates
Minerals
Pharmacological Applications
Wound Healing Activity
Hepatoprotective Activity
Effects of Pinoresinol on Memory Synaptic Plasticity
Effect of Sesame Oil and Sesamol on Heavy Metal Toxicity
Autoimmune Encephalomyelitis
Atherosclerosis
Effect on Serum Cholesterol and in Hypercholesterolemia
Effects on Plasma Tocopherol Levels
Antioxidant and Anticancer Effect of Sesamol
Anticancer Effect of Sesamin
Antihypertensive Effect of Sesamin
Antihypertensive Effect of Sesame Oil
Metabolism of Sesamin
Effect of Sesame Oil during Endotoxemia
Sesame Oil Protects Against Lead-Plus-Lipopolysaccharide-Induced Acute Hepatic Injury
Effect of Sesame Oil on Human Colon Cancer
Antiaging Effect of Sesamol
Modulating Effect of Sesamin
Allergy to Sesame in Humans
Conclusions
References
16 - Kancolla Seeds: High Nutritional Foods With Nutraceutical Properties
Introduction
Nutritional Values of Kancolla Seeds
Proximate Composition
Amino Acid Composition
Dietary Fiber Analysis
Total Carotenoid Content
Total Ascorbic Acid Content
Anion Analysis
Oxalates
Nitrates
Phosphates
Mineral Analysis
Free Sugar Analysis
Phytochemical Composition of Kancolla Seeds
Phytoecdysteroids in Kancolla Seeds
Triterpenoid Saponins
Glycine Betaine, Trigonelline, and Trigonelline Metabolites in Kancolla Seeds
Quaternary Pyridinium Salts in Kancolla Seeds
Polyphenols in Kancolla Seeds
The Antioxidant Activity of Kancolla Seeds Before and After Boiling
References
17 - Health-promoting Potential and Nutritional Value of Madhuca longifolia Seeds
Introduction
Botanical Description and Cultivation
Composition of M. longifolia Seeds and Seed Cake
M. longifolia Butter Content and Composition
Applications of M. longifolia Butter
Applications of M. longifolia Seeds in Health Promotion and Disease Prevention
Antidiabetic and Antihyperglycemic
Anti-inflammatory
Anti-ulcer Activity
Antifertility Activity
Adverse Effects (Allergies and Toxicity)
Conclusion
References
Sec 3
18 - Ginkgo biloba Seeds: Antifungal and Lipid Transfer Proteins from Ginkgo biloba Nuts
Introduction
Lipid Transfer Proteins
Antifungal Proteins
Outlook
References
19 - Mycotoxins in Nuts and Seeds
Introduction
Natural Occurrence
Effect of Heat Processing
Toxicological Effects in Humans
Summary Points
Acknowledgments
References
Sec 4
20 - Lepidium sativum Seeds: Therapeutic Significance and Health-Promoting Potential
Introduction
Botanical Description and Cultivation
Chemical Composition of Lepidium sativum Seeds
Amino Acids Profile
Seed Oil Composition
Edible Applications of Lepidium sativum
Applications of Lepidium sativum to Health Promotion and Disease Prevention
Application As An Excipient in Pharmaceutical Dosage Form
Application As Suspending Agent
Application As Superdisintegrant
Application in Controlled Release System
Therapeutic Applications
Antioxidant Activity
Antimicrobial Potential
Antidiabetic and Hypoglycemic Properties
Anti-inflammation Activity
Antidiarrheal, Antihypertensive, and Diuretic Activities
Prokinetic and Laxative Activities
As Galactogogue and Emmenagogue
As Hematic Agent
Antiasthmatic and Estrogenic Activities
Anticancer Activities
Treatment of Fracture Healing
Adverse Effects (Allergies and Toxicity)
Conclusions
References
21 - The Effects of Nuts on Metabolic Diseases and Disorders
Introduction
Fats
Macrominerals and Micromineral
Phenolic Compounds
Discussion
References
22 - Tea (Camellia oleifera) Seeds: Use of Tea Seeds in Human Health
Introduction
Botanical Description
Historical Cultivation and Usage
Present-day Cultivation and Usage
Applications to Health Promotion and Disease Prevention
Adverse Effects and Reactions (Allergies and Toxicity)
Summary Points
References
23 - Effect of Nigella sativa on Blood Diseases: A Review
Introduction
Effect of Nigella sativa on Hematological Parameters
Induction of Anemia by Chemical Compounds
1,2-Dimethylhydrazine
Cadmium
Chloramphenicol
Dichlorvos
Cisplatin
Streptozotocin
Carbon Tetrachloride
Lead Acetate
Diethylnitrosamine
Phenylhydrazine
Effect of Nigella sativa and Thymoquinone on Platelet Aggregation
Effect of Nigella sativa on Coagulation Parameters
Clinical Studies
Effect of Nigella sativa Seeds and Total Oil in Healthy Women
Effect of Nigella sativa Oil on Lipid Profile in Healthy Volunteers
Effect of Nigella sativa Oil in Obese Women
Administration of Nigella sativa in Diabetic Type 2 Patients
Effect of Nigella sativa on Sickle Cells in vitro
Effect of Nigella sativa on Allergic Patients
Effects of Nigella sativa on Hepatitis C in Egypt
Nigella sativa in Bone Turnover
Conclusion
References
24 - Dermatological Effects of Nigella sativa and Its Constituent, Thymoquinone: A Review
Introduction
Methods
Anti-inflammatory and Immunomodulatory Properties of Nigella sativa and Its Constituent, TQ, Used to Treat Skin Ailments
In vitro and in Vivo Preclinical Studies
Clinical Studies
Anticancer
In vitro Studies
In vivo Studies
Wound Healing Effects
In vivo Studies
In vitro Studies
Antimicrobial Properties Against Skin Relevant Pathogens
Antibacterial
In vitro Studies
In vivo Studies
Antifungal Effects
Antiparasitic Properties
Preclinical in Vitro Studies
Preclinical in Vivo Studies
Clinical Study
Antiviral Effects
Effective in Vitiligo Skin disease
Cosmeceutical Applications
Conclusion
References
25 - Indian Mustard (Brassica juncea L.) Seeds in Health
Introduction
Botanical Descriptions
Historical Cultivation and Usage
Present-Day Cultivation and Usage
Applications to Health Promotion and Disease Prevention
Adverse Effects and Reactions (Allergies and Toxicity)
Summary Points
References
26 - Potential Role of Seeds From India in Diabetes
Introduction
Seeds from Medicinal Plants and Their Role in Diabetes
Abelmoschus moschatus
Geographical Source
Phytochemistry
Abelmoschus moschatus and Diabetes
Abutilon indicum
Geographical Source
Phytochemistry
Traditional Uses
Abutilon indicum and Diabetes
Toxicity Studies
Achyranthes aspera Linn.
Geographical Source
Phytochemistry
Traditional Uses
Achyranthes aspera and Diabetes
Adenanthera pavonina
Geographical Source
Phytochemistry
Traditional Uses
Adenanthera pavonina and Diabetes
Toxicity Studies
Annona muricata Linn.
Geographical Source
Phytochemistry
Traditional Uses
Annona muricata Linn. and Diabetes
Azadirachta indica A. Juss
Geographical Source
Phytochemistry
Traditional Uses
Azadirachta indica A. Juss and Diabetes
Toxicity Studies
Carica papaya Linn.
Geographical Source
Phytochemistry
Traditional Uses
Carica papaya Linn. and Diabetes
Toxicity Studies
Celosia argentea
Geographical Source
Phytochemistry
Traditional Uses
Celosia argentea and Diabetes
Cicer arietinum Linn.
Geographical Source
Phytochemistry
Traditional Uses
Cicer arietinum Linn. and Diabetes
Toxicity Studies
Cyamopsis tetragonoloba Taub
Geographical Source
Phytochemistry
Traditional Uses
Cyamopsis tetragonoloba Taub and Diabetes
Toxicity Studies
Euryale ferox Salisb.
Geographical Source
Phytochemistry
Traditional Uses
Euryale ferox Salisb. and Diabetes
Strychnos potatorum Linn.
Geographical Source
Phytochemistry
Traditional Uses
Strychnos potatorum Linn. and Diabetes
Toxicity Studies
Syzygium cumini Skeels
Geographical Source
Phytochemistry
Traditional Uses
Syzygium cumini Skeels and Diabetes
Toxicity Studies
Tamarindus indica Linn.
Geographical Source
Phytochemistry
Traditional Uses
Tamarindus indica Linn. and Diabetes
Trigonella foenum-Graecum Linn.
Geographical Source
Phytochemistry
Traditional Uses
Trigonella foenum-Graecum Linn. and Diabetes
Toxicity Studies
Conclusion
References
27 - Lupine Seeds (Lupinus spp.): History of Use, Use as An Antihyperglycemic Medicinal, and Use as a Food Plant
Introduction
Botanical Description
Historical Medicinal Use
Current Medicinal Applications
Type 2 Diabetes
Fiber
Alkaloids
DPP-IV Inhibitors
Conglutins
Hyperlipidemia
Hypertension
Adverse Effects
Summary Points
References
28 - Cancer Chemopreventive Potential of Seed Proteins and Peptides
Introduction
Chemopreventive Effects of Seeds
Chemopreventive Role of Bioactive Proteins and Peptides
Protease Inhibitors
Lectins
Lunasin
Protein Hydrolysates/Peptides
Future Prospects
Acknowledgments
References
29 - Use of Red Clover (Trifolium pratense L.) Seeds in Human Therapeutics
Introduction
Botanical Description
Historical Cultivation and Usage
Present-Day Cultivation and Usage
Applications to Health Promotion and Disease Prevention
Adverse Effects and Reactions, Allergies, and Toxicity
Summary Points
References
30 - Milk Thistle Seeds in Health
Introduction
Botanical Descriptions
Historical Cultivation and Usage
Present-day Cultivation and Usage
Applications to Health Promotion and Disease Prevention
Summary Points
References
Sec 5
31 - Nut Consumption and Noncommunicable Diseases: Evidence From Epidemiological Studies
Introduction
Nuts and Metabolic Disorders
Body Weight, Waist Circumference, and Obesity
Glycemia, Insulinemia, and Type 2 Diabetes
Blood Lipids and Dyslipidemia
Blood Pressure and Hypertension
Nuts and Cardiovascular Disease Risk
Nuts and Cancer Risk
Nuts and Affective Disorders
Nuts and Cognitive Disorders
Conclusions
References
32 - Beneficial Effects of Nuts From India in Cardiovascular Disorders
Introduction
Cardiovascular Disorder
Almond
Origin
Morphology
Cultivation
Nutritional Value
Phytochemistry
Almond and Cardiovascular Diseases
Cashews
Origin
Morphology
Cultivation
Nutritional Value
Phytochemistry
Cashew Nuts and Cardiovascular Diseases
Walnut
Origin
Morphology
Cultivation
Nutritional Value
Phytochemistry
Walnut and Cardiovascular Diseases
Pistachios
Origin
Morphology
Cultivation
Nutritional Value
Phytochemistry
Pistachios and Cardiovascular Diseases
Peanuts
Origin
Morphology
Cultivation
Nutritional Value
Phytochemistry
Peanuts and Cardiovascular Diseases
Conclusion
References
33 - Seeds as Herbal Drugs
Introduction
Medicinal Constituents of Seeds
Factors Influencing Medicinal Properties of Seeds
Seeds as Source of Medicinally Important Fixed Oils
Seeds as Herbal Drugs and Source of Medicinally Active Compounds
Summary Points
References
34 - Therapeutic Importance of Caster Seed Oil
Introduction
Castor Oil Is Unique Among all Fats and Oils
Castor Oil and Its Chemistry
Soaps, Waxes, and Greases
Pharmacological and Medicinal Use
Other Health Benefits of Castor Oil
Boosts Immunity
Increases Circulation
Skin Care
Increases Libido
Induces Labor
Treats Fungal Infections
Eases Constipation
Treats Rheumatic Arthritis
Hair Care
Treats Back Pain
Helps in Birth Control
Relieves Menstrual Disorders
Stimulates Lactation
Eye Care
Other Benefits
Uses
Side Effects of Castor Oil
Other Side Effects:
References
35 - Coriandrum sativum L.: Characterization, Biological Activities, and Applications
Coriander Plant
Coriander Oil and Extracts
Extraction Methods
Coriander Essential Oil Composition and Analysis
Factors Affecting Essential Oil Yield and Composition
Coriander Essential Oil Toxicological Data
Coriander Biological Activities
Coriander Antimicrobial Activity
Mode of Action
Uses of Coriander Oil and Extracts
Food Industry
Other Applications
Concluding Remarks
References
36 - Proteinase Inhibitors From Buckwheat (Fagopyrum esculentum Moench) Seeds
Introduction
Botanical Descriptions
History of Cultivation and Usage
Present-day Cultivation and Usage
Applications to Health Promotion and Disease Prevention
Adverse Effects and Reactions (Allergies and Toxicity)
Summary Points
Acknowledgments
References
37 - Pumpkin Seeds: Phenolic Acids in Pumpkin Seed (Cucurbita pepo L.)
Introduction
Historical Cultivation and Usage
Present-Day Cultivation and Usage
Applications to Health Promotion and Disease Prevention
Adverse Effects and Reactions (Allergies and Toxicity)
Summary Points
References
38 - Big Leaf Mahogany Seeds: Swietenia macrophylla Seeds Offer Possible Phytotherapeutic Intervention Against Diabetic Pathophy ...
Introduction
Description and Distribution
Ethnomedicinal Significance of Swietenia macrophylla Seeds
Swietenia macrophylla Seeds as a Potential Phytotherapeutic Agent Against Diabetes
Swietenia macrophylla Seeds Against Ailments Contribute in the Diabetic Pathogenesis
Toxicities and Contraindications of Swietenia macrophylla Seeds
Phytochemicals in Swietenia macrophylla Seeds
Antidiabetic Phytochemicals in Swietenia macrophylla Seeds
Metabolites in Swietenia macrophylla Seeds Against Pathogenesis Contribute in the Complications
Predicted Molecular Interactions of Swietenine with Signal Proteins and Detection of Drug-likeness
Molecular Docking
In silico ADMET Prediction
Conclusion
References
Index
A
B
C
D
E
F
G
H
I
J
K
L
M
N
O
P
Q
R
S
T
U
V
W
Z
Back Cover

Citation preview

NUTS AND SEEDS IN HEALTH AND DISEASE PREVENTION SECOND EDITION Edited by

VICTOR R. PREEDY RONALD ROSS WATSON

Academic Press is an imprint of Elsevier 125 London Wall, London EC2Y 5AS, United Kingdom 525 B Street, Suite 1650, San Diego, CA 92101, United States 50 Hampshire Street, 5th Floor, Cambridge, MA 02139, United States The Boulevard, Langford Lane, Kidlington, Oxford OX5 1GB, United Kingdom Copyright © 2020 Elsevier Inc. All rights reserved. No part of this publication may be reproduced or transmitted in any form or by any means, electronic or mechanical, including photocopying, recording, or any information storage and retrieval system, without permission in writing from the publisher. Details on how to seek permission, further information about the Publisher’s permissions policies and our arrangements with organizations such as the Copyright Clearance Center and the Copyright Licensing Agency, can be found at our website: www.elsevier.com/permissions. This book and the individual contributions contained in it are protected under copyright by the Publisher (other than as may be noted herein). Notices Knowledge and best practice in this field are constantly changing. As new research and experience broaden our understanding, changes in research methods, professional practices, or medical treatment may become necessary. Practitioners and researchers must always rely on their own experience and knowledge in evaluating and using any information, methods, compounds, or experiments described herein. In using such information or methods they should be mindful of their own safety and the safety of others, including parties for whom they have a professional responsibility. To the fullest extent of the law, neither the Publisher nor the authors, contributors, or editors, assume any liability for any injury and/or damage to persons or property as a matter of products liability, negligence or otherwise, or from any use or operation of any methods, products, instructions, or ideas contained in the material herein. Library of Congress Cataloging-in-Publication Data A catalog record for this book is available from the Library of Congress British Library Cataloguing-in-Publication Data A catalogue record for this book is available from the British Library ISBN: 978-0-12-818553-7 For information on all Academic Press publications visit our website at https://www.elsevier.com/books-and-journals

Publisher: Charlotte Cockle Acquisitions Editor: Megan Ball Editorial Project Manager: Lena Sparks Production Project Manager: Prem Kumar Kaliamoorthi Cover Designer: Christian Bilbow Typeset by TNQ Technologies

Contributors Kaveri Mahadev Adki Shobhaben Pratapbhai Patel School of Pharmacy and Technology Management, SVKM’s NMIMS, Mumbai, Maharashtra, India Larissa Lovatto Amorin Universidade Federal de Ouro Preto, Departamento de Ciências Biológicas, Núcleo de Pesquisas em Ciências Biológicas, Laboratório de Enzimologia e Proteômica, Campus Morro do Cruzeiro, Ouro Preto, Minas Gerais, Brazil Franklin Brian Apea-Bah Department of Food and Human Nutritional Sciences, University of Manitoba, Winnipeg, Manitoba, Canada; Richardson Centre for Functional Foods and Nutraceuticals, Smartpak, University of Manitoba, Winnipeg, Manitoba, Canada

Letizia Bresciani Human Nutrition Unit, Department of Veterinary Science, University of Parma, Parma, Italy H.N. Büyükkartal Ankara University, Faculty of Science, Department of Biology, Tando gan, Ankara, Turkey Chanya Chaicharoenpong Institute of Biotechnology and Genetic Engineering, Chulalongkorn University, Bangkok, Thailand Phool Chandra School of Pharmaceutical Sciences, IFTM University, Lodhipur Rajput, Moradabad, Uttar Pradesh, India Hatice Çölgeçen Department of Biology, Faculty of Arts and Sciences, Zonguldak Bülent Ecevit University, Zonguldak, Turkey

Cecilia Baraldi Department of Life Sciences, University of Modena and Reggio Emilia, Modena, Italy

William de Castro Borges Universidade Federal de Ouro Preto, Departamento de Ciências Biológicas, Núcleo de Pesquisas em Ciências Biológicas, Laboratório de Enzimologia e Proteômica, Campus Morro do Cruzeiro, Ouro Preto, Minas Gerais, Brazil

Mikhail A. Belozersky A.N. Belozersky Institute of Physico-Chemical Biology, Moscow State University, Moscow, Russia

Ben O. de Lumen Department of Nutritional Science and Toxicology, University of California Berkeley, Berkeley, California, United States

Trust Beta Department of Food and Human Nutritional Sciences, University of Manitoba, Winnipeg, Manitoba, Canada; Richardson Centre for Functional Foods and Nutraceuticals, Smartpak, University of Manitoba, Winnipeg, Manitoba, Canada

Andressa Jacqueline de Oliveira Department of Biochemistry and Biotechnology, State University of Londrina, Londrina, Paraná, Brazil

Havva Atar Department of Biology, Faculty of Arts and Sciences, Zonguldak Bülent Ecevit University, Zonguldak, Turkey

Sanjib Bhattacharya West Bengal Medical Services Corporation Ltd., Salt Lake City, Kolkata, West Bengal, India Shovonlal Bhowmick Department of Chemical Technology, University of Calcutta, Kolkata, West Bengal, India Anders Borgen Agrologica, Houvej, Mariager, Denmark

Alessandra de Paula Carli Universidade Federal de Ouro Preto, Departamento de Ciências Biológicas, Núcleo de Pesquisas em Ciências Biológicas, Laboratório de Enzimologia e Proteômica, Campus Morro do Cruzeiro, Ouro Preto, Minas Gerais, Brazil; Universidade Federal dos Vales do Jequitinhonha e Mucuri, Instituto de Ciência, Engenharia e Tecnologia, Campus do Mucuri, Teófilo Otoni, Minas Gerais, Brazil

xiii

xiv

CONTRIBUTORS

Marcos Aurélio de Santana Universidade Federal de Ouro Preto, Departamento de Ciências Biológicas, Núcleo de Pesquisas em Ciências Biológicas, Laboratório de Enzimologia e Proteômica, Campus Morro do Cruzeiro, Ouro Preto, Minas Gerais, Brazil Saikat Dewanjee Advanced Pharmacognosy Research Laboratory, Department of Pharmaceutical Technology, Jadavpur University, Kolkata, West Bengal, India Irene Dini Pharmacy Department, “Federico II” University, Naples, Italy Valentina I. Domash V.F. Kuprevich Institute of Experimental Botany of the National Academy of Sciences of Belarus, Minsk, Belarus Celia Domeño Faculty of Veterinary Medicine, University of Zaragoza, Zaragoza, Spain Fernanda C. Domingues CICS-UBI e Health Sciences Research Centre, University of Beira Interior, Covilhã, Portugal Tarun K. Dua Department of Pharmaceutical Technology, University of North Bengal, Darjeeling, West Bengal, India Yakov E. Dunaevsky A.N. Belozersky Institute of Physico-Chemical Biology, Moscow State University, Moscow, Russia María del Carmen Durán-de-Bazúa Chemical Engineering Department, Facultad de Química, Universidad Nacional Autónoma de México, Cd. Universitaria, Ciudad de México, Mexico Giorgia Foca Department of Life Sciences, University of Modena and Reggio Emilia, Reggio Emilia, Italy; Interdepartmental Research Center BIOGEST-SITEIA, University of Modena and Reggio Emilia, Reggio Emilia, Italy

Frixia Galán-Méndez Facultad de Ciencias Químicas, Universidad Veracruzana, Circuito Aguirre Beltrán s/n, Xalapa, Veracruz, Mexico Justyna Godos Troina, Italy

Oasi Research Institute - IRCCS,

Giuseppe Grosso Department of Biomedical and Biotechnological Sciences, University of Catania, Catania, Italy Milton Hércules Guerra de Andrade Universidade Federal de Ouro Preto, Departamento de Ciências Biológicas, Núcleo de Pesquisas em Ciências Biológicas, Laboratório de Enzimologia e Proteômica, Campus Morro do Cruzeiro, Ouro Preto, Minas Gerais, Brazil Ken-ichi Hatano Division of Molecular Science, Faculty of Science and Technology, Gunma University, Kiryu, Gunma, Japan Blanca Hernández-Ledesma Group of Food Proteins, Instituto de Investigación en Ciencias de la Alimentación (CIAL, CSIC-UAM, CEI UAMþCSIC), Madrid, Spain María del Rosario Hernández-Medel Instituto de Ciencias Básicas, Universidad Veracruzana, Avenida Luis Castelazo Ayala s/n, Xalapa, Veracruz, Mexico Melissa Tiemi Hirozawa Department of Biochemistry and Biotechnology, State University of Londrina, Londrina, Paraná, Brazil Azar Hosseini Pharmacological Research Center of Medicinal Plants, Mashhad University of Medical Sciences, Mashhad, Razavi Khorasan Province, Iran

Fatemeh Forouzanfar Neuroscience Research Center, Mashhad University of Medical Sciences, Mashhad, Iran; Department of Neuroscience, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

Hossein Hosseinzadeh Department of Pharmacodynamics and Toxicology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Razavi Khorasan Province, Iran; Pharmaceutical Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, Razavi Khorasan Province, Iran

Anil Bhanudas Gaikwad Laboratory of Molecular Pharmacology, Department of Pharmacy, Birla Institute of Technology and Science, Pilani Campus, Pilani, Rajasthan, India

Chia-Chien Hsieh School of Life Science, Undergraduate and Graduate Programs of Nutrition Science, National Taiwan Normal University, Taipei, Taiwan

CONTRIBUTORS

xv

Ignasius Radix A.P. Jati Department of Food Technology, Faculty of Agricultural Technology, Widya Mandala Catholic University Surabaya, Surabaya, East Java, Indonesia

Daniela Martini DeFENS-Department of Food, Environmental and Nutritional Sciences, Division of Human Nutrition, University of Milan, Milan, Italy

Taiho Kambe Division of Integrated Life Science, Graduate School of Biostudies, Kyoto University, Kyoto, Japan

Takuya Miyakawa Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Bunkyo-ku, Tokyo, Japan

Natalya V. Khadeeva N.I. Vavilov Institute of General Genetics, Russian Academy of Sciences, Moscow, Russia David H. Kinder College of Pharmacy, Ohio Northern University, Ada, OH, United States Kathryn T. Knecht Associate Professor of Pharmaceutical Sciences, School of Pharmacy, Loma Linda University, Loma Linda, CA, United States U. Koca Gazi University, Faculty of Pharmacy, Department of Pharmacognosy, Etiler, Ankara, Turkey Vera Krimer-Malesevic National Laboratories, Belgrade, Serbia

Reference

Yogesh Anant Kulkarni Shobhaben Pratapbhai Patel School of Pharmacy and Technology Management, SVKM’s NMIMS, Mumbai, Maharashtra, India Ankit Pravin Laddha Shobhaben Pratapbhai Patel School of Pharmacy and Technology Management, SVKM’s NMIMS, Mumbai, Maharashtra, India Sonaly Cristine Leal Universidade Federal de Ouro Preto, Departamento de Ciências Biológicas, Núcleo de Pesquisas em Ciências Biológicas, Laboratório de Enzimologia e Proteômica, Campus Morro do Cruzeiro, Ouro Preto, Minas Gerais, Brazil Qin Liu School of Food Science and Engineering, Nanjing University of Finance and Economics, Nanjing, Jiangsu, China Laura Maletti Department of Chemical and Geological Sciences, University of Modena and Reggio Emilia, Modena, Italy Andrea Marchetti Department of Chemical and Geological Sciences, University of Modena and Reggio Emilia, Modena, Italy; Interdepartmental Research Center BIOGEST-SITEIA, University of Modena and Reggio Emilia, Reggio Emilia, Italy

Mohammad Moradzad Master of student of clinical biochemistry, department of clinical Biochemistry, Kurdistan University of medical sciences, Sanandaj, Iran, Kurdistan in Iran Souvik Mukherjee Department of Pharmaceutical Sciences, Guru Ghasidas Vishwavidyalaya (A Central University), Bilaspur, Chhattisgarh, India Aline Myuki Omori Department of Pathological Sciences, State University of Londrina, Londrina, Paraná, Brazil Mario Augusto Ono Department of Pathological Sciences, State University of Londrina, Londrina, Paraná, Brazil Hesham F. Oraby Department of Agronomy, Faculty of Agriculture, Zagazig University, Zagazig, Egypt Folake Lucy Oyetayo Department of Biochemistry, Ekiti State University, Ado-Ekiti State University Ado-Ekiti, Nigeria Victor Olusegun Oyetayo Department of Microbiology, Federal University of Technology, Akure, Nigeria Dilipkumar Pal Department of Pharmaceutical Sciences, Guru Ghasidas Vishwavidyalaya (A Central University), Bilaspur, Chhattisgarh, India Paramita Paul Department of Pharmaceutical Technology, University of North Bengal, Darjeeling, West Bengal, India Yang Qiu Department of Food and Human Nutritional Sciences, University of Manitoba, Winnipeg, Manitoba, Canada Arezoo Rajabian Pharmacological Research Center of Medicinal Plants, Mashhad University of Medical Sciences, Mashhad, Iran

xvi

CONTRIBUTORS

Mohamed Fawzy Ramadan Agricultural Biochemistry Department, Faculty of Agriculture, Zagazig University, Zagazig, Egypt Gianfranco Risuleo Dipartimento di Biologia e Biotecnologie “Charles Darwin”, Sapienza Università di Roma - Piazzale Aldo Moro, Roma Italy Fabrizio Roncaglia Department of Chemical and Geological Sciences, University of Modena and Reggio Emilia, Modena, Italy Neetu Sachan School of Pharmaceutical Sciences, IFTM University, Lodhipur Rajput, Moradabad, Uttar Pradesh, India Achintya Saha Department of Chemical Technology, University of Calcutta, Kolkata, West Bengal, India Patricia Sanchez College of Letters and Science, University of California, Los Angeles, CA, United States Alexandre Gonçalves Santos Universidade Federal de Ouro Preto, Departamento de Ciências Biológicas, Núcleo de Pesquisas em Ciências Biológicas, Laboratório de Enzimologia e Proteômica, Campus Morro do Cruzeiro, Ouro Preto, Minas Gerais, Brazil Elisabete Yurie Sataque Ono Department of Biochemistry and Biotechnology, State University of Londrina, Londrina, Paraná, Brazil Simona Sighinolfi Department of Chemical and Geological Sciences, University of Modena and Reggio Emilia, Modena, Italy Filomena Silva ARAID e Agencia Aragonesa para la Investigación y el Desarrollo, Zaragoza,

Spain; Faculty of Veterinary Medicine, University of Zaragoza, Zaragoza, Spain; CICSUBI e Health Sciences Research Centre, University of Beira Interior, Covilhã, Portugal Julio A. Solís-Fuentes Instituto de Ciencias Básicas, Universidad Veracruzana, Avenida Luis Castelazo Ayala s/n, Xalapa, Veracruz, Mexico Vetriselvan Subramaniyan Associate Professor, Department of Pharmacology, Faculty of Medicine, MAHSA University, Bandar Saujana Putra, Selangor, Malaysia Reka SzTllTsi Department of Plant Biology, University of Szeged, Szeged, Hungary Masakazu Takahashi Department of Bioscience and Biotechnology, Fukui Prefectural University, Fukui, Japan Masaru Tanokura Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Bunkyo-ku, Tokyo, Japan Lorenzo Tassi Department of Chemical and Geological Sciences, University of Modena and Reggio Emilia, Modena, Italy; Interdepartmental Research Center BIOGESTSITEIA, University of Modena and Reggio Emilia, Reggio Emilia, Italy Alexander A. Vassilevski ShemyakinOvchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, Russia Enas Mohamed Wagdi Abdel-Hamed Soil Science Department, Faculty of Agriculture, Zagazig University, Zagazig, Egypt

Preface The objective of this book is to bring together scientific material relating to the health benefits and, where appropriate, adverse effects of nuts and seeds. In general, nuts and seeds are important not only from a nutritional point of view but also in terms of their putative medicinal or pharmacological properties. This book aims to describe these properties in a comprehensive way. However, at the same time, it is recognized that harmful effects also arise. Some “nuts” and “seeds,” for example, are poisonous when ingested in large quantities, but extracts have putative effects on tissues that may offer some therapeutic potential. Many of the nuts and seeds described in this book are components of traditional remedies without any present-day evidence to support their claims; their properties await rigorous elucidation and scientific investigation. Thus, the book embraces nuts and seeds in an unbiased way. The Editors also recognize that there is a wide interpretation of the terms nuts and seeds, and indeed some authorities have claimed that there are at least 12 seed types. The Editors have largely excluded cereals (grains) and other staple food crops, unless there was cause to include them, such as with buckwheat seeds. They have also selected some specific legumes, where there is some therapeutic potential in their extracts or interesting properties. The book Nuts and Seeds in Health and Disease Prevention is divided into two parts. Part I, General Aspects and Overviews, contains holistic information, with sections

on Overviews, Composition, Effects on Health, and Adverse Aspects. In Part II, Effects of Specific Nuts and Seeds, coverage is more specific. Each chapter in Part II contains sections entitled Botanical description, Historical cultivation and usage, Present-day cultivation and usage, Applications to health promotion and disease prevention (the main article), and, finally, Adverse effects and reactions. The Editors were faced with a difficult choice in organizing the chapters in Part II, and this was done using the simplest method available. Thus, in Part II, the nuts and seeds are listed alphabetically in terms of their common names, although each chapter contains full botanical terminology. We realize this is not perfect, for example, there are numerous types of cabbage seeds, and some nuts and seeds may have as many as 20 common names depending on the country where they are grown, but navigation and the retrieval of specific information is aided by a comprehensive index system. This book is designed for health scientists, including nutritionists and dietitians, pharmacologists, public health scientists, those in agricultural departments and colleges, epidemiologists, health workers and practitioners, agriculturists, botanists, health care professionals of various disciplines, policymakers, and marketing and economic strategists. It is designed for teachers and lecturers, undergraduates, and graduates.

xvii

The Editors

Acknowledgments The work of Dr. Watson’s editorial assistant, Bethany L. Stevens, in communicating with authors and editors and working on the manuscripts was critical to the successful completion of the book. It is very much appreciated. Support for Ms. Stevens’ and

Dr. Watson’s editing was graciously provided by Dr. Preedy, Dr. Watson, and Southwest Scientific Editing & Consulting, LLC. Direction and guidance from Elsevier’s staff was critical.

xix

C H A P T E R

1

Rambutan (Nephelium lappaceum L.) Seed and Its Fat

1

Julio A. Solís-Fuentes1, Frixia Galán-Méndez2, María del Rosario Hernández-Medel1, María del Carmen Durán-de-Bazúa3

Instituto de Ciencias Básicas, Universidad Veracruzana, Avenida Luis Castelazo Ayala s/n, Xalapa, Veracruz, Mexico; 2Facultad de Ciencias Químicas, Universidad Veracruzana, Circuito Aguirre Beltrán s/n, Xalapa, Veracruz, Mexico; 3Chemical Engineering Department, Facultad de Química, Universidad Nacional Autónoma de México, Cd. Universitaria, Ciudad de México, Mexico List of Abbreviations ALP Arachidoyl-linoleoyl-palmitoyl-glycerol AOO Arachidoyl-dioleoyl-glycerol AOP Arachidoyl-oleoyl-palmitoyl-glycerol ASO Arachidoyl-stearoyl-oleoyl-glycerol ASP Arachidoyl-stearoyl- palmitoyl-glycerol CB Cocoa butter CBE Cocoa butter equivalent CE Catechin equivalent FA Fatty acid FFAs Free fatty acids FOs Fat and oils GAE Gallic acid equivalent RS Rambutan seed RSF Rambutan seed fat SFC Solid fat content SOS 1,3-distearoyl-2-oleoyl- glycerol TGs Triacylglycerols WHO World Health Organization

Nuts and Seeds in Health and Disease Prevention, Second Edition https://doi.org/10.1016/B978-0-12-818553-7.00001-2

3

Copyright © 2020 Elsevier Inc. All rights reserved.

4

1. Rambutan (Nephelium lappaceum L.) Seed and Its Fat

Introduction Rambutan (Nephelium lappaceum L.) is an exotic plant from Southeast Asian native to the MalaysianeIndonesian region, closely related to fruit species as lychee (Litchi chinensis Sonn.) with whom it shares some morphological features except its long, thick, and soft spines or hairs in its shell; its edible pulp is white, juicy, translucent, subacid-sweet flavored, also similar to litchi. Like other parts of the plant, the seed of rambutan (RS) fruit contains important bioactive components; some researchers have shown that RS extracts can possess hypoglycemic, antinociceptive, analgesic, anti-inflammatory, CNS depressant, antibacterial, and anticancer activity.1 RS has some nutrients and also contains considerable amounts of edible fat with potential uses in the food and pharmaceutical industries, which is important because, currently, a clear trend has been observed toward the use of natural vegetable fats for a total or partial replacement of industrial trans fats obtained by hydrogenation or interesterification of vegetable oils, and, therefore, rambutan seed fat (RSF) properties have become a very important issue for the procurement of nutrition and health of the population.

Botanical Descriptions Rambutan is a perennial tropical tree belonging to the Sapindaceae family, which includes around 37 genera and 72 species. It is a medium size evergreen tree that grows to a height of 12e20 m. The trees can be male, female, or hermaphroditic, with alternate and pinnate leaves (10e30 cm long and 3e11 leaflets) with small apetalous and discoidal flowers (from 2 to 5 cm). Its name is associated with the Malay word “rambut,” which means hairy. Its fruit, in clusters of 15e20 units, sometimes called “hairy lychee” is usually consumed fresh;2,3 it has a single seed, round or ellipsoid in shape; its color varies from pink to deep red or from orange red to yellowish red. There is a translucent and juicy white pulp under the fruit skin whose taste is sweet and pleasant.4 The flowering and fruit production occur between 3 and 5 years after planting. More than 200 varieties of rambutan are cultivated and available throughout tropical Asia.3,5

Historical Cultivation and Usage It is believed that this plant is native to the Malay Archipelago, from where it spread to Thailand, Burma, Sri Lanka, India, Vietnam, the Philippines, and Indonesia. Later, at the beginning of the 20th century, some varieties of rambutan were introduced in the Western Hemisphere due to their high development potential in regions with favorable agroclimatic conditions for their cultivation: between 100 and 1000 m of altitude, temperatures of around 28
C, and blooming during spring and summer. The most extensive cultivation of N. lappaceum is found in the countries of Southeast Asia, especially in Malaysia, the Philippines, and Thailand. The main use of rambutan has been as fresh fruit considering only its pulp as edible, but canned rambutan is also produced and exported by the main producer countries.3,5

I. Overview and General Themes

Applications in Health Promotion and Disease Prevention

5

Present-Day Cultivation and Usage The most important species, from the economic point of view, within the Nephelium genus is the rambutan.3 It is cultivated in Southeast Asia and in Australia, South Africa, and Mexico, and in other tropical places in the world such as Hawaii, the Caribbean islands, Costa Rica, Honduras, and Panama, among many other places. In Thailand, the best known and popular varieties are Rongrien and Chompu, which have crispy arils and are suitable for both fresh and canned consumptions. Data about world cultivated area and production of rambutan are scarce, and estimates for the year 2003 show that its cultivation is on more than 200,000 hectares with a production of between 1.5 and 2 million tons each year. However, considering the tendency in many countries for a greater consumption of healthy fruits, it is feasible that to date such figures are widely exceeded.2 Thailand, Indonesia, and Malaysia are the main producers and exporters of the fruit accounting about 80% of the world production.6 From the fruit, only the pulp is considered edible, generally consumed fresh, although sometimes it is processed industrially to obtain juices, jams, and jellies, among other products, generally in canned presentations.4,7 The rambutan fruit is 34e54% pulp, 37e62% of peel, non-edible until now, and 4e9% of seed, consumed as food in some places and possibly suitable for some applications in foods.8,9 The two main residues of rambutan fruit processing are the peel and seeds. Even though in some Asian countries, RS is eaten after roasting or boiling, having a slight bitter taste, according to popular knowledge, it is known that raw is poisonous.7 RS as a waste in canned manufactures reaches thousands of tons by year in the main producer countries. Only in Thailand it is estimated that the rambutan canning industry produces around 2000 tons of RS each year.10 Chemical composition of RS depends on varietal and phenotypic variability; ranges of some reported values are shown in Table 1.1: moisture 3.31e34.4%, fat 28.2e39.13%, crude protein 7.9e13.7%, with a good quality because of its essential amino acids present around 23.6e27.8%, ash 2.26e2.30%, crude fiber 7.6, and total carbohydrates 28.7e62.4%. Contents of the minerals e K, P, Mg, Mn, Fe, Na, and Zn e have also been reported. Some bioactive compounds are present too: phenolic acid in levels of 4.4e26.7 mg/100g of db seed, with an important presence of tannins, ellagic and gallic acids, geraniin, and corilagin. Thus, RS as a possible food contains important nutrients, as well as other constituents that have recognized bioactivity. Because of its protein and carbohydrate contents, some authors such as Harahap et al.11 have considered that RS is comparable to watermelon seeds (Citrullus lanatus Schrad) or pumpkin seeds (Cucurbita pepo).

Applications in Health Promotion and Disease Prevention RS composition shows important nutritional constituents, with well-known effects in the maintenance of health, if its possibility as food is considered. However, some compounds with an antinutritive and toxic effect have also been identified as being present in RS, so their food safety must be fully evaluated. The bioactive compounds with a known antioxidant

I. Overview and General Themes

6 TABLE 1.1

1. Rambutan (Nephelium lappaceum L.) Seed and Its Fat

Ranges of Rambutan Seed Chemical Composition.

Chemical Constituent

Values

Units

References

Seed in fruit

4.0e9.0

%

8,9

Moisture

3.31e34.40

%

5,19

Protein

7.9e14.1

%

10,24

Essential amino acids

23.63e27.84

g/100g

a

Crude fiber

2.8e8.6

%

10

Crude fat

28.20e39.13

18,24

Ash

1.22e2.30

5,11

Total carbohydrates

28.7e62.4

5,24

Proximal composition

Bioactive compounds mg/100g

18,23

Saponins

2.10e18.96

Alkaloids

1.95

23

Phytates

0.77

23

Oxalates

0.19

23

Flavonoids

1.63

23

Total polyphenolic

7.6

mg GAE/g

b

Tannins (catechin)

13.8

mg CE/g

b

Ellagic acid

461.1

mg/kg

b

Geraniin

423.2

b

Gallic acid

98.0

b

Corilagin

94.5

b

Minerals mg/100g

9

Calcium

9.5

Potassium

84.1

9

Phosphorus

16.6

9

Magnesium

12.3

9

Manganese

1.06

9

Sodium

20.8

9

Iron

0.3

9

Zinc

0.17

9

a

Augustin MA, Chua BC. Composition of rambutan seeds. Pertanika 1988; 11(2):211e215. Mehdizadeh S, Lasekan O, Muhammad K et al. Variability in the fermentation index, polyphenols, and amino acids of seeds of rambutan (Nephelium lappaceum L.) during fermentation. Journal of Food Composition and Analysis 2015; 37:128e135.

b

I. Overview and General Themes

Nephelium lappaceum Seed Fat (RSF) Characteristics, Potential Food, and Therapeutic Uses

7

capacity, present in RS, represent an interesting aspect, since extracts of the seed have shown to possess hypoglycemic, antinociceptive, analgesic, anti-inflammatory, CNS depressant, antibacterial, and anticancer activity.1,12,13 On the other hand, it is well known that the importance of vegetable fat and oils (FOs) in diet and food processing on human health and RS, being a highly available waste, can be used as an unconventional source of natural fats with possible uses in the food industry and as a vehicle for active medicines in the pharmaceutical industries, and in cosmetics. Currently, the identification of new sources of natural dietary fats is an important asset because of their relative scarcity compared to those of vegetable oils and because the demand for edible fats for food applications has been satisfied, in most cases so far, through the partially hydrogenated fats production with high trans fatty acid (FA). The negative effect on health of diets with high levels of trans FA, corroborated in several investigations,14 has been the basis for the approach of one of the global objectives of the World Health Organization to be achieved in the coming years, regarding the elimination of trans fats in processed food products14,15 for being replaced with vegetable oils or with natural fats in agreement with the new approaches reassessing the role of saturated AF on health and on various diseases.16

Nephelium lappaceum Seed Fat (RSF) Characteristics, Potential Food, and Therapeutic Uses RSF Physical and Chemical Properties Table 1.2 shows some physicochemical properties. The RSF is an almost white, semi-solid fat at room temperature, with 31Yþ1.1R color on the Lovibond scale, and a fusion range between 14.5 and 55.8
C. Oleic and arachidic acids are the major FAs; RSF has a saponification and an iodine numbers of 157.0e246.7 and 37.6e50.6, respectively. Chemical composition of RSF shows variability; Table 1.3 presents the ranges of data reported for many authors. The major FAs of RSF are oleic (33.35e46.64%), arachidic (26.03e34.36%), gondoic (5.75e10.55), and stearic acids (5.22e8.97), followed by palmitic and linoleic and in smaller amounts behenic, lignoceric, linoleic, lauric, and myristic acids. Globally, 37.51e46.24% are SFA and 44.19e62.50% are UFA. Triglycerides of RSF correspond to 83.94e95.33%; so far, there are few reports that establish with certainty which are the majority. Harahap et al.11 found that these corresponded to AOO, ASO, AOP, ASP, and ALP with 49.8, 15.0, 12.8, 9.0, and 6.3%, respectively. Other triacylglycerols (TGs) identified by several authors around 3% or less are ALnO, arachidoyl linolenoyl oleoylglycerol; SOO, stearoyl dioleoyl glycerol; OOO, trioleoyl glycerol; and POS, palmitoyl oleoyl stearoyl glycerol, among others.4 In the fraction of unsaponifiable (0.43e0.82%) campesterol, stigmasterol, b-sitosterol, and a-tocopherol have been identified. Additionally, Harahap et al.11 found some metals content in the fat: Ca, Mg, and Zn (at concentrations of 160, 51, and 40 mg/g, respectively) among others.

I. Overview and General Themes

8 TABLE 1.2

1. Rambutan (Nephelium lappaceum L.) Seed and Its Fat

Ranges of Main Physicochemical and Phase Properties of Rambutan Seed Fat.

Properties

Values

Units

References

Physical 5,8

Refractive index

1.468e1.469

Color

31Yþ1.1R

Lovibond

4

Melting onset/offset temperature

14.5/55.8




C

5

DHf

71.2e141.7

J/g

18

Crystallization onset/offset temperature

33.6/45.6




5

Predominant polymorph

B

e

8

Crystal morphology

Spherulites

e

20

DHc

60.4e88.9

J/g

18

10

45.0e47.8

%

4,5

21.1

25.0

5

26.7

19.6

5

33.3

16.2

5

37.8

13.4

5

40.0

11.2

5

Phase

C

Solid fat content (
C)

Chemical Iodine number

37.64e50.61

g I2/100g fat

11,18

Saponification number

157.07e246.73

mg KOH/g fat

11,a

Acid value

2.7e3.9

% As oleic acid

5,19

a Lourith N, Kanlayavattanakul M, Mongkonpaibool K et al. Rambutan seed as a new promising unconventional source of specialty fat for cosmetics. Industrial Crops and Products 2016; 83:149e154.

RSF Phase Behavior RFS has been proposed for being used as zero trans natural margarines and frying shortenings or in the formulation of pharmacological and cosmetic products, and to be used as special high-quality fats, such as those used in confectionery products4,5,8,10, specifically, the latter, as a possible substitute for cocoa butter (CB).9 All these possible applications of RSF derive from the composition and properties including their phase properties. It is known that vegetable FOs are a mixture of different TGs according to the different FAs that constitute them and that, together with the polymorphism, characteristic of these molecules determines their behavior as solideliquid phase,17 and in fact, unlike pure compounds, these mixtures of TGs do not have a single melting or crystallization temperature, but rather a temperature range in which the transition between the solid and liquid phases develops and completes. The transition temperature for the different TGs depends on the position of each I. Overview and General Themes

9

Nephelium lappaceum Seed Fat (RSF) Characteristics, Potential Food, and Therapeutic Uses

TABLE 1.3

Ranges of Chemical Composition of Rambutan Seed Fat.

RSF composition

Values

Units

References

Triacylglycerols

83.94e95.33

%

18

Lauric (C12:0)

0.080e2.476

%

19,a

Myristic (C14:0)

0.13e2.15

4,19

Palmitic (C16:0)

2.40e6.10

5,18

Palmitoleic (C16:1)

0.16e3.27

18

Stearic (C18: 0)

5.22e8.97

18

Oleic (C18:1)

33.35e46.64

18

Linoleic (C18:2)

1.48e3.52

18

Linolenic (C18:3)

0.14e9.90

18

Arachidic (C20:0)

26.03e34.36

11,18

Gondoic (C20:1)

5.75e10.55

18

Behenic (C22:0)

1.64e2.90

5,18

Erucic (C22:1)

0.06e0.73

18

Lignoceric (24:0)

0.12e0.26

18

Saturated FA

37.51e46.24

Unsaturated FA

44.19e62.50

Fatty acids

%

4,18 18

0.37e6.10

%

4,11

AOO

49.84

%

11

AOP

12.82

11

ASO

15.05

11

ASP

9.03

11

ALP

6.32

11

Free fatty acids Main triacylglycerols

Unsaponifiables

0.19e0.82

g/100g

8

Sterol fraction Campesterol

N/D

Stigmasterol

0.32

b-sitosterol

0.61

8

mg/g

8 8

Tocopherol fractions a-tocopherol

8

0.103

mg/g

8

a Winayanuwattikun P, Kaewpiboon C, Piriyakananon K et al. Potential plant oil feedstock for lipase-catalyzed biodiesel production in Thailand. Biomass Bioenergy 2008;32(12):1279e1286.

I. Overview and General Themes

10

1. Rambutan (Nephelium lappaceum L.) Seed and Its Fat

FA in the TG molecule, its length, and the existence or not of insaturations in the chain of each FA. In general, saturated and longer hydrocarbon chains determine SeL transitions at higher temperatures. Thus, the composition in TGs and FAs of RSF explains that this is a semi-solid fat at an ambient temperature around 20
C. Table 1.2 presents the main physicochemical and phase properties of RSF. The solid fat content at several temperatures shows that around 20
C 25% of TGs of RSF are solid. Fig. 1.1 shows the typical fusion curve of RSF in which the temperatures at which the fat begins to melt (onset, 14.7
C) and to which it ends, to be totally liquid (off set, 55.8), can be observed; likewise, its groups of TGs can be identified with lower (29
C), which allow us to perceive the possibility of separating them through fractionation, for example, dry fractionation method. This option will render more specific application alternatives to RSF.5 It is evident that the SeL phase behavior of RSF depends not only on the composition of TGs but also on the variety of N. lappaceum; Chai et al.18 made a comparison of these phase behaviors for 11 Thai varieties, with which the versatility and potential uses of RSF have considerably broadened.

RSF and Its Potential Food Uses for Health Promotion The possibility of using RSF in food is wide due to the numerous applications of vegetable fats in this sector. Nowadays, the most obvious advantages that its alimentary use would bring in the procurement of health is in its contribution to the objective of gradually replacing trans fats in food applications, delineated by different organisms and health institutions at local and international levels. Some of the possible uses of RSF or its fractions, which were studied and reported, are as shortenings and as an additional fat to CB or supplementary CB in confectionary products. Sirisompong et al.8 suggested that the RSF obtained from a Thai variety of rambutan showed chemical and physical characteristics comparable to those of conventional partially hydrogenated edible fats. The use of RSF to replace this type of fats is especially interesting when the oxidation of the product can become an important aspect.

FIGURE 1.1 Typical fusion curve of rambutan seed fat. The figure shows the temperatures at which the process of solideliquid phase transition of rambutan seed fat occurs.

I. Overview and General Themes

Summary Points

11

Mahisanut et al.10 studied the isothermal fractionation of RSF with acetone and with ethanol, obtaining a fraction high in arachidic acid with acetone at 25
C and 24 h of incubation and whose properties were comparable to those of some commercial fully hydrogenated fats. Sonwai and Ponprachanuvut19 evaluated the physicochemical properties, the composition in FAs, and the phase behavior of RSF of varieties grown in Thailand, and their possible industrial applications. They say that the behavior of crystallization or melting turned out to be similar to that presented by CB, with a two-step crystallization curve, and that properties of the RSF may be appropriate in specific applications in various segments of the industry. Febrianto et al.20 evaluated RSF in the development of confectionery products with improved characteristics through a treatment of fermentation and roasting of RS, analyzing the development of flavor compounds similar to cocoa due to the effect of these processes. The results showed that the desired pyrazine compounds were present and that they were up to 42.69% of the total odorant compound in the RSF. The fat from the fermented and roasted RS had similar characteristics with CB increasing its potential to be used as a substitute for CB. In other non-food applications, Uraiwan and Satirapipathkul21 and Witayaudom and Klinkesorn22 reported the feasible use of RSF in the preparation of nanostructured lipid carriers for the delivery of bioactive compounds in cosmeceutical and pharmaceutical products.

Adverse Effects and Reactions, Allergies, and Toxicity The scientific data about RS and RSF safety are scarce. Regarding the seed, it is known that some Asian peoples in rambutan producing regions, as the Philippines, consume it after its roasting.7 The RS has a characteristic bitter taste and has been reported to have some narcotic effects that have been attributed to its contents of tannins, alkaloids, and saponins.18 Fila et al.23 reported the content of some compounds with antinutritional effects in the different parts of rambutan; in the seed, saponins (2.10) and alkaloids (1.95 mg/100 g db) were the most relevant ones, although phytates (0.77), oxalates (0.19), and tannins (0.28), as well as flavonoids (1.63), were also present. The authors concluded that these antinutritional compounds were in RS at tolerable levels. Chai et al.6 evaluated the RS toxicity, as a cocoa-like powder, after the fruit fermentation, drying, and roasted processes of the seed. Their results about brine shrimp lethality tests showed that SR, under these conditions, is not toxic. Eiamwat et al.24 conducted tests to evaluate the toxicity of RSF in oral and dermal intakes in rats and rabbits, finding that a single oral dose of up to 5 and 2 g/kg of body mass in the dermal route was not lethal for rats. The treatment also did not cause skin irritation or signs of toxicity in rabbits. The authors concluded that RSF is a non-toxic fat.

Summary Points • Rambutan is a fruit of Asian origin with a high consumption and industrialization in some regions of the world.

I. Overview and General Themes

12

1. Rambutan (Nephelium lappaceum L.) Seed and Its Fat

• RS is a residue of high availability, whose composition is attractive from the point of view of food, nutrition, and medicine. • RS is a potential source of edible fat whose main FAs are oleic and arachidic acids. • The physicochemical and phase characteristics of RSF make it potentially useable as a substitute for partially hydrogenated fats with high trans FA contents. • Healthy dietary fats, free of trans FA, are of great importance in the promotion and maintenance of people’s health.

References 1. Sukmandari NS, Dash GK, Jusof WHW, Hanafi M. A review on Nephelium lappaceum L. Research Journal of Pharmacy and Technology. 2017;10(8):2819e2827. 2. Paull ER, Duarte O. Tropical Fruit. 2nd ed. Vol. 2. Cammbridge MA, USA: CABI; 2012. 3. Chakraborty B, Mishra DS, Hazarika BN, et al. Rambutan. Chapter 29. In: Ghosh S, ed. Breeding of Underutilized Fruit Crops. Delhi, India: Jaya Publishing House; 2015:425e440. 4. Yanti NAM, Marikkar JMN, Long K, et al. Physico-Chemical characterization of the fat from Red-Skin rambutan (Nephellium lappaceum L.) Seed. Journal of Oleo Science. 2013;62(6):335e343. 5. Solís-Fuentes JA, Camey-Ortíz G, Hernández-Medel MR, et al. Composition, phase behavior and thermal stability of natural edible fat from rambutan (Nephelium lappaceum L.) seed. Bioresource Technology. 2010;101(2):799e803. 6. Chai KF, Chang LS, Adzahan NM, et al. Physicochemical properties and toxicity of cocoa powder-like product from roasted seeds of fermented rambutan (Nephelium lappaceum L.) fruit. Food Chemistry. 2019;271:298e308. 7. Rambutan J Morton. In: Fruits of Warm Climates. Miami, FL, USA: Julia F. Morton; 1987:262e265. 8. Sirisompong W, Jirapakkul W, Klinkesorn U. Response surface optimization and characteristics of rambutan (Nephelium lappaceum L.) kernel fat by hexane extraction. Lebensmittel-Wissenschaft und -Technologie- Food Science and Technology. 2011;44(9):1946e1951. 9. Issara U, Zzaman W, Yang TA. Rambutan seed fat as a potential source of cocoa butter substitute in confectionary product. International Food Research Journal. 2014;21:25e31. 10. Mahisanunt B, Jom KN, Matsukawa S, et al. Solvent fractionation of rambutan (Nephelium lappaceum L.) kernel fat for production of non-hydrogenated solid fat: influence of time and solvent type. Journal of King Saud University Science. 2017;29(1):32e46. 11. Harahap SN, Ramli N, Vafaei N, et al. Physicochemical and nutritional composition of rambutan AnakSekolah (Nephelium lappaceum L.) seed and seed oil. Pakistan Journal of Nutrition. 2012;11(11):1073e1077. 12. Bhat RS, Al-daihan S. Antimicrobial activity of Litchi chinensis and Nephelium lappaceum aqueous seed extracts against some pathogenic bacterial strains. Journal of King Saud University Science. 2014;26(1):79e82. 13. Soeng S. E. Evacuasiany, W. Widowati, et al., Antioxidant and hypoglycemic activities of extract and fractions of Rambutan seeds (Nephelium lappaceum L.). Biomedical Engineering. 2015;1(1):13e18. 14. Uauy R, Aro A, Clarke R, et al. WHO Scientific Update on trans fatty acids: summary and conclusions. European Journal of Clinical Nutrition. 2009;63:S68eS75. 15. WHO. Eliminate Trans-fatty Acids in Global Food Supply. ENS.; June 11, 2018. http://ens-newswire.com/2018/06/ 11/who-eliminate-trans-fatty-acids-in-global-food-supply/ 16. Lawrence GD. Dietary fats and health: dietary recommendations in the context of scientific evidence. Advances in Nutrition. 2013;4(3):294e302. 17. O’Brien RD. Fats and Oils Formulating and Processing for Applications. Boca Raton: CRC Press; 2009. 18. Chai KF, Adzahan NM, Karim R, et al. Characteristics of fat, and saponin and tannin contents of 11 varieties of rambutan (Nephelium lappaceum L.) seed. International Journal of Food Properties. 2018;21(1):1091e1106. 19. Sonwai S, Ponprachanuvut P. Characterization of physicochemical and thermal properties and crystallization behavior of Krabok (Irvingia Malayana) and Rambutan seed fats. Journal of Oleo Science. 2012;61(12):671e679. 20. Febrianto NA, Yang TA, Wan Abdullah WA. Cocoa-like flavor compound development of rambutan seed fat as the effect of fermentation and roasting. International Food Research Journal. 2016;23(5):2166e2174.

I. Overview and General Themes

References

13

21. Uraiwan K, Satirapipathkul C. The entrapment of vitamin E in nanostructured lipid carriers of rambutan seed fat for cosmeceutical uses. In: Key Engineering Materials. Vol. 675. Trans Tech Publications; 2016:77e80. 22. Witayaudom P, Klinkesorn U. Effect of surfactant concentration and solidification temperature on the characteristics and stability of nanostructured lipid carrier (NLC) prepared from rambutan (Nephelium lappaceum L.) kernel fat. Journal of Colloid and Interface Science. 2017;505:1082e1092. 23. Fila WO, Johnson JT, Edem PN, et al. Comparative anti-nutrients assessment of pulp, seed and rind of rambutan (Nephelium lappaceum). Annals of Biological Research. 2012;3(11):5151e5156. 24. Eiamwat J, Reungpatthanaphong S, Laovitthayanggoon S, et al. Toxicity studies on rambutan (Nephelium lappaceum) seed fat and oil extracts using acute oral, dermal and irritation assays. International Journal of Natural Products Research. 2014;4(2):36e39.

I. Overview and General Themes

C H A P T E R

2

Soursop Seed: Soursop (Annona muricata L.) Seed, Therapeutic, and Possible Food Potential Julio A. Solís-Fuentes1, María del Rosario Hernández-Medel1, María del Carmen Durán-de-Bazúa2 1

Instituto de Ciencias Básicas, Universidad Veracruzana, Avenida Luis Castelazo Ayala s/n, Xalapa, Veracruz, Mexico; 2Chemical Engineering Department, Facultad de Química, Universidad Nacional Autónoma de México, Cd. Universitaria, Ciudad de México, Mexico

List of Abbreviations AACGs Annonaceous acetogenins AMS Annona muricata seeds ATP Adenosine triphosphate ED50 Drug dose effective for 50% of exposed population FA Fatty acid Hep 2,2,15 Human hepatoma cells infected with hepatitis B virus Hep G2 Human hepatoma cells IC50 Drug concentration required for 50% inhibition in vitro THF Tetrahydrofuran THP Tetrahydropyran

Introduction Soursop (Annona muricata L) is a plant native to South America that is widely distributed in tropical and subtropical regions of the world; it belongs to the Annonaceae family and its fruits are widely appreciated for their organoleptic characteristics. The family includes several genera characterized by the presence of metabolites with important biological activities. AMS make up approximately 5% of ripe fruit weight. In recent years, scientific

Nuts and Seeds in Health and Disease Prevention, Second Edition https://doi.org/10.1016/B978-0-12-818553-7.00002-4

15

Copyright © 2020 Elsevier Inc. All rights reserved.

16

2. Soursop Seed: Soursop (Annona muricata L.) Seed, Therapeutic, and Possible Food Potential

studies have confirmed the presence of alkaloids, acetogenins, and cyclopeptides, which are compounds of great importance that have attracted pharmacological interest. AMS also have a significant amount of oil whose composition and physical and chemical properties make it potentially attractive in the food sector.

Botanical Descriptions A. muricata is an annonacea belonging to the Magnoliales Order and the Magnoliopsida Class. It is a tree from 4 to 7 m high, with smooth bark. Its leaves are long and pale green and its flowers have yellow-green fleshy petals. Its fruits are large, oval or heart shaped, with small green thin-shelled spines; their size ranges from 10 to 30 cm long and 15 cm wide, and they can weigh up to 2.5 kg. They have white flesh that is creamy and juicy. The seeds are small and numerous, dark-colored, and of compact consistency. They have a rigid coating that contains a kernel-like almond. At present, there is no information on the number of world varieties. Coarsely, soursop types were classified into sweet, subacid, or sour; however, in just one region of Puerto Rico, 14 varieties have been distinguished, and it is believed that in the other producing regions of the world there are many more.1

Historical Cultivation and Usage Cultivated in Mexico already before the Spaniards arrived, the soursop has been highly appreciated for its edible fruit. It was distributed very early in the warm lowlands of eastern and western Africa, southeastern Asia, and China, where it was and continues to be commonly grown on a small scale or as a backyard tree.2 It was first used as fresh fruit and later in various food products processed either by hand or on an industrial scale. Historical information on the medicinal use of AMS is scarce, for it was not until the 20th century when its acaricidal property was first reported.

Present-day Cultivation and Usage The soursop lives in tropical regions with warm weather ranging from sea level to 500 m high. In Mexico, for example, it is cultivated from the state of Sinaloa to the state of Chiapas and from the state of Veracruz to the Yucatan Peninsula along the Gulf of Mexico in small extensions up to 20 ha. In many other parts of the world, it is still grown as backyard trees. There are no data available for world production, imports, and exports. In Surinam, this fruit yields 43 kg/tree and 278 trees/ha. The commercial plantations are limited to the Philippines, the Caribbean, and South America. At this time, it is used mainly as fresh and industrialized fruit. The leaves are used commercially for therapeutic uses. Extracts from the seed powder are presently used as an effective insecticide against lice, worms, and aphids, as well as a larvicide.1,3

I. Overview and General Themes

Applications in Health Promotion and Disease Prevention

17

Applications in Health Promotion and Disease Prevention Annona muricata L. Seed and Its Therapeutic Potential In traditional medicine it is known that the bark, root, leaf, fruit, and seed of the fruit of Annona muricata are used for various medical purposes in the tropics. The crushed seeds are used mainly against internal and external parasites, worms, and lice.1,3 The studies have been carried out in general with crude extracts of organic solvents such as ethanol and n-hexane, and some of them have allowed to identify their bioactivities as well as to evaluate the synergism of their compounds. For example, ethanolic extracts of AMS have demonstrated their power of inhibition on the growth of larvae of Spodoptera litura (Noctuidae),4 Aedes aegypti,5ae5d and Culex quinquefasciatus, a vector of filariasis a set of infectious diseases that affect the lymphatic system and the skin, of great epidemiological importance in lowincome countries.5d,5e Komansilan et al.5c showed that the n-hexane fraction was the most effective and toxic against the mosquito Aedes aegypti, with a LC50 of 73.77 ppm. The extract analyzed by GCMS showed the relative abundance of methyl palmitate (39.93%), methyl oleate (25.05%), and methyl stearate (25.71%). Ranisaharivony et al.5d reported the synergistic activity of AACGs annonacin, murisolin, and annonacinone from AMS extracts and the catalytic hydrogenation of annonacin, which resulted in a more effective mixture of diastereomers than annonacin alone, against the larvae of C. quinquefasciatus. The synergism between the alcoholic extracts of AMS and Piper nigrum in larvicidal evaluations realized by Grzybowski et al.5b resulted in an efficient, simple, and less toxic formula from AMS. In addition, the antimalarial (Plasmodium falciparum) and leishmanicidal activities of the AMS extracts have been evaluated by Boyom et al.5f and Vila-Nova et al., 5g,5h respectively. In other tests conducted by Rizki et al.5i the ethanolic extract of AMS showed an hypoglycemic effect, finding that with a dose of 400 mg/kg the blood glucose levels were significantly decreased in the test subjects. On the other hand, the extracts of AMS have also been evaluated in their effectiveness for the control of pests, which indirectly also affect human health, such is the case of the studies that corroborated the effect against Sitophilus zeamais, which often infest the stored rice,5j and in the cases of cabbage moth Plutella xylostella,5k and the aphid Brevicoryne brassicae, which affects crops of Brassica oleracea.5l The Annonaceae family is characterized, in general, as presenting an exclusive group of secondary metabolites of the family of AACGs, compounds which are also found in large numbers in the AMS.6e8a,8b The AACGs are an important group of long-chain FA derivatives, C-30 to C-37, usually with a terminal g-lactone, saturated or unsaturated. Sometimes the FAs are one to three rings of THF or THP, adjacent or not, which are occasionally replaced with epoxide rings or double bonds near the middle of the aliphatic chain. Accordingly, there can be 7 different types of terminal g-lactones (L-A to L-F), 10 skeletons with THF and/or THP (T-A to T-H, with 3 subtypes of T-G) and mono-, di-, or triepoxides.6-8a,8b Fig. 2.1 shows the general structure of an AACG. The THF ring system can be simple (one ring), adjacent (two rings), or not adjacent, with hydroxyl groups on both sides or not; this creates chiral centers in the molecules, making it very complex to elucidate the structures. Many AACGs isolated as pure compounds are

I. Overview and General Themes

18

2. Soursop Seed: Soursop (Annona muricata L.) Seed, Therapeutic, and Possible Food Potential

FIGURE 2.1 General structure of an AACG. The AACGs are an important group of long-chain FA derivatives, C30, C-32, or C-34, usually with a terminal g-lactone, saturated or unsaturated.

actually a mixture of diastereomers. However, the synthesis of some AACGs makes it possible to assign the chemical configurations of the corresponding natural AACGs unequivocally.8ae8c A little more than 400 compounds have been reported in the literature since the discovery of uvaricin in 1982 all from various genera exclusive to the Annonaceae family,7 although only some types of structures are present in the AACGs isolated from the AMS.3 To date, the AACGs that have been isolated from the seeds of soursop include a terminal g-lactone and THF in their structure; a few have epoxides, yet there are none with adjacent THP or THF.8a,8b More recently, annoreticuin-9-one,8d murisolin, cis-annoreticulin, and sabadelin, as well as a mixture of b-sitosterol and stigmasterol in a ratio of 1:3, were isolated from AMS.8e Various pharmacological investigations have shown that these metabolites possess antitumoral, antidiarrheal, larvicidal, antimalarial, pesticidal, and fungicidal activities, especially with regard to their antitumoral properties in vitro. Some are among the most potent inhibitors of complex I (NADH: ubiquinone oxidoreductase) in the system of mitochondrial electron transport between said complex and the NADH-oxidase in the plasma membrane characteristic of cancer cells; these actions induce apoptosis (programmed cell death), perhaps following the deprivation of ATP. The decrease in ATP is especially toxic to multidrug resistant tumor cells as well as to pesticide-resistant insects that possess ATP-dependent systems of xenobiotic flows, so said metabolites can be regarded as extraordinary antitumor agents and pesticides, especially for inhibiting resistance mechanisms requiring an ATPdependent flow.9a The AACGs may be among the most potent cytotoxic agents known, for example, trilobacin and asiminocin, AACGs isolated from Asimina triloba, have demonstrated values of DE50 < 10
12 mg/mL in several human tumor cell lines.9a A little over 40 AACGs showing cytotoxic activity have been isolated from the AMS, among which cis-annonacin may be mentioned as displaying a selective cytotoxicity toward colon adenocarcinoma cells (HT-29), 10,000 times more potent than that of adriamycin (IC50 ¼ 1.0  10
8 mg/mL, IC50 ¼ 5.1  10
4 mg/mL, respectively); cis-annonacin-10-one showed the same range as adriamycin toward the same type of cell line (IC50 ¼ 9.0  10
4 mg/mL).6 The acetogenin annoreticuin-9-one exhibited cytotoxic activity against human pancreatic tumor PACA-2 cell lines, human prostate adenocarcinoma PC-3, and lung carcinoma A-549.8d In addition, the anticancer and antitumor activity of AMS AACGs has been shown in cases of oral KB cancer, gallbladder tumor, and toxicity against human hepatoma cells.9b A recent study on the phytoestrogenic, hypocholesterolemic, and antioxidant activities demonstrated by the ethanolic extract of AMS suggests the possible utility of AMS to counteract or eliminate cancerous tumors given the close relationship between the levels of estrogen, cholesterol, and, of course, free radicals, with the incidence of cancer.9c I. Overview and General Themes

19

Applications in Health Promotion and Disease Prevention

The bioactivity possessed by these compounds has made it possible to obtain a patent for the isolation, identification, and antitumoral use of AACGs from the AMS, those include muricin A, B, C, D, E, F, and G, among which the most effective were muricin D (IC50 ¼ 6.6  10
4 mg/mL for Hep G2 and IC50 ¼ 4.8  10
2 for Hep 2,2,15); muricin F (IC50 ¼ 4.28  10
2 mg/mL Hep G2 and IC50 ¼ 3.86  10
3 mg/mL for Hep 2,2,15); and longifolicin (IC50 ¼ 4.04  10
4 mg/mL for Hep G2 and IC50 ¼ 4.9  10
3 mg/mL for Hep 2,2,15). However, both muricin A and muricin B showed selective activity toward Hep 2,2,15 cell line with IC50 ¼ 5.13  10
3 mg/mL and IC50 ¼ 4.29  10
3 mg/mL, respectively.10 Table 2.1 shows some of the AACGs isolated from the AMS with cytotoxic activity conform to the structure shown in Fig. 2.2; Table 2.2 presents some of the patented AACGs showing substituents according to Fig. 2.3, and Fig. 2.4 shows the muricin I structure that exemplifies AACGs found with double bonds. TABLE 2.1

Some AACGs With Biological Activity Isolated From AMS. x

y

z

R

R1

*

R2

References

Cis-Annonacin

2

2

5

OH, 4R

OH, 10R

19S

OH, 20S

6

Corossoline

2

2

5

H

OH, 10R

19R

OH, 20R

8b,10

Corossolone

2

2

5

H

¼O

19R

OH, 20R

8b,10

Longifolicin

2

1

6

H

OH, 10R

17R

OH, 18R

8b,10

Murisolin

2

2

5

OH, 4R

H

19S

OH, 20S

8b

Goniothalamicin

2

1

6

OH, 4R

OH, 10R

17S

OH, 18S

6

x, y, z: ethylene number in Fig. 2.2; R, R1, R2: substituents in Fig. 2.2.

FIGURE 2.2 Basic structure for AACG of Table 2.1. In the figure, when is substituted R, R1, and R2 by H/OH; and x, y, and z for 2, 2, and 5, respectively, it has the chemical structure of cis-annonacin. TABLE 2.2

Some Patented AACGs With Biological Activity Isolated From AMS (10). x

y

z

R

R1

R2

References

Muricatetrocin

6

1

9

OH, 19R

OH, 20R

H

8b,10

Muricin A

9

3

2

OH

OH

H

8b,10

Muricin C

11

1

4

OH

OH

H

8b,10

Muricin D

9

1

4

OH

OH

H

8b,10

Muricin E

6

2

5

H

OH

OH

8b,10

x, y, z: methylene and ethylene number in Fig. 2.3; R, R1, R2: substituents in Fig. 2.3.

I. Overview and General Themes

20

2. Soursop Seed: Soursop (Annona muricata L.) Seed, Therapeutic, and Possible Food Potential

FIGURE 2.3 Basic structure for AACG of Table 2.2. In the figure, when is substituted R, R1, and R2 by OH, OH, and H; and x, y, and z for 6, 1, and 9, respectively, it has the muricatetrocin A chemical structure.

FIGURE 2.4 Muricin I structure exemplifies AACGs found with double bonds.

In addition, the cyclopeptides annomuricatin A, annomuricatin B, and annomuricatin C have been isolated from AMS;11a however, X-ray diffraction studies showed that both annomuricatin A and annomuricatin C are the same compound.11b These types of compounds have demonstrated activity in other species of Annona11a and so has lectin that was found to possess inhibitory activity against the fungal pathogens Fusarium oxysporum, F. solana, and Colletotrichum musae, which can produce Fusarium wilt (Panama disease), keratomycosis, and anthracnose, in addition to red cell agglutinating activity.12 Accordingly, the AMS is a source of bioactive compounds (AACGs) with immense possibilities for pharmaceutical uses.

Annona muricata Seed and Its Possible Potential for Food Use The use of soursop fruit in the production of foodstuffs results in various so-called waste materials, one of which is the seeds. Studies of the AMS in its proximal composition have shown that it contains proteins, carbohydrates, ash, and oil in significant amounts. The reports are scarce and contain wide ranges of values that reflect the need for further studies regarding varieties and their degree of variability. Table 2.3 shows the composition and property ranges of AMS and its oil as reported in pertinent literature. There are few studies that have directly explored AMS possibilities as raw material in the food sector, mainly because there is knowledge of the existence of toxic compounds in it. Awan et al.,13 and particularly Fasakin et al.,14 analyzed the AMS with the purpose of highlighting the nutritional potential of the seed; other authors have studied it for the chemical and physical characteristics and the thermal and phase behavior of its oil,15e17a with the objective of contributing to knowledge about the reuse of soursop wastes or of characterizing some of its most important components.

I. Overview and General Themes

21

Annona muricata Seed and Its Possible Potential for Food Use

TABLE 2.3

Reported Composition and Property Ranges of AMS and Oil

Properties

Value Range

Units

References

Seeds in fruit

4.0e5.4

%, as-is basis

14,17a

AMS composition 14,17a,a

Moisture

2.17e34.6

Protein

2.4e27.3

Crude fiber

5.2e43.4

14,15,b

Ash

1.3e13.6

14,15,17c

Raw oil

18.3e37.7

13,17a,17c

Carbohydrates

11.5e34.1

17c,b

Polyphenols

3.1

mg GAE/g

17c

Myristic (C14:0)

0.06

% of total FA

c

Pentadecylic (C15:0)

0.4

d

Palmitic (C16:0)

19e25.5

16,17a

Palmitoleic (C16:1)

1.34e17.29

16,17a,d,e

Margaric (C17:0)

0.07e0.11

c,d

Stearic (C18:0)

3.3e7.90

16,17a,c,e

Oleic (C18:1)

39.18e44.0

16,17a,c

Linoleic (C18:2)

21.74e35.9

16,17ad,e

Linolenic (C18:3)

1.16

c

Arachidic (C20:0)

0.42e1.19

c,e

Gadoleic (C20:1)

0.07e0.16

17c,c

Gandoic (C20:1)

0.16

c

Behenic (C22:0)

0.08e0.10

17c,c

Lignoceric (C24:0)

0.10e0.15

17c,c

SFA

24e31.5

16,17a

UFA

68.5e76.82

16,17a,c

a-Tocopherol

12.5

% of kernel, db

14,15

AMSO composition

mg/kg oil

c

AMSO properties Specific gravity (20 C)

0.9281

c

Refraction index

1.451e1.468

15,17a,c

Saponification index

100e227

13,15 (Continued)

I. Overview and General Themes

22

2. Soursop Seed: Soursop (Annona muricata L.) Seed, Therapeutic, and Possible Food Potential

TABLE 2.3

Reported Composition and Property Ranges of AMS and Oildcont'd

Properties

Value Range

Units

References

Peroxide value

8.5

mEq O2/kg oil

c

Iodine value

87e111

13

3.13

c

Acid value 

SFC at 10 C

1.3

%

17a

Nzekwe ABC, Nzekwe FN. Proximate analysis & characterization of the seed and oil of Annona muricata (soursop). The Nigerian Journal of Research and Production. 2011;18:1e3. b Kimbonguila A, Nzikou JM, Matos L, et al. Proximate composition and physicochemical properties on the seeds and oil of Annona muricata grown in Congo-Brazzaville. Research Journal of Environmental and Earth Sciences. 2010; 2:13e18. c Elagbar ZA, Naik RR, Shakya AK, Bardaweel SK. Fatty acids analysis, antioxidant and biological activity of fixed oil of Annona muricata L. seeds. Journal of Chemistry. 2016; 1e6. https://doi.org/10.1155/2016/6948098. d Navaratne SB, Subasinghe JS. Determination of fatty acid profile and physicochemical properties of watermelon and soursop seed oils. European International Journal of Applied Science and Technology. 2014; 1:26e32. e Nwaehujor IU, Olatunji GA, Afolayan SS, et al. Physicochemical properties of the seed oil of Annona muricata grown in Ilorin, Kwara State. Equijost. 2016; 4:1e4. a

The chemical composition of the kernel seed, according to research conducted to date, may be interesting from the standpoint of food. For example, substances that are important for animal feed, such as protein and fiber, minerals and oil, can be found in the seed at levels from 2.4 to 27.3%, 5.2 to 43.4%, 1.3 to 13.6%, and 18.3 to 37.7%, respectively. In recent times, there has been extensive research to locate and expand nonconventional sources of fats and oils for the purpose of finding applications for them in various fields. The AMS has an important amount of oil. Composed of oleic (39.18e53.92%), linoleic (21.74e35.90%), palmitic (16.39e25.50%), and stearic (3.30e7.90%) as its main FAs, it looks like the oils from conventional oilseeds. The reported values for its refraction index, saponification index, and iodine value range from 1.451 to 1.468, 100 to 227, and 87 to 111, respectively, and studies of its phase and thermal behavior have showed that it has the characteristics of common table or cooking oils; the yellowish oil remains liquid at warm ambient and refrigeration temperatures, its solid fat contents at 10 C being around 1% (see Table 2.3 for references). Even with all these considerations, it is clear that any application of the AMS or its derived components must take into account the characteristic toxicity of raw AMS. Notwithstanding the bromatological characteristics of the seed and its significant oil yield and physicochemical properties, its use for human consumption should be viewed with caution. The usability of AMS and its oil undoubtedly require increased knowledge, particularly in regard to food applications. The reviewed bioactive compounds present in the seed are of toxic and pharmacological importance; consequently, they limit the current use of raw AMS in foods. The processing of AMS to isolate and obtain components of interest to the food industry, such as proteins, oil, or otherwise, also requires more and deeper studies aimed at extraction, purification, toxicological and nutritional evaluations, etc. In this regard, recently AMSO has been used experimentally in the treatment of skin wounds, with interesting results in its positive effect on the healing process.17b Pinto et al.17c studied a method to extract AMSO and reduce its toxicity, using a mixture of chloroformemethanol, of which they were able to obtain a solid fraction of a toxic precipitate, presumably with the majority of AACGs, and a supernatant corresponding to the oil. I. Overview and General Themes

Summary Points

23

The AMSO thus separated from that solid phase and obtained through this method showed no toxicity, but a hepatoprotective effect, through in vivo evaluations with mice. It had an FA profile similar to that reported by other authors in AMS extractions with other organic solvents, maintaining the perspective of its possible use as food with the health benefits because of its high content of monounsaturated and polyunsaturated FA. On the other hand, the possible presence of active compounds such as the AACGs and others from AMS in the crude oil opens the possibility too, to be used in insecticide and therapeutic applications, such as diabetes type 1, among others, in accordance with the proven activity of these compounds. 5l,17c,17d

Adverse Effects and Reactions, Allergies, and Toxicity Soursop seed is not edible and traditionally has been known to be toxic because of the presence of the described compounds; therefore, its components and derivatives destined for human consumption must be subject to careful research protocols to ensure their toxicological safety. Even when it is used as a drug, in its whole form or from crude extracts, it must be handled with care because some of its bioactive components have not been fully evaluated for their allergenic or toxicological cumulative effect. One example is annonacin, which has been isolated from most parts of the plant, including seeds and fruit pulp. It has been shown by clinical studies with rats and epidemiological studies in human populations (like those from the Caribbean island of Guadeloupe and those from New Caledonia) with high and prolonged intakes of soursop fruit pulp that annonacin is associated with the occurrence of a neurodegenerative disease known as atypical parkinsonism. This finding, together with the known effects of the AACG on mitochondrial toxicity, makes the consumption of the AMS, its concentrates, and derivates without purification a risky matter.18e20 The latter, notwithstanding the research efforts to evaluate the safety of some fractions of AMS for food purposes, as in the case of AMSO, it is considered that the results to date are not conclusive, yet.

Summary Points • Soursop, Annona muricata L., is a plant native to South America and is widely cultivated throughout the tropical regions of the world. • A. muricata L. has shown pharmacological activity in its different parts. • Annona muricata seeds make up about 5% of the fruit and are now considered an industrial processing waste. • Annonaceous acetogenins are the most studied bioactive metabolites from Annona muricata seeds and they have high cytotoxic activity with potential therapeutic applications. • Other important components of the Annona muricata seeds have also been studied from a food perspective. • Annona muricata seeds have a significant content of oil which has a composition and properties similar to those oils of conventional sources. • Any component derived from Annona muricata seeds to alimentary consumption must be previously studied about their possible toxicological risk.

I. Overview and General Themes

24

2. Soursop Seed: Soursop (Annona muricata L.) Seed, Therapeutic, and Possible Food Potential

References 1. Morton J. Soursop. In: Curtis FD, Morton JF, eds. Fruits of Warm Climates. Miami, FL: Florida FlairBooks; 1987:75e80. 2. Janick J, Paull RE. The Encyclopedia of Fruit & Nuts. Reading UK: Cambridge University Press; 2008. 3. Johnson HA, Oberlies NH, Alali FQ, McLaughlin JL. Thwarting resistance: Annonaceous acetogenins as new pesticidal and antitumor agents. In: Cutler SJ, Cutler HG, eds. Biologically Active Natural Products: Pharmaceuticals. Boca Raton, FL: CRC Press; 2000:181e191. 4. Leatemia JA, Isman MB. Insecticidal activity of crude seed extracts of Annona spp., Lansium domesticum and Sandoricum koetjape against lepidopteran larvae. Phytoparasitica. 2004;32:30e37. 5a. Bobadilla M, Zavala F, Sisniegas M, et al. Evaluación larvicida de suspensiones acuosas de Annona muricata Linnaeus “guanábana” sobre Aedes aegypti Linnaeus (Diptera, Culicidae). Revista Peruana de Biologia. 2005;12:145e152. 5b. Grzybowski A, Tiboni M, Silva M, et al. Synergistic larvicidal effect and morphological alterations induced by ethanolic extracts of Annona muricata and Piper nigrum against the dengue fever vector Aedes aegypti. Pest Management Science. 2013;69:589e601. 5c. Komansilan A, Abadi AL, Yanuwiadi B, Kaligis DA. Isolation and identification of biolarvicide from soursop (Annona muricata Linn) seeds to Mosquito (Aedes aegypti) larvae. International Journal of Engineering and Technology IJET-IJENS. 2012;12(3):28e32. 5d. Ranisaharivony BG, Ramanandraibe V, Rasoanaivo LH, et al. Separation and potential valorization of chemical constituents of soursop seeds. Journal of Pharmacognosy and Phytochemistry. 2015;4(2):161e171. 5e. Raveloson Ravaomanarivo LH, Razafindraleva HA, Fara Nantenaina Raharimalala FN, et al. Efficacy of seed extracts of Annona squamosa and Annona muricata (Annonaceae) for the control of Aedes albopictus and Culex quinquefasciatus (Culicidae). Asian Pacific Journal of Tropical Biomedicine. 2014;4(10):798e806. 5f. Boyom FF, Fokou PVT, TchokouahaYamthe LR, et al. Potent antiplasmodial extracts from Cameroonian Annonaceae. Journal of Ethnopharmacology. 2011;134:717e724. 5g. Vila-Nova NS, de Morais SM, Cajazeiras Falcão MJ, et al. Leishmanicidal activity and cytotoxicity of compounds from two Annonacea species cultivated in Northeastern Brazil. Revista da Sociedade Brasileira de Medicina Tropical. 2011;44(5):567e571. 5h. Vila-Nova NS, de Morais SM, Cajazeiras Falcão MJ, et al. Different susceptibilities of Leishmania spp. promastigotes to the Annona muricata acetogenins annonacinone and corossolone, and the Platymiscium floribundum coumarin scoparone. Experimental Parasitology. 2013;133:334e338. 5i. Rizki Hasanah RU, Sundhani E, Nurulita NA. Effect of ethanolic extract of Annona muricata L seeds powder to decrease blood glucose level of Wistar male rats with glucose preload. Proceeding ICMHS. 2016:112e115. 5j. Asmazar AD, Idris BA. Evaluation of Jatropha curcas and Annona muricata seed crude extracts again Sitophilus zeamais infesting stored rice. Journal of Entomology. 2012;9(1):13e22. 5k. Barros Gomes I, Predes Trindade RC, Goulart Sant’Ana AE, et al. Bioactivity of microencapsulated soursop seeds extract on Plutella xylostella. Ciência Rural, Santa Maria. 2016;46(5):771e775. 5l. da Paz LC, Leal Soares AM, Oliveira Teixeira RR, et al. Toxicity of the organic extract from Annona muricata L. (Annonaceae) seeds on Brevicoryne brassicae L. (Hemiptera: Aphididae) in cabbage cultivation (Brassica oleracea L.). Ciência Agrícola, Rio Largo. 2018;16(1):55e60. 6. Rieser MJ, Gu Z-M, Fang X-P, et al. Five novel mono-tetrahydrofuran ring acetogenins from the seeds of Annona muricata. Journal of Natural Products. 1996;59:100e108. 7. Zafra-Polo MC, Figadère B, Gallardo T, et al. Natural acetogenins from annonaceae, synthesis and mechanisms of action. Phytochemistry. 1998;48:1087e1117. 8a. Bermejo A, Figadere B, Zafra-Polo MC, et al. Acetogenins from Annonaceae: recent progress in isolation, synthesis and mechanisms of action. Natural Product Reports. 2005;22:269e303. 8b. Liaw C-C, Liou J-R, Wu T-Y, et al. Acetogenins from annonaceae. In: Kinghorn AD, Falk H, Gibbons S, Kobayashi J, eds. Prog Chem Org Nat Prod. Vol. 101. Springer International Publishing Switzerland; 2016:113e280. 8c. Quinn KJ, Islamaj L, Couvertier SM, et al. Convergent total synthesis of Murisolin. European Journal of Organic Chemistry. 2010:5943e5945. 8d. Ragasa CY, Soriano G, Torres OB, et al. Acetogenins from Annona muricata. Pharmacognosy Journal. 2012;4:32e37.

I. Overview and General Themes

References

25

8e. Ragasa CY, Galian RF, Shen C-C. Chemical constituents of Annona muricata. Der Pharma Chemica. 2014;6:382e387. 9a. Alali FQ, Liu X-X, McLaughlin JL. Annonaceous acetogenins: recent progress. Journal of Natural Products. 1999;62:504e540. 9b. Patel MS, Patel JK. A review on a miracle fruits of Annona muricata. Journal of Pharmacognosy and Phytochemistry. 2016;5:137e148. 9c. Abiola T, Kings AT, Akinosho O. Phytochemical screening and evaluation of the phytoestrogenic, hypocholesterolemic and antioxidant activity of ethanolic extract of soursop (Annona muricata) seeds in DMBA-treated female Wistar rats. Biochemistry and Physiology. 2018;7:232. https://doi.org/10.4172/2168-9652.1000232. 10. Wu Y-C. Cytotoxic annonaceous acetogenins from Annona muricata. Patent No. US. 2007, 7,223,792, B2. 11a. Wélé A, Zhang Y, Caux C, et al. Annomuricatin C, a novel cyclohexapeptide from the seeds of Annona muricata. Comptes Rendus Chimie. 2004;7:981e988. 11b. Wu L, Lu Y, Zheng Q-T, et al. Study on the spatial structure of annomuricatin A, a cyclohexapeptide from the seeds of Annona muricata. Journal of Molecular Structure. 2007;827:145e148. 12. Damico DCS, Freire MGM, Gomes VM, et al. Isolation and characterization of a lectin from Annona muricate seeds. Journal of Protein Chemistry. 2003;22:655e661. 13. Awan JA, Kar A, Udoudoh PJ. Preliminary studies on the seeds of Annona muricata. Plant Foods for Human Nutrition. 1981;30:163e168. 14. Fasakin AO, Fehintola EO, Obijole OA, Oseni OA. Compositional analyses of the seed of sour sop, Annona muricata L., as a potential animal feed supplement. Scientific Research and Essays. 2008;3:521e523. 15. Onimawo IA. Proximate composition and selected physicochemical properties of the seed, pulp and oil of sour sop (Annona muricata). Plant Foods for Human Nutrition. 2002;57:165e171. 16. Ocampo S, Diana M, Betancur J, et al. Estudio cromatográfico comparativo de los ácidos grasos presentes en semilla de Annona cherimolioides y Annona muricata L. Vector. 2007;2:103e112. 17a. Solís-Fuentes JA, Amador-Hernández C, Hernández-Medel MR, Durán-de-Bazúa MC. Caracterización fisicoquímica y comportamiento térmico del aceite de “almendra” de guanábana (Annona muricata, L). Grasas y Aceites. 2010;61:58e66. 11b. Cardoso de Sousa R, Teixeira Ferreira T, Lustosa Barros EM, et al. Comparative macroscopic study between oil extracted Annona seed AlGaInP laser and therapeutic ultrasound for skin wounds. International Journal of Pharmaceutical Science Invention. 2016;5:36e41. 11c. Pinto LC, Souza CO, Souza SA, et al. Potential of Annona muricata L. seed oil: phytochemical and nutritional characterization associated with non-toxicity. Grasas y Aceites. 2018;69(1):e234. https://doi.org/10.3989/ gya.0777171. 11d. Pinto LC, Cerqueira-Lima AT, dos Santos Suzarth S, et al. Anonna muricata L. (soursop) seed oil improves type 1 diabetes parameters in vivo and in vitro. Pharma Nutrition. 2018;6(1):1e8. 18. Liaw C-C, Chang F-R, Lin C-Y, et al. New cytotoxic monotetrahydrofuran annonaceous acetogenins from Annona muricata. Journal of Natural Products. 2002;65:470e475. 19. Quispe A, Zavala D, Rojas J, et al. Efecto citotóxico selectivo in vitro de muricin H (acetogenina de Annona muricata) en cultivos celulares de cáncer de pulmón. Revista Peruana de Medicina Experimental y Salud Pública. 2006;23:265e269. 20. Escobar-Khondiker M, Höllerhage M, Muriel MP, et al. Annonacin, a natural mitochondrial complex I inhibitor, causes tau pathology in cultured neurons. Journal of Neuroscience. 2007;27:7827e7837.

I. Overview and General Themes

C H A P T E R

3

Red Horse-Chestnut Seeds of Aesculus 3 Carnea: A New Way for Health and Food Design? Cecilia Baraldi1, Giorgia Foca2, 4, Laura Maletti3, Andrea Marchetti3, 4, Fabrizio Roncaglia3, Simona Sighinolfi3, Lorenzo Tassi3, 4 1

Department of Life Sciences, University of Modena and Reggio Emilia, Modena, Italy; Department of Life Sciences, University of Modena and Reggio Emilia, Reggio Emilia, Italy; 3 Department of Chemical and Geological Sciences, University of Modena and Reggio Emilia, Modena, Italy; 4Interdepartmental Research Center BIOGEST-SITEIA, University of Modena and Reggio Emilia, Reggio Emilia, Italy 2

List of Abbreviations A Aesculus AH Aesculus hippocastanum AP Aesculus pavia AXC Aesculus 3 carnea CWS Cold water solubility SEM-EDS Scanning electron microscopy-energy dispersive X-ray spectroscopy TISS Total inorganic soluble salts

Introduction The information available today on Aesculaceae seeds, and their derived products, seems to be considerable but scattered in various literature frameworks, or often it appears underexploited from chemistry characterization, occurrence in foods, biological activity, and health effects. Bearing this in mind, some years ago we moved and devoted some attention to these very common seeds, produced in large quantities and apparently of little interest to most people.

Nuts and Seeds in Health and Disease Prevention, Second Edition https://doi.org/10.1016/B978-0-12-818553-7.00003-6

27

Copyright © 2020 Elsevier Inc. All rights reserved.

28

3. Red Horse-Chestnut Seeds of Aesculus 3 Carnea: A New Way for Health and Food Design?

In our previous study about chemical composition of the seeds of horse-chestnuts from Aesculus hippocastanum (AH), two specimens were selected: the “Pure” species with white flowers (AHP) and an “Hybrid” one, more diffused in our countries (Modena, Italy), with soft pink flowers (AHH).1,2 The issues seem relevant to some extent for other researchers.3 Looking at the literature, in relation to these topics, it seems to us that there is a lack about analogues investigation on the characteristics of the seeds of another common variety pertaining to the same family of Sapindaceae: Aesculus 3 carnea (AXC). This species gives a spectacular spring flowering with panicle inflorescence of bright red color. For this reason, it is commonly identified as “red horse-chestnut” tree. For example, the flowering of the avenues of the Vienna Prater offers to the visitor an incomparable setting, a magnificent spectacle that cannot be forgotten. His widespread as ornamental shade is mainly because that it is tolerant to drought, wind, and salt and resists very well to the heat of the southern countries. It holds up well in urban areas, even in restricted and compacted soil spaces. The nuts make good nourishment for wildlife and have some other specific uses pertaining to the popular traditions and alternative medicine. So we have looked at the specific literature devoted to herbal medicine and healthy benefits for human consumption of various products as nutritional supplements of vegetable origin. Furthermore, there are many other ethnographic aspects involving different uses as foodstuffs, textiles dyes, crafts materials, etc., as well as tools and technologies related to the socioeconomic and anthropological context in which these plants are used.4 As a consequence, it seems to us that the “state of the art” on the chemical knowledge about the composition of red horse-chestnut can be traced as follows. Some decades ago, Ju-Ying Hsiao et al.5 reported some chromatographic results obtained by studying the tannic fraction related to different vegetative districts, obtained by European AH, the American AP, and AXC, in order to establish some relationships among them. Twodimensional chromatographic separation of the phenolic compounds and characterization of spots was made on leaf and flower material of the three species concerned. Chromatograms of flower extracts strongly supported the idea that AP is one of the parent species. Leaf chromatograms support AH as the other parent species. In fact, the identification of the three major spots of leaves of AH and AXC reveal them as quercetin-3-O-rhamnoside, quercetin-3O-glucoside, and quercetin-3-O-arabinoside. Some very interesting results have been obtained by Foo and Porter,6 who extensively studied the phytochemistry of proanthocyanidin polymers (condensed tannins) in many plants by applying 13C NMR techniques. The proanthocyanidins from AXC and AH unripe fruits both include the unusual doubly linked dimer proanthocyanidin A2 type, and trimers of a similar structure. Furthermore, an average molar mass (MW) for proanthocyanidins of 2200 D (AXC) and 1750 D (AH) for the two cultivars, respectively, was estimated. No products consistent with the presence of proanthocyanidin A2 type structures were detected on acid degradation or by degradation induced by other reagents in mildly acid conditions. Their results suggest that proanthocyanidin A2 type is a metabolic curiosity and its natural formation is subsidiary to the main biosynthetic pathway leading to polymers. As a matter of fact, there are many observations made on the fate of tannins during the aging or maturing of plant tissue, especially with respect to the fruit ripening. It has been observed, quite commonly, that the concentration of proanthocyanidin polymers drops as

I. Overview and General Themes

Historical Cultivation and Usage

29

a fruit matures,7 the MW of tannins increases, and as a consequence, their solubility and hence astringency decreases. Williams et al.8 also investigated the MW profiles of proanthocyanidin polymers from many samples representing a wide range of plant tissues of many different species, by applying an alternative methodology that utilizes gel permeation chromatography of the peracetate derivatives. So the polymers extracted from AXC have been sampled over several seasons at different stages of maturation, and the observed composition (Mn, stereochemistry) has remained remarkably constant, with an average MW y 4200 D, sensibly far from the datum of Foo and Porter related to unripe fruits.7 Some further investigations about amino acids contents and RNA genotypes of Aesculaceae species are due to Fowden et al.,9 while Platt et al.10 commented about the biosynthesis of procyanidins, flavan-3-ols, and other secondary products from the biocycle involving phenylalanine. After this, we can conclude that there is very little information of scientific nature related to this species, so we decided to provide some further investigations on chemical composition of red horse-chestnut seeds.

Botanical Descriptions Division: Magnoliophyta (angiosperms) Class: Magnoliopsida Order: Sapindales Family: Sapindaceae (formerly Hippocastanaceae) Genus: Aesculus Species: Aesculus 3 carnea The subfamily of Hippocastanaceae (genus: Aesculus) comprises 20 species, whose habitat is mainly represented by the temperate zones of the northern hemisphere. Among these species, the A. chinensis and the A. turbinata Blume (Japanese horse-chestnut) seem to be the ones mostly widespread and known in the Eurasian continent, while the A. pavia is probably the most common in North America. From the physiological point of view, AH tolerates low temperatures, prefers fruitful soils, and is less resistant toward the atmospheric contaminants. Unfortunately, a new parasite called Cameraria ohridella (a lepidopter, an exclusive leaf miner for AH) firstly observed in Macedonia about 30 years ago is seriously responsible for the massive attack to AH, in particular toward the white flower species, being the cause of dry leaves and defoliation at the beginning of summer.11

Historical Cultivation and Usage The plant’s origins of AXC, commonly named red horse-chestnut, have been the subject of much academic discussions, heated disputes, and controversies by botanical researchers, but regardless, it remains a wonderful tree for amazing viewings of relaxing landscapes. Most experts believe it to be a hybrid across between common horse-chestnut (AH) originated

I. Overview and General Themes

30

3. Red Horse-Chestnut Seeds of Aesculus 3 Carnea: A New Way for Health and Food Design?

from BalkaneCaucasian regions and red buckeye (AP) originated from North America. The nickname buckeye was attributed by Native Americans because of the similarity of the seeds to the deer’s eyes. Literature and historical notices recognition indicate that the first red horsechestnut appeared in Germany around 1820, from where they spread throughout Europe. The German population of Bavarian region has always shown great passion and respect for the large trees of common AH. In fact, AH trees were planted near the underground caves and cellars where the beer barrels were kept, with the aim of keeping the cells shaded and isolated to refresh the beer especially during the summer. Probably, the Germans who appreciated the rich colored blooms of AP were true pioneers and were the first to hybridize the seeds of AH with those of the ornamental bushes of American buckeye. In this way, it was possible to obtain a new cultivar, the perfect synthesis of genetic characteristics capable of magnifying and enhancing the best qualities of both precursors: giant plants with large canopies and gorgeous clusters of scented-colorful flowers. In 1858, to honor Pierre Louis Briot, a famous French nurseryman, the variety ruby red horse-chestnut AXC ‘Briotii’ was named, probably the most common and widespread, whose flowering is very showy due to the large panicles of deep pink-red color. Although AXC is a hybrid species, the plants produce viable seeds for natural propagation, an oddity for most hybrids.

Present-Day Cultivation and Usage In the European countries, some Aesculaceae varieties comprising AXC are largely diffused and commonly cultivated throughout the territories as ornamental shade trees, up to over 1000 m altitude. However, this genus does not show the tendency to grow wild and do not mix easily with the native flora. It should be planted in large spaces, both as a single specimen and for the formation of avenues. While the precursors AH and AP over the years have suffered the pandemics of leaf miner (Cameraria ohridella) and anthracnose (Guignardia aesculi), it was observed that new plantations made with AXC develop more and more resistance against leaf myopathies and parasitic attacks. This behavior is probably due to the increased content of tannins and other toxic chemical species present in each plant district such as leaves, flowers, fruits, nuts, bark, and wood.11 All the parts of the plants belonging to the genus Aesculus were used for the formulation of medicinal preparations for veterinary use and health products of popular use: decoctions obtained from leaves and seeds as cardiotonic and anti-inflammatory, those from bark and wood chips as febrifuge, and extracts and poultices used to treat dermatitis and psoriasis of various origins. Not only the seeds and the fruits are known for their bitterness but also the leaves and flowers are just as unpleasant. All these vegetative districts contain a great variety of bioactive compounds, called escin, among which a particular glucoside stands out, known as esculin. This compound has been widely characterized and studied from a chemical, clinical, and toxicological point of view, including its mechanism of action at the cellular level.12 If this chemical is ingested in a concentration higher than the useful pharmacological dosage, it may produce some counter-effects that reach the maximum in the case of severe gastroenteritis due to its low absorbability in the blood stream.

I. Overview and General Themes

Red Horse-Chestnuts: Chemical Composition and Characterization

31

However, both esculin (pure compound) and aescin (a multicomponent mixture) are used in homeopathic medicine and in official pharmacology. Esculin is used as a fluidifier of the blood tissue, reducing its viscosity, thus favoring venous transit to the heart. Used in conjunction with escin, it strengthens the venous tissues, preserves their elasticity, and is used to treat varicose veins and other phlebitis-related diseases. It is a drug with pain-relieving effects, and it can prevent the formation of possible clots, but due to the astringent effects on the venous tissues, it could occasionally lead to stroke or other heart problems. To improve tolerability, other natural extracts based on tonic herbs are added to medicinal formulas, in order to minimize and counteract the negative effects. Even today the most significant use is reserved for the seeds of Aesculaceae, with which herbal remedies, pharmaceutical formulations, cosmeceutical preparations, and commodities such as shampoos, shower gel, creams, lotions, sun products, dermoprotectives, and toothpastes are prepared.13 Dried and stored together with common foods, they slowly release aromas that can remove some infesting parasitic forms, such as food moths. Seeds flour is not immediately edible for humans due to the strong bitterness and significant toxicity of the escin fractions. However, substances such as esculin are thermolabile, and after roasting, the seeds can be used as coffee substitutes. The saponins may be extracted from the flour seeds in hot water, being complex mixtures of natural surfactant compounds with marked antimicrobial and bacteriostatic properties. The starchy residue, after multiple extractions and drying, becomes edible.

Red Horse-Chestnuts: Chemical Composition and Characterization A literature survey of the last two decades about the potential applications of the seeds of genus Aesculus provides a number of valuable scientific papers, in many cases, giving standpoints reports of excellent quality. However, in spite of the increasingly interest from the researchers working in different fields, the basic knowledge on chemical characterization is nowadays quite sparse and incomplete. To partly reduce such fragmented information, the AXC nuts harvested in our campus of Modena University (October 2017) have been studied, and a set of new original data will appear on this paper. The composition of AH seeds can be summarized in the following main classes of components: -

starch and non-starch saccharides;14 aminoacids9 and proteins;15 lipids (fats and essential oils); minerals (ashes).

In addition, among other minor compounds, the following classes and species are present, as the most important bioactive ones: - escin (saponin and sapogenin fractions), which is the most abundant;16 - coumarin-derived compounds (esculin, fraxin, scopolin, pavietin, among others);17 - tannins (antocyanidine, leucocyanidine, proanthocyanidin A2);18

I. Overview and General Themes

32

3. Red Horse-Chestnut Seeds of Aesculus 3 Carnea: A New Way for Health and Food Design?

- non-protein nitrogen compounds (i.e., adenine, adenosine, guanine, uric acid); and - vitamin-B complex, methionine, and holine e sterols.19 Escin is a generic mixture of saponins (triterpenoid glycosides)20 and sapogenins, the latter family being more complex, having two molecular groups that show a different basic structure, triterpenic and steroydic ones, variously substituted. Typically, different classes of the main bioactive principles are present in different organs and vegetative districts of Aesculaceae: lipids and escin fractions predominate in the seeds, essential oils in the leaves and flowers, and tannins and coumarin-derived compounds in the bark. In addition, the crude extracts of the seeds contain significant amounts of some other families of different compounds, such as condensed tannins (mainly flavonoids) and sterols.19 A significant literature contribution about the chemical composition of AH chestnut was given by Kapusta et al.,18 tracing the flavonoid profile of the seeds and the wastewater obtained as by-product in the industrial processing of AH seeds. It was concluded that flavonoids present in these fractions can be safely used to obtain quercetine and kaempferol glycoside for cosmetic, nutraceutical, and supplement industries.

Proximate Analysis Red horse-chestnut samples were analyzed for chemical composition (water content of fresh seeds, residual moisture, crude proteins, crude fats, glucides, and ashes) referring to the AOAC procedures.1 In addition to elemental analysis, the total nitrogen content was measured for each sample, by adopting a standard procedure (Kjeldahl method). Lacking specific literature targets, and taking advantage from the analogy with other natural products such as chestnuts, the percentages of nitrogen was transformed into crude protein content by multiplying by a conversion factor of 4.86, in agreement with the data reported in our previous study.1 Cold water solubility (CWS) and the relevant total inorganic soluble salt (TISS) content were determined as previously described.1 The quantitative values of proximate analysis are summarized in Table 3.1, where a stringent comparison has been made with our previous data obtained by studying AHP and AHH species. As can be seen, the water content of horse-chestnut kernels follows the same trend traced by the previous data: all three cultivars had equivalent high moisture content at the harvesting time. The values for residual moisture, after natural desiccation, were 6.59 (AHH), 6.97 (AHP), and 10.4% (AXC), while the values for ash contents were 2.19, 2.51, and 3.14% on dry basis for the three varieties, respectively. As an evidence, there is a straight parallelism between these two last data series, with a significant difference in the residual moisture and ash contents for the three cultivars, with AXC kernels generally over of about þ50% with respect to the other two comparing species, and independently of the nature of the data. The CWS represents an undifferentiated mass of hydrophilic organic molecules, and other inorganic species, swollen by granules of floured sample and transferred to liquid phase as solubleenon-sedimented material. This character should mainly depend on the nature of the starch, others carbohydrates sources (glucides), hydrophilic protein fraction, particulate dimensions, and the granular structure of substrate. CWS data differ significantly for AXC

I. Overview and General Themes

33

Red Horse-Chestnuts: Chemical Composition and Characterization

TABLE 1

Chemical composition data determined on horse-chestnut samples of different botanical origins (AHP, AHH, AXC) from Modena city and comparison with other literature data. a AHP (pure, white flowers)

AHH (hybrid, pink flowers)

AXC (red flowers)

Aesculus indica21

Humidity % (fresh seeds)

50.8  0.6

50.1  0.6

50.2  0.7

50.5

Moisture % (residual)

6.97  0.35

6.59  0.24

10.4  0.31

Proteins % (d.s.)

2.64  0.42

1.82  0.34

3.16  0.40

Lipids % (d.s.)

4.13  0.36

5.10  0.50

4.34  0.42

0.77

Glucids % (d.s.)

15.2  0.66

14.3  0.80

15.6  0.72

11.0

Ashes % (d.s.)

2.51  0.12

2.19  0.09

3.14  0.10

3.83

Cold water solubility % (d.s.)

53.9  0.79

48.6  0.65

55.1  0.62

Total inorganic soluble salts % 2.18  0.10 (d.s.)

1.92  0.11

2.79  0.15

Elemental Analysis N%

0.90  0.10

1.39  0.11

1.49  0.10

C%

43.02  0.27

43.19  0.22

44.82  0.23

H%

5.45  0.12

6.05  0.15

6.25  0.14

S%

0.13  0.06

0.11  0.06

—

(d.s.) ¼ , dry sample. The table summarizes some experimental data representing the main classes of components of Aesculaceae seeds, together with some relevant parameters related to the soluble fractions. a Uncertainties are expressed as standard deviation of five samples (s5).

(55.1%) with respect to AHP (53.9%) and AHH (48.6%) samples. Probably, as a rational hypothesis, the higher solubility of AXC samples could be attributed to some concomitant effects, two of them in particular: i) the higher content of hydrophilic and hydrocolloid species and ii) the less rigid structure of the starch granules related to a greatest swelling ability. In addition, SEM micrograph analyses showed some significant morphological differences on the actual AXC samples, and the previously studied specimens AHP and AHH.1 TISS values are perfectly coherent to the ash content (Table 3.1). Furthermore, we note that TISS on AXC samples represents the 88.9% of the ashes, while the level slightly decreases for AHH (87.7%) and AHP (86.8%) samples. Elemental analysis shows a higher content of N% for AXC (1.49%) with respect to both AHP (0.90%), with an increase equivalent to þ65%, and AHH (1.39%), equivalent to about þ7%. These values follow the trend of the protein fraction, obviously related to N% content, but determined by another way. The C% content also is generally higher for AXC (44.82%) than the other two AHP (43.02%) and AHH (43.19%) samples. Unfortunately, S% is not detectable in AXC specimen. However, this does not imply that S is quite absent in this matrix, but rather that the content should be below the detection limit of instrument (w0.1%).

I. Overview and General Themes

34 TABLE 2

3. Red Horse-Chestnut Seeds of Aesculus 3 Carnea: A New Way for Health and Food Design?

Fatty acid content on horse-chestnut samples of different botanical origin (AHP, AHH, AXC) from Modena city, compared with literature data for other species. a

Acid

AHP (white flowers)

AHH (pink flowers)

AXC (red flowers)

Myristic (C14 : 0)

0.6  0.3

0.6  0.2

—

Myristoleic (C14 : 1)

0.2  0.1

0.4  0.1

—

Pentadecanoic (C15 : 0)

0.1  0.1

0.2  0.1

—

Pentadecenoic (C15 : 1)

0.1 0.1

0.1  0.1

—

Palmitic (C16 : 0)

7.1  0.5

6.8  0.6

7.7  0.5

Palmitoleic (C16 : 1)

0.7  0.3

0.9  0.3

0.1  0.1

Heptadecanoic (C17 : 0)

0.1  0.1

0.4  0.2

0.2  0.1

Heptadecenoic (C17 : 1)

0.1  0.1

0.3  0.1

—

Stearic (C18 : 0)

0.8  0.2

2.9  0.3

1.7  0.4

Oleic (C18 : 1)

43.2  1.1

49.7  1.3

32.6  1.4

Linoleic (C18 :2)

35.2  1.2

23.0  0.8

33.8  1.4

Linolelaidic (C18 : 2)

2.2  0.4

2.1  0.2

—

Linolenic (C18 : 3)

5.9  0.5

7.5  0.5

7.9  0.7

Arachidic (C20 : 0)

0.2  0.1

0.5  0.2

0.7  0.2

Eicosenoic (C20 : 1)

—

—

8.8  0.8

Behenic (C22 : 0)

0.1  0.1

0.3  0.1

—

Erucic (C22 : 1)

2.9  0.3

4.1  0.2

6.5  0.6

Total saturated FA (SFA)

9

11.7

10.3

Total unsaturated FA (UFA)

90.5

88.1

89.7

Total monounsaturated FA (MUFA)

47.2

55.5

48

Total diunsaturated FA (DUFA)

37.4

25.1

33.8

Total three-unsaturated FA (TUFA)

5.9

7.5

7.9

Aesculus indica22

65/70

The table summarizes some experimental data representing the fatty acid content of Aesculaceae seeds of different cultivars, in comparison to relevant parameters from literature, related to another botanical variety. a Uncertainties are here expressed as standard deviation of five samples (s5).

Lipids Analysis The values for crude fats content previously reported for horse-chestnut kernels were 4.13% (AHP) and 5.10% (AHH) on dry sample basis.1 The present study indicates for the cultivar AXC a crude fat content of 4.34%, similar and intermediate to the values found in our previous investigation. However, the current results about the AXC lipid fraction composition (Table 3.2) reveal some peculiarities in comparison with our previous studies on AHP and AHH species

I. Overview and General Themes

Red Horse-Chestnuts: Chemical Composition and Characterization

35

50 45 40 35 30 25 20 15 10 5 tad c Pe ecan nt o ic ad ec en oi Pa c lm iti Pa c lm Ep itol e tad ic e Ep cano tad ic ec en oi c St ea ric Ol ei Li c no l Li no eic lel aid ic Li no len ic Ar ac h Ei idic co se no ic Be he ni c Er uc ic

lei

iri

Pe n

iri M

M

sto

sti c

0

AXC

AHP

AHH

FIGURE 3.1 Fatty acid content on horse-chestnut samples of different botanical origin (AHP, AHH, AXC) from Modena city. For a quick comparison, the figure shows the histogram of the experimental values related to the fatty acid content in the samples extracted from the Aesculaceae seeds of different cultivars.

(Fig. 3.1). We observe that (i) the AXC chromatogram of fatty acids shows fewer peaks others than two varieties of Aesculaceae previously studied; (ii) the small C(14) and C(15) components are quite absent in AXC, while they are present to some extent in AHP and AHH extracts; (iii) on the contrary, AXC contains a significant amount of eicosenoic acid (C20:1), estimated at about 9%, which is quite absent in AHP and AHH; (iv) the saturated fatty acids (SFA) fraction is about 10e11% for the three cultivars, while the total amount of unsaturated species is 89e90%, respectively. Horse-chestnut crude lipids show three predominant unsaturated fatty acids, namely oleic (C18:1), linoleic (C18:2), and linolenic (C18:3) ones, which account for 75e85% of the total fatty acid contents.1 Monounsaturated fatty acid (MUFA) and polyunsaturated fatty acids (PUFA) (Fig. 3.2) have been associated with positive health effects. Literature provides panoramic reviews giving dietary recommendations for u-3 fatty acids, including linolenic acid and parent species, to achieve nutrient adequacy and to prevent and treat cardiovascular

FIGURE 3.2

Total fatty acid fraction content % on horse-chestnut samples of different botanical origin (AHP, AHH, AXC) from Modena city. For a quick comparison, the figure shows the histogram of the experimental values related to the homogeneous fractions of fatty acids content in the samples extracted from the Aesculaceae seeds of different cultivars.

I. Overview and General Themes

36

3. Red Horse-Chestnut Seeds of Aesculus 3 Carnea: A New Way for Health and Food Design?

diseases. The cardioprotective benefits of u-3 fatty acids may be attributed to multiple physiological effects on lipids, blood pressure, vascular function, cardiac rhythms, platelet function, and inflammatory responses. In addition, some data suggest that both u-3 and u-6 PUFA have positive health effects in humans. As an evidence, the seed oil extract from the studied Aesculaceae should be a valuable dietary supplement for the presence of these nutraceutics, being rich on u-3 PUFA almost as soja extract and being poor of SFA of about 50%, respectively.23 In addition, we are obliged to mention that the oils extracted from seeds generally contain both saponifiable and unsaponifiable matter. The unsaponifiable constituents of horsechestnut still remain largely unidentified. Although they represent a very small fraction of the lipid mass, approximately 2e3%, equivalent to 0.08e0.15% on the meal dry basis, they include sterols, triterpenes, aliphatic alcohols, vitamins, chlorophylls, and pigments, among others species.

Glucides Analysis The values obtained for total glucidic content in the AXC extracts, Table 3.3, were within to 15.6%, while the previous results were 14.3% (AHH) and 15.2% (AHP), respectively.1 However, the major evidence, when comparing the results for these three Aesculaceae seeds, is related to the presence of arabinose (8.4%) and rhamnose (4.4%) as greatest components of the glucidic fraction (Fig. 3.3), rather than the two monosaccharides glucose and fructose that were the only glucidic components identified in AHP and AHH extracts.1 In addition, we observe another particular character for AXC: the content ratio glucose/fructose is reversed with respect to AHP and AHH cultivars. Actually, we are unable to determine the presence of other complex sugars (disaccharides or other oligosaccharides, if present) based on monosaccharides condensation. Nevertheless, we mention that the experimental procedure that we adopted for these investigations should be responsible for a more effective “in-column hydrolysis process” during the HPLC determination. Therefore, some further aspects related to these topics will be focused on a further research trial. TABLE 3

Glucidic content on horse-chestnut samples of different botanical origin (AHP, AHH, AXC) from Modena city. a

Component

AHP (white flowers)

AHH (pink flowers)

AXC

Ramnose % (d.s.)

—

—

4.4  0.35

Arabinose % (d.s.)

—

—

8.4  0.61

Glucose % (d.s.)

6.8  0.72

6.9  0.58

2.5  0.17

Fructose % (d.s.)

8.4  0.24

7.4  0.20

0.3  0.09

Total glucids % (d.s.)

15.2  0.75

14.3  0.62

15.6  0.72

The table summarizes some experimental data related to the main glucidic monosaccharide components of Aesculaceae seed extracts from different cultivars. a Uncertainties are here expressed as standard deviation of five samples (s5).

I. Overview and General Themes

Red Horse-Chestnuts: Chemical Composition and Characterization

37

18 16 14 12 10 8 6 4 2 0 ramnose %

arabinose %

glucose % AXC

AHP

fructose %

total Glucids %

AHH

FIGURE 3.3 Glucidic content % of horse-chestnut samples of different botanical origin (AHP, AHH, AXC) from Modena city. For an effective comparison, the figure shows the histogram of the experimental values related to the glucides content in the samples extracts from the Aesculaceae seeds of different cultivars.

However, surveying literature, we observe that the results obtained for AXC glucides seem to partially match the work of Hricoviniova and Babor,16 who analyzed the saccharide constituents of different parts of AH seeds. Even if in absence of any reference to AXC nuts, they reported that starch, arabinans, and glucoarabinans are the main constituents of the cotyledon. A series of monosaccharides (D-glucose, L-arabinose, D-glucuronic acid, D-xylose, D-galactose, and fucose) were found in the hydrolyzed extract of the seeds. Episperm, the protective cellulosic layer, mainly contains xylans, associated to glucoxylans.

Scanning Electron Microscopy Because of the paucity of experimental data about the characterization of Aesculaceae nuts, some specific information on morphological structure of the seeds of the two most common Mediterranean varieties AHP and AHH has also been obtained by applying scanning electron microscopy-energy dispersive X-ray spectroscopy surface analysis techniques.1,2 In continuation with the previous work, we applied the same strategy to investigate the internal morphology of the AXC seeds. These seeds show morphological characteristics different to some extent from those recorded for specimens of AHP and AHH. In fact, the starch component of this matrix exhibits a face constituted by microgranules of pseudoovoidal shape, rather regular dimensions (5 O 10 mm, approximately), confined to clusters in cell walls (50 O 100 mm, approximately), immersed in protoplasmatic material constituted by a hydrocolloid, quite gelatinized in the solid phase (Fig. 3.4). To this respect, we observe some microbubbles, due to the presence of gaseous phases, probably trapped at the time of ripening and/or during the natural drying of the seeds. The presence of this gelled substance in AXC samples, not observed in the previous study, together with the platelet forms of starches of AHP and AHH, is the main morphologic difference detected on these natural specimens. In line with other literature studies about similar seeds,24 we can suppose that the cell walls are probably consisting of cellulose-fibraceous materials and complex polysaccharides

I. Overview and General Themes

38

3. Red Horse-Chestnut Seeds of Aesculus 3 Carnea: A New Way for Health and Food Design?

FIGURE 3.4 SEM images of AXC flour sample (red horse-chestnut, wild type), at different enlargements. SEM images of floured AXC seeds from Modena show particular characteristics of the samples, with pseudoregular ovoid forms for the starch-based particles, located in vacuole cells structure and immersed into a solidegel matrix.

in order to develop mechanical properties necessary to ensure the tightness of the internal total bodies. The mapped elements by EDS spectra reveals the predominant presence of C and O, as well as Na, K, Mg, Ca, Al, Si, P, and Cl in significant concentration.

Applications to Health Promotion and Disease Prevention As previously outlined, AH seed extracts have long been known for their healthpromoting properties. Popular medicine has made extensive use of these natural products to treat various human and animal diseases. However, in recent decades, clinical and methodological research has provided results and evidence that are so relevant to the point of ennobling these herbal preparations as true pharmacological products.25e28 Among others peripheral therapeutic benefits induced by these extracts, it has been recognized that many different bioactive molecules play a role for the treatment and chemiprevention of many and very high concern health injury, such as cancer and cardiovascular diseases. Currently, a similar interest in AXC seed extracts seems rather lacking, or the information provided by the producers of therapeutic preparations is not regular enough to distinguish, at first sight, the cultivar of origin of the seeds or other parts of the plant. The great similarity of the AHP, AHH, and AXC trees present in our Campus, including their fruits, makes it very difficult to recognize the distinctive features, except for the colors of the spring flowering panicles. Therefore, although in presence of some specificity due to some compounds more or less known and described in literature, a certainty remains: whatever the origin of the seeds of the genus Aesculus, the relative extracts should present a wide spectrum of common families of components, whichever can be the cultivar.

I. Overview and General Themes

Applications to Health Promotion and Disease Prevention

39

Escin is the most representative pool of bioactive compounds present in AH seeds extracts, particularly active in the treatment of chronic venous insufficiency, postoperative edema, and so far. In addition, the effectiveness toward health injuries is associated with an excellent tolerability. Generally, this complex mixture contains saponin e sapogenins, proanthocyanidin A2, and esculin as the major constituents, even if it remains formally unresolved with respect to many other components. Some research has shown that saponins, sapogenins and their subfamilies, inhibit the enzymatic activity of hyaluronidase, while rutin, procyanidins, flavonoids, tannins, and other similar compounds inhibit the activity of elastase and collagenase. By attenuating the catabolic activity of hyperactive cellulase enzymes, which break down natural lysosomal defenses (elastin, hyaluronic acid, collagen, etc.), these compounds can shift the competitive equilibria between synthesis and degradation of proteoglycans, in favor of synthesis. The attenuation of the enzymatic activity against the mucopolysaccharides of the capillary walls enhances their resistance and improves the preservation of the perivascular tissue integrity of the capillaries themselves, thus reducing the appearance of edematous pathologies. Among the degradative processes involved in many pathogenic situations, oxidative damage plays a predominant role, since it involves the peroxidation of lipids, the production of free radicals, and the consequent destruction of lipids, proteins, collagen, and other species essential to maintain good health conditions of the tissues. Biomolecules with antioxidant activity by scavenging free radicals can interrupt this cascade of events, preserving cellular tissues and improving their integrity. The horse-chestnut extract has a higher antioxidant activity than vitamin E, having shown one of the highest “active oxygen” scavenging ability compared to other natural products. Moreover, it manifests a potent cell-protective effect toward radicals of any type. These properties are strictly connected to the presence of antioxidant molecules, which develop an antiaging effect.22 Among these molecules, the flavonoids are particularly active species, exerting protective and beneficial effects in several ways. These natural extracts are recognized as particularly active bioformulates, exerting protective and health effects in several ways. Fujimura et al. reported the activity of the AH extracts toward cutaneous tissues, stimulating the contraction of non-muscle cells, such as fibroblasts.29 These contractions play an important role in determining cell morphology, vasoconstriction, and wound healing. The results are firmly encouraging, suggesting that escenic extracts are potent anti-aging compounding. There is some evidence that various escin molecules (saponins and sapogenins subfamilies) show beneficial effects when administered at the right concentration, exhibiting an ethanol absorption inhibitory effect and hypoglycemic activity in the oral glucose tolerance test in vivo.16 These observations are strictly related to the anti-obesity effects of the horsechestnut extracts. Working with the seeds of A. turbinata Blume, Hu et al. give a very good rationale for these evidences, completing the description of the mechanisms of the involved species on the metabolic processes observed both in vitro and in vivo.30 In some recent papers, the apoptotic and antiproliferative activity of b-escin toward some human acute myeloid leukemia cell lines (i.e., HL-60, K562) was examined and evaluated by applying different assays and experimental methodologies.31 The results confirm that b-escin is a potent natural inhibitor of leukemic cell proliferation and an apoptosis inducer, depending on the dose and time of administration. These desirable effects are generally associated to

I. Overview and General Themes

40

3. Red Horse-Chestnut Seeds of Aesculus 3 Carnea: A New Way for Health and Food Design?

a good pharmacological tolerability and give indication that b-escin may be a useful candidate for exploring new potential antileukemic drugs. b-escin from AH extracts was also tested to evaluate the chemopreventive efficacy of its dietary intake on azoxymethane-induced colonic aberrant crypt foci. The cell growth inhibitory effects and the induction of apoptosis in HT-29 human colon carcinoma cell line have also been investigated, giving positive results with either wild-type or mutant p53. These novel features candidate b-escin as an effective agent for both colon cancer chemoprevention and treatment.32 Zhang and Li revealed the presence and confirmed the structures of a number of triterpenoid saponins from the seeds of AP.20 Some differences were observed by comparing these structures with those of saponins isolated from Eurasian AH and A. chinensis in their oligosaccharide moieties, suggesting a different chemotaxonomic feature among the species. A set of triterpenoid saponins was systematically tested in vitro for their activity against 59 cell lines from nine different human cancers, giving, for some of the molecules, an inhibition of all proliferation cell activities. In view of the possible relevance of b-escin in anticancer therapy, some formulations were tested in order to establish their effectiveness on the basis of bioavailability and bioequivalence.33 The positive results strongly encourage pursuing new cure modalities, being the treatments based on well-tolerated natural products. The database that collects the results to support the abovementioned treatments constitutes a growing body of clinical trials. However, many of these studies have used complex formulations. This choice is very good to contrast the pathology, but at the same time, it is always problematic because the well-orchestrated action of different active ingredients produces synergistic behaviors. In these cases, it is almost impossible to extrapolate the effect of a single compound from the overall effect of a complex formulation. Even more so, the effects of other minor constituents cannot be ignored. Therefore, when examining the best evidence obtainable from these treatments, it would be necessary to resort to studies that use and privilege single monopreparations, in order to establish the right relationships between the beneficial effects and the negative ones to be contrasted. In addition to the health benefits for human and animal well-being, many studies have appeared in the literature relating to the antimicrobial and antifungal potential of the Aesculaceae extracts, also with regard to many plants. The family of coumarin derivatives from the genus Aesculus shows some particularly effective compounds, as pavietin from AP, with strong bioactivity of contrast against some microfungal species.17 Furthermore, it has been reported that other molecules, such as protein-1 (Ah-AMP1) isolated from AH seeds, are a very effective antimicrobial plant defense since it inhibits the growth of a broad range of fungine species.34 This knowledge may imply a possible technological transferability, by using natural products into the integrated biological struggle to preserve vegetation and foodstuffs as well. It is well known that natural antibacterials, antimicrobials, antivirals, antifungins, and any other inhibitors of damaging species are generally active at moderate dosages and concentrations, associated to an excellent tolerability for upper organisms. The substitution of synthetic phytotherapeutics by natural products with therapeutic activity in many industrial formulations can provide significant advantages and benefits, especially in terms of biocompatibility, sustainability, and environmental preservation.

I. Overview and General Themes

Summary Points

41

FIGURE 3.5 Molecular structure of esculetin (A) and esculin (B) coumarin derivatives with specific medicinal application as vasoprotective agents. The benzopyranic molecular structure represents the basis of the main components of the coumarins family present in Aesculaceae extracts.

Adverse Effects and Reactions (Allergies and Toxicity) Adjuvant and hemolytic activities of a lot of saponins purified from medicinal and food plants were examined by Oda et al.35 In particular, escins showed a weak adjuvant but a strong hemolytic activity. The oral administration is generally well tolerated because some digestive processes partially destroy the original molecules, forming less toxic species. However, the abuse or misuse of saponins is strongly discouraged. On the contrary, the parenteral administration manifests the most negative effects due to hemolytic activity and can lead to an irreversible destruction of red corpuscles of the hematic tissue. It has been outlined that coumarins (Fig. 3.5) may increase the risk of bleeding if administered to cardiovascular patients treated with anticoagulants, such as heparin and analogues species, or platelet inhibitors such as aspirin. At present, the hemolytic activity seems to be the most important adverse effect associated to therapeutic formulations based on Aesculaceae extracts. However, particular attention is due to esculin, whose major counter-effects are responsible of dangerous gastroenteritis. In the case of oral intake with greater consumption, much more serious problems may arise, such as lack of coordination, spasms, restlessness, depression, vomiting, diarrhea, muscle weakness, occasionally paralysis, and unconsciousness, up to worse evolutionary situations. It is worth noting that both pure compound esculin and the multicomponent mixture aescin are used in official pharmacology. To improve tolerability, other natural extracts based on tonic herbs are added to medicinal formulas, in order to minimize and counteract the negative effects.

Summary Points The Aesculaceae seeds, and their derived products, can be considered as an opencast mine of natural different compounds playing some roles related to several biological activities in many different ways. Recognition about their beneficial effects with health claims is growing worldwide, and some standpoints can be summarized as follows: • Escins, the most important class of bioactive molecules in Aesculaceae seeds, show wide ranging mechanisms of therapeutic activity both in peripheral treatments for clinical disorders and in targeted treatments toward cancerous cells, by inhibiting proliferation or inducing apoptosis. The excellent tolerability of escins indicates that these treatments offer benefits for patients with a case history.

I. Overview and General Themes

42

3. Red Horse-Chestnut Seeds of Aesculus 3 Carnea: A New Way for Health and Food Design?

• The main adverse effects of escins in humans are due to the hemolytic activity. More research efforts along these lines are expected in next future, to improve the selectivity toward aberrant red corpuscles, promoting the b-escin fraction to be a useful candidate agent for exploring new potential antileukemic drugs, among others medicinal targets. • Concerning the nutritional use, fresh or naturally desiccated seeds are usually treated by long leaching with water or wooden ashes to remove harshness and bitterness. These treatments induce a variation on the molecular structures of escin fractions, reducing the toxicity but maintaining their nutraceutical potential and anti-obesity effects. Alternatively, the slow-roasting of nuts would make the escins harmless and the seeds edible. • The Aesculaceae seeds extracts have shown a more potent antioxidant activity than vitamin E. Considering the potent cell-protective effects due to this activity, they can be safely used as radical scavengers, with some anti-aging properties associated to the presence of antioxidant molecules, such as flavonoids. • The claimed toxicity of these extracts makes them natural antibacterials, antimicrobials, antivirals, and antifungins to some extent, also acting as phytotherapeutics biocompatibles. However, much more work is needed to characterize extensively these natural products, to increase the knowledge about the bioavailability and pharmacokinetics of their components, and to fully take advantage about their therapeutic potential.

Acknowledgments This study has been funded by the Italian Ministry of Education, University and Research. LM gratefully acknowledges financial support from POR FSE 2014e20 of Emilia-Romagna region (Italy) for doctoral fellowship. The Interdepartmental Research Center BIOGEST-SITEIA of UNIMORE is also acknowledged for the logistic support of this project.

References 1. Baraldi C, Bodecchi LM, Cocchi M, et al. Chemical composition and characterisation of seeds from two varieties (pure and hybrid) of Aesculus hippocastanum. Food Chemistry. 2007;104:229e236. 2. Cocchi M, Durante C, Foca G, et al. Horse-chestnut seeds: seeds of Aesculus Hippocastanum L. and their possible utilization for human consumption. In: Preedy VR, Watson RR, Patel VB, eds. Nuts and Seeds in Health and Disease Prevention. London: Academic Press; 2011:653e661.  3. Otajagic S, Pinjic D, Cavar S, Vidic D, Maksimovic M. Total phenolic content and antioxidant activity of ethanolic extracts of Aesculus hippocastanum L. Bull of the Chemists and Technologists of Bosnia and Herzegovina. 2012;38:35e39. 4. Gonzàlez-Tejero MR, Casares-Porcel M, Sànchez-Rojas CP, et al. Medicinal plants in the Mediterranean area: synthesis of the results of the project Rubia. Journal of Ethnopharmacology. 2008;116:341e357. 5. Hsiao J-Y, Li H-L. Chromatographic studies on the Red horse-chestnut (Aesculus X carnea) and its putative parent species. Brittonia. 1973;25(1):57e63. 6. Foo LY, Porter LJ. The Phytochemistry of proanthocyanidin polymers. Phytochemistry. 1980;19:1747e1754. 7. Haslam E. Symmetry and promiscuity in procyanidin biochemistry. Phytochemistry. 1977;16:1625e1640. 8. Williams VM, Porter LJ, Hemingway RW. Molecular weight profiles of proanthocyanidin polymers. Phytochemistry. 1983;22(2):569e572. 9. Fowden L, Anderson JW, Smith A. A comparative study of the amino acids and phenylalanyl-tRNA synthetases of Aesculus spp. Phytochemistry. 1970;9:2349e2357. 10. Platt RV, Opie CT, Haslam E. Biosynthesis of flavan-3-ols and other secondary plant products from (2S)-phenylalanine. Phytochemistry. 1984;23(10):2211e2217.

I. Overview and General Themes

References

43

11. Kukuła-Młynarczyk A, Hurej M, Jackowski J. Development of horse chestnut leafminer (Cameraria ohridella deschka & dimic) on red horse chestnut. Journal of Plant Protection Research. 2006;46(1):41e47. 12. Ben Rhouma G, Chebil L, Krifa M, Ghoul M, Chekir-Ghedira L. Evaluation of mutagenic and antimutagenic activities of oligorutin and oligoesculin. Food Chemistry. 2012;135:1700e1707. 13. Thornfeldt C. Cosmeceuticals containing herbs: fact, fiction, and future. Dermatologic Surgery. 2005;31:873e880. 14. Hricoviniova Z, Babor K. Saccharide constituents of horse chestnut (Aesculus-hippocastanum L.) seeds. 1. Monosaccharides and their isolation. Chemical Papers e Chemicke Zvesti. 1991;45:553e558. 15. Azarkovich MI, Gumilevskaya NA. Proteins of cotyledons of mature horse chestnut seeds. Russian Journal of Plant Physiology. 2006;53:629e637. 16. Sirtori CM. Aescin: pharmacology, pharmacokinetics and therapeutic profile. Pharmaceutical Research. 2001;44:183e193. 17. Curir P, Galeotti F, Dolci M, Barile E, Lanzotti V. Pavietin, a coumarin from Aesculus pavia with antifungal activity. Journal of Natural Products. 2007;70:1668e1671. 18. Kapusta I, Janda B, Szajwaj B, et al. Flavonoids in horse chestnut (Aesculus hippocastanum) seeds and powdered waste water byproducts. Journal of Agricultural and Food Chemistry. 2007;55:8485e8490. 19. Stankovic SK, Bastic MB, Jovanovic JA. Composition of the sterol fraction in horse chestnut. Phytochemistry. 1984;23:2677e2679. 20. Zhang Z, Li S. Cytotoxic triterpenoid saponins from the fruits of Aesculus pavia L. Phytochemistry. 2007;68:2075e2086. 21. Parmar C, Kaushal MK. Aesculus indica. In: Wild Fruits. New Delhi: Kalyani Publishers; 1982. 22. Kapoor VK, Dureja J, Chadha R. Herbals in the control of ageing. Drug Discovery Today. 2009;14:992e998. 23. Esteves EA, Martino HSD, Martin BM, Oliveira FCE, Bressan J, Costa NMB. Chemical composition of a soybean cultivar lacking lipoxygenases (LOX2 and LOX3). Food Chemistry. 2010;122:238e242. 24. Mosele MM, Hansen AS, Hansen M, Schulz A, Martens HJ. Proximate composition, histochemical analysis and microstructural localisation of nutrients in immature and mature seeds of marama bean (Tylosema esculentum). Food Chemistry. 2011;127:1555e1561. 25. Salinas FM, Vázquez L, Gentilini MV, et al. Aesculus hippocastanum L. seed extract shows virucidal and antiviral activities against respiratory syncytial virus (RSV) and reduces lung inflammation in vivo. Antiviral Research. 2019;164:1e11. 26. Michelini FM, Alché LE, Bueno CA. Virucidal, antiviral and immunomodulatory activities of ß-escin and Aesculus hippocastanum extract. Journal of Pharmacy and Pharmacology. 2018;70(11):1561e1571. 27. Hassan ST, Masarcíková R, Berchová K. Bioactive natural products with anti herpes simplex virus properties. Journal of Pharmacy and Pharmacology. 2015;67(10):1325e1336. 28. Xin W, Zhang L, Sun F, et al. Escin exerts synergistic anti-inflammatory effects with low doses of glucocorticoids in vivo and in vitro. Phytomedicine. 2011;18(4):272e277. 29. Fujimura T, Tsukahara K, Moriwaki S, Hotta M, Kitahara T, Takema Y. A horse chestnut extract, which induces contraction forces in fibroblasts, is a potent anti-aging ingredient. Journal of Cosmetic Science. 2006;57:369e376. 30. Hu JN, Zhu XM, Han LK, et al. Anti-obesity effects of escins extracted from the seeds of Aesculus turbinata BLUME (Hippocastanaceae). Chemical and Pharmaceutical Bulletin. 2008;56:12e16. 31. Niu YP, Wu LM, Jiang YL, Wang WX, Li LD. b-escin, a natural triterpenoid saponin from Chinese horse chestnut seeds, depresses HL-60 human leukaemia cell proliferation and induces apoptosis. Journal of Pharmacy and Pharmacology. 2008;60:1213e1220. 32. Patlolla JMR, Raju J, Swamy MV, Rao CV. b-Escin inhibits colonic aberrant crypt foci formation in rats and regulates the cell cycle growth by inducing p21(waf1/cip1) in colon cancer cells. Molecular Cancer Therapeutics. 2006;5:1459e1466. 33. Bassler D, Okpanyi S, Schrodter A, Loew D, Schurer M, Schulz HU. Bioavailability of b-aescin from horse chestnut seed extract: comparative clinical studies of two galenic formulations. Advances in Therapy. 2003;20:295e304. 34. Fant F, Vranken WF, Borremans FAM. The three-dimensional solution structure of Aesculus hippocastanum antimicrobial protein 1 determined by H-1 nuclear magnetic resonance. Proteins Structure Function and Genetics. 1999;37:388e403. 35. Oda K, Matsuda H, Murakami T, Katayama S, Ohgitani T, Yoshikawa M. Adjuvant and haemolytic activities of 47 saponins derived from medicinal and food plants. Biological Chemistry. 2000;381:67e74.

I. Overview and General Themes

C H A P T E R

4

The African Breadfruit (Treculia africana) Decne Plant Seed: A Potential Source of Essential Food and Medicinal Phytoconstituents 1

Folake Lucy Oyetayo1, Victor Olusegun Oyetayo2 Department of Biochemistry, Ekiti State University, Ado-Ekiti State University Ado-Ekiti, Nigeria; 2Department of Microbiology, Federal University of Technology, Akure, Nigeria

Introduction The African breadfruit plant (Treculia africana) Decne is an evergreen tropical tree crop believed to be native to a vast area from New Guinea to western Micronesia.1 It grows in the West and tropical African Countries, Nigeria, Ghana, and Sierra Leone, up to 30m in height and bears large seeded fruits2 sought after because of the edible seeds and the seed oil rather than the bitter tasting fleshy pulp in which they are embedded.3 In Southern Nigeria, it is known by various names by different tribes “afon” by the Yorubas, “ukwa” (Ibo), and “ediang” (Efik).4 The plant is of great socioeconomic value especially in Southeast Nigeria where the seed is an important natural resource, contributing to income and cheap additional nutrient sources especially among the poor.5 However, Meregini6 listed the plant as an endangered species of Southern Nigeria due to annual loss of tremendous amount of the seeds.

Botanical Description, Cultivation, and Usage Treculia africana Decne belongs to the family Moraceae and genus Treculia. The family consists of about 50 genera and over 1000 species.7

Nuts and Seeds in Health and Disease Prevention, Second Edition https://doi.org/10.1016/B978-0-12-818553-7.00004-8

45

Copyright © 2020 Elsevier Inc. All rights reserved.

46

4. The African Breadfruit (Treculia africana) Decne Plant Seed

The leaves are large and leathery and flowers are greenish; both sexes are globose with very short-stalked heads. Male heads appear singly in the leaf axils and female heads on the bole and main branches. The plant bears round large seeded yellowish-green fruits with a rough outer surface. The fruit contains numerous, small, ellipsoidal seeds about 1 cm long, each covered with brown shells buried at various depths in the spongy pulp of the fruit. The plant can be propagated from the seed when fairly fresh (as it loses vitality in a few weeks1) and buds. In the wild, the trees are regenerated from seeds of fallen fruits on the forest floor. It fruits up to 2e3 times a year.8 Seed propagated Treculia africana fruits within 4 years.9

Fruit Processing and Seed Production The mature Treculia africana tree produces about 30 fruits per year, yielding an appreciable 3125 seeds per Kg10 after processing (Plate 4.1). After harvesting, the fruits can be sliced and seeds extracted manually, extracting the mucilaginous layer using 1e5% trona and wood ash for 5e25 minutes. Otherwise, harvested fruits are stacked in heaps and left for several days in open air for partial fermentation.2 The fermented mass is shredded/macerated and washed in running water to remove the slimy, jelly-like extraneous substances. The cleaned seeds are then air-dried, dehulled, and converted into several food forms including porridges. Fig. 4.1 below shows various methods involved in the processing of Treculia africana seeds from the matured fruits. In Southeastern Nigeria, the seeds are commonly processed for consumption via fermentation and boiled with “trona” (a food tenderizer and the second most used salt in Nigeria) into a popular food product known as “ukwa.”

PLATE 4.1 The African breadfruit plant (Treculia africana) bearing fruit.

I. Overview and General Themes

Antinutritional Compositions of African Breadfruit Seed

47

Harvested Matured African Breadfruit

Fermentaon

Washing

Air Drying

Dehulling

Processed seeds

FIGURE 4.1

Processing method of African breadfruit (Treculia africana) seeds.

Nutritional Value and Food Uses of Treculia africana Seed The most valuable part of the plant is the edible seeds. The seeds are highly nutritious11 forming an important staple food of high economic value.12 Compositional analyses of African breadfruit fruit seed show it contains a wide array of nutrients: carbohydrates (53.7e62.2%), which form an important part of diets in several developing countries (and can thus serve as substitute for other starchy foods), proteins (13.4e23.2%), fat (10.4e18.9%), and fiber.12,13 It contains a wide array of nutritive elements of which zinc was the most dominant14, while Oyetayo and Oyetayo15 reported calcium concentrations ranging between150e290 (mg/100g) in raw and processed seeds. The seed has significant dietetic values with biological value greater than that of soybeans.16 The seeds may be roasted, dehulled, and eaten as a snack, or they may be parboiled and dehulled prior to cooking, a process which lasts 2e3 hours. Dehulled seeds may be cooked alone or in combination with shelled corn, sorghum, or other vegetables. African breadfruit meals may be garnished with dried fish or meat, resulting in a highly nutritious diet. Apart from these, the seeds are also processed into flour, which has high potential usage for pastries17. Roasted seeds may also be used to thicken soups and gravies. Edible oil can be processed from the seeds18 and used in cooking. Cooked seeds make very delicious meals. Non-alcoholic beverages have also been prepared from the seeds.19 (Plate 4.2)

Antinutritional Compositions of African Breadfruit Seed Despite its dense nutrient composition, the African breadfruit seed is also a source of antinutrients: oxalates, phytates, tannins, and hydrocyanic acid.14,20 Antinutrients, naturally occurring and diverse group of chemical compounds present in plant foods, act to reduce nutrient utilization and food intake resulting in impaired growth in animals.21

I. Overview and General Themes

48

4. The African Breadfruit (Treculia africana) Decne Plant Seed

PLATE 4.2

Seeds of processed African Breadfruit (Treculia africana).

Fasasi et al.14 have reported that processing methods e frying, toasting, and boiling e reduced the antinutritional content of the seed, while Sunday et al.22 reported that boiling was more effective than roasting in reducing the concentrations of trypsin inhibitor, phytic acid, and polyphenols in the seed. Boiling with trona was found to significantly reduce phytate content of the seed better than boiling without trona, improving the zinc and calcium bioavailability, which is nutritionally beneficial.15 Complexes formed between phytic acid and essential divalent minerals interfere with proteolytic digestion making the plant seed nutritionally inferior as a nutritive mineral source.23 Moreover, processing methods such as fermentation and malting have been found to reduce the phytate content of the Treculia africana seed.14

Applications to Health Promotion and Disease Prevention The seed has been used for the treatment of malaria fever and cough.24 In the local setting, water extract of the boiled seed has been claimed to cleanse the stomach.25 Phytochemical screening of the seed water and alcoholic extracts showed the presence of secondary metabolites: flavonoids, polyphenols, anthraquinones, saponins, and cardiac glycosides, which are known to possess antimicrobial and health-promoting activities, and concentration of toxicants in the seed was reported to be lower than levels considered to be critical.26 Oyetayo and Omenwa20 showed that processing of raw breadfruit by fermentation improved the microbial quality of the seeds making them safe for consumption. Consumption of about 300g of fried or boiled breadfruit will supply approximately 17.1e23.1 mg/day of iron to the body while the recommended intake for adults is 18 mg.27 Hence, consumption of the seed can prevent the incidence of anemia. African breadfruit, irrespective of the method of preparation, will serve as a good source of magnesium, an electrolyte, which is also required as cofactor for numerous enzymes involved in nutrient metabolism. Intake of 300g of boiled breadfruit will result the supply of

I. Overview and General Themes

References

49

207e254 mg of magnesium to physiology. Also, intake of breadfruit seeds will provide the body with adequate zinc.28 Lack of zinc results in impaired immunological functions and growth retardation as it is involved in the functioning of neutrophils and natural killer cells.29 Nnorom et al.28 reported trace amounts of heavy metals such as lead, chromium, and nickel in the seed at concentrations that poses no toxicological risk upon consumption of the processed seed. Phytochemical screening of the seed extracts showed that it contains appreciable amounts of secondary metabolites known to have antimicrobial as well as other health-promoting activities. Osabor et al. 26 reported that it contains lower concentration of toxicants, hydrogen cyanide, oxalate, and phytate, than levels considered to be critical.

Future Perspectives Due to the numerous beneficial phytoconstituents, socioeconomic, traditional, and potential industrial uses of the Treculia africana seed, efforts toward the conservation and development of phytochemical dense and early maturing species are necessary. There is also a need for isolation and characterization of various functional constituents of the seed to further elucidate their potential pharmacological activities.

Acknowledgments The authors acknowledge the contributions of Olawale O. and Mercy O. in taking the photographs.

References 1. Morton JF. Breadfruit. In: Morton JF, ed. Fruits of Warm Climates. Miami, Florida: Fairchild Tropical Botanical Garden; 1987:50e58. 2. Enibe SO, Bunso OO, Sefwi AW. African Forest Research Network: African Forest Research Network: Project on Propagation, Early Growth, Nutritional and Development of Treculia Africana Seeds. 2003:231e450. 3. Nwokolo EN, Bragg BB. Influence of phytic acid and crude fibre on the availability of minerals from protein supplements in growing chicks. J. Amin. Sci. 1977;57:477. 4. Onweluzo LJC, Odume L. Method of Extraction and Demucilagination of Treculia Africana: Effect on Composition; 2008. http://www.bioline.org.br/request?nf07008.Assessedon9/1/2008. 5. Baiyeri KP, Mbah BN. Effect of soiless and soil-based nursery media on seedling emergency, growth and response to water stress of african breadfruit (Treculia africana Decne). African Journal of Biotechnology. 2006;5(15):1400e1405. 6. Meregini AOA. Some endangered plants producing edible fruits and seeds in southern Nigeria. Fruits. 2005;60:211e220. 7. Tindal HD. Fruits and Vegetables in West Africa. Food and Agricultural Organization (FAO) Rome; 1965:259. 8. Soetjipo NN, Lubis AS. Vegetables. Vol. 259. Rome: IBPGR Secretariat; 1981. 9. Okafor JC. Promising trees for agro-foresting in southern Nigeria. Journal of the Science of Food and Agriculture. 1980;34:407e415. 10. Oboho EG, Ngalum EL. Germination response of Treculia africana (Decne) seeds in relation to moisture content, storage method and its duration. Journal of Applied and Natural Science. 2016;(1):88e94. 11. Onyekwelu JC, Fayose OJ. Effect of storage methods on the germination and proximate composition of Treculia africana seeds. In: Paper Presented at the Conference on International Agricultural Research for Development. Tropentas, Germany. 2007.

I. Overview and General Themes

50

4. The African Breadfruit (Treculia africana) Decne Plant Seed

12. Edet EE. Chemical evaluation of nutritive value of African bread fruit Treculia africana. Food Chem. 1980;117:47e50. 13. Nwokolo E, Smartt J. Legumes and Oilseeds in Nutrition by. Chapman & Hall; 1996. ISBN 0 412 45930 2. 14. Fasasi SI, Eleyinmi OO, Karim F. Chemical properties of raw and processed Treculia africana seed flour. Journal of Food Agriculture and Environment. 2004;2:145e263. 15. Oyetayo FL, Oyetayo VO. Dynamics of Phytate-Zinc/Calcium Balance of Processed African Breadfruit (Treculia Africana Decne) Seeds. Global science books; 2007:88e90. 16. World Agroforestry Centre (WAC. Treculia africana. In: Agroforestry Database; 2004. http://www. worldagroforestry.org/sea/products/afdbases/af/asp/speciesInfo.asp?SpID¼15. 17. Keay RWJ. In: Trees of Nigeria: A Revised Version of Nigeria Trees. Vols. 1 and 2. Oxford: Stanfield DP, Clarendon Press; 1989. 18. Ugwoke FN, Agbo AE, Ali NC, Attah CP, Ekwueme JI. A note on african breadfruit (Treculia africana Decne). In: Unpublished Report Submitted in Partial Fulfillment of CSC 341. Nsukka, Nigeria: Department of Crop Science, University of Nigeria; 2003:18. 19. Ejiofor MAN. Diversifying utilities of african bread-fruit as food and feed inter. Tree Crops Journal. 1998;5:125e135. 20. Oyetayo VO, Omenwa VC. Microbial and chemical qualities of raw and trona processed african breadfruit (Treculia africana Decne). American Journal of Food Technology. 2006;(1):77e80. 21. Soetan K, Oyewole O. The need for adequate processing to reduce the antinutritional factors in plants used as human foods and animal feeds: a review. African Journal of Food Science. 2009;3(9):223e232. 22. Sunday YG, Matthew NA, Alexandria DH, Emmanuel OO. Effect if heat processing on In vitro protein digestibility and some chemical properties of African breadfruit seeds. Plant food Human Nutrition. 2001;55:357e368. 23. Lonnerdal B Phytic acid-trace element (Zn, Cu, Mn) interaction. International Journal of Food Science and Toxicology. 2002;37:749e758. 24. Irvine JI. Comparative study of the chemical composition and mineral element content of Treculia africana seeds and seed oils. Journal of Food Engineering. 1981;40:241e244. 25. Nuga OO and ofodile EAU potentials of Treculia africana ean endangered species of southwest Nigeria. Journal of Advanced Scientific Research. 2010;10(2). 26. Osabor VN, Ogar DA, Okafor PC. Egbung GE profile of the african bread fruit (Treculia africana). Pakistan Journal of Nutrition. 2009;8:1005e1008. 27. Institute of medicine, food and nutrition board ( IMFN. Dietary Reference Intake for Vitamin A, Silicon, Vanadium and Zinc. Washington DC: National academy press; 2001. 28. Nnorom IC, Ewuzie U, Ogbuagu F, Okereke M. Agwu P and enyinnaya IP mineral contents of Ukwa, african breadfruit (Treculia africana), from south-eastern Nigeria:effect of methods of preparation. International Journal of Physical and Social Sciences. 2015;4(3):230e240. 29. Shankar AH, Prasad AS. Zinc and immunological functions: the biological basis of altered resistance to infection. American Journal of Clinical Nutrition. 1998;68(2):447Se463S.

I. Overview and General Themes

C H A P T E R

5

Perennial Horse Gram (Macrotyloma axillare) Seeds: Biotechnology Applications of Its Peptide and Protein Content e BowmaneBirk Inhibitors and Lectin Marcos Aurélio de Santana1, William de Castro Borges1, Alessandra de Paula Carli1, 2, Larissa Lovatto Amorin1, Alexandre Gonçalves Santos1, Sonaly Cristine Leal1, Milton Hércules Guerra de Andrade1 1

Universidade Federal de Ouro Preto, Departamento de Ciências Biológicas, Núcleo de Pesquisas em Ciências Biológicas, Laboratório de Enzimologia e Proteômica, Campus Morro do Cruzeiro, Ouro Preto, Minas Gerais, Brazil; 2Universidade Federal dos Vales do Jequitinhonha e Mucuri, Instituto de Ciência, Engenharia e Tecnologia, Campus do Mucuri, Teófilo Otoni, Minas Gerais, Brazil

List of Abbreviations BBI BowmaneBirk inhibitors BBIM BowmaneBirk inhibitor concentrate from Macrotyloma axillare BBIS BowmaneBirk inhibitor concentrate from Glycine max DBL Dolichos biflorus seed lectin DMH N-N0 -dimethylhydrazine GalNAc N-Acetyl-a-D-galactosamine MaL Macrotyloma axillare seed lectin

Nuts and Seeds in Health and Disease Prevention, Second Edition https://doi.org/10.1016/B978-0-12-818553-7.00005-X

51

Copyright © 2020 Elsevier Inc. All rights reserved.

52

5. Perennial Horse Gram (Macrotyloma axillare) Seeds

Introduction Macrotyloma axillare is a legume adapted to the tropical and subtropical climate and its main use is associated with green manuring and soil recovery and it is also employed as food for cattle. M. axillare is the source of many biochemically active biomolecules with biotechnology interest, such as BowmaneBirk type inhibitors (BBI) and lectins (anti-A1 seed lectin). BBI are protein molecules with double-head protein domains, one exhibiting anti-trypsin and the other antichymotrypsin activities. Such inhibitors are known to be active toward various enzymatic processes, making them useful to modulate various proteolyticdependent pathways, such as those related to inflammation and neoplastic transformation. Lectins are important biomolecules used on carbohydrate prospection and characterization, given their strict specificities. In this chapter we will describe some strategies for the isolation of BBI and lectin from M. axillare and discuss some of the BBI protective activities, when used as a diet supplement in a mice model of colorectal cancer induced by N-N0 -dimethylhydrazine (DMH). DMH-treated animals presented histopathological dysplasia, elevated peptidase activities from lysosomal fractions, increased proteasome activities (trypsin and chymotrypsin-like), and higher expression of CD44, when animals were treated at a dose of 30 mg/kg, during 12 weeks. In contrast, animal groups receiving a diet supplemented with BBI-enriched fractions from Macrotyloma axillare (BBIM) and soybean (BBIS) at 0.1% w/w of BBI during 8 weeks and treated with DMH displayed decreased proteasome and lysosomal proteolytic activities. In addition, BBI-treated groups did not develop dysplastic alterations. These are pioneering findings on the simultaneous inhibition of both proteasome and lysosomal activities and provided the basis for future investigation on the potential of BBI isolated from M. axillare to be employed in disease processes in which exacerbated proteolysis is enhanced. Our results indicated that the groups that were tested simultaneously with BBI þ DMH presented decrease in chymotrypsin and trypsin-like proteasome activities and lysosome protease activities compare to the controls groups. In addition, such group tests BBI þ DMH did not develop dysplastic alterations. So, we believe that the protective effect provided by BBI fractions tested in such DMH model in mice is associated with the simultaneously inhibitory activity on lysosome proteases and proteasome pathways from isolated small intestine and colon tissues. Such approach was the first time in literature that the proteasome activity was evaluated before an enriched BBI supplemented diet on DMH cancer induced in vivo in mice.

Botanical Description Due to its main use as forage, M. axillare data can be found in specialized web pages of recognized scientific accuracy. The following botanical and morphological description can be found at the Tropical Forages website.1 M. axillare (Fig. 5.1) is a trailing and twining perennial plant, with an erect basal stem usually to about 1 cm diameter, developing to 3 cm in unrestricted stands, and with the ability to climb to >10 m up an appropriate framework such as trees, strong woody taproot, and rootstock. Stems are cylindrical, glabrescent to pubescent with appressed hairs; and no tendency to develop adventitious roots. Leaves

I. Overview and General Themes

Botanical Description

53

FIGURE 5.1

Macrotyloma axillare plant and seeds. Note the trailing pattern of growth. Source: Used with permission from Tropical Forages Tropical Forages: an interactive selection tool.1 Available at: http://www.tropicalforages.info/key/forages/ Media/Images/entities/macrotyloma_axillare/macrotyloma_axillare_02.jpg.

are trifoliolate, with leaflets ovate-lanceolate, to 7.5 cm long and 4 cm across; glabrous to pubescent, slightly glossy on upper surface, paler, matt below; stipules to 5 mm long. Inflorescence is an axillary raceme, comprising 2e4 (sometimes up to 10) whitish to greenish yellow papilionate flowers, with standard oblong-elliptical, 1e2.4 cm long and 0.6e1.5 cm across. Pod is linear oblong, shortly stipulate, laterally flattened, 3e8 cm long, and 5e8 mm broad, glabrous to pubescent, with terminal point up to 7 mm long; containing (5e) 7e8 (e9) seeds. Seeds are subovoid, 3e4 mm long, 2.5e3 mm broad, hard and smooth, buff to reddish brown, with sparse to dense black mottling; 50,000e200,000 seeds/kg.1 Macrotyloma axillare taxonomy1: • • • • • • • • • •

Kingdom: Plantae (plants); Subkingdom: Tracheobionta e vascular plants; Superdivision: Spermatophyta e seed plants; Division: Magnoliophyta e flowering plants; Class: Magnoliopsida e dicotyledons; Subclass: Rosidae; Order: Fabales; Family: Fabaceae e pea family; Genus: Macrotyloma (Wight & Arn.) Verdc. e macrotyloma; Species: Macrotyloma axillare (E. Mey.) Verdc. e perennial horse gram.

I. Overview and General Themes

54

5. Perennial Horse Gram (Macrotyloma axillare) Seeds

Historical Cultivation and Usage As a native plant in Africa, M. axillare has been employed in a variety of veterinary applications. Apart from its main use as forage, M. axillare among other leguminous plants are considered to display antimicrobial, antifungic, and antihelminthic properties.2 In addition, M. axillare has been included in a list of 33 medicinal plants showing that it is also utilized by men for managing sexual impotence and erectile dysfunction in western Uganda.3 Posteriorly, Morris J.B.4 analyzed the anthocyanin content of M. axillare and M. uniflorum and detected the presence of D-pinitol, a carbohydrate that was shown to reduce postprandial glycemic levels in diabetes mellitus type 2 patients. This finding places M. axillare under a category of leguminous plant from which novel and potential biomolecules with pharmacological interest can be isolated. However, despite its potential use, M. axillare is still poorly investigated by the scientific community. In this chapter, we will report on the isolation, structural characterization, and biological activities of two constituents of M. axillare seeds which have constituted the subject of our investigation: the lectin and the BBI (Fig. 5.2).

Present-Day Cultivation and Usage Notwithstanding the biotechnological potential of its seeds, the main use of the Macrotyloma axillare remains in agriculture and plant forage intended to feed the cattle.5 In addition, its use in the recovery of eroded soils has been evaluated.6

Mr

BBl

MaL

MaL*

30 kDa 23 kDa 17 kDa 13 kDa

FIGURE 5.2 1D gel separation of two bioproducts isolated from Macrotyloma axillare. BBI e the BowmaneBirk

inhibitors at approximately 8 kDa after reduction and alkylation of cysteine residues. MaL e M. axillare lectin at approximately 28 kDa (monomer form). BBI and MaL are stained in Coomassie. MaL e M. axillare lectin stained using the PAS method to reveal its carbohydrate content. Source: Original data.

I. Overview and General Themes

Applications to Health Promotion and Disease Prevention e Biotechnology Applications of the Protein Seed Content

55

Applications to Health Promotion and Disease Prevention e Biotechnology Applications of the Protein Seed Content Macrotyloma axillare Seed Lectin (MaL) The availability of a great number of lectins displaying distinct glycidic specificities has resulted in the utilization of such molecules as useful tools in medical and biological research. Lectins can be used to explore the cell surface through its ability to recognize and bind the glycidic moieties of glycoproteins and glycolipids of the cell glycocalyx. Alternatively, immobilization of lectins onto inert matrices offers great potential for affinity chromatography and isolation of glycoproteins, an important primary step toward the characterization of the glycoproteome. Due to their inherent functions, these versatile molecules are also used for blood typing, are employed as mitotic agents (e.g., lectins such as concanavalin A stimulate mitosis in certain types of lymphocytes), and can act as probes to detect the alterations associated with cell development and transformation. Nowadays, lectin derivatives are also commercially available. These include the covalent attachment of fluorescein isothiocyanate, biotin, or radioactive isotopes to lectins to facilitate their detection while preserving their biological activity.7 Lectins from leguminous plants are used as model systems to study the molecular events associated with the interaction between a protein and a specific carbohydrate in biological research. This choice is based on the relative ease to purify and isolate significant amounts of lectins from seed extracts. Once isolated, the lectin specificity can be evaluated by biophysical techniques such as X-ray crystallography, nuclear magnetic resonance, and microcalorimetry. Although a great variety of glycidic specificities have been observed for the isolates, a remarkable conservation of amino acid sequence is reported among leguminous lectins.8 The lectin isolated from M. axillare (MaL) seeds and its counterpart, purified from Dolichos biflorus (DBL) seeds, are specific to the carbohydrate N-Acetyl-a-D-galactosamine (GalNac). This finding offered biotechnological relevance given that this sugar is an antigenic determinant of blood type A from the ABO system. Routinely DBL is used to differentiate subgroups A1 from A2, the difference residing in the number of antigens on the erythrocyte surface which is greater in A1 compared to A2.9 Subgroup A1 is then identified by positive hemagglutination and the test is negative for subgroup A2 under the use of a standard specific activity (Fig. 5.3). Considering that DBL and MaL are extremely related to each other and share the same sugar specificity,10 methodologies to obtain the lectin isolated from M. axillare can be useful as this leguminous plant can be cultivated and obtained in vast amounts in tropical countries. MaL was first isolated by Haylett and Swart10 by affinity chromatography essentially as proposed for the isolation of DBL. The protocol revealed good protein yield; however, the method used for elution involved the utilization of a low pH buffer as an alternative to the use of GalNac as initially described by Etzler and Kabat11 during isolation of DBL. Further studies from our research group, at the Federal University of Ouro Preto, have revealed that a low pH can interfere with MaL activity as a tetramer form, which led us to propose a new protocol for the isolation of MaL. The procedure involved the utilization of

I. Overview and General Themes

56

5. Perennial Horse Gram (Macrotyloma axillare) Seeds

FIGURE 5.3 Macrotyloma axillare seed lectin (MaL) specificity on ABO system. MaL specificity study on native and trypsin-treated erythrocytes for the human ABO antigen system. Source: Reproduced with permission from Santana et al.9

fewer and simple steps resulting in high lectin yields and maximum retention of biological activity therefore suitable to scaling up for industrial production.9 The detailed technique was previously patented.12 The BowmaneBirk Inhibitors from M. axillare e Biochemistry and Particularities of BBI BowmaneBirk type inhibitors, commonly called BBI, are cytoplasmic protein inhibitors of serine proteases. BBI were first isolated from soybean seeds by Bowman in 1946 and then characterized by Birk in 1961. Since then, BBI from other leguminous plants have been isolated and characterized.13 In general, BBI are low molecular mass proteins, of approximately 8 kDa, consisting of a single amino acid chain exhibiting a significant content of cysteine residues which confer a rigid tridimensional folding highly conserved among the different isolates of the Plantae kingdom; from which they are exclusive. The main enzymes inhibited by BBI are trypsin and chymotrypsin. BBI are classified as bivalent inhibitors as their tridimensional folding defines two inhibition loops or “heads” one being specific for binding of trypsin and the other for binding of chymotrypsin.14 Other enzymes reported to be inhibited by BBI are cathepsin G, elastase, and the chymase. Crystallographic analyses have shown that the tridimensional structure of this protein is mainly stabilized by seven disulfide bridges followed by a number of intramolecular hydrogen bonding and a contribution of the hydrophobic core of the molecule.15 As a result, BBI are highly resistant molecules supporting extremes of temperature, ionic strength, and pH, being soluble in the pH range 1.5e12. Such acid-resistant property is very useful considering that BBI can be used as a concentrated preparation for oral administration, with no major side effects. In fact, enriched BBI from soybeans (Glycine max) is already being investigated in accordance with FDA regulations (Investigational New Drug, IND) and many mandatory human studies are in progress to evaluate the therapeutic effect of BBI on various diseases.16

I. Overview and General Themes

Applications to Health Promotion and Disease Prevention e Biotechnology Applications of the Protein Seed Content

57

Serine protease inhibitors react with the protease active site at the serine residue, forming derivatives of inactivated enzymes. This class of proteases are inhibited by Kunitz-type inhibitors, BBI, the potato inhibitors type I and II, and the kazal-type inhibitors.17 In this context, BBI isolated from M. axillare merit attention due to their highly specific inhibitory activity especially considering BBI isolated from germinated seeds.18 The interaction of BBI with either trypsin or chymotrypsin is able to strongly inhibit the activity of these proteases; in addition, simultaneous binding of these two proteases with a single BBI monomer has been reported. The conformational loop of BBI reactive site is fully complementary to the active site of the enzyme to be inhibited, allowing for a strong proteineprotein interaction to take place.15 Although this interaction is reversible, it occurs with an affinity compared to that observed for the formation of the complex resulting from the interaction between the protease and its protein substrate. The perfect docking of the inhibitor at the enzyme active site prevents necessary conformational changes to occur resulting in an unfavorable energetic barrier to hydrolysis. Main Biological Activities Related to BowmaneBirk Inhibitors The BBI antichymotrypsin activity also had been subject of scientific research. The rationale relies on the observation that the inhibition of a cell’s chymotrypsin activity is linked to an anticarcinogenic effect. BBI have also been shown to possess anti-inflammatory and radioprotective properties being also able to inhibit the production of free radicals and some carcinogen-induced transformations. Although a wide spectrum of anticancer activities has been reported, the exact pathway mechanisms, with details, by which these inhibitors promote anticarcinogenic effects, remain to be elucidated.17,19e21 The activity of BBI has been shown to extend beyond their classical chymotrypsin/trypsin targets. In this regard, Chen et al.22 using BBI isolated from soybeans demonstrated that these molecules also inhibited the chymotrypsin-like activity of the 26S proteasome, a major nanomachine involved in intracellular proteolysis. In vitro and in vivo experiments confirmed the specific inhibition of the proteasome’s chymotrypsin-like activity using MCF7 cells isolated from breast cancer tissue. It was also verified that such inhibition is linked to an accumulation of ubiquitylated proteins, the natural proteasome substrates, and a reduction on the levels of regulatory cyclins involved in cell division. Altogether the authors suggested that inhibition of the 26S proteasome activity by BBI could greatly contribute to their preventive effect on cancer. Further evidences of the interaction of BBI and the proteasome have been reported by Saito et al.23 who also showed inhibition of the chymotrypsin-like activity in osteosarcoma cells. More specifically, BBI inhibited the degradation of connexin 43 by the ubiquitinproteasome system in these tumor cells. It was suggested that the antiproliferative effect observed was due to maintenance of connexin function through homeostatic balance promoted by GAP junctions. As BBI are involved in serine proteases inhibition, such activity was explored as anti-inflammatory molecules, given the established role of serine-proteases during inflammation.16 In this context, studies conducted with BBB concentrated (soybean BBIC) were promising for the treatment of ulcerative colitis in humans. The results showed that BBIC-treated groups showed a significant improvement with remission of ulcerative colitis, with no toxic effects being observed compared to the groups receiving a placebo.24

I. Overview and General Themes

58

5. Perennial Horse Gram (Macrotyloma axillare) Seeds

Recently, Tong-Cui Ma et al.,25 in a study with BBIC, demonstrated that these inhibitors block HIV (human immunodeficiency virus) infection in isolated human macrophages. The proposed mechanisms point to the fact that BBI induce the production of extracellular CC chemokines that block the entry of HIV. In addition, BBI activate intracellular restriction factors to block the HIV viral cycle, preventing its replication. However, the molecular mechanisms by which BBI promote this activity require further studies. Anyhow, the obtained data are clinically relevant for prevention and treatment of HIV infection, as BBI fractions can be easily obtained by protein purification techniques. Such inhibitors may be useful as supplementary agents particularly when conventional treatment is not readily available or accessible. BowmaneBirk Inhibitors from Macrotyloma axillare and Its Anticarcinogenic Effect on Colorectal Cancer Given the high abundance of BBI in leguminous plants, these inhibitors are regularly consumed as part of the human diet. As BBI isoforms have shown to be active throughout the extent of the gastrointestinal tract, many studies have been carried out with either natural or synthetic BBI fractions applied to model diseases of the intestines.21,26 Previous studies with BBI fractions isolated from soybean (Glycine max) demonstrated its protective activity on colorectal cancer in mice induced by DMH.27e29 The sequence variations observed for BBI from Glycine max and M. axillare could potentially result in distinct inhibitory potencies. Moreover, the existence of isoforms (DE3 and DE4) for BBI from M. axillare differing only in their antichymotrypsin heads30 prompted us to evaluate its protective activity compared to BBI from soybean (BBIS) in a mice model of colorectal cancer induced by DMH.31 The prospection of proteolytic enzymes was based on previous studies demonstrating that DMH treatment induced enhanced proteasome and lysosomal activities.19,32 Based on previous works developed by our research group,18,33 we standardized a new technique for the isolation of BBI-enriched fractions from M. axillare. This procedure was very reproducible for the isolation of BBI fractions from this species and was patented as a novel procedure for isolating BBI.34 BBI isolation from Glycine max (BBIS) was performed as previously described.35 Test animals were submitted to a diet supplemented with BBI fractions enriched at 0.1% w/w for 8 weeks. To confirm lesions and cancer induction, we employed conventional histology and Western blotting using anti-CD44 as a marker of neoplastic alterations. Chymotrypsin and trypsin-like peptidase assays from both proteasome and lysosomal fractions were evaluated in control and treated animals. As expected, in DMH-treated animals, neoplastic alterations and overexpression of CD44 were found. In contrast, for DMH-treated animals receiving a diet supplemented with BBI from both sources, histopathology and Western blotting approaches revealed tissue morphology and CD44 expression comparable to those observed for control animals. As for the peptidase assays, in DMH-treated mice, the lysosomal trypsin and chymotrypsin-like activities, from both intestine and colon samples, were significantly elevated compared to the control samples (Fig. 5.4). However, when the same activities were measured in the test groups (those under DMH treatment and fed a diet supplemented with BBI-enriched fractions from M. axillare and G. max), significant decreases in these proteolytic activities were observed when compared to the DMH-treated group without BBI supplementation.

I. Overview and General Themes

Applications to Health Promotion and Disease Prevention e Biotechnology Applications of the Protein Seed Content

59

FIGURE 5.4 BBI inhibitory action over the chymotrypsin- and trypsin-like activities recovered from lysosomal fractions. Increased chymotrypsin- and trypsin-like activities were found in the colon and intestines from animals treated with DMH (n ¼ 6). In contrast, animals under DMH treatment and fed a diet supplemented with BBI from either Glycine max (DMHS) or Macrotyloma axillare (DMHM) displayed proteolytic activities comparable to control groups. Results are expressed as arbitrary units of fluorescence/mg of protein. Bars represent the means  SD. Statistically significant differences are indicated by asterisks (*), considering P  .05. Source: Reproduced with permission from Carli et al.31

Analysis of proteasome activities (chymotrypsin- and trypsin-like) from the small intestine and colon samples revealed that diet supplementation with BBI during 8 weeks, applied to healthy animals, resulted in a significant decrease (approximately 50% reduction for both activities) attesting for their internalization and inhibitory activity toward the proteasome (Fig. 5.5). These results are in agreement with those obtained in vivo concerning the biodistribution of BBI previously carried out in our research group.36 When the animals were treated with DMH, the proteasome’s trypsin- and chymotrypsinlike activities were greatly enhanced (around 10-fold higher compared to that of controls). However, in the test groups in which animals had their diet supplemented with BBI fractions (DMHM and DMHS) (Fig. 5.6), proteasome activities decreased significantly and were reduced to values comparable to the respective control groups. To rule out any unrelated proteolytic activity, a proteasome inhibitor (MG132) has been used in these assays.

I. Overview and General Themes

60

5. Perennial Horse Gram (Macrotyloma axillare) Seeds

FIGURE 5.5 BBI inhibitory action over the chymotrypsin- and trypsin-like activities of the proteasome. It is

notable the reduction of approximately 50% in both activities for healthy animals (n ¼ 6) receiving a diet supplemented with BBI from Glycine max (BBIS) or Macrotyloma axillare (BBIM). MG132 was used to control for proteasome’s chymotrypsin-like activity. A group of six animals was used in each treatment group and the results were expressed by arbitrary units of fluorescence/mg protein. Results are expressed as arbitrary units of fluorescence/mg of protein. Bars represent the means  SD. Significantly, different values compared to the control groups were observed at P  .05 using ANOVA/Tukey’s test. Source: Reproduced with permission from Carli et al.31

FIGURE 5.6

BBI inhibitory action over the chymotrypsin- and trypsin-like activities of the proteasome from animals under DMH treatment and diet supplemented with BBI from Glycine max (BBIS) or Macrotyloma axillare (BBIM). DMH treatment greatly increased the trypsin- and chymotrypsin-like activities of the proteasome but they were reduced to control levels when animals were simultaneously fed a diet supplemented with BBI. MG132 was used to control for proteasome’s chymotrypsin-like activity. A group of six animals was used in each treatment group and the results were expressed by arbitrary units of fluorescence/mg protein. Results are expressed as arbitrary units of fluorescence/mg of protein. Bars represent the means  SD. Significantly different values compared to the control groups were observed at P  .05 using ANOVA/Tukey’s test. Source: Reproduced with permission from Carli et al.31

I. Overview and General Themes

Applications to Health Promotion and Disease Prevention e Biotechnology Applications of the Protein Seed Content

61

In summary, our findings corroborate for a cancer suppressive activity of BBI based on their inhibitory actions over the two investigated proteolytic pathways. These results reinforce the possibility of BBI from Macrotyloma axillare to be used as a promising source of molecules for development and formulation of bioproducts aiming to prevent neoplastic transformation.

Macrotyloma axillare Seed Germination and BowmaneBirk Inhibitors The practice of seed germination and consumption as a health-promoting activity has been observed in different cultures. In this context, it was verified that germinated M. axillare seeds exhibit a significant increase in the activity of BBI reaching up to fourfold increase when compared to the activity of the inhibitors isolated from nongerminated seed.18 The activation process requires hydrolysis and reduction of the molecular mass of BBI with the monomer found on 5 days germinated seeds being approximately 6 kDa as opposed to the 8 kDa monomers found on dormant seeds (Fig. 5.7). This reduction in size produces a notable effect on the pharmacokinetic properties of these inhibitors when they are administered to mice. The most evident effect observed was a significant increase in their distribution volume, allowing for a better diffusion of BBI throughout the body (Table 5.1). The higher distribution volume and the increased activity of BBI present in the cotyledon strengthen the possibility of a better efficacy of these inhibitors on cancer prevention. The pharmacokinetic data of tissue distribution revealed a marked accumulation of BBI isolated from M. axillare in the gastric tissue when compared to other organs when BBI are administered orally (Fig. 5.8). This constitutes a striking feature of M. axillare BBI when compared to the ones isolated from soybeans (Glycine max). Although our experimental data on the biodistribution of I125-labeled BBI from Macrotyloma axillare seeds have demonstrated a wide distribution among tissues, oral administration of BBI may be challenging, given its peptide nature. However, recent efforts to obtain BBI

FIGURE 5.7 Inhibitory activity of BBI from Macrotyloma axillare. Relative inhibitory activity of BBI isolated from M. axillare dormant seeds compared to the ones isolated from 5 days germinated seeds. Up to fourfold increase is observed for the antichymotrypsin activity. Source: Original data.

I. Overview and General Themes

62

5. Perennial Horse Gram (Macrotyloma axillare) Seeds

TABLE 5.1

Pharmacokinetic Parameters of BBI Isolated From M. axillare Seeds and Labeled With Radioactive Isotope (I125) Determined After Oral Administration to Swiss Mice

Pharmacokinetic Parameter

Seeds

Cotyledons

a (Distribution constant)

0.0405/minutes

0.0938/minutes

b (Elimination constant)

0.0008/minutes

0.0016/minutes

DV (Distribution volume)

4.76 mL

5.41 mL

t1/2 (Plasma half-life)

14.4 hours

7.2 hours

Source: Data from Patent: Andrade, M.H.G., Santos, A.G. (2007) e Patent number: BRPI0601377 (A).33 Available at: http://v3.espacenet. com/publicationDetails/originalDocument?CC¼BR&NR¼PI0601377A&KC¼A&FT¼D&date¼20071127&DB¼EPODOC&locale¼en_EP.

FIGURE 5.8 Biodistribution of I125-labeled BBI in mice. Specific radioactivity of I125-labeled BBI after oral

administration to mice. The data are represented by average  one standard deviation (n ¼ 6). Note that apart from the blood compartment, the stomach is the preferential site of accumulation of BBI isolated from Macrotyloma axillare seeds. Source: Data from Patent: Andrade, M.H.G., Santos, A.G. (2007) e Patent number: BRPI0601377 (A).33 Available at: http://v3.espacenet.com/publicationDetails/originalDocument? CC¼BR&NR¼PI0601377A&KC¼A&FT¼D&date¼20071127&DB¼EPODOC&locale¼en_EP.

delivery systems have been developed using liposome platforms as alternative routes for their administration.37 Such approaches can be very useful to optimize the BBI pharmacokinetic parameters for their future use as therapeutic biomolecules.

Adverse Effects and Reactions e Toxicity of Macrotyloma axillare To our knowledge, no specific study on the toxicity and adverse effects promoted by M. axillare seeds has been reported to date. Beyond this, it is worth emphasizing that BBI and lectin molecules are consumed daily as constituents of a variety of food and grains belonging to the animal and human diet. However, the two aforementioned constituents

I. Overview and General Themes

References

63

isolated from this plant when administered to animals (lectin and BBI content), particularly at high doses, are expected to provoke adverse effects because of their intrinsic biochemical function. Firstly, the lectin by binding to specific cell receptors might interfere with normal cell physiology, and in this context, there are a number of reports demonstrating the harmful effects of lectins especially in the gastrointestinal tract of recipients.17,38 In parallel, purified BBI or concentrated samples (BBIC) are expected to produce some degree of antinutritional effect by inhibiting the enzymes trypsin and chymotrypsin as they play a key role in protein digestion and therefore amino acid utilization. However, as discussed earlier, many formulations of BBIC (from soybean) have been used as biomolecules in several experiments and have not produced significant adverse effects.16,24 Anyhow, new studies are needed to fully evaluate the therapeutic potential of BBI isolated from Macrotyloma axillare aiming at a better understanding of their benefits and risks to the animal and human health.

Summary Points • M. axillare is a leguminous plant widely found in tropical areas of the world. • Presently the main utilization of M. axillare includes forage to cattle, recovery of eroded soil, and a primary source of biomolecules from its seeds, such as D-pinitol, the anti-A1 lectin (MaL), and BBI. • The 28 kDa lectin monomer isolated from M. axillare (MaL) is specific to the carbohydrate N-Acetyl-a-D-galactosamine (GalNac, anti-A1 antigen). • MaL constitutes a relevant clinical tool as it allows discrimination between blood groups A1 and A2 of the ABO system. • M. axillare seeds are a source of the classic known trypsin and chymotrypsin plant inhibitors called BBI. • BBI-enriched fractions from Macrotyloma axillare revealed to be protective in a colorectal cancer model induced by DMH in mice, based on their inhibitory activities over the proteasome and lysosomal proteolytic pathways. • Pharmacokinetic data obtained after oral administration of BBI isolated from M. axillare revealed considerable accumulation of the inhibitors in the stomach. • BBI isolated from 5 days germinated seeds of M. axillare display increased inhibitory activity over trypsin and chymotrypsin.

References 1. Cook BG, Pengelly BC, Brown SD, et al. Tropical Forages: An Interactive Selection Tool; 2005 [CD-ROM], CSIRO, DPI&F(Qld), CIAT ILRI, Brisbane, Aust. https://cgspace.cgiar.org/handle/10568/49072. Accessed April 11, 2019 2. McGaw LJ, Eloff JN. Ethnoveterinary use of southern African plants and scientific evaluation of their medicinal properties. Journal of Ethnopharmacology. 2008;119(3):559e574. 3. Kamatenesi-Mugisha M, Oryem-Origa H. Traditional herbal remedies used in the management of sexual impotence and erectile dysfunction in western Uganda. African Health Sciences. 2005;5(1):40e49.

I. Overview and General Themes

64

5. Perennial Horse Gram (Macrotyloma axillare) Seeds

4. Morris JB. Macrotyloma axillare and M. uniflorum: descriptor analysis, anthocyanin indexes, and potential uses. Genetic Resources and Crop Evolution. 2008;55(1):5e8. 5. Blumenthal MJ, SNSW. Origin, evaluation and use of Macrotyloma as forage - a review. Tropical Grasslands. 1993;27:16e29. 6. Gomes Confessor J, Fernandes D, Machado T, Rodrigues SC. Avaliação dos resultados obtidos após a implementação de técnicas voltadas ao controle de erosão e recuperação de área degradada no município de uberlândiaMG. In: I Simpósio Mineiro de Geografia - Universidade Federal de Alfenas. Alfenas; 2014. https://www.unifal-mg. edu.br/simgeo/system/files/anexos/Jefferson GomesConfessor.pdf. Accessed April 16, 2019. 7. Kennedy JF, Palva PMG, Corella MTS, Cavalcanti MSM, Coelho LCBB. Lectins, versatile proteins of recognition: a review. Carbohydrate Polymers. 1995;26(3):219e230. 8. Loris R, Hamelryck T, Bouckaert J, Wyns L. Legume lectin structure. Biochimica et Biophysica Acta - Protein Structure and Molecular Enzymology. 1998;1383(1):9e36. 9. Santana MA, Santos AMC, Oliveira ME, et al. A novel and efficient and low-cost methodology for purification of Macrotyloma axillare (Leguminosae) seed lectin. International Journal of Biological Macromolecules. 2008;43(4):352e358. 10. Haylett T, Swart L. Isolation and characterization of an anti-A1 lectin from Macrotyloma axillare. South African Journal of Chemistry. 1982;35(1):33e36. 11. Etzler ME, Kabat EA. Purification and characterization of a lectin (plant hemagglutinin) with blood group A specificity from Dolichos biflorus. Biochemistry. 1970;9(4):869e877. 12. Guerra de Andrade MH, Baba EH, Santana MA. Purification and Isolation of Lectin from Macrotyloma axillare by Thermally Treating Brute Extract of Leguminous Plant Seeds; 2004. Patent number BR200401517-A. https:// worldwide.espacenet.com/publicationDetails/biblio?FT¼D&date¼20051219&DB¼&locale¼en_EP&CC¼BR&NR¼ PI0401517A&KC¼A&ND¼4. 13. Norioka S, Ikenaka T. Amino acid sequences of trypsin-chymotrypsin inhibitors (A-I, A-II, B-I, and B-II) from peanut (Arachis hypogaea): a discussion on the molecular evolution of legume Bowman-Birk type inhibitors. Journal of Biochemistry. 1983;94(2):589e599. 14. Prakash B, Selvaraj S, Murthy MR, Sreerama YN, Rao DR, Gowda LR. Analysis of the amino acid sequences of plant Bowman-Birk inhibitors. Journal of Molecular Evolution. 1996;42(5):560e569. 15. Chen P, Rose J, Love R, H Wei C, Wang B-C. Reactive sites of an anticarcinogenic Bowman-Birk proteinase inhibitor are similar to other trypsin inhibitors. The Journal of Biological Chemistry. 1992;267. 16. Safavi F, Rostami A. Role of serine proteases in inflammation: BowmaneBirk protease inhibitor (BBI) as a potential therapy for autoimmune diseases. Experimental and Molecular Pathology. 2012;93(3):428e433. 17. Losso JN. The biochemical and functional food properties of the Bowman-Birk inhibitor. Critical Reviews in Food Science and Nutrition. 2008;48(1):94e118. 18. Cesar JJ, Santana MA, Oliveira ME, et al. A new extraction and purification methodology of Bowman-Birk inhibitors from seeds and germinated seeds of Macrotyloma axillare. Chromatographia. 2009;69(3e4):357e360. 19. Billings PC, Newberne PM, Kennedy AR. Protease inhibitor suppression of colon and anal gland carcinogenesis induced by dimethylhydrazine. Carcinogenesis. 1990;11(7):1083e1086. 20. Kennedy AR. Prevention of carcinogenesis by protease inhibitors. Cancer Research. 1994;54(7 Suppl.):1999se2005s. 21. Clemente A, Del M, Arques C. Bowman-Birk inhibitors from legumes as colorectal chemopreventive agents. World Journal of Gastroenterology. 2014;20(30):10305e10315. 22. Chen Y-W, Huang S-C, Lin-Shiau S-Y, Lin J-K. BowmaneBirk inhibitor abates proteasome function and suppresses the proliferation of MCF7 breast cancer cells through accumulation of MAP kinase phosphatase-1. Carcinogenesis. 2005;26(7):1296e1306. 23. Saito T, Sato H, Virgona N, et al. Negative growth control of osteosarcoma cell by Bowman-Birk protease inhibitor from soybean; involvement of connexin 43. Cancer Letters. 2007;253(2):249e257. 24. Lichtenstein GR, Deren JJ, Katz S, Lewis JD, Kennedy AR, Ware JH. Bowman-Birk inhibitor concentrate: a novel therapeutic agent for patients with active ulcerative colitis. Digestive Diseases and Sciences. 2008;53(1):175e180. 25. Ma T-C, Guo Le, Zhou R-H, et al. Soybean-derived Bowman-Birk inhibitor (BBI) blocks HIV entry into macrophages. Virology. 2018;513:91e97.

I. Overview and General Themes

References

65

26. Clemente A, Moreno FJ, Marín-Manzano M del C, Jiménez E, Domoney C. The cytotoxic effect of Bowman-Birk isoinhibitors, IBB1 and IBBD2, from soybean ( Glycine max ) on HT29 human colorectal cancer cells is related to their intrinsic ability to inhibit serine proteases. Molecular Nutrition and Food Research. 2010;54(3):396e405. 27. Kennedy AR. The Bowman-Birk inhibitor from soybeans as an anticarcinogenic agent. American Journal of Clinical Nutrition. 1998;68(6 Suppl.):1406e1412. 28. Kennedy AR, Billings PC, Wan XS, Newberne PM. Effects of Bowman-Birk inhibitor on rat colon carcinogenesis. Nutrition and Cancer. 2002;43(2):174e186. 29. Kennedy AR, Beazer-Barclay Y, Kinzler KW, Newberne PM. Suppression of carcinogenesis in the intestines of min mice by the soybean-derived Bowman-Birk inhibitor. Cancer Research. 1996;56(4):679e682. 30. Joubert FJ, Kruger H, Townshend GS, Botes DP. Purification, some properties and the complete primary structures of two protease inhibitors (DE-3 and DE-4) from Macrotyloma axillare seed. European Journal of Biochemistry. 1979;97(1):85e91. 31. de Paula Carli A, de Abreu Vieira PM, Silva KTS, et al. Bowman-Birk inhibitors, proteasome peptidase activities and colorectal pre neoplasias induced by 1,2-dimethylhydrazine in Swiss mice. Food and Chemical Toxicology. 2012;50(5):1405e1412. 32. St Clair WH, Billings PC, Kennedy AR. The effects of the Bowman-Birk protease inhibitor on c-myc expression and cell proliferation in the unirradiated and irradiated mouse colon. Cancer Letters. 1990;52(2):145e152. 33. de Andrade MHG, M.E. Oliveira, Santos AG. Preparation Process of Activated Bowman-Byrk Inhibitors Raw Extracts from Macrotyloma axillare with Higher Activity than the Inhibitor from Soy Bean Seeds; 2006. Patent number: BRPI0601377 (A). Available at: http://v3.espacenet.com/publicationDetails/originalDocument? CC¼BR&NR¼PI0601377A&KC¼A&FT¼D&date¼20071127&DB¼EPODOC&locale¼en_EP. 34. de Andrade MHG, de Castro Borges W, Silva KTS, de Faria FMT, Leal SC, de Santana MA. Purification Process and Isolation of Bowman-Byrk Inhibitors from Glycine max and Macrotyloma axillare; 2010. Patent number: BRPI1005871 (A2). Available at: https://worldwide.espacenet.com/publicationDetails/biblio? FT¼D&date¼20130401&DB¼&locale¼en_EP&CC¼BR&NR¼PI1005871A2&KC¼A2&ND¼4. 35. Yavelow J, Collins M, Birk Y, Troll W, Kennedy a R. Nanomolar concentrations of Bowman-Birk soybean protease inhibitor suppress x-ray-induced transformation in vitro. Proceedings of the National Academy of Sciences of the United States of America. 1985;82(16):5395e5399. 36. Chen C, Kong A-NT. Dietary cancer-chemopreventive compounds: from signaling and gene expression to pharmacological effects. Trends in Pharmacological Sciences. 2005;26(6):318e326. 37. Joanitti GA, Sawant RS, Torchilin VP, de Freitas SM, Azevedo RB. Optimizing liposomes for delivery of Bowman-Birk protease inhibitors d platforms for multiple biomedical applications. Colloids and Surfaces B: Biointerfaces. 2018;167:474e482. 38. Vasconcelos IM, Oliveira JTA. Antinutritional properties of plant lectins. Toxicon. 2004;44(4):385e403.

I. Overview and General Themes

C H A P T E R

6

Biological Properties of a Partially Purified Component of Neem Oil: An Updated and Revised Work Gianfranco Risuleo* Dipartimento di Biologia e Biotecnologie “Charles Darwin”, Sapienza Università di Roma Piazzale Aldo Moro, Roma Italy

Introduction Neem oil is a natural mixture obtained from the seeds of Azadirachta indica (A. Juss), a plant also known as neem tree. Its composition is heterogeneous1 but contains high levels of many organic different compounds. The best-known and studied component is azadirachtin, a tetra-nor-triterpenoid active as pest and insect control. The neem tree Azadirachta indica belongs to the mahogany family of Meliaceae and thrives best in subtropical semi-arid to arid areas. It is originally native of Myanmar but is now spread to India, Africa, Central America, Caribbean area, and the Philippines. Neem is an evergreen plant, its crown has a round shape, and the leaves are rather large typically pinnate with a size of about 30 cm. The efflorescence is represented by small scented white flowers and is abundant in springtime. At 5 years of age, the tree starts producing yellow smooth olive-like fruits of about 2 cm. Within a white hard capsule, one or two seeds resembling almonds are enclosed. The seeds contain high concentrations of azadirachtin, the best characterized and used active principle of the neem tree.1 In India, magic properties are traditionally attributed to the tree’s wood. In any case, the termite-resistant wood is used for housing facilities, furniture, and art items. The bark contains high levels of tannins and a rubbery amber-colored lattice used to dye clothes, which is also used in popular medicine ointments. The oil prepared from the seeds has been extensively used in ayurveda, unani, and homeopathic medicines for centuries.1,2 The Sanskrit

*

The Author is presently on retirement.

Nuts and Seeds in Health and Disease Prevention, Second Edition https://doi.org/10.1016/B978-0-12-818553-7.00006-1

67

Copyright © 2020 Elsevier Inc. All rights reserved.

68

6. Biological Properties of a Partially Purified Component of Neem Oil: An Updated and Revised Work

name sarva roga nivarini means “universal healer of all illnesses.” Therefore, the common Indian name for the tree is “the village pharmacy”; as matter of fact, fruits, seeds, leaves, and roots are used as a remedy against many ill-being predicaments3,4 (see also the web site http://www.natureneem.com/index.htm). The actual situation at the present time is that a few compounds have been purified from the whole oil, e.g., nimbin, nimbinin, and nimbidin, but certainly the secondary metabolite, azadirachtin, constitutes a very important tool in agriculture as biological pest control. Furthermore, neem-derived formulations are used worldwide to control many pests including insects and nematodes. These preparations do not kill but repel parasites, inhibit growth, and alter their behavior and physiology; thus, pests become unable to feed, breed, or metamorphose. In addition, being of low cost, neem products are ideal multipurpose means in disadvantaged countries. In conclusion, neem products find usages in fields as diverse as agriculture, cosmetic industry, and even nutrition since shoots, flowers, and leaves are eaten in India and Southeast Asia.5,6 A number of beneficial effects for human health have been attributed to neem compounds; for instance, fruit, leaves, bark, and roots of A. indica have been reported to combat fungal infections, inflammation, and viral and bacterial infections.7,8 Antitumor and antiproliferative activities have been also ascribed to extracts of A. indica.9,10 Neem tree extracts seem also to exert a negative control on type II diabetes.11 Neem products have become increasingly popular since they have shown positive biological responses with apparently relatively negligible undesired side effects. As a matter of fact, concerning relevant adverse effects and reactions scant data are available in literature. In any case, leaves and bark of the tree do not seem to have a significant toxicity. However, seeds may be poisonous in large doses, and oral administration of 5e30 mL of oil was reported to be toxic in 13 infants. Coma, hepato- and encephalopathy, metabolic acidosis, and death were also episodically reported.2,12 The effects are symptomatic without a specific antidote. In adults, neem oil is relatively safe, but the oral LD50 is 14 mL/kg and 24 mL/kg in rats and rabbits, respectively. High dosages in rats cause also diarrhea, convulsions, motor and respiratory problem, and, eventually, death. In humans, possible allergic reactions should be tested prior to use of neem products. The use as pesticide (limited to non-food crops) was approved by the Environmental Protection Agency. Azadirachtin, one of the most used neem products, is biodegradable, non-mutagenic, and non-toxic to mammals, fish, and birds.13 In conclusion, neem products can be considered safe if used at low dosage and for short time. As far as the work carried out in our laboratory is concerning, we prepared and characterized a methanolic extract of the whole oil. This product was deprived of the terpenoid/ limonoid moiety, and its bioactivity was tested. A critical review of the results obtained with this neem derivative is the focus of this work.

State of the Art in Our Laboratory at the First Edition of the Book The work in our laboratory has been focused on a derivative of the whole seed neem oil. The partially purified derivative was obtained by methanol extraction. This extract defined as

I. Overview and General Themes

Introduction

69

MEx, exhibits a number of biological activities. Here, we summarize significant results obtained by our research team: • The biological effects exhibited by MEx are not attributable to azadirachtin, the best investigated component of the whole neem oil, but rather to other bioactive molecules14; • The differential cytotoxicity was monitored on tumor cells as compared to normal fibroblasts in culture, with the tumor line being more sensitive to administration of the derivative15; • The antiviral action of MEx was shown on murine polyomavirus, whose de novo DNA synthesis is significantly inhibited16; • The neem product MEx increases the membrane fluidity; therefore, its action is mainly exerted at cell membrane level controlling the differential entry of exogenous material17; • The effect on cell survival/proliferation was assessed by membrane lipoperoxidation and synthesis of proliferation gene markers. Results indicated that administration of MEx triggered the apoptotic death.18,19 The data briefly summarized above were extensively presented and critically discussed in a chapter of the book previously published: Nuts and Seeds in Health and Disease Prevention (first ed.).20 The reader is addressed to that work for an extensive and detailed review. The chemical characterization and bioactivity of MEx can be found in14 and reviewed in the study by Berardi and co-workers.21 These studies are supplied with abundant iconographic material and literature data, which will provide full information about the state of the art.

Neem Products: an Update In our laboratory, an attempt was made to further fractionate and isolate a single pure component of the MEx endowed of the biological properties discussed above. To have a starting informative basis, the first series of experiments was carried out to assay the biological activity and properties of the unfractionated oil. However, these trials produced conflicting results because of their low solubility in the aqueous cell culture medium and the different composition that depended on commercial origin of the material under investigation. Methanol extracts, whole MEx, and HT-MEx, this latter was obtained by heating of whole MEx in order to eliminate azadirachtin, showed high cytotoxicity at the investigated concentration range. By definition, this cytotoxic activity was not attributable to azadirachtin since this molecule, due to its thermolabile character, had been inactivated by the heat treatment.14 The fraction defined as “h”14,21 seemed the one endowed of the highest inhibitory effect on cell growth. Therefore, also in the light of the parallel results on the differential cytotoxicity on normal and tumor cells, we attempted a subfractionation of component “h.” We could obtain, by HPLC, four different compounds from this fraction, but, unfortunately, none of them seemed to exhibit a substantial cytotoxic effect (unpublished results). This was rationalized as the existence of a synergic effect of the different compounds present in this fraction. This effectively ruled out the possibility that a single component could be identified as the one endowed of an antiproliferative action per se. This synergic effect is not uncommon in the case of complex natural substances (see for instance 22,23). This makes their analytical investigation very often frustrating if not impossible. Therefore, in spite of the stimulating

I. Overview and General Themes

70

6. Biological Properties of a Partially Purified Component of Neem Oil: An Updated and Revised Work

results in basic research, the elucidation of the molecular mechanisms underlying the multifaceted biological activities of neem remains elusive. In the author’s opinion, the antiviral activity, the apoptosis inducing MEx, and the membrane fluidifying of MEx are unlikely to be attributable to a single component. Rather, as stated above, more chemicals concur to the bioactivity of the mixture. In conclusion, in our laboratory, pursuing a pure active principle endowed of one of these properties was established as unlikely. Bioactive principles for therapeutic purposes must be pure, in the largest majority of cases; consequently, the research on neem compounds in our laboratory was closed. In any case, the work on the bioproperties of neem does not seem to have made a very great progress during the last years: a few statistical data may help to corroborate this statement. As of now (July 2019), works on neem products have produced, during the last 10 years, only 11 review articles and about 115 research papers in English, all over the world. For instance, the use of nanocomposites and nanotechnologies in general, to deliver exogenous agents within a cell, is largely absent from neem-related literature. We would like stress that this specific subject is in extremely rapid expansion in many fields of both basic science and technology. In particular, it is strongly boosting in biomedically-oriented applications (see for instance24e26). Possibly, this derives from the fact that we stress this point again: no single pure compound utilizable in advanced biomedicine has been yet identified. Concerning neem oil and its products, some literature data about putative applications in human health care do exist; nevertheless, to our knowledge, no experimental or clinical trial is yet in progress.27e30 Finally, more “exotic” nondirectly biorelated applications do exist.31,32 However, the use of neem oil and its derivatives seems to find its best fortune as insecticide and in agricultural environments.33e37 Reports have also appeared on the usage of neem in a veterinary context, though it is used in concomitance with other active principle.38e40 This supports our idea that in spite of the undisputable and many-fold biological activities as well as the good biocompatibility of neem, this highly complex natural mixture needs synergic actions exerted by other substances to be fully effective. In conclusion, although neem oil and its derivatives may claim a centuries-long history of use with beneficial effects, their application to the human therapy and advanced biomedicine is still lagging behind. This most likely derives from the very complex composition of the whole oil, which renders the identification and purification of a single active component highly difficult. Of course, this does not mean that reaching a successful goal also in human therapy is impossible, but, as things stand now, the road seems to be very long to go and punctuated by difficulties and traps.

References 1. Schmutterer H. The Neem Tree and Other Meliaceous Plants. Mumbai, India: Neem Foundation; 2002. 2. Brahmachari G. Neem-tree an omnipotent plant: a retrospection. ChemBioChem. 2004;5:408e421 (2004). 3. Subapriya R, Nagini S. Medicinal properties of neem leaves: a review. Current Medicinal Chemistry e Anti-Cancer Agents. March 2005;5(2), 149-6. PMID: 15777222. 4. Gupta SC, Prasad S, Tyagi AK, Kunnumakkara AB, Aggarwal BB. Neem (Azadirachta indica): an indian traditional panacea with modern molecular basis. Phytomedicine. October 15, 2017;34:14e20. https://doi.org/ 10.1016/j.phymed.2017.07.001. Epub 2017 Jul 3.

I. Overview and General Themes

References

71

5. Puri HS. The Divine Tree Azadirachta indica. Amsterdam e The Netherlands: OPA Overseas Publishers Association, published by Harwood Academic Publishers; 1999. 6. Koul O, Wahab S. In: Koul, Wahab, eds. Neem: Today and in New Millennium. Dordrecht e The Netherlands: Kluwer Academic Publishers; 2004. 7. Badam L, Joshi SP, Bedekar SS. In vitro’ antiviral activity of neem (Azadirachta indica. A. Juss) leaf extract against group B coxsackieviruses. Journal of Communicable Diseases. 1999;31:79e90. 8. Parida MM, Upadhyay C, Pandya G, Jana AM. Inhibitory potential of neem (Azadirachta indica Juss) leaves on dengue virus type-2 replication. Journal of Ethnopharmacology. 2002;79, 273-227. 9. Bose A, Haque E, Baral R. Neem leaf preparation induces apoptosis of tumor cells by releasing cytotoxic cytokines from human peripheral blood mononuclear cells. Phytotherapy Research. 2007;21:914e920. 10. Harish Kumar G, Chandra Mohan KV, Jagannadha Rao A, Nagini S. Nimbolide a limonoid from Azadirachta indica inhibits proliferation and induces apoptosis of human choriocarcinoma (BeWo) cells. Investigational New Drugs. 2009;27:246e252. 11. Saxena A, Vikram NK. Role of selected Indian plants in management of type 2 diabetes: a review. Journal of Alternative & Complementary Medicine. 2004;10:369e378. 12. Dhongade RK, Kavade SG, Damle RS. Neem oil poisoning. Indian Pediatrics. 2008;45:56e57. 13. Gandhi M, Lal R, Sankaranarayanan A, Banerjee CK, Sharma PL. Acute toxicity study of the oil from Azadirachta indica seed (neem oil). Journal of Ethnopharmacology. 1988;23(1):39e51. 14. Di Ilio V, Pasquariello N, van der Esch SA, Cristofaro M, Scarsella G, Risuleo G. Cytotoxic and antiproliferative effects induced by a non-terpenoid polar extract of A. indica seeds on 3T6 murine fibroblasts in culture. Molecular and Cellular Biochemistry. 2006;287:69e77. 15. Ricci F, Berardi V, Risuleo G. Differential cytotoxicity of MEX: a component of Neem oil whose action is exerted at the cell membrane level. Molecules. 2008;14:122e132. 16. Berardi V, Aiello C, Bonincontro A, Risuleo G. Alterations of the plasma membrane caused by murine polyomavirus proliferation: an electrorotation study. Journal of Membrane Biology. 2009;229:19e25. 17. Bonincontro A, Di Ilio V, Pedata O, Risuleo G. Dielectric properties of the plasma membrane of cultured murine fibroblasts treated with a nonterpenoid extract of Azadirachta indica seeds. Journal of Membrane Biology. 2007;215:75e79. 18. Mattetti A, Risuleo G. Apoptosis: a mode of cell death. September 2014 Biochemistry & Molecular Biology. 2014;2:34e39. https://doi.org/10.12966/bmb.09.02.2014. Print ISSN: 2331-8252 Online ISSN: 2331-8260. 19. Aiello C, Andreozzi P, La Mesa C, Risuleo G. Biological activity of SDS-CTAB cat-anionic vesicles in cultured cells and assessment of their cytotoxicity ending in apoptosis. Journal of Colloids and Surfaces B: Biointerfaces. 2010;78:149e154. 20. Aiello C, Berardi V, Ricci F, Risuleo G. In: Preedy VR, Watson RR, Patel VB, eds. Biological properties of a methanolic extract of neem oil, a natural oil from the seeds of the Neem Tree (Azadirachta indica var. A. Juss). London, Burlington, San Diego: Academic Press is an imprint of Elsevier; 2011:813e821. ISBN 978-0-12-375688-6 (Chapter 96), pg. 813-821.1517. 21. Berardi V, Galati G, Risuleo G. Bioactivity of MEx: a derivative of whole neem oil obtained by methanol extraction. J. Biological Medicine. 2011;1:23e2918. 22. Pezzani R, Salehi B, Vitalini S, et al. Synergistic effects of plant derivatives and conventional chemotherapeutic agents: an update on the cancer perspective. Medicina. April 17, 2019;55(4):E110. https://doi.org/10.3390/ medicina55040110. PubMed PMID:30999703; PubMed Central PMCID: PMC6524059. 23. Yang Y, Zhang Z, Li S, Ye X, Li X, He K. Synergy effects of herb extracts:pharmacokinetics and pharmacodynamic basis. Fitoterapia. January 2014;92:133e147. https://doi.org/10.1016/j.fitote.2013.10.010. Epub 2013 Oct 28. PubMed PMID: 24177191. 24. Muzi L, Cadarsi S, Mouchet F, et al. Examining the impact of few-layer graphene using cellular and amphibian models. 2D Materials. 2016;3:1e10. https://doi.org/10.1088/2053-1583/3/2/025009, 025009. 25. Bamburowicz-Klimkowska M, Poplawska M, Grudzinski IP. Nanocomposites asbiomolecules delivery agents in nanomedicine. Journal of Nanobiotechnology. April 3, 2019;17(1):48. https://doi.org/10.1186/s12951-019-0479-x. Review. PubMed PMID: 30943985; PubMed Central PMCID: PMC6448271. 26. Risuleo G, La Mesa C. Nanoparticles and molecular delivery: state of the art and future perspectives. In: Gupta R, ed. Nutraceuticals in Veterinary Medicine. Nature Publishing Group, Palgrave Macmillan, Macmillan Education and Springer Science; 2019 (Press).

I. Overview and General Themes

72

6. Biological Properties of a Partially Purified Component of Neem Oil: An Updated and Revised Work

27. Tharmarajah L, Samarakoon SR, Ediriweera MK, et al. In vitro anticancer effect of gedunin on human teratocarcinomal (NTERA-2) cancer stem-like cells. BioMed Research International. 2017;2017:2413197. https://doi.org/ 10.1155/2017/2413197. Epub 2017 Jun 7. PubMed PMID:28680880; PubMed Central PMCID: PMC5478822. 28. Avinash B, Venu R, Prasad TNVKV, Alpha Raj M, Srinivasa Rao K, Srilatha C. Synthesis and characterisation of neem leaf extract, 2, 3-dehydrosalanol and quercetin dihydrate mediated silver nano particles for therapeutic applications. IET Nanobiotechnology. June 2017;11(4):383e389. https://doi.org/10.1049/iet-nbt.2016.0095. PubMed PMID: 28530186. 29. Patel SM, Nagulapalli Venkata KC, Bhattacharyya P, Sethi G, Bishayee A. Potential of neem (Azadirachta indica L.) for prevention and treatment of oncologic diseases. Seminars in Cancer Biology. October 2016;40e41:100e115. https://doi.org/10.1016/j.semcancer.2016.03.002. Epub 2016 Mar 24. Review. PubMed PMID: 27019417. 30. Moga MA, Balan A, Anastasiu CV, Dimienescu OG, Neculoiu CD, Gavriș C. An overview on the anticancer activity of Azadirachta indica (neem) in gynecological cancers. International Journal of Molecular Sciences. December 5, 2018;19(12):E3898. https://doi.org/10.3390/ijms19123898. PubMed PMID: 30563141; PubMed Central PMCID: PMC6321405. 31. Tayyab Z, Safi SZ, Rahim A, et al. Preparation of cellulosic Ag-nanocomposites using an ionic liquid. Journal of Biomaterials Science, Polymer Edition. 2019 May e June;30(9):785e796. https://doi.org/10.1080/09205063.2019.1605869. Epub 2019 Apr 24. PubMed PMID: 31018777. 32. Devarajan Y, Munuswamy DB, Mahalingam A. Influence of nano-additive on performance and emission characteristics of a diesel engine running on neat neem oil biodiesel. Environmental Science and Pollution Research International. September 2018;25(26):26167e26172. https://doi.org/10.1007/s11356-018-2618-6. Epub 2018 Jul 4. PubMed PMID: 29974438. 33. Saleem S, Muhammad G, Hussain MA, Bukhari SNA. A comprehensive review of phytochemical profile, bioactives for pharmaceuticals, and pharmacological attributes of Azadirachta indica. Phytotherapy Research. July 2018;32(7):1241e1272. https://doi.org/10.1002/ptr.6076. Epub 2018 Apr 19. PMID: 29671907. 34. Benelli G, Canale A, Toniolo C, et al. Neem (Azadirachta indica): towards the ideal insecticide? Natural Product Research. February 2017;31(4):369e386. https://doi.org/10.1080/14786419.2016.1214834. Epub 2016 Aug 12. Review. PubMed PMID: 27687478. 35. Benelli G, Murugan K, Panneerselvam C, Madhiyazhagan P, Conti B, Nicoletti M. Old ingredients for a new recipe? Neem cake, a low-cost botanical by-product in the fight against mosquito-borne diseases. Parasitology Research. February 2015;114(2):391e397. https://doi.org/10.1007/s00436-014-4286-x. Epub 2015 Jan 7. PubMed PMID: 25563612. 36. Senthil-Nathan S. Physiological and biochemical effect of neem and other Meliaceae plants secondary metabolites against Lepidopteran insects. Frontiers in Physiology. December 20, 2013;4:359. https://doi.org/10.3389/ fphys.2013.00359. Review. PubMed PMID:24391591; PubMed Central PMCID: PMC3868951. 37. Chaudhary S, Kanwar RK, Sehgal A, et al. Progress on Azadirachta indica based biopesticides in replacing synthetic toxic pesticides. Frontiers of Plant Science. May 8, 2017;8:610. https://doi.org/10.3389/fpls.2017.00610. eCollection 2017. PMID: 28533783. 38. Fiorella C, Delia F, Domenico O, et al. A formulation of neem and hypericum oily extract for the treatment of the wound myiasis by Wohlfahrtia magnifica in domestic animals. Parasitology Research. June 19, 2019. https:// doi.org/10.1007/s00436-019-06375-x [Epub ahead of print] PubMed PMID: 31218416. 39. Sayed-Ahmed MZ, Ahdy AM, Younis EE, El-Khodery SA, Baraka HN. Comparative effectiveness of Sumaq and Neem extract cream, Eniloconazole and glycerine iodine on dermatophytosis in Arabian horses: a randomized clinical trial. Tropical Animal Health and Production. May 2019;51(4):905e910. https://doi.org/10.1007/s11250018-1773-6. Epub 2018 Dec 15. PubMed PMID: 30554365. 40. Attia MM, Khalifa MM, Mahdy OA. The prevalence of Gasterophilus intestinalis (Diptera: oestridae) in donkeys (Equus asinus) in Egypt with special reference to larvicidal effects of neem seed oil extract (Azadirachta indica) on third stage larvae. Open Veterinary Journal. 2018;8(4):423e431. https://doi.org/10.4314/ovj.v8i4.12. Epub 2018 Nov 15. PubMed PMID: 30538934; PubMed Central PMCID: PMC6243205.

I. Overview and General Themes

C H A P T E R

7

Bioactive Compounds of Oregano Seeds Havva Atar, Hatice Çölgeçen Department of Biology, Faculty of Arts and Sciences, Zonguldak Bülent Ecevit University, Zonguldak, Turkey List of Abbreviation EO Essential oil

Introduction Medicinal plants have highly valuable natural compounds because of their biological activities. Among many medicinal plants worldwide, Origanum, also known as oregano, is the most widespread species in Turkey. There, it is consumed as a spice as well as a medicinal plant because of its appetizing smell and taste. It is an aromatic plant, as well. In Turkey, oregano production started in 2004 at 7,000 tons from a nearly 5.5-hectare field. Since then, it has been increasingly cultivated in Turkey and across other continents.1,2 A main reason for this rise in the production of Origanum is its rich bioactive compounds: essential oil (EO), phenolic compounds, flavonoids, flavanones, tocopherols, carvacrol, benzoic acid, rosmarinic acid, and cinnamic acid derivatives.3e5 These various bioactive compounds serve as an important herbal cure for several therapeutic indications but also for some other purposes. Hence, it is high time that we know more about this different medicinal plant, Origanum sp.

Nuts and Seeds in Health and Disease Prevention, Second Edition https://doi.org/10.1016/B978-0-12-818553-7.00007-3

73

Copyright © 2020 Elsevier Inc. All rights reserved.

74

7. Bioactive Compounds of Oregano Seeds

Botanical Description The most important species of the Lamiaceae family in Turkey is Origanum. Within the genus Origanum L. are 41 species and 52 taxa, 21 of which are endemic species.1 However, in Turkey, the genus Origanum L. consists of 23 species and 27 taxa, 16 of which are endemic species.6,7 Among these, Origanum species cultivated for medicinal purposes are O. onites, O. vulgare, O. majorana, O. minutiflorum (endemic), and O. syriacum.8 Although it is assumed to be a plant indigenous to the Mediterranean region of Turkey, surprisingly, it can withstand different and severe conditions such as cold or drought. Also, it is not selective in terms of soil, so it can be cultivated in other regions such as Aegean and southeast Anatolia.7

Historical Cultivation and Use Origanum had an important place in ancient pre-Judaic religions and mythology. It is related that Aphrodite, Venus, and the ancient Egyptian god Osiris used to wear this plant during rites. The Greek goddess Artemis and the goddess of Rome, Diana, were also associated with the plant, and both Artemis and Lucina worn a crown made of it.9 Besides, according to legends, some kings used it as a perfume after a bath.9,10 Since antiquity, people around the world have used plants to treat a variety of diseases. For instance, they have added plants to salads or dishes, made herbal teas, and turned them into creams. That ancient people called plants miraculous has brought to mind a question for today’s scientists: what is therapeutic in plants? Ethnobotany, ethnopharmacology, and medical studies have proven that plants are effective in treating diseases such as cancer, AIDS, Alzheimer, alcoholism, microbial infections, and premature aging.11 Therefore, interest in medicinal plants has increased. In recent years, Turkey has increased Origanum cultivation because of its commercial, economic, and medicinal value. Commercially, Origanum species are used as a spice in sausages, soup, salads, and snack foods.12 In particular, Origanum onites (Syn. O. smyrnaeum L.), and O. vulgare are at the top of the list of commercial Origanum species in Turkey. Origanum onites is mainly a wild plant still cultivated.13,14

Current Cultivation and Use The seeds and aerial parts of Origanum species have a high percentage of EO, a secondary metabolite formed in aromatic plants.15 This is why Origanum species are used as spices and condiments and in herbal teas and cosmetics for their aromatic taste and scent.2,3 Also, there are studies on using as seed conservation.16 To prolong the expiration date of many packaged products such as fish or meat, the EO of this plant is commonly used because it is environment friendly; it is also used to reduce weight loss, microbial proliferation, oxidation, and respiration rate in products.16,17 Moreover, several technologies have attracted the attention of researchers worldwide to lengthen the shelf life of fruits. For example, Hashemi et al. coated fresh-cut apricots successfully using O. vulgare subsp. viride EO. In another study, Jouki et al. described an application of edible films containing oregano EO for the same purpose.18,19

I. Overview and General Themes

Adverse Effects and Reactions, Allergies, and Toxicity

75

Applications for Health Promotion and Disease Prevention All plants are rich in bioactive compounds. The flowers, buds, seeds, leaves, branches, bark, fruits, and roots of Origanum have EO. Its leaves and flowering stems are naturally antiseptic and antioxidant because of the high thymol content in one type of EO.3,20,21 Tameme et al. reported that the aerial flowering parts of O. vulgare subsp. viride are used in Iranian traditional medicine as a diuretic, stomachic, antineuralgic, antitussive, and expectorant. Origanum species also have a lot of benefits for respiratory diseases (e.g., asthma, cough, and chest pain); abnormal menstrual cycles; kidney and liver diseases; metabolic, hormonal, and neuronal disorders; and skin and urogenital system diseases.22 Moreover, it is known that oregano water is imbibed for gastrointestinal disorders to reduce blood cholesterol and glucose levels.3 The seeds of Origanum majorana include catechin, cinnamic acid, gallic acid, and ascorbic acid.5 Also, Marin et al. determined the seed oil composition of O. vulgare to be 4.2% palmitic, 1.3% stearic, 6.1% oleic, 23.8% linoleic, and 64.6% linolenic acids.23 In addition, in studies on the seeds of Origanum tytthanthum Gontsch, fatty acids and triacylglycerides were identified, whereas in the aboveground part, EO was found.24 In O. vulgare and O. onites seeds, a fatty acid composition and planteose25 were revealed. Because of its various bioactive compounds, this plant has many applications ranging from promoting health to preventing disease. Origanum seed and oils extracted from its leaves are sold as a food additive or a strong antioxidant and antimicrobial agent. In addition, the oil of Origanum is rich in carvacrol, thymol, p-cymene and g-terpinene, which is used to lower blood cholesterol and a painkiller in rheumatism or toothache.14,26 The EO also has antimicrobial, antioxidant, antiviral, and anticancer activity.18,22,27,28 As claimed by researchers, Origanum oil has traditionally been used as a treatment for indigestion, cough, painful or late menstruation, respiratory disorders, indigestion, dental caries, rheumatoid arthritis, urinary tract disorders, dyspepsia, scrofulosis, and a spasmolytic effect on smooth muscle.2,16,29 Samarth et al. also reported that oregano extract reduced micronuclei frequencies in human lymphocytes and mouse bone marrow. Furthermore, oregano extract is radioprotective.30

Adverse Effects and Reactions, Allergies, and Toxicity Although Origanum species have a medicinal effect, they include EOs as a potential source of functional biocompounds. Thus, when combined with other compounds, they are likely to cause an allergic reaction. For example, if Origanum oil is used at a higher dose, it can cause symptoms such as nausea, vomiting, gastric distress, and central hyperactivity. Especial in infants, children, pregnant or breastfeeding women, and people with diabetes, the prescribed dose of commercial oregano oil should not be exceeded. Guldiken et al. reported that high doses of phytochemicals in herbs and spices can result in toxic effects including carcinogenic, neurotoxic, genotoxic, teratogenic, cytotoxic, nephrotoxic, hepatotoxic, and gastrointestinal.28 Moreover, it has been proved that large doses of phytochemicals in herbs and spices have fumigant toxicity and an insecticidal effect on insects.2

I. Overview and General Themes

76

7. Bioactive Compounds of Oregano Seeds

Summary Points • Origanum is a plant that has been grown in the Mediterranean region since ancient times. • Origanum is domesticated in Turkey, so its seeds are cultivated. • Origanum has a great importance in the pharmaceutical, medical, and agricultural industries. Moreover, researchers are interested in preparing functional foods from them. In addition, Origanum seeds have important biological activity. • Origanum seeds contain a medicinally important EO, fatty acids, and a terpenoid composition. • Owing to their EO, Origanum seed has naturally therapeutic effects. • An edible film coating has been improved because of the bioactive compounds of Origanum. Because of this, the shelf life of foods has been extended.

References 1. Davis PH. Flora of Turkey and the East Aegean Islands. Vol. 7. Edinburgh: Edinburgh University Press; 1982. 2. Tameme HJ, Hameed IH, Idan SA, Hadi MH. Biochemical analysis of Origanum vulgare seeds by fouriertransform infrared (FT-IR) spectroscopy and gas chromatography-mass spectrometry (GC-MS). Journal of Pharmacognosy and Phytotherapy. 2015;7(9):221e237. 3. Demirci F, Paper DH, Franz G, Husnu K, Baser C. Investigation of the Origanum onites L. Essential oil using the chorioallantoic membrane (CAM) assay. Journal of Agricultural and Food Chemistry. 2004;52:251e254. 4. Suhaj M. Spice antioxidants isolation and their antiradical activity: a review. Journal of Food Composition and Analysis. 2006;19:531e537. 5. Dhull SB, Kaur P, Purewal SS. Phytochemical analysis, phenolic compounds, condensed tannin content and antioxidant potential in Marwa (Origanum majorana) seed extracts. Resource-Efficient Technologies. 2016;2:168e174. 6. Güner A, Özhatay N, Ekim T, Baser KHC. Flora of Turkey and the East Aegean Islands, Supplement 2, Volume 11. Edinburgh: Edinburgh University Press; 2000. _ 7. Gürbüz B, Ipek A, Ayvaz N. Türkiye Florasındaki Origanum Türlerinin yayılıs alanları ve Ticareti. Türk Bilimsel Derlemeler Dergisi. 2011;4(2):55e58. 8. Azcan N, Kara M, Demirci B, Baser KHC. Fatty acids of the seeds of Origanum onites L. and O. vulgare L. Lipids. 2004;39:487e489. 9. Adams J. Oregano and Marjoram an Herb Society of America Guide to the Genus Origanum. Kirtland, OH: Herb Society of America-9019 Kirtland Chardon Rd.; 2005. 10. Alakbarov F. Aromatic herbal baths of the ancients. The Journal of the American Botanical Council. 2003;57:40e49. 11. Heinrich M, Bremner P. Ethnobotany and ethnopharmacy-their role for anti-cancer drug development. Current Drug Targets. 2006;7:239e245. 12. Novak JN, Langbehn J, Pank F, Franz CM. Essential oil compounds in a historical sample of marjoram (Origanum majorana L., Lamiaceae). Flavour and Fragrance Journal. 2002;17:175e180. 13. Aydın S, Baser KHC, Oztürk Y. The chemistry and pharmacology of Origanum (Kekik) water. In: Essential Oils: Basic and Applied Research, 27th International Symposium on Essential Oils, 8-11 September, Vienna, Austria, 52e60. 1997. 14. Baser KHC. The Turkish Origanum species. In: Kintzios SE, ed. Oregano, the Genera Origanum and Lippia. London: Taylor and Francis; 2002:108e126. 15. Bakkali F, Averbeck S, Averbeck D, Idaomar M. Biological effects of essential oils e a review. Food and Chemical Toxicology. 2008;46:446e475. _ Basil-seed gum containing Origanum vulgare subsp. 16. Hashemi SMG, Khaneghah AM, Ghahfarrokhi MG, Es I. viride essential oil as edible coating for fresh cut apricots. Postharvest Biology and Technology. 2017;125:26e34.

I. Overview and General Themes

References

77

17. Okcul Z, Yavuz Y, Kerse S. Edible film and coating applications in fruits and vegetables. Alinteri Journal of Agricultural Sciences. 2018;33(2):221e226. 18. Jouki M, Yazdi FT, Mortazavi SA, Koocheki A, Khazaei N. Effect of quince seed mucilage edible films incorporated with oregano or thyme essential oil on shelf life extension of refrigerated rainbow trout fillets. International Journal of Food Microbiology. 2014a;174:88e97. 19. Jouki M, Yazdi FT, Mortazavi SA, Koocheki A, Khazaei N. Quince seed mucilage films incorporated with oregano essential oil: physical, thermal, barrier, antioxidant and antibacterial properties. Food Hydrocolloids. 2014b;36:9e19. 20. Cervato G, Carabelli M, Gervasio S, Cittera A, Cazzola R, Cestaro B. Antioxidant properties of oregano (Origanum vulgare) leaf extracts. Journal of Food Biochemistry. 2000;24:453e465. 21. Bendini A, Toschı TG, Lercker G. Antioxidant activity of oregano (Origanum vulgare L.) leaves. Italian Journal of Food Science. 2002;14(1):17e24. 22. Beltran JMG, Esteban MA. Properties and applications of plants of Origanum sp. Genus. SM Journal of Biology. 2016;2(1):1006. 23. Marin PD, Sajdl V, Kapor S, Tatic B, Petkovic B. Fatty acids of the saturejoideae, ajugoideae and scutellarioideae (Lamiaceae). Phytochemistry. 1991;30:2979e2982. 24. Asilbekova DT, Glushenkova AI, Azcan N, Özek T, Baser KHC. Lipids of Origanum tytthanthum. Khimija Prirodnykh Soyedineniy. 2000;2:100e102. 25. Ietswaart JH. A Taxonomic Revision of the Genus Origanum (Labiatae). Leiden university press the Hague/Boston/ London; 1980. 26. Lemioglu F, Bagci S. Evaluation of the long-term effects of Oleum origani on the toxicity induced by administration of streptozotocin in rats. Journal of Pharmacy and Pharmacology. 1997;49:1157e1161. 27. Kaefer CM, Milner JA. The role of herbs and spices in cancer prevention. Journal of Nutritional Biochemistry. 2008;19:347e361. _ Capanoglu E. Phytochemicals of herbs and spices: 28. Guldiken B, Ozkan G, Catalkaya G, Ceylan FD, Yalcinkaya IE, health versus toxicological effects. Food and Chemical Toxicology. 2018;119:37e49. 29. Farashah HD, Afshari RT, Sharifzadeh F, Chavoshinasab S. Germination improvement and a-amylase and b-1,3glucanase activity in dormant and nondormant seeds of Oregano (Origanum vulgare). Australian Journal of Crop Science. 2011;5(4):421e427. 30. Samarth RM, Samarth M, Matsumoto Y. Medicinally important aromatic plants with radioprotective activity. Future Science OA. 2017;3(4).

I. Overview and General Themes

C H A P T E R

8

Mango Seed: Mango (Mangifera indica L.) Seed and Its Fats Julio A. Solís-Fuentes1, María del Carmen Durán-de-Bazúa2 1

Instituto de Ciencias Básicas, Universidad Veracruzana, Avenida Luis Castelazo Ayala s/n, Xalapa, Veracruz, Mexico; 2Chemical Engineering Department, Facultad de Química, Universidad Nacional Autónoma de México, Cd. Universitaria, Ciudad de México, Mexico

List of Abbreviation CB Cocoa butter CBA Cocoa butter alternative CBE Cocoa butter equivalent FA Fatty acid FFA Free fatty acids FO Fat and oils GAE Gallic acid equivalent MKF Mango kernel fat MSK Mango seed kernel POP 2-Oleodipalmitin POS 2-oleopalmitostearin SFC Solid fat content SOO 1-Stearoyl-2,3-dioleoyl glycerol SOS 2-Oleodistearin TG Triacylglycerol WHO World Health Organization

Introduction Lipids, and particularly FOs, are a large group of important compounds in the structure and functioning of cells, essential in the diet for their nutritional value and highly desirable for their effect on the functional properties of food. Unlike oil, natural vegetable fats are

Nuts and Seeds in Health and Disease Prevention, Second Edition https://doi.org/10.1016/B978-0-12-818553-7.00008-5

79

Copyright © 2020 Elsevier Inc. All rights reserved.

80

8. Mango Seed: Mango (Mangifera indica L.) Seed and Its Fats

scarce and have multiple applications. Mango kernel fat (MKF) has a composition and physical characteristics that make it a consumer alternative to processed semi-solid edible fats high in artificial trans-FA content, with serious adverse effects on human health maintenance, inclusive with an increased mortality rate in several diseases.

Botanical Description Economically speaking, the mango is the most important fruit crop of the family Anacardiaceae (cashew or poison ivy family) in the order of Sapindales. The family contains 73 genera and between 600 and 700 species, well-known for the presence of caustic resins in the leaves, bark, and fruits. Several of these, including mango, may cause some type of dermatitis in humans. The genus Mangifera contains about 60 species, of which about 15 produce edible fruits, among them M. sylvatica, a possible ancestor of M. indica. Currently there are over 1000 known varieties of mango, whose nomenclature is sometimes complicated because of certain regionalisms. In the world, only a few varieties are grown on a commercial scale and traded. The fruit has a large central stone, flattened, with a woody cover containing a nucleus or kernel with a single embryo or two to five embryos (Hindu and Indo-Chinese varieties).1,2

Historical Cultivation and Usage Mango has been cultivated since prehistoric times. Apparently, it is endemic to northeastern India, where it is estimated to be cultivated from 4000 to 6000 years ago, and Myanmar (Burma), possibly also to Ceylon. It was distributed toward China in the seventh century and throughout Southeast Asia and the Malay Archipelago, from where it spread, centuries later, to Africa and the New World through the first Portuguese and Spanish sea routes and colonization. The geographical distribution of mango in the world was completed with the first permanent plantation in Florida dating from the 1860s.2,2a Historically, its flesh has been used almost exclusively as fresh and processed fruit. Various plant parts have been used in traditional medicine as a cure for a number of diseases. The kernel seed has been edible for human people and animals in some Asian groups and, in different preparations, has been used as a vermifuge and as an astringent in diarrhea, hemorrhages, and bleeding hemorrhoids. The kernel fat has been administered in cases of gastritis.1

Present-Day Cultivation and Usage Mango fruit is one of the most important crops. At present, the mango is cultivated on a commercial scale all over the tropical and subtropical regions over 4.2 million hectares in around 100 countries, covering about 50% of the cultivated area for the tropical fruits worldwide. From the year 1997 to 2017, mango production grew 2.1 times, going from 18 million to 38 million tons, represented close to 40% of the aggregate production of the main tropical fruits. Asia concentrates 74.4%, Africa 13.1%, and America 12.4% of mango production in the world.3,3a Up to date, mango is still mainly used as food; however, extensive research is realized about its nutritional properties, bioactive constituents, and the lipids from MSK. MKF is one of the vegetable fats allowed in the manufacture of chocolate as CBE, and it has been subjected to an increasing revaluation with promising prospects toward wider uses.4e6,6a I. Overview and General Themes

Applications to Health Promotion and Disease Prevention

81

Applications to Health Promotion and Disease Prevention The MSK is being revalued not only for its quality as a natural food e edible, nontoxic, and nutritious with high-quality protein and amino acid score e but also for its bioactive compounds which have shown positive impacts, directly or indirectly, on health and nutrition.6b The bioactive constituents polyphenols and different lipid fractions, some of which are also present in MKF, have been evaluated in their protective action for the prevention of different diseases and health disorders. MKF has shown significant functional and physicochemical characteristics that make it possible to replace other fats, mainly those trans-fats widely used as shortenings, margarines, ingredients, or fat matrices in the food, pharmaceutical, or cosmetic industries. Such industrial trans-fats are now facing serious objections because of their adverse effects on health.6c,6d MKF has shown great resistance to self-oxidation and to be able to replace synthetic antioxidants in food stabilization which can contribute to reach more natural and healthy elaborated foods and dietary patterns.6b

The Chemical Composition and Lipids of the Mango Seed Kernel By its biological nature, MSK has a composition that responds to varietal and phenotypic variations. The available data are for a substantial but still minority number of commercially important varieties from major producing regions. Table 8.1 gives the range of values most often reported for the relevant chemical constituents of the kernel. The seed, depending on the variety, can constitute between 3 and 25% of the total mass of the fruit and the kernel occupies from 24 to 85% of the seed; it has a moisture content between 33 and 86% and the solid dried matter has proteins (4.0e8.1%), crude fiber (1.7e7.6%), ash (1.0e3.7%) with several identified minerals: K, P, Mg, Na, Ca, Fe, Zn, and Cu; total carbohydrates (70e76%), around 0.1e8.6% of phenolic compounds (although values of up to 29.2% db have been reported in a variety of Thai mango) and between 3.7 and 13.7% of crude fat. MSK proteins have high scores of essential amino acids (78) and protein quality (177e189 adults) and an in vitro digestibility between 26.7 and 29.8%.6,7,7a,7b Although phenolic compounds act as antinutritive factors, they have become the subject of interest because of their high bioactivities; MSK is comparable to parts of other plants considered to have of greater antioxidant properties.7a Up to a total of around 20 different phenolic compounds have been identified in the MSK, and many of them have shown antioxidant, antimicrobial, and antiproliferative activities.4,6,7b,8,9,9a Lipids are important components of food and also basic structural and functional constituents of cells; therefore, they are decisive in states of health and illness of individuals. MKF has been studied on its yield during the extraction, its toxicological safety, its composition of fatty acid, glycerides, unsaponifiable compounds, and complex lipids, and its chemical, physical, thermal, and phase properties. As edible fat, such aspects are important to evaluate their effect on health promotion as well as its applicability as a natural substitute or a supplement to fats from other sources. Fat yields from the MSK fluctuate in a wide range among varieties. Van Pee et al.10 reported values between 6.8 and 12.6% (db) for African varieties; others such as Lakshminarayanan

I. Overview and General Themes

82 TABLE 8.1

8. Mango Seed: Mango (Mangifera indica L.) Seed and Its Fats

Ranges of Main Physicochemical Properties of Mango (Mangifera indica) Kernel Seed (MSK) and Its Fats (MKF)

Kernel Composition

Values

Units

References

Seed in fruit

3e25

%

11

Kernel in seed

24e85

11,15d

Moisture

33e86

11

Protein

4.0e10.0

11,21

Essential amino acids

25.2e37.3

g amino acid/100 g

21

Crude fiber

1.7e7.6

%

7,17

Crude fat

3.7e13.7

11,22

Ash

1.0e3.7

11

Total carbohydrates

70e76

17

Proximal composition

Bioactive and minerals Phenolic compounds

0.1e8.6

g GAE/100 g db

6,23

Potassium

60e365

mg/100 g

7a,24

Phosphorus

20e230

7a,24

Magnesium

22e980

7a,24

Sodium

2.6e150

7a,24

Calcium

10e450

7a,24

Iron

.05). Significant differences in TEAC and FRAP were detected among purple wheat milling fractions, with the highest value observed for the bran and shorts fraction. Reprinted (adapted) with permission from (Siebenhandl, Grausgruber, Pellegrini, Rio, Fogliano, Pernice, Berghofer31). Copyright (2007) American Chemical Society.

to 1920 mmol TE/100 g (Charcoal). Wheat genotype, growing environment, and extraction solvents may cause differences in ORAC values among samples.46 The DPPH RSA assay is a method used to evaluate the antiradical activity of antioxidants such as dietary phenolic compounds in plants.48 The assay follows the ET mechanism46 although the stable DPPH• radical has an unpaired valence electron at one of its nitrogen bridge atoms.49 This is because the rate-limiting step for the reaction involves a rapid ET from the antioxidant (e.g., phenoxide anion) to DPPH. The hydrogen atom abstracted from the neutral phenolic compound by DPPH• is marginal because it occurs very slowly in alcohols such as methanol and ethanol.46 Fig. 10.6 shows the DPPH RSA of different purple wheat extracts, and the results are expressed as mmol of Trolox equivalent (TE) per 100 g of grain on dry weight basis.8 Consistent with the ORAC results, the acidified methanol extracts (715e857 mmol TE/100 g) exhibited higher DPPH RSA than the acetone extracts (492e616 mmol TE/100 g), but with smaller differences (1.2e1.5-fold). In both extracts, the RSA decreased in the order Charcoal > Konini > Indigo.8 Yu, Beta9 compared the DPPH RSA between Konini and Indigo wholemeal flours and observed no difference between the varieties, both for the soluble free and insoluble-bound extracts. However, DPPH RSA of the insoluble-bound extracts was higher than that of the corresponding soluble free extracts.9 While significant RSA was measured in all the purple wheat samples tested, the variety Charcoal had the highest antioxidant activity using both ORAC and DPPH assays.8 This indicates that among the three varieties (Charcoal, Konini, and Indigo), Charcoal may provide the highest antioxidant health benefits if grown in the right environment. Between Konini and Indigo, however, while Liu, Qiu, Beta8 observed Konini to have higher antioxidant activity (ORAC and DPPH) than Indigo, Yu, Beta9 found their DPPH RSA to be comparable. It is noteworthy that the insoluble-bound phenolic extracts of the two purple wheat varieties Konini and Indigo had higher DPPH RSA than that of the soluble free phenolic extracts. This trend was observed also in the TPCs of the varieties.9 This, together with the other observations, demonstrates relationships between the total phenolic and anthocyanin contents and the antioxidant activities and implies that the antioxidant activities measured using the radical scavenging activities are due to the constituent phenolic compounds in the purple wheat grains.

II. Role of Seeds in Nutrition and Antioxidant Activities

122

10. Purple Wheat (Triticum sp.) Seeds: Phenolic Composition and Antioxidant Properties

Trolox equivalent antioxidant capacity (TEAC) is based on the ET mechanism and measures the scavenging ability of antioxidants against the 2,20 -azinobis-(3-ethylbenzothiazoline-6sulfonate) (ABTS þ) radical cation.46 Siebenhandl, Grausgruber, Pellegrini, Rio, Fogliano, Pernice, Berghofer31 measured the TEAC of Ethiopian purple wheat wholemeal and its milled fractions (Table 10.3) and reported the TEAC of wholemeal to be 0.63 mmol/100 g. Among the milled fractions, the bran and shorts showed the highest TEAC (1.91 mmol/100 g), followed by the middling fraction (0.72 mmol/100 g). Purple wheat flour had the lowest TEAC, approximately fourfold less than the bran and shorts fraction. According to Siebenhandl, Grausgruber, Pellegrini, Rio, Fogliano, Pernice, Berghofer,31 the purple wheat produced 0.9, 11.8, 44.6, and 42.7% of bran, shorts, middlings, and flour, when compared to the whole grain. Therefore, the contribution of each milled fraction to the total antioxidant activity of purple wheat whole grain is important. For instance, the middling fraction made the most contribution (42%) to the total antioxidant activity of purple wheat whole grains measured by the TEAC assay. Yu, Beta9 reported the TEAC value of insoluble-bound phenolic extract from Konini to be higher than that of Indigo while TEAC of the soluble free extracts from the two varieties were not different from each other. Similar to the DPPH RSA, the TEAC of the insoluble-bound extracts of Konini and Indigo was higher than TEAC values of the soluble free extracts.9 Ferric reducing antioxidant power (FRAP) assay measures the ability of phenolics to reduce the yellow ferric 2,4,6-tripyridyl-s-triazine (Fe(III)-TPTZ) to a blue ferrous complex (Fe(II)-TPTZ). Similar to the TEAC results, the bran and shorts fraction exhibited the greatest FRAP (Table 10.3), approximately 3.5-fold higher than middling and flour fractions.31 The significant differences in FRAP among the milled fractions indicate that the antioxidants are not evenly distributed in purple wheat grain, but instead are predominantly located in the pericarp and aleurone layers.6 

Adverse Effects and Reactions (Allergies and Toxicity) No reports are available on allergies and toxicity resulting from purple wheat products. However, it can be speculated that adverse effects and reactions associated with common wheat may also be applicable to purple wheat. Indeed, some peptides derived from gluten proteins present in wheat are known to be responsible for celiac disease, an intestinal disorder caused by T-cell responses to these peptides.50 Secondary intolerances including viral hepatitis and intestinal infections may also occur in predisposed individuals. However, there is potential for selection of nontoxic varieties for celiac disease patients.50 High levels of wheat-specific immunoglobulin E (IgE) have been reported in patients with anaphylaxis.51

Summary Points and Future Perspectives • Extracts from purple wheat seeds differ in their antioxidant capacities, depending on the extraction methods employed as well as genotypic and environmental effects. • Acidified methanol appears to recover more extractable phenolic antioxidants, including anthocyanins, from purple wheat than aqueous acetone.

II. Role of Seeds in Nutrition and Antioxidant Activities

References

123

• Among the purple wheat varieties Charcoal, Konini, and Indigo, Charcoal has the highest antioxidant capacity because of its high total phenolics, anthocyanins, and flavonoids contents. • Thermal processing causes reduction in the total phenolic and anthocyanin contents of purple wheat. However, the antioxidant capacities of the processed products either remain unchanged or increase after processing, presumably due to Maillard reaction products that exhibit antioxidant activity. • Knowledge of the antioxidant effects of several purple wheat varieties would be useful for screening and selecting genotypes with higher antioxidant activity and potential health-promoting properties. • There is a need to elucidate the effect of thermal processing on the structural chemistry of purple wheat phenolic compounds, such as anthocyanins, to enable an understanding of the effect of thermal processing on their antioxidant properties. • There is a need to study the bioaccessibility and bioavailability of phenolic compounds, including anthocyanins present in purple wheat food products to enable evidence-based health benefit claims on the consumption of purple wheat foods. • The adverse effects and reactions associated with common wheat may also be applicable to purple wheat.

References 1. Nelson JH. Wheat: its processing and utilization. American Journal of Clinical Nutrition. 1985;41:1070e1076. 2. FAO. Food Outlook-Biannual Report on Global Food Markets. Rome, Italy: Food and Agriculture Organization of the United Nations; 2019. 3. Zeven AC. Wheats with purple and blue grains: a review. Euphytica. 1991;56(3):243e258. 4. Abdel-Aal E-SM, Young JC, Rabalski I. Anthocyanin composition in black, blue, pink, purple, and red cereal grains. Journal of Agricultural and Food Chemistry. 2006;54(13):4696e4704. 5. Adom KK, Sorrells ME, Liu RH. Phytochemical profiles and antioxidant activity of wheat varieties. Journal of Agricultural and Food Chemistry. 2003;51(26):7825e7834. 6. Beta T, Nam S, Dexter JE, Sapirstein HD. Phenolic content and antioxidant activity of pearled wheat and rollermilled fractions. Cereal Chemistry. 2005;82(4):390e393. 7. Mpofu A, Sapirstein HD, Beta T. Genotype and environmental variation in phenolic content, phenolic acid composition, and antioxidant activity of hard spring wheat. Journal of Agricultural and Food Chemistry. 2006;54(4):1265e1270. 8. Liu Q, Qiu Y, Beta T. Comparison of antioxidant activities of different colored wheat grains and analysis of phenolic compounds. Journal of Agricultural and Food Chemistry. 2010;58(16):9235e9241. 9. Yu L, Beta T. Identification and antioxidant properties of phenolic compounds during production of bread from purple wheat grains. Molecules. 2015;20:15525e15549. 10. Belay G, Tesemma T, Bechere E, Mitiku D. Natural and human selection for purple-grain tetraploid wheats in the Ethiopian highlands. Genetic Resources and Crop Evolution. 1995;42(4):387e391. 11. Li W, Shan F, Sun S, Corke H, Beta T. Free radical scavenging properties and phenolic content of Chinese blackgrained wheat. Journal of Agricultural and Food Chemistry. 2005;53(22):8533e8536. 12. Tesemma T, Belay G. Aspects of Ethiopian tetraploid wheats with emphasis on durum wheat genetics and breeding research. In: Gebre Mariam H, Tanner DG, Hulluka M, eds. Wheat Research in Ethiopia: A Historical Perspective. Addis Ababa, Ethiopia. IAR/CIMMYT; 1991:47e71.

II. Role of Seeds in Nutrition and Antioxidant Activities

124

10. Purple Wheat (Triticum sp.) Seeds: Phenolic Composition and Antioxidant Properties

13. Dykes L, Rooney LW. Phenolic compounds in cereal grains and their health benefits. Cereal Foods World. 2007;52(3):105e111. 14. Pietta P-G. Flavonoids as antioxidants. Journal of Natural Products. 2000;63(7):1035e1042. 15. Pasqualone A, Bianco AM, Paradiso VM, et al. Production and characterization of functional biscuits obtained from purple wheat. Food Chemistry. 2015;180:64e70. 16. Hosseinian FS, Li W, Beta T. Measurement of anthocyanins and other phytochemicals in purple wheat. Food Chemistry. 2008;109(4):916e924. 17. Lu Y, Luthria D, Fuerst EP, Kiszonas AM, Yu L, Morris CF. Effect of processing on phenolic composition of dough and bread fractions made from refined and whole wheat flour of three wheat varieties. Journal of Agricultural and Food Chemistry. 2014;62(43):10431e10436. 18. Qiu Y, Liu Q, Beta T. Antioxidant activity of commercial wild rice and identification of flavonoid compounds in active fractions. Journal of Agricultural and Food Chemistry. 2009;57(16):7543e7551. 19. Khoo HE, Azlan A, Tang ST, Lim SM. Anthocyanidins and anthocyanins: colored pigments as food, pharmaceutical ingredients, and the potential health benefits. Food and Nutrition Research. 2017;61(1). https://doi.org/ 10.1080/16546628.16542017.11361779. 20. Singleton VL, Orthofer R, Lamuela-Raventós RM. Analysis of total phenols and other oxidation substrates and antioxidants by means of Folin-Ciocalteu reagent. Methods in Enzymology. 1999;299:152e178. 21. Waterhouse AL. Determination of total phenolics. Current Protocols in Food Analytical Chemistry. 2002;I(1.1):I1.1.1eI1.1.8. 22. Ainsworth EA, Gillespie KM. Estimation of total phenolic content and other oxidation substrates in plant tissues using FolineCiocalteu reagent. Nature Protocols. 2007;2(4):875e877. 23. Basílio N, Pina F. Chemistry and photochemistry of anthocyanins and related compounds: a thermodynamic and kinetic approach. Molecules. 2016;21(11):1502. 24. Giusti MM, Wrolstad RE. Characterization and measurement of anthocyanins by UV-visible spectroscopy. Current Protocols in Food Analytical Chemistry. 2001;F1.2:F1.2.1eF1.2.13. 25. Malesev D, Kuntic V. Investigation of metal-flavonoid chelates and the determination of flavonoids via metalflavonoid complexing reactions. Journal of the Serbian Chemical Society. 2007;72(10):921e939. 26. Chang C-C, Yang MH, Wen HM, Chern JC. Estimation of total flavonoid content in propolis by two complementary colorimetric methods. Journal of Food and Drug Analysis. 2002;10(3):178e182. 27. Herald TJ, Gadgil P, Tilley M. High-throughput micro plate assays for screening flavonoid content and DPPHscavenging activity in sorghum bran and flour. Journal of the Science of Food and Agriculture. 2012;92(11):2326e2331. 28. Hassan SM, Al Aqil AA, Attimarad M. Determination of crude saponin and total flavonoids content in guar meal. Advancement in Medicinal Plant Research. 2013;1(2):24e28. 29. Herald TJ, Gadgil P, Perumal R, Bean SR, Wilson JD. High-throughput micro-plate HCl-vanillin assay for screening tannin content in sorghum grain. Journal of the Science of Food and Agriculture. 2014;94(10):2133e2136. 30. Proestos C, Sereli D, Komaitis M. Determination of phenolic compounds in aromatic plants by RP-HPLC and GC-MS. Food Chemistry. 2006;95(1):44e52. 31. Siebenhandl S, Grausgruber H, Pellegrini N, et al. Phytochemical profile of main antioxidants in different fractions of purple and blue wheat, and black barley. Journal of Agricultural and Food Chemistry. 2007;55(21):8541e8547. 32. Guo W, Beta T. Phenolic acid composition and antioxidant potential of insoluble and soluble dietary fibre extracts derived from select whole-grain cereals. Food Research International. 2013;51(2):518e525. 33. Abdel-Aal E-SM, Hucl P. Composition and stability of anthocyanins in blue-grained wheat. Journal of Agricultural and Food Chemistry. 2003;51(8):2174e2180. 34. McCallum J, Walker J. Proanthocyanidins in wheat bran. Cereal Chemistry. 1990;67(3):282e285. 35. Dinelli G, Carretero AS, Di Silvestro R, et al. Determination of phenolic compounds in modern and old varieties of durum wheat using liquid chromatography coupled with time-of-flight mass spectrometry. Journal of Chromatography A. 2009;1216(43):7229e7240. 36. Li W, Pickard MD, Beta T. Effect of thermal processing on antioxidant properties of purple wheat bran. Food Chemistry. 2007;104(3):1080e1086. 37. Li W, Pickard MD, Beta T. Evaluation of antioxidant activity and electronic taste and aroma properties of anthobeers from purple wheat grain. Journal of Agricultural and Food Chemistry. 2007;55(22):8958e8966.

II. Role of Seeds in Nutrition and Antioxidant Activities

References

125

38. Yilmaz Y, Toledo R. Antioxidant activity of water-soluble Maillard reaction products. Food Chemistry. 2005;93(2):273e278. 39. Ficco DBM, De Simone V, De Leonardis AM, et al. Use of purple durum wheat to produce naturally functional fresh and dry pasta. Food Chemistry. 2016;205:187e195. 40. Parthasarathy S, Khan-Merchant N, Penumetcha M, Santanam N. Oxidative stress in cardiovascular disease. Journal of Nuclear Cardiology. 2001;8(3):379e389. 41. Li J, Wuliji O, Li W, Jiang Z-G, Ghanbari HA. Oxidative stress and neurodegenerative disorders. International Journal of Molecular Sciences. 2013;14:24438e24475. 42. Dayem AA, Choi H-Y, Kim J-H, Cho S-G. Role of oxidative stress in stem, cancer, and cancer stem cells. Cancers. 2010;2(2):859e884. 43. Dalle-Donne I, Rossi R, Giustarini D, Milzani A, Colombo R. Protein carbonyl groups as biomarkers of oxidative stress. Clinica Chimica Acta. 2003;329(1):23e38. 44. Parthasarathy S, Rankin SM. Role of oxidized low density lipoprotein in atherogenesis. Progress in Lipid Research. 1992;31(2):127e143. 45. Dizdaroglu M, Jaruga P. Mechanisms of free radical-induced damage to DNA. Free Radical Research. 2012;46(4):382e419. 46. Huang D, Ou B, Prior RL. The chemistry behind antioxidant capacity assays. Journal of Agricultural and Food Chemistry. 2005;53(6):1841e1856. 47. Davies MJ. Protein oxidation and peroxidation. Biochemical Journal. 2016;473(7):805e825. 48. Brand-Williams W, Cuvelier M, Berset C. Use of a free radical method to evaluate antioxidant activity. Lebensmittel-Wissenschaft und -Technologie- Food Science and Technology. 1995;28(1):25e30. 49. Arteaga JF, Ruiz-Montoya M, Palma A, Alonso-Garrido G, Pintado S, Rodríguez-Mellado JM. Comparison of the simple cyclic voltammetry (CV) and DPPH assays for the determination of antioxidant capacity of active principles. Molecules. 2012;17(5):5126e5138. 50. SpaenijeDekking L, KooyeWinkelaar Y, van Veelen P, et al. Natural variation in toxicity of wheat: potential for selection of nontoxic varieties for celiac disease patients. Gastroenterology. 2005;129(3):797e806. 51. Pourpak Z, Mansouri M, Mesdaghi M, Kazemnejad A, Farhoudi A. Wheat allergy: clinical and laboratory findings. International Archives of Allergy and Immunology. 2004;133(2):168e173. 52. Awika JM, Rooney LW. Sorghum phytochemicals and their potential impact on human health. Phytochemistry. 2004;65(9):1199e1221. 53. Bunzel M. Chemistry and occurrence of hydroxycinnamate oligomers. Phytochemistry Reviews. 2010;9(1):47e64.

II. Role of Seeds in Nutrition and Antioxidant Activities

C H A P T E R

11

Protective Role of Nigella sativa and Thymoquinone in Oxidative Stress: A Review 1

Fatemeh Forouzanfar1, 2, Hossein Hosseinzadeh3, 4

Neuroscience Research Center, Mashhad University of Medical Sciences, Mashhad, Iran; Department of Neuroscience, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran; 3Department of Pharmacodynamics and Toxicology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Razavi Khorasan Province, Iran; 4 Pharmaceutical Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, Razavi Khorasan Province, Iran 2

Introduction Oxidative stress occurs when there is an imbalance between production of reactive oxygen species (ROS) and antioxidant homeostasis system.1 ROS such as superoxide anion ( ), hydroxyl radical (OH
), hydrogen peroxide (H2O2), and singlet oxygen (1O2) are highly reactive and unstable.1 Free radicals possess one or more unpaired electrons in their outer electronic orbits, those are potentially toxic for neuronal cells, and excessive production of those leads to oxidative stress that have been implicated to play an important role in the etiology of various diseases.1,2

Free Radicals and Antioxidant Defense , a major cellular free radical, is involved in a large number of deleterious changes often related with an enhancement in peroxidative processes and related to a low antioxidant concentration. While itself is not so reactive to biomolecules, but it helps in generation of more powerful$OH and peroxonitrite (ONOO
). In phagocytes, is produced in large

Nuts and Seeds in Health and Disease Prevention, Second Edition https://doi.org/10.1016/B978-0-12-818553-7.00011-5

127

Copyright © 2020 Elsevier Inc. All rights reserved.

128

11. Protective Role of Nigella sativa and Thymoquinone in Oxidative Stress: A Review

amounts by the enzyme NADPH oxidase for killing certain pathogens. Moreover, is a byproduct of mitochondrial respiration, as well as numerous other enzymes, for instance, NADH oxidase, xanthine oxidase, cyclooxygenases (COX), and monooxygenase.3 Despite all the considerable advancements in modern medicine, there has been a growing interest and demand in using medicinal plants for treating and preventing different diseases.4,5 Nigella sativa Linneaus (N. sativa) is an annual flowering plant that belongs to the botanical family Ranunculaceae, which is popularly called with different names such as shonaiz (Persian), black caraway seeds (USA), black cumin (English), and kalajira (Bangali).6,7

Chemical Constituents Black cumin seeds contain numerous esters of structurally unusual unsaturated fatty acids with terpene alcohols (7%); besides, traces of alkaloids are found, which belong in two different types: isoquinoline alkaloids that are represented by nigellimin and nigellimin-Noxide, and pyrazol alkaloids that include nigellidin and nigellicin. In the essential oil (average 0.5%, maximum 1.5%), thymoquinone (TQ) was identified as the main constituent (up to 50%) as well as p-cymene (40%), pinene (up to 15%), dithymoquinone and thymohydroquinone. Other terpene derivatives that were found only in trace amounts are carvacrol, carvone, limonene, 4-terpineol, and citronellol.7,8 The essential oil has considerable (10%) amounts of fatty acid ethyl esters, the seeds have a fatty oil rich in unsaturated fatty acids, mainly linoleic acid (50, 60%), oleic acid (20%), eicosadienoic acid (3%), and dihomolinoleic acid (10%). N. sativa seeds contain 36e38% fixed oils, saponin, proteins, alkaloids, and 0.4e2.5% essential oil. TQ, thymohydroquinone, dithymoquinone, and thymol are considered the main active constituents of N. sativa essential oil.9

Traditional Medicine N. sativa has been used for several diseases for centuries, which are related to respiratory, stomach and intestinal health, liver and kidney function, circulatory and immune system support, and for the general well-being.7,10 The seeds were used as pungent, expectorant, appetizer, aromatic, sudoriferous, thermogenic, diuretic, purgative, stimulant, sedative, and carminative.11e13

Pharmacological Properties N. sativa has been broadly studied in recent years and research projects have reported that it possesses several medicinal properties including antioxidant,14,15 protective effects on lipid peroxidation,16 antitussive,17 analgesic,18,19 gastroprotective,20 anti-asthmatic,21 anticancer, anti-inflammatory, immunomodulatory, and antitumor properties,22e24 also gastric ulcer healing,25 tumor growth suppression,26 memory improvement,27 anti-anxiety,28 stimulate milk production,29 men infertility improvement,30 cardiovascular disorders,31 antiviral activity against cytomegalovirus,32 antibacterial activity,33 antidermatophyte,34 effective in metabolic syndrome35 have been reported for this therapeutic plant. II. Role of Seeds in Nutrition and Antioxidant Activities

In vitro Antioxidant Activity of Nigella sativa

129

This chapter would explain the antioxidant effects of N. sativa and its constituents as well as the possible mechanisms of actions underlying the effects on which various research projects have been done.

In vitro Antioxidant Activity of Nigella sativa The most pharmacologically active constituent of the volatile oil of N. sativa seeds is TQ, which has strong antioxidant potentials via scavenging ability of different free radicals. TQ has a potent superoxide anion scavenger activity.36 By thin-layer chromatography analysis, it has been shown that the compounds that isolated from N. sativa (including TQ, carvacrol, t-anethole, and 4-terpineol) have significant free radical scavenging properties.37,38 These four constituents had antioxidant activity when tested in the diphenylpicrylhydrazyl assay for non-specific hydrogen atom or electron-donating activity. In addition, they were effective OH radical scavenging agents in non-enzymatic lipid peroxidation assay in liposomes and in the deoxyribose degradation assay.38 In an in vitro assay, the ethanol extracted of N. sativa and TQ exhibited a reduction of DNA fragmentation even more than g-tocopherol, which may be mediated through the antioxidant and anti-apoptotic effects of N. sativa and TQ.39 Toluene induces oxyradical and eventually causes oxidative stress and damage to cells. N. sativa extracts prevented the depletion of intracellular glutathione (GSH) in fibroblasts that were exposed to toluene.40 Mousavi et al. showed that pretreatment with the ethanolic extract of N. sativa and TQ reversed the increased ROS production and reduced serum/glucose deprivation-induced cytotoxicity in PC12 cells.41 1,1-Diphenyl-2-picrylhydrazyl (DPPH) radical has been widely used to test the free radical scavenging ability.42 DPPH scavenging activities of crude methanolic extract, ethyl acetate fraction, hexane fraction, and water fraction (WF) of black cumin were 27.8, 78.8, 12.1, and 32.1 mg gallic acid equivalents (GAE)/g, respectively. The extract and fractions exhibited high effect on reducing the oxidation of b-carotene. The predominant phenolic compounds identified by high-performance liquid chromatography with diode array detection in crude methanolic extract and WF of black cumin were hydroxybenzoic, syringic, and p-coumaric acids.43 In another research, chemiluminescence and spectrophotometry methods were used to evaluate the free radical scavenging effects of TQ, dithymoquinone, and thymol. The results showed that thymol acted as singlet oxygen quencher, whereas TQ and dithymoquinone showed superoxide dismutase (SOD)-like activity.44 Also, a study was carried out by Mansour et al.45 revealed that both TQ and its metabolite dithymoquinone are potent superoxide anion scavengers and general free radical scavengers that have half maximal inhibitory concentration (IC50) in the nanomolar and micromolar ranges, respectively. The following result suggested the importance of such free radical scavenging compounds in the treatment of hypertension, which is closely related with oxidative stress.45 Depletion in free radicals with TQ can diminish the risk of free radicals attacking to DNA, therefore leading to reduction in the risk of cancers. In an in vitro assay, TQ, in a dose-dependent manner, inhibits the activity of hepatic CYP1A1/A2 isozymes involved in biotransformation of many xenobiotics in to reactive genotoxic radical derivatives.46

II. Role of Seeds in Nutrition and Antioxidant Activities

130

11. Protective Role of Nigella sativa and Thymoquinone in Oxidative Stress: A Review

In vivo Antioxidant Activity of Nigella sativa N. sativa protected many organs against oxidative stress induced by a variety of diseases that described in detail in the following titles (Fig. 11.1, Table 11.1):

Acetaminophen-Induced Hepatotoxicity Acetaminophen-induced liver injury has been confirmed in experimental animal models as well as clinical cases.47 The in vitro and in vivo findings in a recent study showed that the N. sativa seed extract (100 and 900 mg/kg) has protective effects against acetaminopheninduced hepatotoxicity and metabolic disturbances. The protective effects might be related to improving antioxidant activities and attenuation of lipid peroxidation and ROS generation.48

Aging Aging is a continuous process that can lead to changes in biological systems. Induction of oxidative stress and apoptosis, hepatotoxicity, and neurotoxicity are the processes involved in aging.49 In a study conducted by Shahroudi et al., the anti-aging effect of N. sativa oil in

Nigella sava

Schistosomiasis

Diabetes mellitus

Neurotoxicity

Ischemic/reperfusion (I/R) injury

Arthritis

Hematotoxicity

Subarachnoid haemorrhage

Plasmodium yoelli infection

Necrosis and fibrosis of liver

Ulcerative colitis Aging Seizure

Fatty liver

Nephrotoxicity Hypercholesterolemia

FIGURE 11.1

Thymoquinone

Encephalomyelitis Cardiotoxicity

Inhibitory effects of Nigella sativa and its constituent, thymoquinone, on some diseases by anti-

oxidant activities.

II. Role of Seeds in Nutrition and Antioxidant Activities

131

In vivo Antioxidant Activity of Nigella sativa

TABLE 11.1

Selected in vivo Studies of Significant Antioxidant Effects of N. Sativa and Its Constituents.

Study Model

Treatment

Laboratory Findings

References

Diabetes mellitus

Aqueous extract of N. sativa and TQ

Decreased MDA levels and increased SOD enzyme, diminished COX-2 mRNA expression

75

Diabetes mellitus

Aqueous extract of N. sativa, TQ, and oil

Decreased the MDA and serum glucose levels, increased 78 the serum insulin and tissue SOD

Diabetes mellitus

N. sativa oil

Decreased morphological changes and preserved pancreatic b-cells integrity

Diabetes mellitus

N. sativa oil and TQ Increased the area of insulin-immunoreactive b-cells

80

Patients with type 2 diabetes

N. sativa capsule

Improves glycemic control and ameliorates oxidative stress

73

Patients with type 2 diabetes

N. sativa oil

Improvement of glycemic status and lipid profile

81

Hepatic ischemia

N. sativa

Increment of total antioxidant capacity, decrement of TOS and OSI and MPO in hepatic tissue as well as histological liver tissue damage

99

Muscle ischemic injury

TQ

Increment of total SH contents and antioxidant capacity and decrement of the MDA level

100

Renal ischemia/ reperfusion injury

Hydroalcoholic extract of N. sativa

Diminished MDA level and DNA damage, increased the 101 renal thiol content

Plasmodium yoeliiinfected mice

Methanolic extract of N. sativa seeds

Decreased MDA levels, increased the levels of GSH content, ameliorated the activities of red cell SOD and CAT, hepatic SOD, and GST

113

Colitis

N. sativa oil

Increased SOD activity

126

Epilepsy

N. sativa oil

Decreased NO levels and increased GSH levels

120

Nephrotoxicity

N. sativa oil

Decreased the MDA and NO generation and increased SOD and GPx activities

106

Nephrotoxicity

TQ

Decreased TBARS and NOx levels and increased GSH, GPx, CAT, and ATP activities

107

Nephrotoxicity

TQ

Increased GSH and SOD levels and decreased the MDA level

108

Hypercholesterolemia TQ

Increased SOD1, CAT level, and the expression of liver antioxidant genes (GPX2)

95

Schistosoma mansoniinfected mice

N. sativa oil

Decreased the MDA level and increased the GSH levels

117

Aging

N. sativa oil

Decreased Bax/Bcl2, caspase-3 protein levels, and lipid peroxidation, recovered the GSH content in brain and liver tissues

49

77

(Continued)

II. Role of Seeds in Nutrition and Antioxidant Activities

132 TABLE 11.1

11. Protective Role of Nigella sativa and Thymoquinone in Oxidative Stress: A Review

Selected in vivo Studies of Significant Antioxidant Effects of N. Sativa and Its Constituents.dcont'd

Study Model

Treatment

Laboratory Findings

References

Arthritis

TQ

Decreased TBARS level, restored the GSH level and SOD 50 activity

Ferric nitrilotriacetate- N. sativa induced oxidative stress

Diminished H2O2 generation and lipid peroxidation. Renal glutathione content, glutathione-metabolizing enzymes, and antioxidant enzymes were also improved

52

1,2TQ Dimethylhydrazine (DMH)-induced colon carcinogenesis

Restored the MDA and conjugated diene levels and enzyme activities like CAT, GPX, and SOD activities above to near-normal values

56

DENA-induced initiation of liver cancer

Decreased levels of total nitrate/nitrite, total bilirubin, TBARS, and increased GSH, GPx, CAT, and GST as well as increased the gene expression of GST, GPx, and CAT

58

TQ

Liver damage TQ induced by tamoxifen

Inhibited TAM-induced hepatic GSH depletion and LPO 60 accumulation, normalized the activity of SOD

Carbon tetrachloride as a hepatotoxic agent

N. sativa oil

Increased the reduced antioxidant enzyme levels (GSH) and decreased the lipid peroxidation

61

Acrylamide induced neurotoxicity

TQ

Decreased MDA levels

63

Thioacetamide induced liver cirrhosis

N. sativa oil

Improvement in the altered levels of antioxidant enzymes such as CAT, SOD, GPx, TBARS, and reduced GSH

64

Restored the activities of SOD, CAT, GPx, and acetylcholine esterase (AChE) and decreased the generation of lipid peroxidation and protein carbonyl content

65

Propoxur-intoxicated N. sativa oil rats

Subarachnoid hemorrhage

N. sativa oil

Decreased MDA level, increased GSH level. Naþ/Kþ-ATPase activity was increased

125

Subarachnoid hemorrhage

TQ

Increased the activities of non-enzymatic (GSH and vitamin C) and enzymatic (SOD, CAT, GPx, and GST) antioxidants as well by reducing the levels of MDA

124

HCV patients

Ethanolic extract capsules

Exhibited potential therapeutic benefits via diminishing viral load and alleviating the altered liver function

93

Fatty liver

N. sativa oil

Decreased MDA level

88

II. Role of Seeds in Nutrition and Antioxidant Activities

133

In vivo Antioxidant Activity of Nigella sativa

TABLE 11.1

Selected in vivo Studies of Significant Antioxidant Effects of N. Sativa and Its Constituents.dcont'd

Study Model

Treatment

Laboratory Findings

References

Experimental autoimmune encephalomyelitis

TQ

Increased the glutathione content as well as inhibited the activation of NF-kB

82

Neurotoxicity

N. sativa oil

Increased antioxidant enzymes and decreased the levels of free radicals

111

Haematotoxicity

N. sativa

Decreased malondialdehyde and increased total antioxidant capacity

91

Cardiotoxicity

N. sativa oil

Decreased lipid peroxidation, levels of protein carbonyl and nitric oxide, and improvement in antioxidant enzyme status and increased activity of glutathione peroxidase

70

a mouse model of aging induced with D-galactose was assessed. N. sativa oil (0.1 mL/kg) decreased the expressions of procaspase-3, and caspase-3 cleaved, in addition to the Bax/Bcl2 ratio, in liver and brain tissues. Moreover, administration of N. sativa oil elevated the GSH content, and reduced lipid peroxidation in brain and liver tissues was seen.49

Arthritis The antioxidant and anti-arthritic activity of TQ by collagen-induced arthritis in Wistar rat was evaluated. Administration of TQ decreased thiobarbituric acid reactive substances (TBARS) level, restored the GSH level and SOD activity, also significantly decreased the levels of pro-inflammatory mediators (IL-1b, IL-6, TNF-a, IFN-g, and PGE2), and increased the level of IL-10.50 Tekeoglu et al. showed that administration of TQ, clinically and radiologically, suppressed Freund’s incomplete adjuvant-induced arthritis in rats.51

Carcinogenes Ferric nitrilotriacetate is a strong oxidant that generates extremely reactive hydroxyl radical and induces injuries of various organs that include the kidney and liver. Oral administration with N. sativa showed a potent chemopreventive agent and suppresses ferric nitrilotriacetate-induced oxidative stress diminished in g-glutamyltranspeptidase, xanthine oxidase, H2O2 generation, lipid peroxidation, blood urea nitrogen (BUN), serum creatinine, renal ornithine decarboxylase activity, and DNA synthesis. Renal glutathione content, glutathione-metabolizing enzymes, and antioxidant enzymes were also improved to significant levels in Wistar rats.52 The dietary supplement of N. sativa suppressed the oxidative stress caused by oxidized oil in rats.53 The oral feeding of the diet containing N. sativa inhibited the oxidative stress (reduced GSH and nitric oxide [NO]) that was induced by hepatocarcinogens such as NaNO3 and dibutylamine in rats.54

II. Role of Seeds in Nutrition and Antioxidant Activities

134

11. Protective Role of Nigella sativa and Thymoquinone in Oxidative Stress: A Review

1,2-Dimethylhydrazine (DMH), as a potent colon carcinogen, in oxidative condition produces electrophilic diazonium ion thus elicit oxidative stress in the RBCs.55,56 The potential protective effect of TQ on erythrocyte lipid peroxidation and antioxidant status during DMH-induced colon carcinogenesis in rats was investigated. Pretreatment with TQ restored the malondialdehyde (MDA) and conjugated diene levels and enzyme activities such as catalase (CAT), glutathione peroxidase (GPx), and SOD activities above to near-normal values.56 Diethylnitrosamine (DENA) is known as a hepatocarcinogenic agent.57,58 In the liver tissue, DENA induced severe histopathological lesions and increased the levels of total nitrate/nitrite, total bilirubin, TBARS, alanine transaminase (ALT), and alkaline phosphatase (ALP) and decreased GSH, GPx, CAT, and glutathione S-transferase (GST), as well as reduced the gene expression of GST, GPx, and CAT. These changes have been significantly corrected by TQ supplementation in DENA-induced initiation of liver cancer in rats.58 Tamoxifen (TAM) is a non-steroidal anti-estrogen drug that is used in treatment of breast cancer. On the other hand, TAM is a hepatocarcinogen that is due to overproduction of ROS content59,60 Suddek et al. showed that the hepatotoxicity induced by TAM resulted in the elevation of serum levels of liver enzymes including ALT, ALP, lactate dehydrogenase (LDH), g-glutamyltransferase (GGT), and total bilirubin, plus reduction of reduced GSH in the liver, diminished SOD activity, and accumulation of lipid peroxides. Pretreatment with TQ significantly prevented the elevation in serum activity of the assessed enzymes and significantly inhibited TAM-induced hepatic GSH depletion. This constituent of N. sativa normalized the activity of SOD and inhibited the rise in TNFa. It also ameliorated the histopathological changes.60 Carbon tetrachloride (CCl4) is a hepatotoxic agent that induces the generation of free radicals that initiate cell damage. N. sativa treatment increased the reduced antioxidant enzyme levels and decreased the elevated lipid peroxidation as well as liver enzymes in carbon tetrachloride-treated rats.61 The inhibition of neurotransmission via the disruption of presynaptic NO signaling, nerve terminal, and axonal degeneration, increasing lipid peroxidation, diminution of antioxidant capacity of nervous system, and induction of apoptosis signaling are different mechanisms that are mediated by acrylamide (ACR) neurotoxicity.62,63 Administration of TQ significantly and dose-dependently decreased MDA and gait abnormalities in ACR-induced neurotoxicity in Wistar rats.63 A study was carried out to evaluate the ameliorating effect of N. sativa seed oil on the liver damage induced by thioacetamide in albino rats for a period of 8 weeks. Treatment with 10 mL/kg body weight of N. sativa oil leads to a remarkable improvement in the altered levels of albumin, bilirubin, total protein, GGT, ALT, ALP, and improvement in the altered levels of antioxidant enzymes such as CAT, SOD, GPx, TBARS, and reduced GSH.64 Propoxur is a carbamate insecticide that has an inhibitory effect on acetylcholine esterase enzyme and induces oxidative stress that leads to generation of free radicals. It has been suggested that lipid peroxidation is one of the molecular mechanisms involved in carbamate induced toxicity. Antioxidants and glutathione metabolism-regulating enzymes may protect the cellular system against various harmful effects of free radicals that resulted by pesticides.65,66 After administration of propoxur, the enzymatic antioxidant (SOD, CAT, GPx, and GST) activities and non-enzymatic antioxidant (GSH) levels were significantly diminished, and lipid peroxidation and protein carbonyl content increased. Administration of

II. Role of Seeds in Nutrition and Antioxidant Activities

In vivo Antioxidant Activity of Nigella sativa

135

N. sativa oil to propoxur-intoxicated rats restored the activities of SOD, CAT, GPx, and acetylcholinesterase and decrease the generation of lipid peroxidation and protein carbonyl content in the different brain regions of rats to normal levels.65

Cardiotoxicity Oxidative stress plays an important role in the development of myocardial infarction (MI).67 Ethanol extract of N. sativa restored lipid peroxidation levels, myocardial endogenous antioxidants, and cardiac biomarker enzymes in isoproterenol-induced MI rats.68 Cyclosporine A is a well-known immunosuppressor agent used in transplant surgery and in the treatment of autoimmune diseases, and it causes unwanted side effects in many organs such as the heart.69 Pretreatment with N. sativa oil decreased the subsequent cyclosporine A injury in the rat heart was demonstrated by decreased connective tissue among myocardial fibers, mild myocardial disorganization, diminution in lipid peroxidation, levels of protein carbonyl and NO, and improvement in antioxidant enzyme status and increased activity of GPx.70

Chemotherapeutic Agent Cisplatin is an effective chemotherapeutic agent that leads to gastrointestinal toxicity.71 Administration of N. sativa oil to cisplatin-treated rats significantly increased brush border membrane enzyme activities in intestinal homogenates, and brush border membrane vesicles also improved the activities of carbohydrate metabolism enzymes and the enzymatic and non-enzymatic antioxidant parameters in the intestine of cisplatin-treated rats.71

Diabetes Mellitus Oxidative stress plays a pivotal role in pathogenesis of diabetes mellitus and its complications. Oxidative stress and ROS have been proposed to be involved in the development of insulin resistance, loss of b-cell function, and diabetic complications. The burden of production of these free radicals is mostly counteracted by the antioxidant defense system, including the enzymatic scavenger SOD, GPx, and CAT.72,73 COX-2 is a complex enzyme that have both COX and peroxidase activities. Both activities have been shown to increase the ROS formation.74,75 Free radicals, mainly ROS evidenced by the formation of lipid peroxidation products, have been implicated in the cytokine-mediated islet cell injury, mostly based on the protective influence of antioxidants in various models of diabetes.74,75 Streptozotocin (STZ) is an antibiotic which produced by Streptomyces achromogenes and experimental evidence has demonstrated that STZ induced metabolic processes, leading an increment in generation of ROS followed by DNA damage, including alkylation and DNA strand breaks and inhibits free radical scavenger-enzymes.76,77 Administration of aqueous extract of N. sativa and TQ for 30 days in treatment of STZ-induced diabetic rats significantly suppressed pancreatic tissue lipid peroxidation (MDA levels) and increased the level of SOD antioxidant enzyme correlated with the diminution in COX-2 mRNA expression.75 The aqueous extract and oil of N. sativa as well as TQ decreased the MDA and serum

II. Role of Seeds in Nutrition and Antioxidant Activities

136

11. Protective Role of Nigella sativa and Thymoquinone in Oxidative Stress: A Review

glucose levels, as well as increased the serum insulin and tissue SOD in STZ-induced diabetic rats.78 The volatile oil of N. sativa decreased morphological changes and preserved pancreatic b-cells integrity in STZ-induced diabetic rats.77 Persistent hyperglycemia has been considered as a primary risk factor for neuropathy. Long-term hyperglycaemia leads to subsequent enhanced oxidative stress.79,80 Treatment with N. sativa oil and TQ significantly increased the area of insulin-immunoreactive b cells. TQ and especially N. sativa exhibited less morphologic changes in sciatic nerves. Also, myelin breakdown significantly decreased in STZinduced diabetic rats.80 Moreover, one-year administration of N. sativa (as powder in capsules of 500 mg in a dose of 2 g/day) improved glycemic control and ameliorated oxidative stress in type 2 diabetes mellitus patients.73 The result of a double-blind, randomized controlled clinical trial showed that treatment with N. sativa oil leads to improvement of glycemic status and lipid profile in patients with type 2 diabetes.81

Encephalomyelitis A study was carried out to evaluate the possible effects of TQ on the inhibition of activation of NF-kB in experimental autoimmune encephalomyelitis in the rat model of multiple sclerosis. Administration of TQ (1 mg/kg/day) concomitant to myelin basic protein prevented and ameliorated experimental autoimmune encephalomyelitis. TQ was also able to counter perivascular cuffing and infiltration of mononuclear cells in the brain and spinal cord. TQ increased the red blood cell GSH content as well as inhibited the activation of NF-kB in the brain and spinal cord.82 Other study showed that administration of N. sativa oil protected brain and medulla spinalis tissues against oxidative stress induced by experimental autoimmune encephalomyelitis. The protective effect may be due the antioxidant effect.83

Ethanol Toxicity Ethanol induced hepatotoxicity and nephrotoxicity that are demonstrated by histopathological damages in addition to elevated levels of MDA and reduction of GSH content in the liver and kidney. Also, ALT, aspartate aminotransferase (AST), and ALP levels increased in the liver.84 N. sativa fixed oil attenuated ethanol-induced increased levels of MDA, as well as histopathological changes in liver and kidney tissues. Furthermore, N. sativa fixed oil improved the level of ALT, AST, and ALP in the liver and GSH level in liver and kidney tissues.84 N. sativa or TQ administration protected gastric mucosa from acute alcohol-induced mucosal injury and led to decrement of the gastric tissue histamine levels and myeloperoxidase (MPO) activities in ethanol-treated rats.85 Recently, Hosseini et al. showed that TQ has preventive effects against ethanol toxicity in the liver and kidney tissues through increased antioxidant capacity and reduction inflammation and also severity of apoptosis.86

Fatty Liver Accumulation of fat in the liver can lead to steatosis and steatohepatitis. At later stage, this condition can progress to cirrhosis. Up to 10% of cirrhotic fatty liver diseases develop hepatocellular carcinoma.87,88 Free fatty acids are the main sources of production of free radicals, and as the disease progresses, oxidative stress and MDA levels are increased in the body.88 II. Role of Seeds in Nutrition and Antioxidant Activities

In vivo Antioxidant Activity of Nigella sativa

137

Induction of non-alcoholic fatty liver with inflammation in rats with high fructose diet resulted in significant dyslipidemia, increment in TNF-a and MDA levels with significant high liver triglycerides and cholesterol levels, and liver dysfunction. Oral administration of N. sativa crude oils led to a significant improvement of all parameters.88

Haematotoxicity Carbon tetrachloride (CCl4) is a haloalkane that has immunosuppressive effect; it has been extensively used for its solvent possessions;89 administrations of an aqueous suspension of N. sativa protected the hematopoietic cells from the damaging effects of exposure to CCl4 and the protective effect might be related to the antioxidative effect of N. sativa.90 4-Nonylphenol (NP) is an emerging concern contaminant that is highly spread in the aquatic ecosystem,91 exposed to NP elevated the total peroxide and MDA and depleted total antioxidant capacity of blood lysate in Clarias gariepinus. Supplementation of N. sativa seed (at the doses of 25 and 50 g/kg feed) ameliorated the previously listed manifestations; the -authors suggested that N. sativa seed protected Clarias gariepinus against NP haematotoxicity by improving the oxidative stress.91

HCV-Related Fibrosis Marked induction of ROS in infected cells leads to oxidative stress that is responsible for induced HCV-related fibrosis, cirrhosis, and liver failure.92 Abdel-Moneim and co-workers showed that administration of the ethanolic extracts capsules (each containing 500 mg of N. sativa) in HCV patients exhibited potential therapeutic benefits via diminishing viral load and alleviating the altered liver function.93 In another study, administration of N. sativa in patients with HCV was tolerable and safe and led to reduction of viral load, improvement of oxidative stress, clinical condition, and glycemic control in diabetic patients.94

Hypercholesterolemia Cholesterol-rich diets increased free radical production, followed by oxidative stress that induced hypercholesterolemia.95 Oxidative stress contributes to the development of atherosclerosis in the vascular wall via the formation of ROS.95,96 TQ significantly enhanced the plasma and liver antioxidant capacity (SOD1, CAT) and increased the expression of liver antioxidant genes (GPx2). Besides, TQ effectively decreased low-density lipoprotein cholesterol (LDLC) levels in hypercholesterolemic rats.95 Ahmad et al. showed that pretreatment of hyperlipidemic rats with N. sativa oil reduced the level of lipid peroxidation markers in plasma. Furthermore, administration of N. sativa oil elevated the antioxidant enzymes activities in erythrocytes and the liver of hyperlipidemic rats.97

Ischemic/Reperfusion Injury ROS is produced on reperfusion and induced tissue injury by activating some mediators and plays an important role in the injury caused by ischemia/reperfusion (I/R).98,99 The effect of N. sativa on liver ischemia/reperfusion injury was evaluated in rat. Rats underwent II. Role of Seeds in Nutrition and Antioxidant Activities

138

11. Protective Role of Nigella sativa and Thymoquinone in Oxidative Stress: A Review

hepatic ischemia for 45 min followed by 60 min period of reperfusion. Administration of N. sativa before ischemia and reperfusion lead to decrement of plasma alanine transaminase, AST, and LDH levels, increment of total antioxidant capacity as well as total oxidative status, oxidative stress index (OSI), MPO in hepatic tissue decreased and histological liver tissue damage reduced.99 Hosseinzadeh et al. showed that TQ has marked protective effects against muscle ischemic injury. After TQ was administrated, antioxidant capacity increased in muscle flap, and the MDA level was significantly decreased.100 Moreover, treatment with N. sativa hydroalcoholic extract resulted in a significant diminish in MDA level as well as DNA damage. This plant also increased the renal thiol content caused by renal I/R injury in rats.101

Morphine-Induced Oxidative Stress Morphine induces oxidative stress in the brain.102 N. sativa oil via inhibition of morphineinduced NO overproduction, oxidative stress, and via maintenance of the cellular antioxidant status has benefitted effects in prevention of opioid tolerance and dependence in mice.103 L-Arginine is the only natural substrate for all isoforms of NO synthase and the sole metabolic precursor for molecule NO biosynthesis.104 The concurrent administration of L-arginine antagonized the inhibitory effects of N. sativa oil on the development of morphine tolerance and dependence.103 Administration of TQ attenuated the development of tolerance and dependence to morphine in mice.105

Nephrotoxicity Gentamicin sulfate (GS) is believed to generate ROS in the kidney.106 Plasma MDA and NO levels increased and erythrocyte SOD, and GPx activities decreased significantly in GS-induced nephrotoxicity in rats.106 Administration of N. sativa oil with GS injection significantly decreased the MDA and NO generation and increased SOD and GPx activities as compared with GS group.106 Similar experiment was carried out with TQ against GS-induced nephrotoxicity. GS resulted in a significant increase in TBARS, serum creatinine, BUN, and total nitrate/nitrite (NOx), and a significant decrease in CAT reduced GSH, GPx, and adenosine triphosphate (ATP) levels in the kidney tissues.107 Oral supplementation of TQ resulted in a complete reversal of the GM-induced increase in BUN, creatinine, TBARS, and NOx and decrease in GSH, GPx, CAT, and ATP to control levels. Also, histopathological examination of kidney tissues confirmed the biochemical data, wherein TQ supplementation prevents GM-induced degenerative changes in kidney tissues.107 The most frequent adverse effect of cyclosporine A is the occurrence of nephrotoxicity, while hepatic injury also limits its clinical application.108 Treatment of rats with TQ significantly prevented cyclosporine A-induced changes in the levels of GSH, SOD, and MDA in kidney and liver tissues and attenuated the hepatic morphological changes that are detected in histological examination.108 Sodium nitrite, a food preservative, has been reported to exert deleterious toxic effects, which can affect different organs such as the kidney.109 N. sativa oil showed dose-dependent amelioration of sodium nitrite-induced nephrotoxicity in rats via blocking oxidative stress, reduction of fibrosis/inflammation, restoration of glycogen level, improvement of cytochrome C oxidase, and inhibition of apoptosis.109

II. Role of Seeds in Nutrition and Antioxidant Activities

In vivo Antioxidant Activity of Nigella sativa

139

Neurotoxicity Lead, a known neurotoxin, produces adverse effects on the brain via increased production of ROS that can lead to oxidative stress.110 The administration of 250 and 500 mg/kg ethanolic N. sativa extract reversed the adverse effects of lead-induced neurotoxicity in mice by significantly increasing the expression of SOD1, peroxiredoxin (Prdx6), and amyloid precursor protein (APP695) and reducing the expression of APP770 in cortex and hippocampus regions.110 Chlorpyrifos is an insecticide;111 the neurotoxic effects of chlorpyrifos are associated to the ability of its metabolite (Chlorpyrifos-oxon) to bind and irreversibly inhibit acetylcholinesterase, and administration of N. sativa oil improved semen picture, antioxidant enzymes, and moderate chlorpyrifos-induced reproductive toxicity in male rats.111

Plasmodium yoelii Infection Plasmodium infection leads to overproduction of free radicals and impairs antioxidant system.112 Administration 1.25 g/kg methanolic extract of N. sativa seeds (MENS) during 5 days to the P. yoelii-infected mice significantly attenuated the serum and hepatic MDA levels. Also, MENS significantly ameliorated the activities of red cell SOD and CAT as well as the hepatic SOD and GST. MENS treatment significantly increased the levels of both red cell and hepatic GSH content of P. yoelii-infected mice.113

Radiotherapy Radiotherapy is known as a type of main methods for cancer treatment, and evidence has accumulated to suggest that radiation-mediated oxidative stress can induce apoptosis.114 Intragastric administration of N. sativa in the amount of 400 mg/kg significantly decreased the elevated tissue MDA levels and increased the reduced GPx and SOD activity in intestinal tissues samples on radiation-induced intestinal injury in rats.115 Besides, N. sativa led to decrement of radiation-induced morphological changes in the irradiated rat jejunal mucosa.115 Demir et al. showed that N. sativa oil and TQ prevented cataractogenesis in ionizing radiation-induced cataracts in the lenses of rats.116

Schistosomiasis Schistosomiasis decreased the levels of protective endogenous antioxidants and increased the generation of free radicals. Administration of N. sativa oil significantly improved the antioxidant capacity (decreased the elevated MDA level, also increased the GSH levels) also led to a significant reduction in the percentage of mature eggs in Schistosoma mansoni-infected mice.117 Administration of N. sativa oil succeeded partially to correct the changes in ALT, GGT, AP activity, and the albumin content in serum of S. mansoni-infected mice (118). Furthermore, it produced an effective property against the hepatosplenic damaging effect caused by S. mansoni infection.118

II. Role of Seeds in Nutrition and Antioxidant Activities

140

11. Protective Role of Nigella sativa and Thymoquinone in Oxidative Stress: A Review

Seizure Oxidative stress can dramatically change the cell function, and an excessive production of these compounds has been correlated to seizure-induced neuronal death.119,120 Treatment with N. sativa oil induced non-significant changes in MDA but slightly attenuated the increased NO levels and decreased GSH levels by 22.02% as compared to control in animal model of epilepsy.120 In other study, N. sativa oil significantly prevented pentylenetetrazol-kindled seizures in mice and decreased the oxidative stress induced by pentylenetetrazol kindling.121 Hosseinzadeh et al. demonstrated that TQ suppresses epileptic seizures in rats, and the protective effects may be via an opioid receptor-mediated increase in GABAergic tone.122

Subarachnoid Hemorrhage Subarachnoid hemorrhage (SAH) is a common cerebrovascular event associated to various etiologies, primarily aneurysmal rupture. Cerebral vasospasm is the main complication of SAH leading to significant morbidity and mortality. Vasoconstriction secondary to SAH is induced by oxidative stress.123,124 N. sativa oil-inhibited SAH induced lipid peroxidation and neutrophil infiltration of the brain tissue in rat via reduction in the reactive hydroxyl, peroxyl, and superoxide radical’s levels. The oil also increased the levels of GSH to normal level. The Naþ/Kþ-ATPase is responsible in the generation of the membrane potential via the active transport of sodium and potassium ions in cellular membrane. N. sativa oil treatment was able to restore Naþ/ K þ -ATPase activity to normal levels. It maintains neuronal excitability and controls cellular volume in the central nervous system. Furthermore bloodebrain barrier integrity was preserved, brain edema was decreased, and neurological symptoms were improved in the rats with SAH.125 Administration of TQ counteracted the induced oxidative stress in rat’s brain tissue by enhancing the activities of non-enzymatic (GSH and vitamin C) and enzymatic (SOD, CAT, GPx, and GST) antioxidants as well by reducing the levels of MDA in the rat brain to normal levels.124

Ulcerative Colitis The excessive production of ROS by inflamed mucosa contributes significantly to the development of tissue injury in ulcerative colitis.126 Administration of black cumin oil significantly increased SOD activity but did not alter GSH level on trinitrobenzene sulfonic acid-induced experimental colitis in rats.126 TQ acts as SOD-like substances to scavenge O
2 that is produced by the xanthine/xanthine oxidase system. TQ has a scavenging effect on superoxide generated in cellular and noncellular biological systems with the IC50 of 8e20 and 60 mM, respectively.36,126

Conclusion The present review shows that black seeds and its constituent, TQ, have a lot of medicinal properties due to their antioxidant effects, and these medicinal properties were used for

II. Role of Seeds in Nutrition and Antioxidant Activities

References

141

treating many diseases such as diabetes mellitus, I/R injury, hepatotoxicity, ulcerative colitis, seizure, nephrotoxicity, hematotoxicity, arthritis, SAH, and encephalomyelitis. The antioxidant effects of N. sativa and TQ are possibly contributed by their multiple actions such as upregulating the activities of enzymatic antioxidants (CAT, SOD, GPx, and GST), preventing depletion of non-enzymatic antioxidants (GSH), suppression of the oxidative stress and inhibition of COX, inflammation, and diminution of lipid peroxidation.

References 1. Uttara B, Singh AV, Zamboni P, Mahajan R. Oxidative stress and neurodegenerative diseases: a review of upstream and downstream antioxidant therapeutic options. Current Neuropharmacology. 2009;7:65e74. 2. Leong X-F, Rais Mustafa M, Jaarin K. Nigella sativa and its protective role in oxidative stress and hypertension. Evidence-Based Complementary and Alternative Medicine. 2013;2013. 3. Lü JM, Lin PH, Yao Q, Chen C. Chemical and molecular mechanisms of antioxidants: experimental approaches and model systems. Journal of Cellular and Molecular Medicine. 2010;14:840e860. 4. Shaikh BT, Hatcher J. Complementary and alternative medicine in Pakistan: prospects and limitations. Evidence-Based Complementary and Alternative Medicine. 2005;2:139e142. 5. Shayganni E, Bahmani M, Asgary S, Rafieian-Kopaei M. Inflammaging and cardiovascular disease: management by medicinal plants. Phytomedicine. 2016;23:1119e1126. 6. Khan MA. Chemical composition and medicinal properties of Nigella sativa Linn. Inflammopharmacology. 1999;7:15e35. 7. Shrivastava R, Agrawal R, Parveen Z. A review on therapeutic applications of Nigella sativa. Journal of Chemical Sciences. 2011;1:241e248. 8. Javidi S, Razavi BM, Hosseinzadeh H. A review of neuropharmacology effects of Nigella sativa and its main component, thymoquinone. Phytotherapy Research. 2016;30:1219e1229. 9. El-Tahir KE-DH, Bakeet DM. The black seed Nigella sativa Linnaeus-A mine for multi cures: a plea for urgent clinical evaluation of its volatile oil. Journal of Taibah University for Science. 2006;1:1e19. 10. Hanafy M, Hatem M. Studies on the antimicrobial activity of Nigella sativa seed (black cumin). Journal of Ethnopharmacology. 1991;34:275e278. 11. Forouzanfar F, Bazzaz BSF, Hosseinzadeh H. Black cumin (Nigella sativa) and its constituent (thymoquinone): a review on antimicrobial effects. Iranian Journal of Basic Medical Sciences. 2014;17:929e938. 12. Badary OA, Al-Shabanah OA, Nagi MN, Al-Bekairi AM, Elmazar M. Acute and subchronic toxicity of thymoquinone in mice. Drug Development Research. 1998;44:56e61. 13. Paarakh PM. Nigella sativa Linn.eA comprehensive review. Indian Journal of Natural Products and Resources. 2010;1:409e429. 14. Hosseinzadeh H, Moghim FF, Mansouri SMT. Effect of Nigella sativa seed extracts on ischemia-reperfusion in rat skeletal muscle. Pharmacologyonline. 2007;2:326e335. 15. Alenzi F, Altamimi MA, Kujan O, et al. Antioxidant properties of Nigella sativa. Journal of Molecular and Genetic Medicine. 2013;7, 1747-0862. 16. Hosseinzadeh H, Parvardeh S, Asl MN, Sadeghnia HR, Ziaee T. Effect of thymoquinone and Nigella sativa seeds oil on lipid peroxidation level during global cerebral ischemia-reperfusion injury in rat hippocampus. Phytomedicine. 2007;14:621e627. 17. Hosseinzadeh H, Eskandari M, Ziaee T. Antitussive effect of thymoquinone, a constituent of Nigella sativa seeds, in Guinea pigs. Pharmacologyonline. 2008;2:480e484. 18. Amin B, Hosseinzadeh H. Black cumin (Nigella sativa) and its active constituent, thymoquinone: an overview on the analgesic and anti-inflammatory effects. Planta Medica. 2016;82:8e16. 19. Amin B, Taheri MMH, Hosseinzadeh H. Effects of intraperitoneal thymoquinone on chronic neuropathic pain in rats. Planta Medica. 2014;80:1269e1277. 20. Abdel-Sater KA. Gastroprotective effects of Nigella Sativa oil on the formation of stress gastritis in hypothyroidal rats. International Journal of Physiology, Pathophysiology and Pharmacology. 2009;1:143e149. 21. Boskabady M, Mohsenpoor N, Takaloo L. Antiasthmatic effect of Nigella sativa in airways of asthmatic patients. Phytomedicine. 2010;17:707e713.

II. Role of Seeds in Nutrition and Antioxidant Activities

142

11. Protective Role of Nigella sativa and Thymoquinone in Oxidative Stress: A Review

22. Khan A, Chen H, Tania M, Zhang D. Anticancer activities of Nigella sativa (black cumin). African Journal of Traditional, Complementary and Alternative Medicines. 2011;8:226e232. 23. Salem ML. Immunomodulatory and therapeutic properties of the Nigella sativa L. seed. International Immunopharmacology. 2005;5:1749e1770. 24. Majdalawieh AF, Hmaidan R, Carr RI. Nigella sativa modulates splenocyte proliferation, Th1/Th2 cytokine profile, macrophage function and NK anti-tumor activity. Journal of Ethnopharmacology. 2010;131:268e275. 25. Bukhari MH, Khalil J, Qamar S, et al. Comparative gastroprotective effects of natural honey, Nigella sativa and cimetidine against acetylsalicylic acid induced gastric ulcer in albino rats. Journal of College of Physicians and Surgeons Pakistan. 2011;21:151e156. 26. Salim EI. Cancer chemopreventive potential of volatile oil from black cumin seeds, Nigella sativa L., in a rat multi-organ carcinogenesis bioassay. Oncology Letters. 2010;1:913e924. 27. Hosseini M, Mohammadpour T, Karami R, Rajaei Z, Sadeghnia HR, Soukhtanloo M. Effects of the hydroalcoholic extract of Nigella Sativa on scopolamine-induced spatial memory impairment in rats and its possible mechanism. Chinese Journal of Integrative Medicine. 2015;21:438e444. 28. Sayeed MSB, Shams T, Hossain SF, et al. Nigella sativa L. seeds modulate mood, anxiety and cognition in healthy adolescent males. Journal of Ethnopharmacology. 2014;152:156e162. 29. Hosseinzadeh H, Tafaghodi M, Mosavi MJ, Taghiabadi E. Effect of aqueous and ethanolic extracts of Nigella sativa seeds on milk production in rats. Journal of Acupuncture and Meridian Studies. 2013;6:18e23. 30. Kolahdooz M, Nasri S, Modarres SZ, Kianbakht S, Huseini HF. Effects of Nigella sativa L. seed oil on abnormal semen quality in infertile men: a randomized, double-blind, placebo-controlled clinical trial. Phytomedicine. 2014;21:901e905. 31. Sultan MT, Butt MS, Anjum FM, Jamil A. Influence of black cumin fixed and essential oil supplementation on markers of myocardial necrosis in normal and diabetic rats. Pakistan Journal of Nutrition. 2009;8:1450e1455. 32. Salem ML, Hossain MS. Protective effect of black seed oil from Nigella sativa against murine cytomegalovirus infection. International Journal of Immunopharmacology. 2000;22:729e740. 33. Hosseinzadeh H, Fazly Bazzaz B, Haghi MM. Antibacterial activity of total extracts and essential oil of Nigella sativa L. seeds in mice. Pharmacologyonline. 2007;2:429e435. 34. Aljabre SHM, Randhawa MA, Akhtar N, Alakloby OM, Alqurashi AM, Aldossary A. Antidermatophyte activity of ether extract of Nigella sativa and its active principle, thymoquinone. Journal of Ethnopharmacology. 2005;101:116e119. 35. Razavi B, Hosseinzadeh H. A review of the effects of Nigella sativa L. and its constituent, thymoquinone, in metabolic syndrome. Journal of Endocrinological Investigation. 2014;37:1031e1040. 36. Badary OA, Taha RA, Gamal El-Din AM, Abdel-Wahab MH. Thymoquinone is a potent superoxide anion scavenger. Drug and Chemical Toxicology. 2003;26:87e98. 37. Ali B, Blunden G. Pharmacological and toxicological properties of Nigella sativa. Phytotherapy Research. 2003;17:299e305. 38. Burits M, Bucar F. Antioxidant activity of Nigella sativa essential oil. Phytotherapy Research. 2000;14:323e328. 39. Babazadeh B, Sadeghnia HR, Kapurchal ES, Parsaee H, Nasri S, Tayarani-Najaran Z. Protective effect of Nigella sativa and thymoquinone on serum/glucose deprivation-induced DNA damage in PC12 cells. Avicenna Journal of Phytomedicine. 2012;2:125e132. 40. Ashraf SS, Rao MV, Kaneez FS, et al. Nigella sativa extract as a potent antioxidant for petrochemical-induced oxidative stress. Journal of Chromatographic Science. 2011;49:321e326. 41. Mousavi S, Tayarani-Najaran Z, Asghari M, Sadeghnia H. Protective effect of Nigella sativa extract and thymoquinone on serum/glucose deprivation-induced PC12 cells death. Cellular and Molecular Neurobiology. 2010;30:591e598. 42. Miliauskas G, Venskutonis P, Van Beek T. Screening of radical scavenging activity of some medicinal and aromatic plant extracts. Food Chemistry. 2004;85:231e237. 43. Mariod AA, Ibrahim RM, Ismail M, Ismail N. Antioxidant activity and phenolic content of phenolic rich fractions obtained from black cumin (Nigella sativa) seed cake. Food Chemistry. 2009;116:306e312. 44. Kruk I, Michalska T, Lichszteld K, Kładna A, Aboul-Enein HY. The effect of thymol and its derivatives on reactions generating reactive oxygen species. Chemosphere. 2000;41:1059e1064.

II. Role of Seeds in Nutrition and Antioxidant Activities

References

143

45. Mansour MA, Nagi MN, El-Khatib AS, Al-Bekairi AM. Effects of thymoquinone on antioxidant enzyme activities, lipid peroxidation and DT-diaphorase in different tissues of mice: a possible mechanism of action. Cell Biochemistry and Function. 2002;20:143e151. 46. Fouda A-MM, Daba M-HY, Yousef Ahmed A. Antigenotoxic effects of thymoquinone against benzo [a] pyrene and mitomycin C-induced genotoxicity in cultured human lymphocytes. Research in Immunology: International Journal. 2014;2014. 47. McGill MR, Williams CD, Xie Y, Ramachandran A, Jaeschke H. Acetaminophen-induced liver injury in rats and mice: comparison of protein adducts, mitochondrial dysfunction, and oxidative stress in the mechanism of toxicity. Toxicology and Applied Pharmacology. 2012;264:387e394. 48. Adam GO, Rahman MM, Lee S-J, et al. Hepatoprotective effects of Nigella sativa seed extract against acetaminophen-induced oxidative stress. Asian Pacific Journal of Tropical Disease. 2016;9:221e227. 49. Shahroudi MJ, Mehri S, Hosseinzadeh H. Anti-aging effect of Nigella sativa fixed oil on D-galactose-induced aging in mice. Journal of Pharmacopuncture. 2017;20:29e35. 50. Bourgou S, Pichette A, Marzouk B, Legault J. Antioxidant, anti-inflammatory, anticancer and antibacterial activities of extracts from Nigella sativa (black cumin) plant parts. Journal of Food Biochemistry. 2012;36:539e546. 51. Tekeoglu I, Dogan A, Ediz L, Budancamanak M, Demirel A. Effects of thymoquinone (volatile oil of black cumin) on rheumatoid arthritis in rat models. Phytotherapy Research. 2007;21:895e897. 52. Khan N, Sultana S. Inhibition of two stage renal carcinogenesis, oxidative damage and hyperproliferative response by Nigella sativa. European Journal of Cancer Prevention. 2005;14:159e168. 53. Al-Othman AM, Ahmad F, Al-Orf S, Al-Murshed KS, Arif Z. Effect of dietary supplementation of Ellataria cardamomum and Nigella sativa on the toxicity of rancid corn oil in Rats. International Journal of Pharmacology. 2006;2:60e65. 54. El Gendy S, Hessien M, Salam IA, et al. Evaluation of the possible antioxidant effects of Soybean and Nigella Sativa during experimental hepatocarcinogenesis by nitrosamine Precursors. Turkish Journal of Biochemistry. 2007;32:5e11. 55. Devasena T, Menon VP, Rajasekharan KN. Prevention of 1, 2-dimethylhydrazine-induced circulatory oxidative stress by bis-1, 7-(2-hydroxyphenyl)-hepta-1, 6-diene-3, 5-dione during colon carcinogenesis. Pharmacological Reports. 2006;58:229e235. 56. Jrah Harzallah H, Grayaa R, Kharoubi W, Maaloul A, Hammami M, Mahjoub T. Thymoquinone, the Nigella sativa bioactive compound, prevents circulatory oxidative stress caused by 1, 2-dimethylhydrazine in erythrocyte during colon postinitiation carcinogenesis. Oxidative Medicine and Cellular Longevity. 2012;2012. 57. Sullivan B, Meyer T, Stershic M, Keefer L. Acceleration of N-nitrosation reactions by electrophiles. IARC Scientific Publications. 1991;105:370e374. 58. Sayed-Ahmed MM, Aleisa AM, Al-Rejaie SS, et al. Thymoquinone attenuates diethylnitrosamine induction of hepatic carcinogenesis through antioxidant signaling. Oxidative Medicine and Cellular Longevity. 2010;3:254e261. 59. Caballero F, Gerez E, Oliveri L, Falcoff N, Batlle A, Vazquez E. On the promoting action of tamoxifen in a model of hepatocarcinogenesis induced by p-dimethylaminoazobenzene in CF1 mice. The International Journal of Biochemistry & Cell Biology. 2001;33:681e690. 60. Suddek GM. Protective role of thymoquinone against liver damage induced by tamoxifen in female rats. Canadian Journal of Physiology and Pharmacology. 2014;92:640e644. 61. Kanter M, Coskun O, Budancamanak M. Hepatoprotective effects of Nigella sativa L and Urtica dioica L on lipid peroxidation, antioxidant enzyme systems and liver enzymes in carbon tetrachloride-treated rats. World Journal of Gastroenterology. 2005;11:6684e6688. 62. LoPachin RM. Acrylamide neurotoxicity: neurological, morhological and molecular endpoints in animal models. Chemistry and Safety of Acrylamide in Food. 2005:21e37. 63. Mehri S, Shahi M, Razavi BM, Hassani FV, Hosseinzadeh H. Neuroprotective effect of thymoquinone in acrylamide-induced neurotoxicity in Wistar rats. Iranian Journal of Basic Medical Sciences. 2014;17:1007e1011. 64. Nehar S, Kumari M. Ameliorating effect of Nigella sativa oil in thioacetamide-induced liver cirrhosis in albino rats. Indian Journal of Pharmaceutical Education and Research. 2013;47:135e139. 65. Mohamadin AM, Sheikh B, El-Aal AAA, Elberry AA, Al-Abbasi FA. Protective effects of Nigella sativa oil on propoxur-induced toxicity and oxidative stress in rat brain regions. Pesticide Biochemistry and Physiology. 2010;98:128e134.

II. Role of Seeds in Nutrition and Antioxidant Activities

144

11. Protective Role of Nigella sativa and Thymoquinone in Oxidative Stress: A Review

66. Banerjee B, Seth V, Bhattacharya A, Pasha S, Chakraborty A. Biochemical effects of some pesticides on lipid peroxidation and free-radical scavengers. Toxicology Letters. 1999;107:33e47. 67. Walters JW, Amos D, Ray K, Santanam N. Mitochondrial redox status as a target for cardiovascular disease. Current Opinion in Pharmacology. 2016;27:50e55. 68. Hassan MQ, Akhtar M, Ahmed S, Ahmad A, Najmi AK. Nigella sativa protects against isoproterenol-induced myocardial infarction by alleviating oxidative stress, biochemical alterations and histological damage. Asian Pacific Journal of Tropical Biomedicine. 2017;7:294e299. 69. Rezzani R, Angoscini P, Rodella L, Bianchi R. Alterations induced by cyclosporine A in myocardial fibers and extracelular matrix in rat. Histology & Histopathology. 2002;17:761e766. 70. Ebru U, Burak U, Yusuf S, et al. Cardioprotective effects of Nigella sativa oil on cyclosporine A-induced cardiotoxicity in rats. Basic and Clinical Pharmacology and Toxicology. 2008;103:574e580. 71. Shahid F, Farooqui Z, Rizwan S, Abidi S, Parwez I, Khan F. Oral administration of Nigella sativa oil ameliorates the effect of cisplatin on brush border membrane enzymes, carbohydrate metabolism and antioxidant system in rat intestine. Experimental & Toxicologic Pathology. 2017;69:299e306. 72. Shi Y, Vanhoutte PM. Reactive oxygen-derived free radicals are key to the endothelial dysfunction of diabetes. Journal of Diabetes. 2009;1:151e162. 73. Kaatabi H, Bamosa AO, Badar A, et al. Nigella sativa improves glycemic control and ameliorates oxidative stress in patients with type 2 diabetes mellitus: placebo controlled participant blinded clinical trial. PLoS One. 2015;10:e0113486. 74. Marnett LJ. Cyclooxygenase mechanisms. Current Opinion in Chemical Biology. 2000:545e552. 75. Al Wafai RJ. Nigella sativa and thymoquinone suppress cyclooxygenase-2 and oxidative stress in pancreatic tissue of streptozotocin-induced diabetic rats. Pancreas. 2013;42:841e849. 76. Kröncke K-D, Fehsel K, Sommer A, Rodriguez M-L, Kolb-Bachofen V. Nitric oxide generation during cellular metabolization of the diabetogenic N-mefhyl-N-Nitroso-Urea streptozotozin contributes to islet cell DNA damage. Journal of Biological Chemistry. 1995;376:179e186. 77. Kanter M, Akpolat M, Aktas C. Protective effects of the volatile oil of Nigella sativa seeds on b-cell damage in streptozotocin-induced diabetic rats: a light and electron microscopic study. Journal of Molecular Histology. 2009;40:379e385. 78. Abdelmeguid NE, Fakhoury R, Kamal SM, Al Wafai RJ. Effects of Nigella sativa and thymoquinone on biochemical and subcellular changes in pancreatic b-cells of streptozotocin-induced diabetic rats. Journal of Diabetes. 2010;2:256e266. 79. Cameron N, Cotter M, Basso M, Hohman T. Comparison of the effects of inhibitors of aldose reductase and sorbitol dehydrogenase on neurovascular function, nerve conduction and tissue polyol pathway metabolites in streptozotocin-diabetic rats. Diabetologia. 1997;40:271e281. 80. Kanter M. Effects of Nigella sativa and its major constituent, thymoquinone on sciatic nerves in experimental diabetic neuropathy. Neurochemical Research. 2008;33:87e96. 81. Heshmati J, Namazi N, Memarzadeh M-R, Taghizadeh M, Kolahdooz F. Nigella sativa oil affects glucose metabolism and lipid concentrations in patients with type 2 diabetes: a randomized, double-blind, placebo-controlled trial. Food Research International. 2015;70:87e93. 82. Mohamed A, Afridi D, Garani O, Tucci M. Thymoquinone inhibits the activation of NF-kappaB in the brain and spinal cord of experimental autoimmune encephalomyelitis. Biomedical Sciences Instrumentation. 2005;41:388e393. 83. Ozugurlu F, Sahin S, Idiz N, et al. The effect of Nigella sativa oil against experimental allergic encephalomyelitis via nitric oxide and other oxidative stress parameters. Cellular and Molecular Biology. 2005;51:337e342. 84. Pourbakhsh H, Taghiabadi E, Abnous K, Hariri AT, Hosseini SM, Hosseinzadeh H. Effect of Nigella sativa fixed oil on ethanol toxicity in rats. Iranian Journal of Basic Medical Sciences. 2014;17:1020e1031. 85. Kanter M, Coskun O, Uysal H. The antioxidative and antihistaminic effect of Nigella sativa and its major constituent, thymoquinone on ethanol-induced gastric mucosal damage. Archives of Toxicology. 2006;80:217e224. 86. Hosseini SM, Taghiabadi E, Abnous K, Timcheh Hariri A, Pourbakhsh H, Hosseinzadeh H. Protective effect of thymoquinone, the active constituent of Nigella sativa fixed oil, against ethanol toxicity in rats. Iranian Journal of Basic Medical Sciences. 2017;20:927e939. 87. Qian Y, Fan J-G. Obesity, fatty liver and liver cancer. Hepatobiliary and Pancreatic Diseases International. 2005;4:173e177.

II. Role of Seeds in Nutrition and Antioxidant Activities

References

145

88. Al-Okbi SY, Mohamed DA, Hamed TE, Edris AE. Potential protective effect of Nigella sativa crude oils towards fatty liver in rats. European Journal of Lipid Science and Technology. 2013;115:774e782. 89. Guo T, McCay J, Brown R, et al. Carbon tetrachloride is immunosuppressive and decreases host resistance to Listeria monocytogenes and Streptococcus pneumoniae in female B6C3F1 mice. Toxicology. 2000;154:85e101. 90. Essawy AE, Hamed SS, Abdel-Moneim AM, Abou-Gabal AA, Alzergy AA. Role of black seeds (Nigella sativa) in ameliorating carbon tetrachloride induced haematotoxicity in Swiss albino mice. Journal of Medicinal Plants Research. 2010;4:1977e1986. 91. Abou Khalil N, Abd-Elkareem M, Sayed A. Nigella sativa seed protects against 4-nonylphenol-induced haematotoxicity in Clarias gariepinus (Burchell, 1822): oxidant/antioxidant rebalance. Aquaculture Nutrition. 2017;23:1467e1474. 92. Cesaratto L, Vascotto C, Calligaris S, Tell G. The importance of redox state in liver damage. Annals of Hepatology. 2004;3:86e92. 93. Abdel-Moneim A, Morsy BM, Mahmoud AM, Abo-Seif MA, Zanaty MI. Beneficial therapeutic effects of Nigella sativa and/or Zingiber officinale in HCV patients in Egypt. EXCLI Journal. 2013;12:943e955. 94. Barakat EMF, El Wakeel LM, Hagag RS. Effects of Nigella sativa on outcome of hepatitis C in Egypt. World Journal of Gastroenterology. 2013;19:2529e2536. 95. Ismail M, Al-Naqeep G, Chan KW. Nigella sativa thymoquinone-rich fraction greatly improves plasma antioxidant capacity and expression of antioxidant genes in hypercholesterolemic rats. Free Radical Biology and Medicine. 2010;48:664e672. 96. Shi W, Haberland ME, Jien M-L, Shih DM, Lusis AJ. Endothelial responses to oxidized lipoproteins determine genetic susceptibility to atherosclerosis in mice. Circulation. 2000;102:75e81. 97. Ahmad S, Beg ZH. Evaluation of therapeutic effect of omega-6 linoleic acid and thymoquinone enriched extracts from Nigella sativa oil in the mitigation of lipidemic oxidative stress in rats. Nutrition. 2016;32:649e655. 98. Hassan-Khabbar S, Cottart CH, Wendum D, et al. Postischemic treatment by trans-resveratrol in rat liver ischemia-reperfusion: a possible strategy in liver surgery. Liver Transplantation. 2008;14:451e459. 99. Yildiz F, Coban S, Terzi A, et al. Nigella sativa relieves the deleterious effects of ischemia reperfusion injury on liver. World Journal of Gastroenterology. 2008 7;14:5204e5209. 100. Hosseinzadeh H, Taiari S, Nassiri-Asl M. Effect of thymoquinone, a constituent of Nigella sativa L., on ischemiaereperfusion in rat skeletal muscle. Naunyn Schmiedebergs Arch Pharmacol. 2012;385:503e508. 101. Havakhah S, Sadeghnia HR, Mosa-Al-Reza Hajzadeh NM, et al. Effect of Nigella sativa on ischemia-reperfusion induced rat kidney damage. Iranian Journal of Basic Medical Sciences. 2014;17:986e992. _ Nazıro 102. Özmen I, glu M, Alici HA, Sahin F, Cengiz M, Eren I. Spinal morphine administration reduces the fatty acid contents in spinal cord and brain by increasing oxidative stress. Neurochemical Research. 2007;32:19e25. 103. Abdel-Zaher AO, Abdel-Rahman MS, ELwasei FM. Blockade of nitric oxide overproduction and oxidative stress by Nigella sativa oil attenuates morphine-induced tolerance and dependence in mice. Neurochemical Research. 2010;35:1557e1565. 104. Brosnan CF, Battistini L, Raine CS, Dickson DW, Casadevall A, Lee SC. Reactive nitrogen intermediates in human neuropathology: an overview. Developmental Neuroscience. 1994;16:152e161. 105. Hosseinzadeh H, Parvardeh S, Masoudi A, Moghimi M, Mahboobifard F. Attenuation of morphine tolerance and dependence by thymoquinone in mice. Avicenna Journal of Phytomedicine. 2016;6:55e66. _ Balikci E. Protective effects of Nigella sativa against gentamicin-induced nephrotoxicity in rats. 106. Yaman I, Experimental & Toxicologic Pathology. 2010;62:183e190. 107. Sayed-Ahmed MM, Nagi MN. Thymoquinone supplementation prevents the development of gentamicininduced acute renal toxicity in rats. Clinical and Experimental Pharmacology and Physiology. 2007;34:399e405. 108. Farag MM, Ahmed GO, Shehata RR, Kazem AH. Thymoquinone improves the kidney and liver changes induced by chronic cyclosporine A treatment and acute renal ischaemia/reperfusion in rats. Journal of Pharmacy and Pharmacology. 2015;67:731e739. 109. Al-Gayyar MM, Hassan HM, Alyoussef A, Abbas A, Darweish MM, El-Hawwary AA. Nigella sativa oil attenuates chronic nephrotoxicity induced by oral sodium nitrite: effects on tissue fibrosis and apoptosis. Redox Report. 2016;21:50e60. 110. Butt UJ, Shah SAA, Ahmed T, Zahid S. Protective effects of Nigella sativa Lin. seed extract on Lead induced neurotoxicity during development and early life in mice model. Toxicol Res. 2017;7:32e40.

II. Role of Seeds in Nutrition and Antioxidant Activities

146

11. Protective Role of Nigella sativa and Thymoquinone in Oxidative Stress: A Review

111. Mosbah R, Yousef MI, Maranghi F, Mantovani A. Protective role of Nigella sativa oil against reproductive toxicity, hormonal alterations, and oxidative damage induced by chlorpyrifos in male rats. Toxicology and Industrial Health. 2016;7:1266e1277. 112. Rodrigues JR, Gamboa ND. Effect of dequalinium on the oxidative stress in Plasmodium berghei-infected erythrocytes. Journal of Parasitology Research. 2009;104:1491e1496. 113. Okeola VO, Adaramoye OA, Nneji CM, Falade CO, Farombi EO, Ademowo OG. Antimalarial and antioxidant activities of methanolic extract of Nigella sativa seeds (black cumin) in mice infected with Plasmodium yoelli nigeriensis. Parasitology Research. 2011;108:1507e1512. 114. Lee K, Park J-S, Kim Y-J, et al. Differential expression of Prx I and II in mouse testis and their up-regulation by radiation. Biochemical and Biophysical Research Communications. 2002;296:337e342. 115. Orhon ZN, Uzal C, Kanter M, Erboga M, Demiroglu M. Protective effects of Nigella sativa on gamma radiationinduced jejunal mucosal damage in rats. Pathology, Research & Practice. 2016;212:437e443. 116. Demir E, Taysi S, Al B, et al. The effects of Nigella sativa oil, thymoquinone, propolis, and caffeic acid phenethyl ester on radiation-induced cataract. Wiener Klinische Wochenschrift. 2016;128:587e595. 117. Shenawy E, Nahla S, Soliman MF, Reyad SI. The effect of antioxidant properties of aqueous garlic extract and Nigella sativa as anti-schistosomiasis agents in mice. Revista do Instituto de Medicina Tropical de Sao Paulo. 2008;50:29e36. 118. Mahmoud M, El-Abhar H, Saleh S. The effect of Nigella sativa oil against the liver damage induced by Schistosoma mansoni infection in mice. Journal of Ethnopharmacology. 2002;79:1e11. 119. Frantseva MV, Velazquez JLP, Hwang PA, Carlen PL. Free radical production correlates with cell death in an in vitro model of epilepsy. European Journal of Neuroscience. 2000;12:1431e1439. 120. Ezz HSA, Khadrawy YA, Noor NA. The neuroprotective effect of curcumin and Nigella sativa oil against oxidative stress in the pilocarpine model of epilepsy: a comparison with valproate. Neurochemical Research. 2011;36:2195e2204. 121. Ilhan A, Gurel A, Armutcu F, Kamisli S, Iraz M. Antiepileptogenic and antioxidant effects of Nigella sativa oil against pentylenetetrazol-induced kindling in mice. Neuropharmacology. 2005;49:456e464. 122. Hosseinzadeh H, Parvardeh S, Nassiri-Asl M, Mansouri M-T. Intracerebroventricular administration of thymoquinone, the major constituent of Nigella sativa seeds, suppresses epileptic seizures in rats. Medical Science Monitor. 2005;11:BR106eB110. 123. Ayer R, Zhang J. Oxidative stress in subarachnoid haemorrhage: significance in acute brain injury and vasospasm. Acta Neurochirurgica Supplement. 2008;108:33e41. 124. Sheikh BY, Mohamadin AM. Thymoquinone a potential therapy for cerebral oxidative stress. Asian Journal of Natural & Applied Sciences. 2012;1:76e92. 125. Ersahin M, Toklu HZ, Akakin D, Yuksel M, Ye gen BÇ, Sener G. The effects of Nigella sativa against oxidative injury in a rat model of subarachnoid hemorrhage. Acta Neurochirurgica. 2011;153:333e341. 126. Isik F, Akbay TT, Yarat A, et al. Protective effects of black cumin (Nigella sativa) oil on TNBS-induced experimental colitis in rats. Digestive Diseases and Sciences. 2011;56:721e730.

II. Role of Seeds in Nutrition and Antioxidant Activities

C H A P T E R

12

Black Soybean Seed: Black Soybean Seed Antioxidant Capacity Ignasius Radix A.P. Jati Department of Food Technology, Faculty of Agricultural Technology, Widya Mandala Catholic University Surabaya, Surabaya, East Java, Indonesia List of Abbreviations CAM Cellular adhesion molecule CHD Coronary heart disease DNA Deoxyribonucleic acid DPPH Diphenyl-b-picrylhydrazyl FRAP Ferric reducing antioxidant power IRF-1 Interferon regulatory transcription factor-1 LDL Low-density lipoprotein LPS Lipopolysaccharide mRNA Messenger RNA PPARg Peroxisome proliferator-activated receptor g ROS Reactive oxygen species TBARS Thiobarbituric acid reactive substance TNF Tumor necrosis factor UVB Ultraviolet B VCAM Vascular cell adhesion molecule

Introduction Soybeans have been consumed in Asian countries since ancient times, especially in China, Japan, Korea, and Indonesia. Foods based on soybeans, such as tofu, natto, and tempeh, are an integral part of the Asian diet, contributing a high amount of protein intake along with meat-based foods. For centuries, black soybeans have been known and used as traditional

Nuts and Seeds in Health and Disease Prevention, Second Edition https://doi.org/10.1016/B978-0-12-818553-7.00012-7

147

Copyright © 2020 Elsevier Inc. All rights reserved.

148

12. Black Soybean Seed: Black Soybean Seed Antioxidant Capacity

remedies to treat common colds and fevers along with their symptoms such as headaches. Soybeans are also used for people who have irritable bowel syndrome and experience an uncomfortable sensation in the chest.1 Investigations into the biological activities of black soybeans are rapidly increasing because of reports revealing that black soybeans have a high content of anthocyanin in their seed coat.2 Anthocyanin is a secondary metabolite of plants that is responsible for the formation of their black, purple, and red color. Studies on anthocyanin in black soybean seed coats also show that black soybean seed coat extract has antioxidant activities postulated to contribute to the prevention of degenerative diseases such as cancer, coronary heart disease, and diabetes because of their ability to inhibit the rate of oxidation in human metabolism.3 Studies on elucidating the mechanism of anthocyanin in preventing and treating such diseases have become the focus of scientists. This chapter provides information on black soybeans as a plant and a source of food products and describes the ability of the extract of anthocyanin of soybeans and its food products to prevent diseases and promote human health.

History, cultivation, and use According to ancient scripture, black soybeans are believed to have been planted and cultivated in Asia. The period of the Shang Dynasty (1700e1100 BC) was postulated to be the earliest time of black soybean cultivation, especially in northern China.4 Together with rice, wheat, millet, and adzuki beans, black soybeans were named as one of the five sacred grains mainly owing to their importance in the daily life of the Chinese, although black soybeans were rarely consumed as a staple food but were commonly used as a medicinal food and remedies. It is believed that the inclusion of black soybeans as a sacred grain was for mythological and supernatural reasons as a grains from God that has the ability to cure numerous diseases. The increased amount of international trading in the 16th century led to the spread of black soybean cultivation in other Asia regions such as Japan, Korea, and Indonesia. Beside its use as a food, black soybeans have become an essential part of many traditional ceremonies. In Indonesia, black soybeans are used in traditional ceremonies such as weddings and funerals.5 The soybean (Glycine max L. Merril) is a species of plant belonging to the Leguminoseae group also known as soja max and glycine soja. There are numerous varieties of soybean, based on the color of their seed coat. The most common are yellow, green, and black seed coat soybeans. In addition to the black seed coat, black soybeans have a yellow seed interior (cotyledons) and a near-spherical shape. To obtain anthocyanin, the seed coat should be removed from the cotyledons. This can be done by soaking, boiling, and then peeling or drying followed by coarse milling of the black soybean. Compared with other beans, the hilum of soybeans is longer and thinner. Soybean species are considered a short season crop that usually needs 3e5 months of growth from germination until harvest. As a rainfed bush crop, an average of 350e450 mm rainfall is needed for the black soybean plant for optimal growth and yield.6 Despite its demand for a sufficient amount of water, excess water can have a detrimental effects on the plant, such as impaired germination, leading to anaerobic respiration, and an increase in the incidence of pathogenic activity. Although

II. Role of Seeds in Nutrition and Antioxidant Activities

149

Anthocyanin

TABLE 12.1

Black Soybean-Based Food Products.

Product

Description

Country of Production

In si, tau si

Dried by-product of mashed black soybean sauce fermented with Aspergillus oryzae

China, the Philippines

Tempeh

Traditional food from black or yellow soybean fermented with Rhizopus oligosporus

Indonesia

Tofu

Protein gel-like product from soybeans

Asian countries

Soy sauce

Sauces fermented with A. oryzae and Aspergillus sojae, used as condiment

Asian countries

Natto

Traditional Japanese soybean product fermented with Bacillus subtilis

Japan

Chungkookjang

Steamed black soybeans fermented with Bacillus species

Korea

black soybeans are commonly found in tropical areas, based on their nature, they can be cultivated within a wide range of temperatures. The development of fermentation methods has led to an in black soybean-based food products.7e10 An example of black soybean-based food products is presented in Table 12.1. Although black soybeans are popular as a food with medicinal properties, their cultivation and use are limited compared with yellow soybean. There are few large-scale farming areas as well as food industries for black soybeans. A report from Indonesia revealed that there is cooperation between large-scale industry and farmers to plant black soybeans for use in soy sauce, which is a popular condiment among Indonesians.11 This cooperation has become part of community empowerment to increase the population’s livelihood and reduce the poverty of traditional farmers who have only a small farm, by providing the seeds and purchasing the yield at a fair price. There is still much future potency that can be optimized from black soybean seeds especially related to their health-promoting properties. For example, industries could extract anthocyanin from the black soybean seed coat. From the seeds, bioactive peptides could be isolated, and both could potentially be used as an alternative medicine.

Anthocyanin Black soybeans are not as popular as yellow soybeans, but researchers have investigated the ability of black soybeans to provide health-promoting properties as conceived of in several Asian countries for centuries. One factor with a key role in the ability of black soybean to be used as a remedy is the anthocyanin content in the seed coat. Anthocyanin is part of phenolic substances, which are the secondary metabolites of plants. Numerous studies reported that polyphenol substances exhibit significant antioxidative activities; they are suggested to be responsible for preventing several degenerative diseases.12 Anthocyanin is a class of polyphenols that is a water-soluble pigment responsible for the red, blue, and black colors of flowers and plants. Anthocyanin is widely known to have bioactive properties and substantial pharmaceutical activity. Fig. 12.1 shows the chemical structure of anthocyanin.

II. Role of Seeds in Nutrition and Antioxidant Activities

150

12. Black Soybean Seed: Black Soybean Seed Antioxidant Capacity

FIGURE 12.1

Chemical structure of anthocyanin.

A study of the anthocyanin content of black soybeans was first reported in 1921 by Nagai regarding its formation in plants.13 Research on anthocyanin in plants including black soybeans has been growing rapidly. These findings includes the elucidation of individual anthocyanins and their biological activities.14 Information about individual anthocyanins in black soybeans is presented in Table 12.2. As shown in Table 12.2, the major anthocyanin found in black soybean seed coat is cyanidin-3-glucoside. Other individual anthocyanins such as malvidin, delphinidin, and petunidin-3-glycoside, which are considered new anthocyanin, are also presented. There are variations in anthocyanin content in the black soybean seed coat. This could be because of the species, climatic conditions, and the geographical location.15 However, compared with other plant foods such as rice, sorghum, berries, and grapes, the anthocyanin content of black soybean seed coats is relatively high. The anthocyanin contents of different varieties of black soybean are illustrated in Table 12.3.

Antioxidant activity of black soybeans and black soybeanebased food products Studies on the antioxidant activity of plant foods are rapidly increasing. In the human system, antioxidants are is believed to have the important function of stabilizing free radicals, whereas free radicals are substances commonly found in humans as a result of metabolism.16 TABLE 12.2

Individual Anthocyanin of Black Soybeans.

Black Soybean varieties/Sources

Individual Anthocyanins

References

Cheongja 3/Korea

Cyanidin-3-O-glucoside, petunidin-3-O-glucoside, delphinidin-3-O-glucoside

Jang et al.

A3/Sichuan, China

Cyanidin 3 glucoside, petunidin 3 glucoside, delphinidin 3 glucoside, peonidin 3 glucoside

Wu et al.

Black Tokyo/Serbia

Cyanidin 3 glucoside, pelargonidin 3 glucoside, delphinidin 3 glucoside

Kalusevic et al.

Cheongja 4 ho/Miryang, Korea

Cyanidin-3-O-glucoside, petunidin-3-O-glucoside, delphinidin-3-O-glucoside

Ryu and Koh

852/Heilongjiang, China

Cyanidin 3 glucoside

Xie et al.

II. Role of Seeds in Nutrition and Antioxidant Activities

61

62

65

64

63

151

Antioxidant activity of black soybeans and black soybeanebased food products

TABLE 12.3

Anthocyanin Content of Different Varieties of Black Soybeans.

Black Soybean varieties/Sources

Total Anthocyanin content (mg/G)

References

Mallika

13.63

Astadi et al.

46

Cikuray

14.68

Astadi et al.

46

Cheongja 3/Korea

12.11

Jang et al.

A3/Sichuan, China

3.95

Wu et al.

62

QWT31/Yunnan, China

4.96

Wu et al.

62

QWT5/Guizhou, China

3.01

Wu et al.

62

JJ16/Chongqing. China

3.62

Wu et al.

62

Black Tokyo/Serbia

1.92

Kalusevic et al.

Cheongja4ho/Miryang, Korea

1.68

Ryu and Koh

852/Heilongjiang, China

6.96

Xie et al.

61

63

64

65

The few free radicals in a normal condition usually can be neutralized by antioxidants synthesized by the human body, such as superoxide dismutase or glutathione peroxidase. However, pollution, an unbalanced diet, excessive exposure to sunlight, and smoke could multiply the number of free radicals, which need additional antioxidants from the diet to prevent excessive oxidation.17 Free radicals attack lipids, protein, and DNA, leading to the development of various diseases such as atherosclerosis, cancer, and coronary heart disease.18 Rapid progress in research on antioxidant activity is supported by the development of antioxidant activity assays. In vitro procedures such as diphenyl-b-picrylhydrazyl (DPPH), ferric reducing antioxidant power (FRAP), oxygen radical absorbance capacity, thiobarbituric acid reactive substance (TBARS), linoleic bleaching system, superoxide anion, hydroxyl radical antioxidant capacity, total reactive antioxidant potential, potassium ferricyanide reducing power, and cupric reducing antioxidant capacity were created to mimic biological processes in the human body.19e21 These procedures could assist researchers in the early stages of exploring the potency of samples of antioxidants. Among a number of procedures, DPPH is the most commonly used method for examining the antioxidant activity of black soybean seed coat extract. Results showed that black soybean seed coat extract possesses high antioxidant activity owing to anthocyanin and other phenolic content that could act as hydrogen donors that donating hydrogen to stabilize free radicals.22e24 The 20 -azinobis(3-ethylbenzothiazoline-6-sulphonic acid) method is also widely used to determine black soybean antioxidant activity. The results are in line with the DPPH method, which shows that black soybean extract is able to donate a hydrogen atom.25,26 Another commonly method used is FRAP. Reports suggest that black soybean seed coat extract significantly reduces iron, which means that it has high antioxidant content.27,28 The antioxidant activity of black soybeans can also be examined in a lipid system using linoleic acid or other lipids. It was proven that black soybean extract was able to reduce lipid oxidation, as measured by the inhibition of TBARS formation.29,30 This simple in vitro technique assists in the research of

II. Role of Seeds in Nutrition and Antioxidant Activities

152

12. Black Soybean Seed: Black Soybean Seed Antioxidant Capacity

the biological activity of black soybeans using cell cultures, animal experiments, and human subjects to explore the mechanisms of health-promoting and disease-preventing properties of black soybean seeds. Beside the investigation into the antioxidant activity of raw black soybean seed extract, there are also reports on the ability of black soybean seed food products to act as antioxidants. A study on the antioxidant activity of tofu as the most popular soybean-based product was conducted using black bean tofu. Tofu from black bean showed relatively high antioxidant activity when it was examined using the thiocyanate method. Moreover, from the lipid peroxidation assay, black bean tofu inhibited the rate of lipid peroxidation, which affected the shelf life of tofu to be longer than that of yellow soybean tofu.31 Meanwhile, research on chungkookjang, a traditional Korean paste made of fermented black soybeans, revealed that fermented black soybeans could scavenge DPPH radicals better than unfermented ones. It was also postulated from an in vivo study that a diet of chungkookjang could increase superoxide dismutase and catalase activity as an antioxidant within the body, and thus could stabilize free radicals. Moreover, hepatic TBARS was also reduced. The higher antioxidant activity of fermented black soybeans results from the increase in polyphenol content caused by partial cleavage of the glycosides by fermentation.32 Research on food products shows that the antioxidant activity does not significantly decrease with processing.33e35 This finding can be seen in Fig. 12.2, which shows that different processing times and temperatures did not significantly decrease the antioxidant activity of black and yellow soybean crackers. This provides a promising future for the development of functional foods from black soybean seeds. Thus, consuming black soybean seed products should be promoted to increase the intake of healthy food.

Health-promoting and disease-preventing effects of black soybean seed Black soybean seed has been used as a medicinal food and remedy for centuries. However, there is only limited scientific research to support such claims. Much research has been done to elucidate the mechanism of black soybean seed for promoting health and preventing disease. Factors related to the development of many diseases have been found, and the researchers elaborated on the results to investigate the health-enhancing properties of black soybean seed against inflammatory disease, atherosclerosis, diabetes, obesity, coronary heart disease, cancer, and so on.

FIGURE 12.2 Effects of different types of processing on antioxidant activity of soybean crackers. ABTS, 2,2’azino-bis(3-ethylbenzothiazoline-6-sulphonic acid; TE, Trolox Equivalent; UC, Unbaked Control. II. Role of Seeds in Nutrition and Antioxidant Activities

Health-promoting and disease-preventing effects of black soybean seed

153

Atherosclerosis and coronary heart disease Atherosclerosis is a condition in which the blood vessels are partially or fully blocked by the accumulation of plaque; thus, blood cannot circulate throughout the body. The development of atherosclerosis is believed to be caused by the oxidation of low-density lipoprotein (LD)L. As a transport mechanism for cholesterol, LDL is potentially oxidized and may accumulate in the lining of blood vessels. This condition leads to the development of diseases such as coronary heart disease (CHD). The beneficial properties of anthocyanin in reducing CHD risk have been reported.36e38 The ability of black soybeans to prevent the oxidation of LDL is proposed to be related to the delay of plaque formation. An early report also suggested that the daily intake of soybean protein is associated with a decrease in cardiovascular disease risk.39 Other studies showed that the polyphenol content of soybean seed coats could prolong the lag time of LDL oxidation and that the ability of black soybean for this activity compared with yellow soybeans.40 Research on the Malika and Cikuray varieties of black soybean in Indonesia revealed that the seed coat extract had the ability to prevent isolated human LDL oxidation. This beneficial properties could be due to the ability of anthocyanin in the extract to scavenge free radicals and thus inhibit the reaction between LDL and free radicals.41 By using an in vitro monocyte-endothelial cell adhesion assay, researchers proved that black soybean extract had the ability to prevent atherosclerosis. This method is usually used to examine the potency of a sample to inhibit the development of atherosclerosis by mimicking the first phase of atherosclerosis.42 In the early stage of atherosclerosis, cellular adhesion molecules (CAMs) on the vascular endothelial cells are activated by different factors, especially inflammatory conditions. After that, CAMs are bound by leukocytes such as monocytes.43 Both the seed coat extract and the embryo extract of Yak Kong black soybean from Korea were able to attenuate the adhesion of THP-1 monocytes to LPSinduced human umbilical vein endothelial cells by up to 40% compared with the lipopolysaccharide (LPS)-stimulated control group. This research was done within a nontoxic dose of extract (5e20 mg/mL).44

Obesity and diabetes Obesity has become an enormous problem in both developed and developing countries. Moreover, the rate of obesity in children is increasing rapidly. Obesity is believed to have an important role as a risk factor for diabetes.45 A study on the effect of consuming black soybean anthocyanin in rats revealed that anthocyanin is suggested to have antiobesity properties as well as provide hypolipidemic effects. Supplementation of black soybean anthocyanin in high-fat diet rats could moderate weight gain in the liver and decrease epididymal and perirenal fat pads. Moreover, black soybean anthocyanin supplementation improved the lipid profile of rats by decreasing cholesterol and triglyceride serum levels and increasing the high-density lipoprotein cholesterol concentration.46 Research on the antiobesity properties of Monascus pilosus fermented black soybean was reported. Using adipocytes and high-fat diet-induced obese mice, the research revealed that lipid accumulation in 3T3-L1 adipocytes was inhibited by fermented black soybeans. Consuming fermented black soybeans could decrease body weight gain in the mice. Meanwhile, consuming fermented black soybeans significantly lowered the messenger RNA

II. Role of Seeds in Nutrition and Antioxidant Activities

154

12. Black Soybean Seed: Black Soybean Seed Antioxidant Capacity

(mRNA) levels of adipogenesis-related genes such as peroxisome proliferator activated receptor, fatty acid synthase, and fatty acid binding protein.47Another report on the relation between consuming black soybeans and obesity prevention was reported in Korea. Consuming black soybeans decreased the intake of food, fat accumulation, and lipogenesis gene expression such as acetyl CoA carboxylase and CCAAT-enhancer-binding protein a. Fig. 12.3 shows the fat weight and adipocyte cell size in control and high-fat diet mice treated with black soybeans, and high-fat diet mice treated with orlistat, respectively. The results revealed that black soybean treatment decreased the fat weight and adipocyte cell size of high-fat diet rats. Black soybean intake was also closely related to the increase level of lipoprotein lipase, hormone-sensitive lipase, and adenosine monophosphateeactivated protein kinase as a lipolysis protein. Although strong evidence of the effect of black soybean on lipogenesis gene expression and a decrease in fat accumulation was clearly observed, the mechanism of action of black soybean seed coat extract remains unclear.48 Adipocyte differentiation is closely related to the incidence of diabetes. A study on the antidiabetes effect of black soybean and its anthocyanin cyanidin 3 glucoside was done on the adipocyte differentiation of mice. Intake of black soybean and its anthocyanin decreased

(A)

(B)

CON

HF

d

5

c

4 3 b

1 0

BBC

O

a

CON

HF

BBC

O

b

1600

P < 0.001

6

2

HF

O

Adipocyte size (µm)

Mesenteric fat i 100g bw (g)

BBC

CON

1400 1200

P < 0.05 a

a

a

1000 800 600 400 200 0

CON

HF

FIGURE 12.3

BBC

O

Mesenteric fat weight and adipocytes cell size of mice. BBC, high-fat diet with black soybean treatment; CON, control; HF, high-fat diet; O, high-fat diet with orlistat treatment.

II. Role of Seeds in Nutrition and Antioxidant Activities

Health-promoting and disease-preventing effects of black soybean seed

155

body weight as well as the weight of white adipose tissue and the size of adipocytes in white adipose tissue. Moreover, smaller adipocytes were observed on 3T3-LI cells treated with black soybean extract. This result correlated with the increase in peroxisome proliferatoractivated receptor g and C/enhancer binding protein a gene expression. Moreover, it was suggested that adiponectin secretion increased, tumor necrosis factor (TNF) decreased, insulin signaling was activated, and the uptake of glucose increased.49

Inflammation and cancer Inflammation has long been implicated in the development of cancer. Studies of inflammation related to the onset of various diseases have been widely performed, including the antiinflammatory effect of black soybean anthocyanin.50e52 A study showed that anthocyanin from black soybeans could inhibit the antigen-induced TNF-a stimulation of vascular cell adhesion molecule-1 (VCAM-1) by regulating DNA sequence GATAs and interferon regulatory transcription factor-1 (IRF-1). VCAM-1 is believed to be a target for highly metastatic human melanoma cells. These cells have a high-affinity conformation at their cell surface, facilitating adherence and subsequent migration. The IRF-1 and transcription factor genes bind to GATA in the VCAM-1 gene promoter region. These metastatic cells have a pathological role in inflammatory processes that eventually lead to cancer and atherosclerosis. Stimulation of cells with TNF-a increases VCAM-1 expression. Pretreatment of cells with anthocyanins inhibited VCAM-1 expression and reduced the nuclear levels of GATAs and IRF-1.53 Antiinflammatory and antifibrotic activities of Cheongja 3 black soybean was reported using an animal model for the treatment of Peyronie disease. The result showed that Peyronie disease plaque formation was reduced. Moreover, strong transforming growth factor-b1 immunoreactivity was observed with the increased expression in the collagenous connective tissue and fibroblast.54 Several epidemiological studies consistently suggested that the risk for cancer can be reduced by consuming soybean-based foods containing an antioxidant compound such as anthocyanin.55,56 A case-control report from Korea revealed that the risk for breast cancer in Korean women was reduced by consuming black soybeans.57 This is attributed to the anthocyanin content of black soybeans. Research on the antiinflammatory and antiproliferative effects of black soybean anthocyanin was also done using HT-29 human colon adenocarcinoma cells, which showed that cyanidin and delphinidin significantly inhibited cell growth and suppressed cyclooxygenase-2 and inducible nitric oxide synthase mRNA production.58 Anthocyanin was also reported to have the ability to protect skin as an anticancer and antiaging agent.59 It was reported that black soybean anthocyanin defended keratinocytes from ultraviolet B (UVB)-induced cytotoxicity and apoptosis. The mechanism behind this ability of anthocyanin is inhibition of the caspase 3 pathway and Bax protein level reduction as a proapoptotic. A study on mouse skin revealed that anthocyanin can prevent apoptotic cell death. This probably results from the ability of anthocyanin to modulate UVB-mediated reactive oxygen species (ROS) production after UVB exposure. ROS has a key role in the apoptosis pathway. Therefore, by modulating ROS production, lipid peroxidation could be reduced, and the oxidative damage of DNA and cellular protein could be obviated.60

II. Role of Seeds in Nutrition and Antioxidant Activities

156

12. Black Soybean Seed: Black Soybean Seed Antioxidant Capacity

Summary Points Black soybeans originated from China and are widely cultivated in Indonesia, Japan, and Korea. Black soybeans are usually used for traditional medicine, herbs, and remedies. Black soybeans can be used as ingredients to produce tempeh, natto, miso, sweet soy sauce, tofu, and soy milk. Black soybeans contain a high amount of anthocyanin in their seed coat, especially cyanidin 3 glucoside, which possesses high antioxidant activity. Processing does not significantly decrease the antioxidant activity of black soybeans. Black soybeans have antiobesity, antidiabetes, anticancer, and antiatherosclerotic properties.

References 1. Furuta S, Takahashi M, Takahata Y, et al. Radical-scavenging activities of soybean cultivars with black seed coats. Food Science and Technology Research. 2007. https://doi.org/10.3136/fstr.9.73. 2. Choung MG, Baek IY, Kang ST, et al. Isolation and determination of anthocyanins in seed coats of black soybean (Glycine max (L.) Merr.). Journal of Agricultural and Food Chemistry. 2001. https://doi.org/10.1021/jf010550w. 3. Mohamed S. Functional foods against metabolic syndrome (obesity, diabetes, hypertension and dyslipidemia) and cardiovascular disease. Trends in Food Science & Technology. 2014. https://doi.org/10.1016/j.tifs.2013.11.001. 4. Radix Astadi I, Paice AG. Black soybean (Glycine max L: Merril) seeds’ antioxidant capacity. In: Nuts and Seeds in Health and Disease Prevention. 2011. https://doi.org/10.1016/B978-0-12-375688-6.10027-1. 5. Sjauw-Koen-Fa AR, Blok V, Omta O. Exploring the applicability of a sustainable smallholder sourcing model in the black soybean case in Java. The International Food and Agribusiness Management Review. 2017. https://doi.org/ 10.22434/IFAMR2016.0171. 6. Dashti NH, Cherian VM, Smith DL, Mcgill J. Soybean production. Abiotic Biot Stress Soybean Prod. 2016. https:// doi.org/10.1016/B978-0-12-801536-0.00010-4. 7. Iwasaki K, Okazaki K. Brewing of functional soybean paste (miso) using black soybean. Journal of the Brewing Society of Japan. 2014. https://doi.org/10.6013/jbrewsocjapan1988.103.17. 8. Chen KI, Erh MH, Su NW, Liu WH, Chou CC, Cheng KC. Soyfoods and soybean products: from traditional use to modern applications. Applied Microbiology and Biotechnology. 2012. https://doi.org/10.1007/s00253-012-43307. 9. Murooka Y, Yamshita M. Traditional healthful fermented products of Japan. Journal of Industrial Microbiology and Biotechnology. 2008. https://doi.org/10.1007/s10295-008-0362-5. 10. Shahidi F, Ho C-T. Flavor chemistry of ethnic foods. In: Flavor Chemistry of Ethnic Foods. 2011. https://doi.org/ 10.1007/978-1-4615-4783-9_1. 11. Apriyantono A, Setyaningsih D, Hariyadi P, Nuraida L. Sensory and peptides characteristics of soy sauce fractions obtained by ultrafiltration. Advances in Experimental Medicine and Biology. 2004. https://doi.org/10.1007/ 978-1-4419-9090-7_15. 12. Potì F, Santi D, Spaggiari G, Zimetti F, Zanotti I. Polyphenol health effects on cardiovascular and neurodegenerative disorders: a review and meta-analysis. International Journal of Molecular Sciences. 2019. https://doi.org/ 10.3390/ijms20020351. 13. Kim SM, Chung MJ, Ha TJ, et al. Neuroprotective effects of black soybean anthocyanins via inactivation of ASK1-JNK/p38 pathways and mobilization of cellular sialic acids. Life Sciences. 2012. https://doi.org/ 10.1016/j.lfs.2012.04.025. 14. Lee JH, Kang NS, Shin SO, et al. Characterisation of anthocyanins in the black soybean (Glycine max L.) by HPLC-DAD-ESI/MS analysis. Food Chemistry. 2009. https://doi.org/10.1016/j.foodchem.2008.05.056. 15. Kim EH, Lee OK, Kim JK, et al. Isoflavones and anthocyanins analysis in soybean (Glycine max (L.) Merill) from three different planting locations in Korea. Field Crops Research. 2014. https://doi.org/10.1016/j.fcr.2013.10.020.

II. Role of Seeds in Nutrition and Antioxidant Activities

References

157

16. Cadenas E, Davies KJA. Mitochondrial free radical generation, oxidative stress, and aging. Free Radical Biology and Medicine. 2000. https://doi.org/10.1016/S0891-5849(00)00317-8. 17. Pisoschi AM, Pop A. The role of antioxidants in the chemistry of oxidative stress: a review. European Journal of Medicinal Chemistry. 2015. https://doi.org/10.1016/j.ejmech.2015.04.040. 18. Mayne ST. Antioxidant nutrients and chronic disease: use of biomarkers of exposure and oxidative stress status in epidemiologic research. Journal of Nutrition. 2018. https://doi.org/10.1093/jn/133.3.933s. 19. Magalhães LM, Segundo MA, Reis S, Lima JLFC. Methodological aspects about in vitro evaluation of antioxidant properties. Analytica Chimica Acta. 2008. https://doi.org/10.1016/j.aca.2008.02.047. 20. Gülçin I. Antioxidant activity of food constituents: an overview. Archives of Toxicology. 2012. https://doi.org/ 10.1007/s00204-011-0774-2. 21. Moon JK, Shibamoto T. Antioxidant assays for plant and food components. Journal of Agricultural and Food Chemistry. 2009. https://doi.org/10.1021/jf803537k. 22. Xu BJ, Chang SKC. A comparative study on phenolic profiles and antioxidant activities of legumes as affected by extraction solvents. Journal of Food Science. 2007. https://doi.org/10.1111/j.1750-3841.2006.00260.x. 23. Xu B, Chang SKC. Antioxidant capacity of seed coat, dehulled bean, and whole black soybeans in relation to their distributions of total phenolics, phenolic acids, anthocyanins, and isoflavones. Journal of Agricultural and Food Chemistry. 2008. https://doi.org/10.1021/jf801196d. 24. Juan MY, Chou CC. Enhancement of antioxidant activity, total phenolic and flavonoid content of black soybeans by solid state fermentation with Bacillus subtilis BCRC 14715. Food Microbiology. 2010. https://doi.org/10.1016/ j.fm.2009.11.002. 25. Peng H, Li W, Li H, Deng Z, Zhang B. Extractable and non-extractable bound phenolic compositions and their antioxidant properties in seed coat and cotyledon of black soybean (Glycinemax (L.) merr). Journal of Functional Foods. 2017. https://doi.org/10.1016/j.jff.2017.03.003. 26. Xu JL, Shin JS, Park SK, et al. Differences in the metabolic profiles and antioxidant activities of wild and cultivated black soybeans evaluated by correlation analysis. Food Research International. 2017. https://doi.org/ 10.1016/j.foodres.2017.08.026. 27. Kumar V, Rani A, Dixit AK, Pratap D, Bhatnagar D. A comparative assessment of total phenolic content, ferric reducing-anti-oxidative power, free radical-scavenging activity, vitamin C and isoflavones content in soybean with varying seed coat colour. Food Research International. 2010. https://doi.org/10.1016/j.foodres.2009.10.019. 28. Dajanta K, Janpum P, Leksing W. Antioxidant capacities, total phenolics and flavonoids in black and yellow soybeans fermented by Bacillus subtilis: a comparative study of Thai fermented soybeans (thua nao). International Food Research Journal. 2013;20(6):3125e3132. 29. Byun JS, Han YS, Lee SS. The effects of yellow soybean, black soybean, and sword bean on lipid levels and oxidative stress in ovariectomized rats. International Journal for Vitamin and Nutrition Research. 2010. https://doi.org/ 10.1024/0300-9831/a000010. 30. Puvaca N, Kostadinovic L, Popovic S, et al. Proximate composition, cholesterol concentration and lipid oxidation of meat from chickens fed dietary spice addition (Allium sativum, Piper nigrum, Capsicum annuum). Animal Production Science. 2016. https://doi.org/10.1071/an15115. 31. Shih MC, Yang KT, Kuo SJ. Quality and antioxidative activity of black soybean tofu as affected by bean cultivar. Journal of Food Science. 2002. https://doi.org/10.1111/j.1365-2621.2002.tb10623.x. 32. P S, K DS, K S, M BR. Fermented soybeans, Chungkookjang, prevent hippocampal cell death and b-cell apoptosis by decreasing pro-inflammatory cytokines in gerbils with transient artery occlusion. Experimental Biology and Medicine. 2016. https://doi.org/10.1177/1535370215606811. 33. Lee N-R, Woo G-Y, Jang J-H, et al. Antioxidant production by Bacillus methylotrophicus isolated from chungkookjang, Korean traditional fermented food. International Journal of Environmental Science and Technology. 2013. https://doi.org/10.5322/jesi.2013.22.7.855. 34. Slavin M, Lu Y, Kaplan N, Yu L. Effects of baking on cyanidin-3-glucoside content and antioxidant properties of black and yellow soybean crackers. Food Chemistry. 2013. https://doi.org/10.1016/j.foodchem.2013.04.039. 35. Dixit AK, Bhatnagar D, Kumar V, Rani A, Manjaya JG, Bhatnagar D. Gamma irradiation induced enhancement in isoflavones, total phenol, anthocyanin and antioxidant properties of varying seed coat colored soybean. Journal of Agricultural and Food Chemistry. 2010. https://doi.org/10.1021/jf904228e.

II. Role of Seeds in Nutrition and Antioxidant Activities

158

12. Black Soybean Seed: Black Soybean Seed Antioxidant Capacity

36. Kimble R, Keane KM, Lodge JK, Howatson G. Dietary intake of anthocyanins and risk of cardiovascular disease: a systematic review and meta-analysis of prospective cohort studies. Critical Reviews in Food Science and Nutrition. 2018. https://doi.org/10.1080/10408398.2018.1509835. 37. Cassidy A, Bertoia M, Chiuve S, Flint A, Forman J, Rimm EB. Habitual intake of anthocyanins and flavanones and risk of cardiovascular disease in men. American Journal of Clinical Nutrition. 2016. https://doi.org/10.3945/ ajcn.116.133132. 38. Amiot MJ, Riva C, Vinet A. Effects of dietary polyphenols on metabolic syndrome features in humans: a systematic review. Obesity Reviews. 2016. https://doi.org/10.1111/obr.12409. 39. Messina M. Insights gained from 20 Years of soy research. Journal of Nutrition. 2010. https://doi.org/10.3945/ jn.110.124107. 40. Takahashi R, Ohmori R, Kiyose C, Momiyama Y, Ohsuzu F, Kondo K. Antioxidant activities of black and yellow soybeans against low density lipoprotein oxidation. Journal of Agricultural and Food Chemistry. 2005. https:// doi.org/10.1021/jf048062m. 41. Astadi IR, Astuti M, Santoso U, Nugraheni PS. In vitro antioxidant activity of anthocyanins of black soybean seed coat in human low density lipoprotein (LDL). Food Chemistry. 2009;112(3). https://doi.org/10.1016/ j.foodchem.2008.06.034. 42. Moore KJ, Tabas I. Macrophages in the pathogenesis of atherosclerosis. Cell. 2011. https://doi.org/10.1016/ j.cell.2011.04.005. 43. Wang H, Patterson C, Praticò D, et al. Atherosclerosis: Risks, Mechanisms, and Therapies. 2015. https://doi.org/ 10.1002/9781118828533. 44. Lee CC, Dudonné S, Dubé P, et al. Comprehensive phenolic composition analysis and evaluation of Yak-Kong soybean (Glycine max) for the prevention of atherosclerosis. Food Chemistry. 2017. https://doi.org/10.1016/ j.foodchem.2017.05.012. 45. Guzzardi MA, Iozzo P. Obesity and diabetes. In: Interdisciplinary Concepts in Cardiovascular Health: Volume II: Secondary Risk Factors. 2013. https://doi.org/10.1007/978-3-319-01050-2_2. 46. Kim J, Lee HJ, Kim JY, Kim MK, Kwon O. Plant proteins differently affect body fat reduction in high-fat fed rats. Preventive Nutrition and Food Science. 2012. https://doi.org/10.3746/pnf.2012.17.3.223. 47. Lee YS, Choi BK, Lee HJ, et al. Monascus pilosus-fermented black soybean inhibits lipid accumulation in adipocytes and in high-fat diet-induced obese mice. Asian Pacific Journal of Tropical Medicine. 2015;8(4):276e282. https://doi.org/10.1016/S1995-7645(14)60330-8. 48. Kim SY, Wi HR, Choi S, Ha TJ, Lee BW, Lee M. Inhibitory effect of anthocyanin-rich black soybean testa (Glycine max (L.) Merr.) on the inflammation-induced adipogenesis in a DIO mouse model. Journal of Functional Foods. 2015;14:623e633. https://doi.org/10.1016/j.jff.2015.02.030. 49. Matsukawa T, Inaguma T, Han J, Villareal MO, Isoda H. Cyanidin-3-glucoside derived from black soybeans ameliorate type 2 diabetes through the induction of differentiation of preadipocytes into smaller and insulinsensitive adipocytes. The Journal of Nutritional Biochemistry. 2015;26(8):860e867. https://doi.org/10.1016/ j.jnutbio.2015.03.006. 50. Xu L, Choi TH, Kim S, et al. Anthocyanins from black soybean seed coat enhance wound healing. Annals of Plastic Surgery. 2013. https://doi.org/10.1097/SAP.0b013e31824ca62b. 51. Mueller M, Hobiger S, Jungbauer A. Anti-inflammatory activity of extracts from fruits, herbs and spices. Food Chemistry. 2010. https://doi.org/10.1016/j.foodchem.2010.03.041. 52. Lin TK, Zhong L, Santiago JL. Anti-inflammatory and skin barrier repair effects of topical application of some plant oils. International Journal of Molecular Sciences. 2018. https://doi.org/10.3390/ijms19010070. 53. Nizamutdinova IT, Jin YC, Chung J, et al. The anti-diabetic effect of anthocyanins in streptozotocin-induced diabetic rats through glucose transporter 4 regulation and prevention of insulin resistance and pancreatic apoptosis. Molecular Nutrition & Food Research. 2009. https://doi.org/10.1002/mnfr.200800526. 54. Kim SW, Sohn DW, Bae WJ, Kim HS, Kim SW. The Anti-inflammatory and antifibrosis effects of anthocyanin extracted from black soybean on a peyronie disease rat model. Urology. 2014;84(5):1112e1116. https:// doi.org/10.1016/j.urology.2014.06.026. 55. Ko K-P, Park SK, Yang JJ, et al. Intake of soy products and other foods and gastric cancer risk: a prospective study. Journal of Epidemiology. 2013. https://doi.org/10.2188/jea.je20120232. 56. De Pascual-Teresa S, Sanchez-Ballesta MT. Anthocyanins: from plant to health. Phytochemistry Reviews. 2008. https://doi.org/10.1007/s11101-007-9074-0.

II. Role of Seeds in Nutrition and Antioxidant Activities

References

159

57. Do MH, Lee SS, Jung PJ, Lee MH. Intake of fruits, vegetables, and soy foods in relation to breast cancer risk in Korean women: a case-control study. Nutrition and Cancer. 2007. https://doi.org/10.1080/01635580701268063. 58. Kim Y-H, Kim DS, Woo SS, et al. Antioxidant activity and cytotoxicity on human cancer cells of anthocynanin extraction from black soybean. Korean Journal of Crop Science. 2008;53(4):407e412. 59. Tsoyi K, Park H, Fau - Kim YM, Kim Y, Fau - Chung JI, et al. Anthocyanins from black soybean seed coats inhibit UVB-induced inflammatory. Journal of Agricultural and Food Chemistry. 2008;56(19):8969e8974. 60. Gorrini C, Harris IS, Mak TW. Modulation of oxidative stress as an anticancer strategy. Nature Reviews Drug Discovery. 2013. https://doi.org/10.1038/nrd4002. 61. Jang H, Ha US, Kim SJ, Yoon BI, Han DS, Yuk SM, Kim SW. Anthocyanin extracted from black soybean reduces prostate weight and promotes apoptosis in the prostatic hyperplasia-induced rat model. Journal of agricultural and food chemistry. 2010;58(24):12686e12691. https://doi.org/10.1021/jf102688g. 62. Wu HJ, Deng JC, Yang CQ, Zhang J, Zhang Q, Wang XC, Yang F, Yang WY, Liu J. Metabolite profiling of isoflavones and anthocyanins in black soybean [Glycine max (L.) Merr.] seeds by HPLC-MS and geographical differentiation analysis in Southwest China. Analytical Methods. 2017;9(5):792e802.   c S, Nedovic V. Micro63. Kalusevic A, Levic S, Calija B, Pantic M, Belovic M, Pavlovic V, Bugarski B, Milic J, Zili encapsulation of anthocyanin-rich black soybean coat extract by spray drying using maltodextrin, gum Arabic and skimmed milk powder. Journal of microencapsulation. 2017;34(5):475e487. https://doi.org/10.1080/ 02652048.2017.1354939. 64. Ryu D, Koh E. Application of response surface methodology to acidified water extraction of black soybeans for improving anthocyanin content, total phenols content and antioxidant activity. Food chemistry. 2018;261:260e266. https://doi.org/10.1016/j.foodchem.2018.04.061. 65. Xie Y, Zhu X, Li Y, Wang C. Analysis of the ph-dependent fe (iii) ion chelating activity of anthocyanin extracted from black soybean [glycine max (l.) merr.] coats. Journal of agricultural and food chemistry. 2018;66(5):1131e1139. https://doi.org/10.1021/acs.jafc.7b04719.

II. Role of Seeds in Nutrition and Antioxidant Activities

C H A P T E R

13

Fenugreek (Trigonella foenum) Seeds in Health and Nutrition Dilipkumar Pal, Souvik Mukherjee Department of Pharmaceutical Sciences, Guru Ghasidas Vishwavidyalaya (A Central University), Bilaspur, Chhattisgarh, India

Introduction Trigonella foenum-graecum L (fenugreek), commonly known as methi (in Hindi), has been used as a culinary spice and flavoring agent and as a medicinal plant from ancient times. Fenugreek is a leguminous, herbaceous, rain-fed crop included among the seed spices. It is about 30e60 cm tall. The leaflets are about 2e2.5 cm long and the flowers are 1e2 cm long, axillary, and sessile. It is cultivated throughout the country. Among the spices, fenugreek is used as an esoteric food to enhance the flavor and color of food; it is also used to modify the texture of food. Therapeutic utility indicates fenugreek as a medicinal plant.1 Medicinal plants are employed in pharmaceuticals, nutraceutical, cosmetics, food supplements, and so on. They are also used as traditional sources of medicine. Among the list of important medicinal plants may be found fenugreek. The seeds and plants are basically hot and dry; they are also suppurative, aperient, and diuretic. They have some useful aspects for dropsy, chronic cough, and enlargement of the liver and spleen.2 The leaves of fenugreek are beneficial for both internal and external swelling and burns and also applied to prevent the hair from falling out. The seeds are considered to be carminative, tonic, and aphrodisiac. Fenugreek is employed to ease childbirth; moreover, it increases the milk flow of mothers. Egyptian women take fenugreek for menstrual pain and tourists use it as a hilba tea for stomach problems. Not only that, the plant is recommended for use in dyspepsia for loss of appetite, for diarrhea of puerperal women, and for rheumatism.3 An infusion of seeds is given to smallpox patients as a cooling drink. Fenugreek seed contains various bioactive compounds such as flavonoids (quercetin, rutin, and vitexin), saponins (graecunin, fenugrin B, and fenugreekine), and amino acids (isoleucine, 4-hydroxyisoleucine, histidine, leucine, and lysine). As a medicinal plant it shows activity against allergies, appetite or loss of catarrh, bronchial problems, cholesterol, diabetic retinopathy, gas, gastric disorders, lung infections, excessive

Nuts and Seeds in Health and Disease Prevention, Second Edition https://doi.org/10.1016/B978-0-12-818553-7.00013-9

161

Copyright © 2020 Elsevier Inc. All rights reserved.

162

13. Fenugreek (Trigonella foenum) Seeds in Health and Nutrition

mucus, sore throat, abscesses, anemia, asthma, boils, body odor, bronchitis, cancer, swollen eyes, fever, gallbladder problems, heartburn, inflammation, sinus problems, ulcers, uterine problems, and so on. A study in India showed that fenugreek seed is used to reduce blood sugar and harmful fats. Not only therapeutic, fenugreek is used as a spice worldwide. The leaves are eaten as a green leafy vegetable in the diet. Fenugreek seeds are bitter and have been in use for over 2500 years. Fenugreek is a leguminous herb belonging to Fabaceae. It is cultivated throughout the world, especially in Asian and North African countries. According to the language, it has different names, such as Fenugrec (French), Methi (Hindi), Bockshorklee (German), Fienogreco (Italian), Pazhitnik (Russian), Alholva (Spanish), Koroha (Japanese), Hulba (Arabian), Halba (Malaya), and K’u-Tou (China). India is the major producer of fenugreek. Its main consumers are culinary and medicinal users. In the indigenous system, it is effective against anorexia, and as gastric stimulant.3e8

Scientific Classification Kingdom: Plantae Division: Magnoliophyta Class: Magnoliopsida Order: Fabales Family: Fabaceae

Morphology of Seed 1. 2. 3. 4.

Appearance: solid rhomboidal seeds, 3e5 mm long, 2 mm thick. Hard, pebble-like. Color: yellowish brown or light brown. Odor: spicy. Taste: bitter.9,10

Earlier Cultivation of Fenugreek Seed Fenugreek is believed to have been brought into cultivation within the nearest East. It is unclear whether a wild strain of the rosid dicot genus gave rise to domesticated fenugreek. Burned fenugreek seeds were recovered from Tell Halal, Iraq (carbon dated to 4000 BC), and Bronze Age levels of Lachish and desiccated seeds from the grave of a pharaoh of Egypt. Cato the Elder lists fenugreek as an herb and leguminous plant as a crops grown to feed cows. In an AD 1 recipe, the Romans added wine and fenugreek. In the first century in Galilee, it was grown as a staple food11 (Figs. 13.1e13.4).

Current Cultivation Fenugreek is an annual legume, diploid (2n ¼ 16) plant with no abnormality. Morphologically, it is an erect, aromatic annual closely resembling giant trefoil. The stem is cylindrical,

II. Role of Seeds in Nutrition and Antioxidant Activities

Current Cultivation

FIGURE 13.1 Fenugreek seeds.

FIGURE 13.2

Phytoconstituents of fenugreek seeds.

II. Role of Seeds in Nutrition and Antioxidant Activities

163

164

13. Fenugreek (Trigonella foenum) Seeds in Health and Nutrition

Moisture 9.0% Ash 3.0% Starch 6.0%

Total Fiber 48.0%

Lipids 8.0%

Fenugreek seed

Protein 76.0% Gum 20.0%

Neutral detergent fiber 28.0%

FIGURE 13.3 Other constituents of fenugreek seeds. By Inhibiting the Secretion of Gastric Acid

By Increasing the Mucus Production

By Altering the Urease Activity of H. Pylori

Antiulcer Activity of Trigonella foenumgraecum

By Increasing the Level of Glutathione

FIGURE 13.4

By Disruption of Membrane Proton Motive Force

By Interfering with the Synthesis of prostaglandin

Gastroprotective activity of fenugreek seeds.

30e60 cm long, and chromatic in color; its roots are huge, finger-like structures. Fenugreek has pinnate, trifoliate, long, pediculate compound leaves with toothed, lanceolate, stipules that are triangular and subdivided to simple leaflets. It blooms white to yellow-white axillary and sessile flowers that are hermaphroditic and pollinated by bugs.12

II. Role of Seeds in Nutrition and Antioxidant Activities

Therapeutic Potential of Fenugreekenugreek Seed

165

The flowers have five petals referred to as banner, wing, and keel. The ovary is deeply immature and aldohexose whereas the spore grains are oval to circular. Fenugreek flowers produce two to eight brown, 15-cm-long pods. Each pod contains 10e20 seeds, which are tiny (5 metric linear units long), hard, and smooth; they are an uninteresting yellow. Fenugreek needs 5e10 days for germination whereas the primary trifoliate leaf takes 5e8 days after germination.13 It is an aggressive plant that can grow on dry grasslands, cultivated or uncultivated lands, hillsides, and planes in addition to field edges. However, it needs a good quantity of daylight.14 Fenugreek desires 4e7 months to succeed to maturity. Flowering occurs near the summer solstice (June to August) and seeds ripen throughout late summer (August to September). It a drought-tolerant plant that grows well in tropical climates with a gentle winter and humid summer, but its leaf and flower development is temperature dependent.15

Phytochemical Constituents The main chemical components of Trigonella foenum-graceum are fiber, flavonoids, polysaccharides, saponins, fixed oils, and some identified alkaloids.16 Mature seeds mainly contain amino acids, fatty acids, vitamins, saponins, and a large quantity of folic acid (84 mg/100 g). They also contain triethylamine, neurin, trigonelline, choline, gentianine, carpine, betaine, isoleucine, 4-hydroxyisoleucine, histidine, leucine, lysine, L-tryptophan, argenine,17 gum, neutral detergent, fiber, quercetin, rutin, vitexin, isovitexin, graecunins, fenugrin B, fenugreekine, trigofoenosides A-G, yamogenin, diosgenin, smilagenin, sarsasapogenin, tigogenin, neotigogenin, gitogenin, neotigogenin, yuccagenin, saponaretin, coumarin, lipids, vitamins, 28% mucilage, 22% proteins, and 5% of a strong-smelling, bitter fixed oil.18

Therapeutic Potential of Fenugreekenugreek Seed Treatment of Diabetes In animal and human trials, fenugreek seeds were found to lower fasting liquid body substance aldohexose levels. Fenugreek is also used as antidiabetic remedy for I and II polygenic I and II disease.19 Saponins and diosgenin, two main constituents of fenugreek, are answerable for hypo-lipidemic and anti-diabetic action.20 Fenugreek is an antihyperglycemic herb in humans and laboratory animals.

In Cancer Therapy Fenugreek may be a healthful herb to aid cancer patients in therapy interventions. Fenugreek extract has a protective result by modifying cyclophosphamide-induced cell death and free radical-mediated lipid peroxidation within the albino mice model. The same result also indicates that fenugreek seed is also important for cancer treatment.21e25 Flavonoids and catechin were shown to be apoptotic in human malignant neoplastic disease cells. Diosgenin

II. Role of Seeds in Nutrition and Antioxidant Activities

166

13. Fenugreek (Trigonella foenum) Seeds in Health and Nutrition

in fenugreek prevents cell growth and induced cell death within the H-29 human carcinoma cell line. The herb has hepatoprotective properties, and a polyphenol extract acts as a protective agent against fermentation alcohol-induced abnormalities in the liver.26

Fenugreek As Antioxidant Fenugreek contains a property as an associate inhibitor owing to the presence of flavonoids and polyphenols. Fenugreek seeds are rich in polyphenol and have protective effects against H peroxide-induced oxidization by protecting erythrocytes from hemolysis and macromolecule peroxidation. An in vitro study reported that the fenugreek extract inhibit g-radiation iatrogenic strand break formation in inclusion body pBR322 deoxyribonucleic acid.27

Fenugreek Results in Cholesterol An abnormal deficiency of cholesterol within the blood is known as hypocholesterolemia. Fenugreek is also responsible for the fecal steroid and cholesteric excretion. It results from action between the bile acids and saponins inflicting the formation of micelles too large for the channel to soak up. Another effect is that the fiber-rich gum portion of the seed reduces the rate of the internal organ synthesis of cholesterol. Each mechanism contributes to lowering cholesterol.28

Anthelmintic Seeds of fenugreek have marked and potent anthelmintic activity. During this action, the alcoholic extracts have promising results. In comparison, a water extract showed less activity.

Fenugreek in Bactericide Activity The seed extracts are effective against Escherichia coli, Salmonella typhi, and Staphylococcus aureus. To form this binary compound, extract seeds are stewed in water. Fenugreek has bactericide activity. Synthetic a-glycosidase inhibitors such as acarbose have side effects on the abdomen, including abdominal distention, as a result of the excessive inhibition of duct gland enzymes that ends up in abnormal microorganism fermentation of undigested carbohydrates in the colon. Analysis of the use of antidiabetic drug plants that delicately inhibit duct gland enzymes is useful.29 The glycolytic activity of a-amylase could occur through the direct blockage of the active center at many allosteric sites of the catalyst as conjointly urged for different inhibitors. Fenugreek extract contains a-amylase repressing factors that most likely act with the active sites of the catalyst in a substrate-specific manner. For inhibiting the expansion of Pseudomonas spp., E. coli, enteric bacteria dysenteriae, and Salmonella typhosa, fenugreek is effective.

II. Role of Seeds in Nutrition and Antioxidant Activities

Therapeutic Potential of Fenugreekenugreek Seed

167

Fenugreek in Obesity Some research indicates that fenugreek extract supplementation reduces body and animal tissue weight. The probable mechanism of fenugreek decreasing overall body and adipose tissue weight is that it flushes carbohydrates from the body before they enter the bloodstream, resulting in weight loss. Also, fenugreek seeds contain a high proportion (40%) of soluble fiber.30 These fibers form a gelatin-like structure that may retard the digestion and absorption of food from the gut and build fullness within the abdomen, suppressing appetite and promoting weight loss.31

Fenugreek in Gastroprotection Fenugreek seeds are effective for peptic ulceration. The aqueous extract and a gel fraction isolated from the seeds of fenugreek showed significant lesion-protective effects. The cytoprotective result of the seeds occurs because of the antisecretory action and effects on mucosal glycoproteins. The increase in lipid peroxidation evoked by alcohol is also prevented by fenugreek seeds. It also enhances the inhibitor potential of the internal organ membrane and thus can lower membrane injury. Numerous research showed that the soluble gel fraction derived from the seeds was simpler than omeprazole in preventing lesion formation. These observations show that fenugreek seeds possess anti-ulcer potential.32

Fenugreek’s Influence on Digestion Spices consumed in the diet influence duct gland digestive enzymes. Fenugreek conspicuously increased duct gland enzyme activity in rats fed spicy diets for 8 weeks. The high fiber of fenugreek relieves constipation.33

Fenugreek in Inflammation A 100- and 200-mg/kg dose of fenugreek reduced carrageenan-induced paw swelling in rats. Fenugreek extract contains an alkaloid and it was reported to provide an antiinflammatory property by reducing formalin-induced swelling in rats and an antipyretic property by considerably reducing an physiological state evoked by brewer’s yeast. The anti-inflammatory property of fenugreek may be because of the presence of saponins and flavonoids.34

Fenugreek in Cardiovascular Disease Endothelial dysfunction is a devastating condition associated with numerous disorders such as hardening of the arteries, hypertension, diabetes mellitus, and so on. Oil obtained from fenugreek, together with different essential oils, was employed to lower pressure in spontaneously hypertensive rats. The binary compound and benzene extract of fenugreek indicated drug activity in a dose-dependent manner by increasing the amount of body waste and natriuretic activity by increasing the quantitative relation of levels of Naþ/Kþ ions in Wistar rats.35

II. Role of Seeds in Nutrition and Antioxidant Activities

168

13. Fenugreek (Trigonella foenum) Seeds in Health and Nutrition

Adverse Effects Fenugreek has some side effects. It may increase the risk for bleeding, reduce potassium levels in the blood, and cause loose stools in some women, as well as the possibility of facial swelling or breathing difficulties. It also produces uterine contractions and hypoglycemia in some mothers.36e38

Conclusion Fenugreek is used as a spice in the preparation of various dishes, as well as to cure many diseases. Trigonella foenum-graecum is established as a medicinal plant because of different properties such as anticancer, anti-inflammatory, antiseptic, aphrodisiac, astringent, bitter, demulcent, emollient, expectorant, anthelmintic, wound healing and gastroprotective. Moreover, it is a primary supplement used for type 2 diabetes or noninsulin-dependent diabetes mellitus. Fenugreek is a rich source of polysaccharide galactomannan and also a source of saponins such as diosgenin, yamogenin, gitogenin, tigogenin, and neotigogens. It also contains flavonoids, amino acid, alkaloids, and other bioactive constituents such as mucilage, and volatile oils. However, it has some side effects, too, because it may increase the risk for bleeding and may reduce potassium levels in the blood. It may cause numbness, facial swelling, breathing problem, and fainting. It may induce an allergic reaction, as well as dizziness, diarrhea, and gastric problems. Consumption of fenugreek has proved safe for human life and may easily be implemented for health benefits through its rich fiber content and other bioactive components. Fenugreek seed reduce low-density cholesterol and triacylglycerols and is also used to reduce blood sugar levels with its high concentration of phytochemicals.

References 1. Pal D. A review on Cyperus rotundus as a tremendous source of pharmacologically active herbal medicine. International Journal of Green Pharmacy. 2015;9(4). 2. Nayak AK, Pal D, Das S. Calcium pectinate-fenugreek seed mucilage mucoadhesive beads for controlled delivery of metformin HCl. Carbohydrate Polymers. 2013;96(1):349e357. 3. Nayak AK, Pal D, Santra K. Screening of polysaccharides from tamarind, fenugreek and jackfruit seeds as pharmaceutical excipients. International Journal of Biological Macromolecules. 2015;79:756e760. 4. Nayak AK, Pal D. Trigonella foenum-graecum L. seed mucilage-gellan mucoadhesive beads for controlled release of metformin HCl. Carbohydrate Polymers. 2014;107:31e40. 5. Nayak AK, Pal D, Santra K. Swelling and drug release behavior of metformin HCl-loaded tamarind seed polysaccharide-alginate beads. International Journal of Biological Macromolecules. 2016;82:1023e1027. 6. Nayak AK, Pal D, Santra K. Development of pectinate-ispagula mucilage mucoadhesive beads of metformin HCl by central composite design. International Journal of Biological Macromolecules. 2014;66:203e211. 7. Preedy VR, Watson RR, Patel VB, eds. Nuts and Seeds in Health and Disease Prevention. Academicpress; 2011. 8. Pal D, Nayak AK. Plant polysaccharides-blended ionotropically-gelled alginate multiple-unit systems for sustained drug release. Handbook of Composites From Renewable Materials. 2017;6:399e400. 9. Nayak AK, Pal D. Sterculia gum-based hydrogels for drug delivery applications. In: Polymeric Hydrogels as Smart Biomaterials. Cham: Springer; 2016:105e151.

II. Role of Seeds in Nutrition and Antioxidant Activities

References

169

10. Nayak AK, Hasnain MS, Pal D. Gelled microparticles/beads of sterculia gum and tamarind gum for sustained drug release. In: Polymer Gels. Singapore: Springer; 2018:361e414. 11. Nayak AK, Bera H, Hasnain MS, Pal D. Synthesis and characterization of graft copolymers of plant polysaccharides. In: Biopolymer Grafting. Elsevier; 2018:1e62. 12. Nayak AK, Pal D. Functionalization of tamarind gum for drug delivery. In: Functional Biopolymers. Cham: Springer; 2018:25e56. 13. Pal D, Nayak AK, Saha S. Interpenetrating polymer network hydrogels of chitosan: applications in controlling drug release. Cellulose-Based Superabsorbent Hydrogels. 2018:1e41. 14. Sharma RD, Raghuram TC, Rao NS. Effect of fenugreek seeds on blood glucose and serum lipids in type I diabetes. European Journal of Clinical Nutrition. 1990;44(4):301e306. 15. Smith M. Therapeutic applications of fenugreek. Alternative Medicine Review. 2003;8(1):20e27. 16. Altuntas E, Özgöz E, Taser ÖF. Some physical properties of fenugreek (Trigonella foenum-graceum L.) seeds. Journal of Food Engineering. 2005;71(1):37e43. 17. Dixit P, Ghaskadbi S, Mohan H, Devasagayam TP. Antioxidant properties of germinated fenugreek seeds. Phytotherapy Research. 2005;19(11):977e983. 18. Valette G, Sauvaire Y, Baccou JC, Ribes G. Hypocholesterolaemic effect of fenugreek seeds in dogs. Atherosclerosis. 1984;50(1):105e111. 19. Kaviarasan S, Naik GH, Gangabhagirathi R, Anuradha CV, Priyadarsini KI. In vitro studies on antiradical and antioxidant activities of fenugreek (Trigonella foenum graecum) seeds. Food Chemistry. 2007;103(1):31e37. 20. Raghuram TC, Sharma RD, Sivakumar B, Sahay BK. Effect of fenugreek seeds on intravenous glucose disposition in non-insulin dependent diabetic patients. Phytotherapy Research. 1994;8(2):83e86. 21. Sowmya P, Rajyalakshmi P. Hypocholesterolemic effect of germinated fenugreek seeds in human subjects. Plant Foods for Human Nutrition. 1999;53(4):359e365. 22. Petropoulos GA, ed. Fenugreek: The Genus Trigonella. CRC Press; 2003. 23. Petit PR, Sauvaire YD, Hillaire-Buys DM, et al. Steroid saponins from fenugreek seeds: extraction, purification, and pharmacological investigation on feeding behavior and plasma cholesterol. Steroids. 1995;60(10):674e680. 24. Ravikumar P, Anuradha CV. Effect of fenugreek seeds on blood lipid peroxidation and antioxidants in diabetic rats. Phytotherapy Research. 1999;13(3):197e201. 25. Bhanger MI, Bukhari SB, Memon S. Antioxidative activity of extracts from a Fenugreek seeds (Trigonella foenum-graecum). Pakistan Journal of Analytical & Environmental Chemistry. 2008;9(2):6. 26. Kaviarasan S, Vijayalakshmi K, Anuradha CV. Polyphenol-rich extract of fenugreek seeds protect erythrocytes from oxidative damage. Plant Foods for Human Nutrition. 2004;59(4):143e147. 27. Raju J, Gupta D, Rao AR, Yadava PK, Baquer NZ. Trigonella foenum graecum (fenugreek) seed powder improves glucose homeostasis in alloxan diabetic rat tissues by reversing the altered glycolytic, gluconeogenic and lipogenic enzymes. Molecular and Cellular Biochemistry. 2001;224(1e2):45e51. 28. Thirunavukkarasu V, Anuradha CV, Viswanathan P. Protective effect of fenugreek (Trigonella foenum graecum) seeds in experimental ethanol toxicity. Phytotherapy Research. 2003;17(7):737e743. 29. Sharma RD, Raghuram TC, Rao VD. Hypolipidaemic effect of fenugreek seeds. A clinical study. Phytotherapy Research. 1991;5(3):145e147. 30. Pal D, Panda C, Sinhababu S, Dutta A, Bhattacharya S. Evaluation of psychopharmacological effects of petroleum ether extract of Cuscuta reflexa Roxb. stem in mice. Acta Poloniae Pharmaceutica. 2003;60(6):481e486. 31. Gupta M, Mazumder UK, Pal D, Bhattacharya S, Chakrabarty S. Studies on brain biogenic amines in methanolic extract of Cuscuta reflexa Roxb. and Corchorus olitorius Linn. seed treated mice. Acta Poloniae Pharmaceutica. 2003;60(3):207e210. 32. Pal D, Sannigrahi S, Mazumder UK. Analgesic and anticonvulsant effects of saponin isolated from the leaves of Clerodendrum infortunatum Linn. in mice. Indian J Exp Biol. 2009. 33. Mazumder UK, Gupta M, Pal D, Bhattacharya SA. Chemical and toxicological evaluation of methanol extract of Cuscuta reflexa Roxb. stem and Corchorus olitorius Linn. seed on hematological parameters and hepatorenal functions in mice. Acta Poloniae Pharmaceutica. 2003;60(4):317e324. 34. Pal D, Mishra P, Sachan N, Ghosh AK. Biological activities and medicinal properties of Cajanus cajan (L) Millsp. Journal of Advanced Pharmaceutical Technology & Research. 2011;2(4):207. 35. Pal D, Balasaheb NS, Khatun S, Bandyopadhyay PK. CNS activities of the aqueous extract of Hydrilla verticillata in mice. Natural Product Sciences. 2006;12(1):44.

II. Role of Seeds in Nutrition and Antioxidant Activities

170

13. Fenugreek (Trigonella foenum) Seeds in Health and Nutrition

36. Srinivasan K. Fenugreek (Trigonella foenum-graecum): a review of health beneficial physiological effects. Food Reviews International. 2006;22(2):203e224. 37. Bahmani M, Shirzad H, Mirhosseini M, Mesripour A, Rafieian-Kopaei M. A review on ethnobotanical and therapeutic uses of fenugreek (Trigonella foenum-graceum L). Journal of Evidence-Based Complementary & Alternative Mdicine. 2016;21(1):53e62. 38. Ulbricht C, Basch E, Burke D, et al. Fenugreek (Trigonella foenum-graecum L. Leguminosae): an evidence-based systematic review by the natural standard research collaboration. Journal of Herbal Pharmacotherapy. 2008;7(3e4):143e177.

II. Role of Seeds in Nutrition and Antioxidant Activities

C H A P T E R

14

Tamarind (Tamarindus indica) Seeds in Health and Nutrition Dilipkumar Pal, Souvik Mukherjee Department of Pharmaceutical Sciences, Guru Ghasidas Vishwavidyalaya (A Central University), Bilaspur, Chhattisgarh, India List of Abbreviations DPPH 2,2-Diphenyl-1-picrylhydrazyl NO Nitrous oxide PG Prostaglandin WHO World Health Organization

Introduction The integral role of seeds in preagricultural diets is understandable given their high energy and nutrient density.1 Seeds are also particularly important in human nutrition because of their unique composition in bioactive compounds.2 Of note, in the last decade, a large body of scientific evidence has been built on the beneficial effects of increasing consumption of plant seeds3 and derived products on various health outcomes.4 Tamarind (Tamarindus indica L.) is a member of the dicotyledonous family Fabaceae (Leguminosae). It grows in more than 50 countries of the world. The major areas of production are in Asian countries such as India, Bangladesh, Sri Lanka, Thailand, and Indonesia and in the African and the American continents. The tamarind tree is a long-lived, large evergreen, or semi-green tree and grows wild, although cultivated to a limited extent. Tamarind is a multipurpose tree species; almost every part of it finds some use.5 The fruit contains about 55% pulp, 34% seed, and 11% shell and the fiber in a pod. The fruit is pendulous, and the pods are oblong or sausage shaped, curved or straight, with rounded ends. Pods contain 1e10 seeds, irregularly shaped, flattened, rhomboid, with the center of each flat side of the seed marked with a large center

Nuts and Seeds in Health and Disease Prevention, Second Edition https://doi.org/10.1016/B978-0-12-818553-7.00014-0

171

Copyright © 2020 Elsevier Inc. All rights reserved.

172

14. Tamarind (Tamarindus indica) Seeds in Health and Nutrition

depression.6 The seeds are very hard, shiny, reddish, or purplish brown. They are embedded in the pulp, lined with a tough parched like membrane, and joined to each other with tough fibers.7 Tamarind seed is a by-product of the tamarind pulp industry. India produces about 0.3 million tons of tamarind yearly, of which the seed constitutes about 30e34% of the whole fruit. In this chapter, we have discussed about the health and nutrition of tamarind seed. It is available all over the country.8 Tamarindus indica is having some reported activities like antidiabetic, hypolipidemic, hepatoprotective, and antimicrobial properties, antiproliferative properties, etc. This plant is consumed by rural people as vegetable.9

Botanical Description Tamarindus indica is of moderate to large in size, evergreen tree, up to 24 m in height, and 7 m in girth. The latest morphologic and molecular analyses and continued study will clarify the exact positioning of Tamarindus in relation to its putatively related genera. It is a large evergreen tree with an exceptionally beautiful spreading crown and is cultivated throughout almost the whole country, except in the Himalayas and western dry regions.10 Leaves are alternate, compound, with 10e18 pairs of opposite leaflets; leaflets are narrowly oblong, 12e32
3e11 mm, petiole and rachis finely haired, midrib and net veining more or less conspicuous on both surface.11 Flowers are attractive, pale yellow or pinkish, small, lax spikes about 2.5 cm in width. Flower buds are completely enclosed by two bracteoles, which fall very early; sepals 4, petals 5, the upper 3 are well developed and the lower 2 are minute.12 Fruit is a pod, indehiscent, subcylindrical, 10e18
4 cm, straight or curved, velvety, rustybrown; the shell of the pod is brittle and the seeds are embedded in a sticky edible pulp. Seeds are 3e10, approximately 1.6 cm long, and irregularly shaped, and testa is hard, shiny, and smooth13 (Figs. 14.1e14.4).

FIGURE 14.1

Tamarind seed.

II. Role of Seeds in Nutrition and Antioxidant Activities

173

Vernacular Names

Tyrosine 287 mg/gm. Threonine 200 mg/gm. Valine 306 mg/gm. Isoleucine 313 mg/gm. Glycine 331 mg/gm.

Histidine 143 mg/gm.

Serine 350 mg/gm. Leucine 531 mg/gm. Phenyl alanine 318 mg/gm.

Alanine 312 mg/gm. Arginine 450 mg/gm.

Glutamic 1056 mg/gm.

Aspartic 768 mg/gm. Lysine 475 mg/gm.

Cysteine 106 mg/gm.

Proline 287 mg/gm. Methionine 113 mg/gm.

FIGURE 14.2

Amino acid profile of tamarind seed.

O

O

OH Palmitic acid(14-20%)

OH Stearic acid(6-7%)

O O

OH

OH Arachidonic acid(2-4%)

Oleic acid(15-27%)

O OH Linoleic acid(36-49%)

FIGURE 14.3 Fatty acid profile of tamarind seed.

Vernacular Names Assam: Teteli Bengal: Ambli, tentul, tinturi, nuli English: Tamarind tree Gujarat: Ambli Hindi: Imli, amli

II. Role of Seeds in Nutrition and Antioxidant Activities

174

14. Tamarind (Tamarindus indica) Seeds in Health and Nutrition

OH

O OH

HO HO

Taxifolin

Lupanone

OH

OH OH

O

O

O HO

HO

O Apigenin

O

OH

HO

OH

O

HO Eriodicytol OH

OH

OH OH

OH

O

OH

HO

O

HO

Naringenin

O

OH OH OH OH

O HO

FIGURE 14.4

Procyanidin

Phytochemicals of tamarind seed.

Malayalam: Amlam Oriya: Tentuli Punjab: Imli Tamil: Ambilam, amilam Telugu: Amlika, chinta, sinja, sinta Urdu: Imli Nepal: Titri14

Taxonomical Classification Kingdom: Plantae Unranked: Angiosperms Unranked: Eudicots Unranked: Roside Order: Fabales Family: Fabaceae Subfamily: Detarioideae Genus: Tamarindus Species: indica15

Historical Cultivation and Usage Tamarindus indica is probably indigenous to tropical Africa but has been cultivated for so long on the Indian subcontinent that it is sometimes reported to be indigenous there, where it

II. Role of Seeds in Nutrition and Antioxidant Activities

Present-Day Cultivation and Usage

175

is known as imli in Hindi and Urdu. It grows wild in Africa in locales as diverse as Sudan, Cameroon, Nigeria, Zambia, and Tanzania. In Arabia, it is found growing wild in Oman, especially Dhofar, where it grows on the sea-facing slopes of mountains. It reached South Asia likely through human transportation and cultivation several thousand years BC.16 It is widely distributed throughout the tropical belt, from Africa to South Asia, northern Australia, and throughout Oceania, Southeast Asia, Taiwan, and China. In the 16th century, it was introduced to Mexico, and to a lesser degree to South America, by Spanish and Portuguese colonists, to the degree that it became a staple ingredient in the region’s cuisine.17 It grows well in both semiarid and humid monsoon climates and can grow on a wide range of soil types. It is a tree of the tropics; it can tolerate temperatures up to 47 C but is very sensitive to frost. It is mainly grown in areas with 500e1500 mm rain/year but tolerates down to 350 mm if irrigated at the time of establishment. In the wet tropics with over 4000 mm rain, flowering and fruit setting is significantly reduced and in India it is not grown in areas receiving more than 1900 mm rain/year. Regardless of total annual rainfall, it produces more fruit when subjected to a fairly long dry period. Throughout Southeast Asia, the fruit of the tamarind is used as a poultice applied to foreheads of fever sufferers.18 The fruit exhibits laxative effects because of its high quantities of malic acid, tartaric acid, and potassium tartrate. Its use for the relief of constipation has been documented throughout the world. At homes and temples, especially in Buddhist Asian countries, the fruit pulp is used to polish brass shrine statues and lamps and copper, brass, and bronze utensils.19 The copper alone or in brass reacts with moist carbon dioxide to gain a green coat of copper carbonate. Tamarind contains tartaric acid, a weak acid that can remove the coat of copper carbonate. Hence, tarnished copper utensils are cleaned with tamarind or lime, another acidic fruit. Throughout South Asia and the tropical world, tamarind trees are used as ornamental, garden, and cash crop plantings. Commonly used as a bonsai species in many Asian countries, it is also grown as an indoor bonsai in temperate parts of the world.20

Present-Day Cultivation and Usage Seeds can be scarified or briefly boiled to enhance germination. They retain their germination capability for several months if kept dry. The tamarind has long been naturalized in Indonesia, Malaysia, Sri Lanka, Philippines, the Caribbean, and the Pacific Islands. Thailand has the largest plantations of the ASEAN nations, followed by Indonesia, Myanmar, and the Philippines.21 In parts of Southeast Asia, tamarind is called asam. It is cultivated all over India, especially in Maharashtra, Chhattisgarh, Karnataka, Telangana, Andhra Pradesh, and Tamil Nadu. Extensive tamarind orchards in India produce 275,500 tons (250,000 MT) annually.22 In the United States, it is a large-scale crop introduced for commercial use, second in net production quantity only to India, mainly in the southern states, notably south Florida (due to tropical and semitropical climates), and as a shade tree, along roadsides, in dooryards and in parks. As a traditional food plant in Africa, tamarind has the potential to improve nutrition, boost food security, foster rural development, and support sustainable land care. In Madagascar, its fruit and leaves are a well-known favorite of the ring-tailed lemur, providing as much as 50% of their food resources during the year if available.23

II. Role of Seeds in Nutrition and Antioxidant Activities

176

14. Tamarind (Tamarindus indica) Seeds in Health and Nutrition

Nutritional Characterization of Tamarind Seeds Whole tamarind seed and seed kernel are rich sources of protein. Fat or oil comprise of 4.5e16.2% of total composition. Crude fiber percentage is very less in whole seed while the seed coat is rich in fiber (20%) and tannins (20%). Remaining 50e57% is carbohydrate. Amino acid profile from the chemical composition, it can be seen that tamarind seeds are a good source of protein24. Amino acid profiles of tamarind reveal that the proteins contain fairly balanced essential amino acid levels. Except a few, all the amino acids such as isoleucine, leucine, lysine, methionine, phenylalanine, and valine contents are considerably high in seed. In terms of protein content and WHO standards, tamarind seeds score well for three of the eight essential amino acids. However, for each of the eight essential amino acids, score is close to or above the 100% mark, except tryptophan.25 Fatty acid profile: Tamarind seeds give amber-colored oil, free of smell and sweet in taste, which resembles linseed oil. Tamarind oil has iodine value below 100 mg/g, which places it in the nondrying oil group. The low levels of percent of free fatty acids in tamarind oil indicate that the oil may be good edible oil with an extended shelf-life without spoilage via oxidative rancidity.26 Tamarind seed is a good source of fatty acids. It contained between 1 and 2 mg/g dry weight linoleic acid. Tamarind seeds have a higher percentage of unsaturated (55.6%) fatty acids than saturated (44.4%) fatty acids. Linoleic acid, present in tamarind seed oil, is one of the most important polyunsaturated acids in human food because of its association in the reduction or prevention of cardiovascular diseases.27 Mineral composition: Tamarind seeds appear to be a good source of different mineral elements such as calcium, phosphorus, magnesium, and potassium. Of all the minerals studied, potassium is the element in highest concentration, with the values for the trace mineral copper also relatively high. The high concentration of potassium is nutritionally significant by playing a key role in neuromuscular function (Table 14.1).28

TABLE 14.1

Minerals Composition of Tamarind Seed

Minerals (mg/100 g)

Seeds

Calcium

9.3e786.0

Phosphorus

68.4e165.0

Magnesium

17.5e118.3

Potassium

272.8e610.0

Copper

1.6e19.0

Iron

6.5

Zinc

2.8

Manganese

0.9

II. Role of Seeds in Nutrition and Antioxidant Activities

Biological Activities

177

Phytochemicals Composition of Tamarind Seed Phytochemical investigation carried out on T. indica seed revealed the presence of many active constituents, such as phenolic compounds, cardiac glycosides, L-()-malic acid, tartaric acid, the mucilage and pectin, arabinose, xylose, galactose, glucose, and uronic acid.28 The ethanolic extract of T. indica seed showed presence of fatty acids and various essential elements such as arsenic, calcium, cadmium, copper, iron, sodium, manganese, magnesium, potassium, phosphorus, lead, and zinc. The seed polysaccharides are found with a main chain consisting of b-1,4-connected glucose molecules together with xylose (a-1,6) and galactose; total protein; lipids with fatty oils; and some keto acids. Two triterpenes such as lupanone and lupeol are present in tamarind seed. Proanthocyanidins in various forms, such as apigenin, catechin, procyanidin B2, epicatechin, procyanidin dimer, procyanidin trimer, along with taxifolin, eriodictyol, naringenin, of total phenols, are preset in tamarind seed.29

Biological Activities Antioxidant properties: The seed and pericarp of T. indica contain phenolic antioxidant compound. Soxhlet methanolic extract of T. indica may be an important source of cancer chemopreventive. All extracts of T. indica exhibited good antioxidant activity (64.5e71.7%) against the linoleic acid emulsion system and the values were lower and higher than the synthetic antioxidant, butylated hydroxyanisole, and ascorbic acid. Thai tamarind seed coat using solvent extraction with ethanol was found to be the most active in terms of peroxide value. Ethanolic extract of fruit pulp of T. indica showed significant antioxidant and hypolipidemic activity in hypercholesterolemia hamsters and antioxidant activity of ethanolic extract of seed coat of T. indica by DPPH free radical scavenging method using ascorbic acid as a standard. This activity of T. indica extract may be attributed to its free radical scavenging ability. Ethanol extract prepared from the seed coat of T. indica exhibited antioxidant activity as measured by the thiocyanate and thiobarbituric method. Ethyl acetate extracts prepared from the seed coat also had strong antioxidant activity. T. indica seed coat, a by-product of the tamarind gum industry, could be used as a safe and low-cost source of antioxidant, although other herbals may be more effective.30 Antidiabetic activity: An aqueous extract from T. indica seeds had a potent antidiabetogenic activity in streptozotocin-induced diabetic male rats. The aqueous extract of T. indica seeds was given to mild diabetic and severe diabetic rats, and hyperglycemia was significantly reduced, measured by fasting blood glucose levels. Similarly, hyperlipidemia was found to be reduced, measured by different contents of cholesterol.31 This rat model may shed some light on the basis of ancient herbal therapy in India.32 Antivenom activities: In the Indian traditional medicine, various plants have been used widely as a remedy against snake bite. In a study, the effect of T. indica seed extract was investigated for its pharmacologic and enzymatic activity. Tamarind seed extract inhibited phospholipase A, protease, hyaluronidase, L-amino acid oxidase, and 50 -nucleotidase enzyme activities of venom in a dose-dependent manner. The extract of T. indica neutralized the

II. Role of Seeds in Nutrition and Antioxidant Activities

178

14. Tamarind (Tamarindus indica) Seeds in Health and Nutrition

degradation of the b-chain of the human fibrinogen and the indirect hemolysis caused by the venom.33 The extract prolonged the clotting time moderately, and myotoxic effects, such as edema and hemorrhage, induced by the venom were neutralized significantly when different doses of the extract were administered, and hence T. indica extract is an alternative for the serum therapy.34 Anticancer activity: Ameliorative effect of T. indica seed extract has been shown in chemical-induced acute nephrotoxicity and renal cell carcinoma. This effect can be explained by antioxidant effect. However, oxidative damage is strongly associated with cancer; polyphenol compounds (2-hydroxydihydroxyacetophenone, methyl 3,4-dihydroxybenzoate, 3,4dihydroxyphenylacetate, epicatechin, tannin, anthocyanidins, and oligomeric proanthocyanidin) in T. indica seed extract have antioxidant enzyme induction properties and cancer-related signal pathway blockage effect.35 Anti-inflammatory effect: Anti-inflammatory effects of leaves, seeds, and other parts of T. indica have been shown, but this effect is not as strong as acetylsalicylic acid. Analgesic effect also has been shown in mechanic, chemical, and thermal pain models. It stabilizes the red blood cell membrane and prevents the damage. In addition, it shows anti-inflammatory effect and inhibits the release of PG and NO (diclofenac-like effect).36 When lysosomal damage occurs, phospholipase A2 appears and stimulates the production of inflammatory agents via hydrolysis of phospholipids. Prevention of cell damage causes cytoplasm content preservation and decreases inflammatory response. Polyphenols and flavonoid content of T. indica associated with anti-inflammatory and antinociceptive effects.37 Activity on eye: T. indica seed polysaccharide is used in eye drops to increase its effective time period because of its mucoadhesive properties. The mixture with hyaluronic acid is used in xerophthalmia, and with the aid of Timolol, it decreases intraocular pressure. Studies reported the effect of it in corneal wound healing especially after surgical procedures.38 Activity on nerve repair: It has been shown that xyloglucan obtained from T. indica seed serves suitable media for degenerated nerves and aids nerve regeneration.39

Tamarind Seed Polysaccharide: a Promising Natural Excipient for Pharmaceuticals The natural polymers always have exceptional properties which make them distinct from the synthetic polymers, and tamarind seed polysaccharide (TSP) is one such example which shows more valuable properties making it a useful excipient for a wide range of applications. TSP is a natural polysaccharide obtained from the seeds of Tamarindus indica, recently gaining a wide potential in the field of pharmaceutical and cosmetic industries. Its isolation and characterization involve simple techniques resulting in cost-effective yield in its production. TSP shows uniqueness in its high drug holding capacity, high swelling index, and high thermal stability, especially necessary for various novel drug delivery systems. It also plays the role of stabilizer, thickener, binder, release retardant, modifier, suspending agent, viscosity enhancer, emulsifying agent, as a carrier for novel drug delivery systems in oral, buccal, colon, ocular systems, nanofabrication, and wound dressing and is also becoming an important part of food, cosmetics, confectionery, and bakery. Various studies and experiments have

II. Role of Seeds in Nutrition and Antioxidant Activities

Tamarind Seed Polysaccharide: a Promising Natural Excipient for Pharmaceuticals

179

been carried out to prove its multifunctional potentiality, from which it can be concluded that TSP can be a promising natural polysaccharide having enormous applications. TSP is a multifunctional polymer, which plays the role of stabilizer, thickener, binder, release retardant, modifier, suspending agent, viscosity enhancer, emulsifying agent, as a carrier for novel drug delivery systems for oral, buccal, colon, ocular systems, nanofabrication, wound dressing, food, cosmetics, confectionery, bakery, etc. Attempts made to study the use of TSP as a suspending agent in the formulation of nimesulide suspension showed that TSP acts as a stable suspending agent that reduced the rate of settling and permitted in the easy redispersion of any settled particulate matter. The comparative studies on castor oil emulsions with TSP and gum acacia have shown that 2% w/v of TSP was more effective than using 10% w/v of gum acacia. TSP was also compared with other natural suspending agents using a pharmaceutical formulation of paracetamol suspension. The newly developed pH-sensitive composite beads of diclofenac sodium by ionotropic gelation method using TSPealginate was suitable for the controlled delivery for a prolonged period. The sodium alginate TSP ratio and cross-linker (CaCl2) concentration influenced the drug encapsulation efficiency and drug release. The swelling and degradation of the developed beads were affected by different pH values of the test medium. TSP was used in combination with carbopol, HPMC K4M, and CMC for the fabrication of buccal mucoadhesive tablets of nifedipine for avoiding first-pass metabolism and prolonging the duration of action. Using fresh goat buccal mucosa as the model tissue, the modified in vitro assembly was measured to evaluate its bioadhesive strength. The best mucoadhesive performance and in vitro drug release profile was exhibited by the tablet containing carbopol and TSP in the ratio of 1:1. This formulation was considered to be more useable as it showed less erosion, faster hydration rate, and optimum pH of the surrounding medium. Tamarind gum was also employed as a novel bioadhesive material in the delivery of pilocarpine by ophthalmic in situ gelling systems. The combination of alginate, tamarind gum, and chitosan was identified to the most successful means for sustained delivery of 80% drug for 12 hour. In vivo mitotic study and ocular irritation studies showed significant long-lasting decrease in pupil diameter of rabbits and well-tolerated nonirritating effect with tamarind gum-based formulation. The mucoadhesive property of TSP was successfully employed for ocular administration of hydrophilic and hydrophobic antibiotics such as gentamicin, ofloxacin, etc. The TSP viscosified solutions of the drug instilled into rabbit showed that the aqueous humor and corneal concentration of the dose was remarkably higher than the drug alone. The absorption and drug elimination was prolonged by TSP; the concentration of drug in cornea exceeded the minimum inhibitory concentration studied from the cases of keratoconjunctivitis. The mucoadhesive polymer extracted from tamarind seeds proved as an effective candidate for ocular delivery of antibiotics, rufloxacin, and ofloxacin, for the treatment of bacterial keratitis experimentally induced by Pseudomonas aeruginosa and Staphylococcus aureus in rabbits. The polysaccharide significantly increased the intra-aqueous penetration of the drugs in both infected and uninfected eyes. The effect of TSP delivery of rufloxacin for substantial reduction of bacteria in the cornea was at a higher rate. The natural polymers always have unique properties which make them distinct from the synthetic polymers, and TSP is also not an exemption from this, as it shows a wide range of properties making it a potent polymer not only in the field of food industries but also in the field of pharmaceutical industries. TSP is insoluble in organic solvents such as ethanol,

II. Role of Seeds in Nutrition and Antioxidant Activities

180

14. Tamarind (Tamarindus indica) Seeds in Health and Nutrition

methanol, acetone, and ether and in cold water, but it gets dissolved completely in hot water at temperatures above 85 C, yielding a highly viscous colloidal solution or a mucilaginous gel showing typical non-Newtonian rheologic behavior and pseudoplastic properties. TSP possesses various properties such as high viscosity, adhesivity, noncarcinogenicity, broad pH tolerance, and biocompatibility. It is also found to be a potential emulsifier, nontoxic, and nonirritant with hemostatic activity. Other distinguishable properties of TSP have also been identified, which include the high drug holding capacity, high swelling index, and high thermal stability, making it a suitable excipient for drug delivery system. Apart from this, it is an excellent viscosity enhancer showing mucomimetic, mucoadhesive, and bioadhesive activities. Recent studies on TSP for various drug formulations revealed other unique properties with wide applications in the pharmaceutical area, which include its potent antidiabetic activity that reduces blood sugar level. In addition to this, the property of forming films with high tensile strength and flexibility makes it a good excipient for ocular preparations. This film is transparent, nonhygroscopic, and nonsticky and retains its form even on rough handling, and the Ferning pattern is similar to natural tear film. The powder and the solution properties of TSP, such as density, flow, compressibility, melting point, moisture content, water retention, swelling index, pH, and surface tension, are evidently proved to be satisfactory for its applications in various fields, especially for pharmaceutical formulation development.

Possible Adverse Effects and Reaction(s) No health risk or side effects exist if there is proper administration of tamarind seed. Therapeutic dosages have not been recorded. Tamarind might lower blood sugar levels. There is a concern that it might interfere with blood sugar control. If you have diabetes and use tamarind, monitor your blood sugar levels closely. Dosing adjustments for diabetes medications might be needed. There is a concern that it might interfere with blood sugar control during and after surgery. Stop using tamarind at least 2 weeks before a scheduled surgery. Taking tamarind with aspirin might increase how much aspirin the body absorbs. This could increase the amount of aspirin in the body and might increase the chance of aspirin side effects. Taking tamarind with ibuprofen might increase how much ibuprofen the body absorbs. This could increase the amount of ibuprofen in the body and might increase the chance of ibuprofen side effects.40

Summary of Key Point(s) * * * *

T. indica is a cheap and easily available plant. It is a rich source of essential amino acids, phytochemicals, and vitamins. In traditional medicine, it has so many well-known health benefits. With the aid of modern techniques, it could be that there is a need of further investigation about this plant and its potential antioxidant and anti-inflammatory properties that can help in many of the diseases.

II. Role of Seeds in Nutrition and Antioxidant Activities

References

181

References 1. Bemiller JN, Whistler RL, Barkalow DG, Chen C-C. Aloe, chia, flaxseed, okra, psyllium seed, quince seed, and tamarind gums. Industrial Gums. 1993:227e256 (Elsevier). 2. Bewley JD, Black M. Seeds: physiology of development and germination. NY: Plenum Press; 1994:1e33. 3. Bhadoriya SS, Ganeshpurkar A, Narwaria J, Rai G, Jain AP. Tamarindus indica: extent of explored potential. Pharmacognosy Reviews. 2011;5(9):73. 4. Bhavin P, Piyush P, Ashok B, Shrawaree H, Swati M, Ganesh C. Evaluation of tamarind seed polysaccharide (TSP) as a mucoadhesive and sustained release component of nifedipine buccoadhesive tablet & comparison with HPMC and Na CMC. International Journal of PharmTech Research. 2009;1(3):404e410. 5. bin Mohamad MY, Akram HB, Bero DN, Rahman MT. Tamarind seed extract enhances epidermal wound healing. International Journal of Biology. 2012;4(1):81. 6. Bruneton J. Pharmacognosy, Phytochemistry, Medicinal Plants. Lavoisier publishing; 1995. 7. Chungsamarnyart N, Jansawan W. Effect of Tamarindus indicus L. against the Boophilus microplus. Kasetsart Journal (Natural Sciences). 2001;35:34e39. 8. Cui SW, Ikeda S, Eskin MN. Seed polysaccharide gums. Functional Food Carbohydrates. 2006:139e177 (CRC Press). 9. El-Siddig K. Tamarind: Tamarindus Indica L. Vol. 1. 2006 (Crops for the Future). 10. Evans WC. Trease and Evans Pharmacognosy. International Edition. E-Book: Elsevier Health Sciences; 2009. 11. Heinrich M, Barnes J, Gibbons S, Williamson EM. Fundamentals of Pharmacognosy and Phytotherapy E-Book. Elsevier Health Sciences; 2012. 12. Hemshekhar M, Kemparaju K, Girish KS. Tamarind (Tamarindus indica) seeds: an overview on remedial qualities. Nuts and Seeds in Health and Disease Prevention. 2011:1107e1114 (Elsevier). 13. Huang L-q. Molecular Pharmacognosy. Springer Science & Business Media; 2012. 14. Karan SK, Mishra SK, Pal D, Mondal A. Isolation of b-sitosterol and evaluation of antidiabetic activity of Aristolochia indica in alloxan-induced diabetic mice with a reference to in-vitro antioxidant activity. Journal of Medicinal Plants Research. 2012;6(7):1219e1223. 15. Karan SK, Mondal A, Mishra SK, Pal D, Rout KK. Antidiabetic effect of Streblus asper in streptozotocin-induced diabetic rats. Pharmaceutical Biology. 2013;51(3):369e375. 16. Minu V, Harsh V, Ravikant T, Paridhi J, Noopur S. Medicinal plants of Chhattisgarh with anti-snake venom property. International Journal of Current Pharmaceutical Review and Research. 2012;3(2):1e10. 17. Mukherjee S, Nag M, Roy D, Paul RK, Mir SA. Extraction and chemical tests on Cicer arietinum seed collected from North Bengal region of West Bengal, India. PharmaTutor. 2018;6(10):1e4. 18. Nayak AK, Hasnain MS, Pal D. Gelled microparticles/beads of sterculia gum and tamarind gum for sustained drug release. In: Polymer Gels. Springer; 2018:361e414. 19. Nayak AK, Malakar J, Pal D, Hasnain MS, Beg S. Soluble starch-blended Ca2þ-Zn2þ-alginate composites-based microparticles of aceclofenac: formulation development and in vitro characterization. Future Journal of Pharmaceutical Sciences. 2018;4(1):63e70. 20. Nayak AK, Pal D. Tamarind seed polysaccharide: an emerging excipient for pharmaceutical use. Indian Journal of Pharmaceutical Education and Research. 2017;51:S136eS146. 21. Nayak AK, Pal D. Functionalization of tamarind gum for drug delivery. In: Functional Biopolymers. Springer; 2018:25e56. 22. Nayak AK, Pal D, Das S. Calcium pectinate-fenugreek seed mucilage mucoadhesive beads for controlled delivery of metformin HCl. Carbohydrate Polymers. 2013;96(1):349e357. 23. Nayak AK, Pal D, Pany DR, Mohanty B. Evaluation of Spinacia oleracea L. leaves mucilage as an innovative suspending agent. Journal of Advanced Pharmaceutical Technology and Research. 2010;1(3):338. 24. Nayak AK, Pal D, Santra K. Screening of polysaccharides from tamarind, fenugreek and jackfruit seeds as pharmaceutical excipients. International Journal of Biological Macromolecules. 2015;79:756e760. 25. Nayak AK, Pal D, Santra K. Swelling and drug release behavior of metformin HCl-loaded tamarind seed polysaccharide-alginate beads. International Journal of Biological Macromolecules. 2016;82:1023e1027. 26. Nimse SB, Pal D, Mazumder A, Mazumder R. Synthesis of cinnamanilide derivatives and their antioxidant and antimicrobial activity. Journal of Chemistry. 2015;2015. 27. Pal D. Sunflower (Helianthus annuus L.) Seeds in health and nutrition. Nuts and Seeds in Health and Disease Prevention. 2011:1097e1105 (Elsevier).

II. Role of Seeds in Nutrition and Antioxidant Activities

182

14. Tamarind (Tamarindus indica) Seeds in Health and Nutrition

28. Pal D, Mitra S. A preliminary study on the in vitro antioxidant activity of the stems of Opuntia vulgaris. Journal of Advanced Pharmaceutical Technology and Research. 2010;1(2):268. 29. Patil DN, Datta M, Chaudhary A, Tomar S, Kumar Sharma A, Kumar P. Isolation, purification, crystallization and preliminary crystallographic studies of chitinase from tamarind (Tamarindus indica) seeds. Acta Crystallographica Section F. 2009;65(4):343e345. 30. Pazhanivelan S, Amanullah MM, Vaiyapuri K, Athyamoorthi K, Radhamani S. Influence of planting techniques and amendments on the performance of tamarind (Tamarindus indicus) and changes in soil properties in rainfed alkali soil. Research Journal of Agriculture and Biological Sciences. 2008;4(4):285e288. 31. Rani P, Pal D, Hegde RR, Hashim SR. Anticancer, anti-inflammatory, and analgesic activities of synthesized 2(substituted phenoxy) acetamide derivatives. BioMed Research International. 2014;2014. 32. Rao YS, Mathew KM. Tamarind. Handbook of Herbs and Spices. 2012:512e533. Elsevier. 33. Sachan N, Chandra P, Pal D. Assessment of gastroprotective potential of Delonix regia (Boj Ex Hook) Raf against ethanol and cold restrain stress-induced ulcer in rats. Tropical Journal of Pharmaceutical Research. 2015;14(6):1063e1070. 34. Sachan N, Shrivastav A, Pal D. Evaluation of antidepressant activity of ethanolic extract of Abies webbiana and Berberis aristata in laboratory animals. Journal of Drug Delivery and Therapeutics. 2019;9(1):244e247. 35. Saha S, Pal D, Kumar S. Hydroxyacetamide Derivatives: Cytotoxicity, Genotoxicity, Antioxidative and Metal Chelating Studies. 2017. 36. Samajdar S, Mukherjee S, Das PP. Seed germination inhibitors: molecular and phytochemical aspects. International Journal of Applied Pharmaceutical Sciences and Research. 2018;3(2):12e23. 37. Samuelsson G, Bohlin L. Drugs of Natural Origin: A Treatise of Pharmacognosy. CRC Press Inc; 2017. 38. Sannigrahi S, Mazumder UK, Pal D, Mishra SL, Maity S. Flavonoids of Enhydra Fluctuans exhibits analgesic and anti-inflammatory activity in different animal models. Pakistan Journal of Pharmaceutical Sciences. 2011;24(3). 39. Ushanandini S, Nagaraju S, Harish Kumar K, et al. The anti-snake venom properties of Tamarindus indica (leguminosae) seed extract. Phytotherapy Research. 2006;20(10):851e858. 40. Youngken HW. A Text Book of Pharmacognosy: P. Blakiston’s Son & Company; 1921.

II. Role of Seeds in Nutrition and Antioxidant Activities

C H A P T E R

15

Sesame Seed in Controlling Human Health and Nutrition 1

Dilipkumar Pal1, Phool Chandra2, Neetu Sachan2

Department of Pharmaceutical Sciences, Guru Ghasidas Vishwavidyalaya (A Central University), Bilaspur, Chhattisgarh, India; 2School of Pharmaceutical Sciences, IFTM University, Lodhipur Rajput, Moradabad, Uttar Pradesh, India List of Abbreviations % Percentage AchE Acetylcholinesterase cm Centimeter CNS Central nervous system DNA Deoxyribonucleic acid DNZ Donepezil hydrochloride monohydrate DOCA Deoxycorticosterone acetate ELISA Enzyme-linked immunosorbent assay eNOS Endothelial nitric oxide synthase IFN Interferon IgE Immunoglobulin E IL Interleukin JNK c-Jun N-terminal kinase kDa Kilodaltons LDL Low-density lipoprotein LDLR Low-density lipoprotein receptor LOX Lipoxygenase LPS Lipopolysaccharide LTP Long-term potentiation m Meter MAPK Mitogen-activated protein kinase mL Milliliter mRNA Messenger ribonucleic acid NADPH Nicotinamide adenine dinucleotide phosphate hydrogen NF-k Nuclear factor kappa NOARG NG-nitro-L-arginine NOS Nitric oxide synthase Nrf2 Nuclear factor erythroid 2-related factor 2 Pb Lead

Nuts and Seeds in Health and Disease Prevention, Second Edition https://doi.org/10.1016/B978-0-12-818553-7.00015-2

183

Copyright © 2020 Elsevier Inc. All rights reserved.

184

15. Sesame Seed in Controlling Human Health and Nutrition

PI3K Phosphatidylinositol 3-kinase PPARg Peroxisome proliferator-activated receptor gamma ROS Reactive oxygen species SSHD Sesame seed high dose SSLD Sesame seed low dose TNF Tumor necrosis factor UDP Uridine diphosphate UV Ultraviolet XO Xanthine oxidase mg Microgram mM Micrometer

Introduction Sesamum indicum L. (family: Pedaliaceae) is thought as the oldest oilseed and is used by human beings; it is commonly known as beniseed, sesame, sesamum, gingelly, simsim, and til.1 Sesame, an important oilseed crop of the world, is produced in India. Sesame seed provides nutritious protein, highly stable oil, and meals, is used in confectionery foods, and possesses varieties of therapeutic medicinal properties.2 Sesame was widely dispersed by people both westward and eastward, reaching China and Japan, which themselves converted secondary distribution hubs.1 In old literature, the sesame seed was referred to as the “queen of oilseeds.”3 Further, in traditional Indian medicine, Ayurveda, sesame oil has been used as the basal oil for human body massage since 700-1, 100 B.C.4

Plant Profile Vernacular Names Sesamum indicum (til) is recognized by altered names in different areas of the globe along with regions of India. Commonly, it is known as gingelly, sesame, and til.5 Country

Names

Afghanistan

Konjele, kunjit, til

Arabian countries

Djyldiylan, duban, samusam, simsim

Africa

Juljulyn

Persia

Kumjad

Japan

Goma

China

Hu ma zhi or hei zhi ma or hu-ma

II. Role of Seeds in Nutrition and Antioxidant Activities

Plant Profile

Country

185

Names

In India Hindi and Bengali

Til

Malayalam

Achelu, ellu, woellu, yallu, chitelu, chitrallu, ellu, karellu, scetel

Sanskrit

Til

Tamil

Ellu, hllu, nuvvulu, yellu-chedi

Telugu

Tilmi, nuvvu, nuvvulu, polla-nuvullu

Geographical Distribution Commonly, the sesame seeds of white color are found in the Americas, Europe, the Indian subcontinent, and West Asia. The countries with higher production of sesame seeds include Japan, China, the United States, Canada, the Netherlands, Turkey, France, Tanzania, Myanmar, India, Sudan, Ethiopia, and Myanmar, etc., and are presented in Fig. 15.1.

Morphology Cultivated sesame is normally erect, branched annual, occasionally perennial, 0.5e2 m in height, with a well-developed root system, and multiflowered, whose fruit is a capsule containing a number of small oleaginous seeds. It has a tap root system with profuse lateral branches. Long-season types, occasionally treated as perennials, have an extensive and penetrating root system, and short-season types have less extensive and shallower roots.

FIGURE 15.1 Distribution of sesame seed around the word.

II. Role of Seeds in Nutrition and Antioxidant Activities

186

15. Sesame Seed in Controlling Human Health and Nutrition

The stem is erect, normally square in section with definite longitudinal furrows, although rectangular and abnormally wide, and flat shapes occur. It can be smooth, slightly hairy, or very hairy, and these characteristics are used to differentiate the types. The stem is light green to purple, branching angular, and straight with an average height of 1e1.5 m and sometimes up to 3 m. Lower leaves are broad, sometimes lobal, and margins often prominently toothed with the teeth diverted outwards. Intermediate leaves are entire, lanceolate, and sometimes slightly separated. The upper leaves are more narrow and lanceolate. Leaf size varies from 3 to 17.5 cm in length and 1 to 7 cm in width, with a petiole of 1 to 5 cm in length. The surface of leaves is generally glabrous but in some types may be pubescent. Generally of a dull, darkish-green, leaves can be much lighter with occasionally a yellowish tint or bluish when leaves are very hairy. Flowers arise in the axils of leaves and on the upper portion of the stem and branches, and the node number on the main shoot at which the first flower is produced is a varietal characteristic and highly heritable. Flowers occur singly on the lower leaf axils with multiple flowers on the upper stem or branches. Calyx lobes are short, velvety, narrow, acuminate, and united at the base. The five lobes are of variable sizes, the lower one being the longest and upper one the shortest. The flower is zygomorphic with a slightly bilabiate tubular corolla of five lobes. The upper lip of the corolla is entire and the lower divided into three, of which the central division is the longest. Stamens are attached to the tube of the corolla. Of the five stamens, four are functional and the fifth is either sterile or completely lacking. The four greenish white functional stamens are arranged in pairs, one pair being shorter than the other. There are two anther cells, opening longitudinally, connected usually gland-tipped.1 The ovary is superior, usually two-celled, and cells are often completely or partially divided by false septa. The style is terminal, filiform, and simple. The stigma is usually two-lobed and hairy (Fig. 15.2).

(A)

(B)

Leaves FIGURE 15.2

(C)

Seed capsule

Flower

Different parts of the Sesamum indicum L.

II. Role of Seeds in Nutrition and Antioxidant Activities

Chemical Composition

187

Chemical Composition Sesame seed has high food value due to its high content of oil and protein. The composition is markedly influenced by genetic and environmental factors.6e8 The proximate composition of whole sesame seeds is given in Table 15.1.

Lipids Content Sesame seeds contain more oil than many other oilseeds. A wide range of the oil content, from 37 to 63%, has been reported in sesame seeds.6,9 The glycerides of sesame oil, therefore, are mostly triunsaturated (58 mol%) and diunsaturated (36 mol%) with small quantities (6 mol%) of TABLE 15.1

Chemical Composition of Raw Sesame Seeds.

Component

RS

Dry matter (%)

95.29B
0.19

Oila

52.24C
0.34

Proteina

25.77C
1.02

Total fibersa

19.33A
1.97

Insoluble fibersa

13.96A
1.62

Soluble fibersa

5.37A
0.28

Asha

4.68A
0.20

Calciuma

1.03A
0.04

Potassiumb

525.9B
17.90

Magnesiumb

349.9A
39.32

Phosphorusb

516C
26.89

Sodiumb

15.28A
1.63

Ironb

11.39A
0.27

Copperb

2.15A
0.06

Zincb

8.87B
0.26

Manganeseb

3.46A
0.43

Soluble sugarsa

2.48C
0.09

Starcha

0.88B
0.01

Polyphenolsb

87.77C
3.15

All the given values are means of tree determinations
standard deviation. Mean in a row followed by the same uppercase letters is not significantly different (P < 0.05). RS: raw sesame seed. A P < 0.05 B P < 0.01 C P < 0.001 a In % dry matter. b In mg/100 g dry matter With Copyright permission Elleuch M., et al. Quality characteristics of sesame seeds and by-products. Food Chemistry. 2007;103(2):641e650.

II. Role of Seeds in Nutrition and Antioxidant Activities

188

15. Sesame Seed in Controlling Human Health and Nutrition

monounsaturated glycerides. The unsaponifiable matters in sesame oil include sterols (b-sitosterol, compesterol, and stigmasterol), triterpenes (triterpene alcohols that include at least six compounds, of which three were identified viz., cycloartanol, 24-methylene cycloartanol, and amyrin), pigments, tocopherols, and two compounds that are not found in any other oil, namely sesamin and sesamolin. Sesame seed contains high levels of sesamin and g-tocopherol compared with a- and d-tocopherol, and their concentration is influenced by genetic, environmental, and geographical factors.10 Sesamin is one of the major precursors of mammalian lignans in sesame seed (Sesamum indicum) as observed in vitro and in rats.11

Fatty Acid Composition Sesame oil contains about 80% unsaturated fatty acids.6 The saturated fatty acids account for less than 20% of the total fatty acids. About 44 and 42% of linoleic and oleic acids and 13% saturated fatty acids are found in sesame oil.12 A group of scientist8,13 reported the fatty acid composition of sesame seed and sesame oil that is presented in Tables 15.2e15.4.

Endogenous Antioxidants Sesame oil is thought to be most resistant to oxidative rancidity among the commonly used vegetable oils.14 It exhibits greater resistance to autoxidation due to presence of tocopherols (vitamin E). The unsaponifiable matter responsible for the high stability includes sesamol and phytosterol that are generally not found in other oils. Sesamolin, upon hydrolysis, yields sesamol. Sesame oil contains 0.5e1.0% sesamin15 and 0.3e0.5% sesamolin14 with only traces of free sesamol.16,17 Sesamol is released from sesamolin by hydrogenation, acid or acid bleaching earth, or other conditions of processing and storage.16,18 Endogenous antioxidants and stability of sesame oil affect processing and storage.19 Structures of natural antioxidants found in sesame oil are depicted in Fig. 15.3. TABLE 15.2

Physicochemical Characterization of Raw Sesame Seeds.

Component

RS 41.46C
1.52

a

Oil (%)

Free fatty acids (as oleic acid %)

2.37A
0.41

Iodine value (g of I2/100 g of oil)

99.08A
0.73 0.18A
0.04

Chlorophyll (mg/kg) Viscosity (mPa s)

12.93A
0.05

Refractive index (at 40  C)

1.470A
0.001

Polyphenols (as mg gallic acid/kg of oil)

23.06A
4.41

Sesamol (mg/kg of oil) Induction period (h)

8.11A
0.70 28.23C
0.73

All the given values are means of tree determinations
standard deviation. Mean in a row followed by the same uppercase letters is not significantly different (P > .05). RS, raw sesame seed. a In % dry matter A P< 0.05 C P < 0.001 With Copyright permission Elleuch M., et al. Quality characteristics of sesame seeds and by-products. Food Chemistry. 2007;103(2):641e650.

II. Role of Seeds in Nutrition and Antioxidant Activities

Chemical Composition

TABLE 15.3

189

Fatty Acid Composition (%) of Raw Sesame Seeds.

Component

RS

Palmitic C16:0

11.18A
0.76

Palmitoleic C16:1

0.21A
0.005

Stearic C18:0

6.40A
0.17

Oleic C18:1(n-9) cis-Vaccenic C18:1(n-11)

44.06A
0.58 0.97A
0.002

Linoleic C18:2

35.56A
0.07

Linolenic C18:3

0.50A
0.03

Arachidic C20:0

0.70A
0.04

Eicosenoic C20:1

0.18A
0.01

Lignoceric C24:0

0.20A
0.005

SAFA

18.49
0.98

MUFA

45.44
0.60

PUFA

36.06
0.11

All the given values are means of tree determinations
standard deviation. Mean in a row followed by the same uppercase letters is not significantly different (P > .05). SAFA, saturated fatty acids, MUFA, monounsaturated fatty acids, PUFA, polyunsaturated fatty acid, RS, raw sesame seed. A P < 0.001 With Copyright permission Elleuch M., et al. Quality characteristics of sesame seeds and by-products. Food Chemistry. 2007;103(2):641e650.

TABLE 15.4 Fatty Acid

Fatty Acids Composition of Sesame Oil. % of Total Fatty Acids

Palmitic

7.8e9.1

Stearic

3.6e4.7

Arachidic

0.4e1.1

Oleic

45.3e49.4

Linoleic

37.7e41.2

Proteins Sesame seed contains 17e32% protein with an average of about 25%.6 Protein content tends to decline with increase in productivity level.1 The proteins in the seed are located mostly in the outer layers of the seed. Based on their solubility, sesame proteins have been classified and depicted in Table 15.3 as albumin (8.6%), globulins (67.3%), prolamin (1.3%), and glutelin (6.9%) fractions.20 Essential amino acid composition of sesame meal proteins1 is presented in Table 15.5 and Fig. 15.4.

II. Role of Seeds in Nutrition and Antioxidant Activities

190

15. Sesame Seed in Controlling Human Health and Nutrition

Vitamin E

Sesamol

Sesamolin

Sesamin

Sesangolin FIGURE 15.3 TABLE 15.5

Natural antioxidants found in oil from sesame.

Type of Proteins Present in the Sesame Seed.

Type of Protein

%

Albumin

8.6

Globulins

67.3

a-Globulin

60e70

b-Globulin

25

Prolamin

1.3

Glutelin

6.9

II. Role of Seeds in Nutrition and Antioxidant Activities

Chemical Composition

Arginine

Histidine

Isoleucine

Leucine

Lysine

Methionine

Cystine

Phenylalanine

Tyrosine

Threonine

Tryptophan

Valine FIGURE 15.4

Essential amino acid composition of sesame meal proteins.

II. Role of Seeds in Nutrition and Antioxidant Activities

191

192

15. Sesame Seed in Controlling Human Health and Nutrition

Carbohydrates The carbohydrate content of sesame seeds is higher than that of soybean seeds and is comparable to that of groundnut seeds. Sesame seeds contain 14e25% carbohydrates. The seeds contain about 5% sugars, most of which are of reducing type. The sugars present in sesame seeds and defatted flour are given in Fig. 15.5. Sesame seeds are reported to contain 3e6% crude fiber.1,21 The crude fiber is present mostly in husk or seed coat. A team of scientist reported 0.58e2.34% and 0.71e2.59% hemicellulose A and B, respectively, in defatted flour. Hemicellulose A was found to contain galacturonic acid and glucose in the ratio of 1:12.9, while hemicellulose B contained galacturonic acid, glucose, arabinose, and xylose in the ratio of 1:3.8:3.8:3.1.1,22

D-glucose

D-fructose

D-galactose

D-fructose

Raffinose

Stachyose

Sesamose

Planteose

FIGURE 15.5

Sugars present in sesame seeds and defatted flour.

II. Role of Seeds in Nutrition and Antioxidant Activities

Pharmacological Applications

193

Minerals Sesame seed is a good source of certain minerals, particularly calcium, phosphorus, iron sodium, and potassium, and selenium.23e25

Pharmacological Applications Wound Healing Activity The seed of S. indicum was evaluated for wound healing activity on excision, incision burn, and dead space modes in experimental rats. They have selected sesame seed low dose (SSLD 2.5%) and sesame seed high dose (SSHD 5%) for topical application, whereas for oral administration, they selected the crushed seed suspension (250 mg/kg and 500 mg/kg). They found that SSLD and SSHD showed significant (P < 0.001) decrease in wound contraction in excision and incision wound model. Also, they reported that breaking strength of a 10-day-old wound was increased. In burn wound model, application of SSLD and SSHD shortened the period of epithelization significantly (P < 0.001) and produced a significant decrease (P < 0.001) in wound contraction. In dead space model, breaking strength of a 10-day-old granulation tissue was significantly promoted by both SSLD and SSHD. The dry tissue weight and hydroxyproline content were significantly (P < 0.001) increased.26

Hepatoprotective Activity Scientist investigated the protective effects of Sesamum indicum seed extract (100e300 mg/ mL) against oxidative stress induced by vanadium on isolated rat hepatocytes. They resulted quite similar to a-tocopherol (100 mM), and different concentrations of extract (100e300 mg/ mL) protected the isolated hepatocyte against all oxidative stress/cytotoxicity markers induced by vanadium including cell lysis, ROS generation, mitochondrial membrane potential decrease, and lysosomal membrane damage. Besides, vanadium-induced mitochondrial/ lysosomal toxic interaction and vanadium reductive activation mediated by glutathione in vanadium toxicity were significantly (P < 0.05) ameliorated by Sesamum indicum extracts. On the basis of findings, they suggested a hepatoprotective role for extracts against liver injury resulted from vanadium toxicity.27

Effects of Pinoresinol on Memory Synaptic Plasticity Pinoresinol is a lignan found in sesame seed and olive oil. Previous studies have reported that pinoresinol inhibits a-glucosidase and, therefore, act as a hypoglycemic agent.28 Synaptic plasticity is the capability of synapses to strengthen or deteriorate over time in response to changes in their activity. Functional synaptic plasticity is regulated by and regulates various neurological phenomena including learning and memory.29,30 Yu et al. (2019) explore the utility of pinoresinol obtained from the sesame seed in the memory impairment in dementia utilizing cholinergic model. They found that in comparison with scopolamine alone, a statistically significant increase was observed between the groups receiving 25 mg/kg of

II. Role of Seeds in Nutrition and Antioxidant Activities

194

15. Sesame Seed in Controlling Human Health and Nutrition

FIGURE 15.6 The effect of pinoresinol on AChE activity. (A) The effect of pinoresinol on ChE from mouse hippocampus. (B) The effect of pinoresinol on AChE from mouse hippocampus. To block BuChE, ZINC12613047 (50 mM) was added into brain homogenates. (C) The effect of pinoresinol on AChE from human erythrocyte. Data represent mean
SEM (n ¼ 3/group). With Copyright permission Yu J, et al. The effects of pinoresinol on cholinergic dysfunction-induced memory impairments and synaptic plasticity in mice. Food and Chemical Toxicology. 2019;125: 376e382.

pinoresinol and donepezil hydrochloride monohydrate-positive group after administration of scopolamine (P < 0.05). Moreover, pinoresinol (50 mM) expedited induction of hippocampal long-term potentiation (LTP), a cellular model of learning and memory. It blocked acetylcholinesterase (AchE), an acetylcholine-degrading enzyme, activity in a concentration-dependent manner (Fig. 15.6). Also, it facilitated calcium influx into neuro2a cell. Finally, a team of scientist29 suggested that pinoresinol improves memory impairment and facilitates hippocampal LTP induction and might be connected to the consequence of pinoresinol on AChE and calcium influx.

Effect of Sesame Oil and Sesamol on Heavy Metal Toxicity Heavy metals are harmful to the body if they are not eliminated from the body and their deposition takes place in the soft tissue. It is documented that sesame and sesame oil are powerful agents to nullify the hepatic and renal toxicity without adverse effects. Ironpersuaded lipid peroxidation is potentially inhibited by the sesame oil and sesamol by inhibiting the hydroxyl radical generation, nitric oxide, superoxide anion, and xanthine oxidase. Also, sesame oil, a significant inhibitor of proinflammatory mediators, thereby weakens the lead-induced hepatic destruction by inhibiting IL-1b levels, TNF-a, and nitric oxide. It is always found that metal-chelating therapy includes adverse reactions; therefore, treating toxicity with sesame oil and sesamol could be a better alternative.31

Autoimmune Encephalomyelitis Autoimmune encephalomyelitis is an inflammatory demyelinating disease of the CNS that has related aspects to multiple sclerosis.32 Ghazavi and Mosayebi in 2012 investigated the effectiveness of treatment with sesame oil on the experimentally induced autoimmune encephalomyelitis. Autoimmune encephalomyelitis was induced in 8-week-old C57BL/6 mice with 0.1 mL of an emulsion containing 200 mg of the encephalitogenic peptide of

II. Role of Seeds in Nutrition and Antioxidant Activities

Pharmacological Applications

195

MOG35-55 5 (MEVGWYRSPFSRVVHLYRNGK; Diapharm Ltd., Russia). Sesame oil (4 mL/ kg/day as oral gavage) obtained from sesame seeds was started on day 3 before the immunization and continued to day 25 postimmunization. ELISA method was utilized for determination of IFN-gamma and IL-10 production from cultured spleen supernatants. The outcomes showed that daily oral gavage of sesame oil significantly reduced the clinical symptoms of autoimmune encephalomyelitis in C57BL/6 mice. In addition, sesame oil-treated mice showed a significantly delayed disease onset. Mononuclear cells isolated from the spleen of sesame oil-treated mice showed a significant decrease in the production of IFN-gamma compared with control mice. IL-10 production was also enhanced in splenic mononuclear cells in sesame oil-treated mice. The ratio of IFN-gamma to IL-10 in sesame oil-treated autoimmune encephalomyelitis-induced mice was significantly less than control group of mice. They reported that sesame oil therapy protected mice from developing autoimmune encephalomyelitis by reducing IFN-gamma secretion. Therefore, sesame oil treatment may be effective in multiple sclerosis patients by immunomodulating the Th1 immune response.33e35

Atherosclerosis Diet has intense effects on the progress of atherosclerosis. Fatty acid composition, antioxidants, and other components such as lignans have major effects on the atherosclerotic process. Sesame oil has both mono- and polyunsaturated fatty acid constituents in equal proportions. In addition, it also has high levels of numerous antioxidants and inducers of peroxisome proliferator-activated receptor. In this study, male low-density lipoprotein (LDL) receptor (LDLR)/mice was fed atherogenic diet or atherogenic diet reformulated with the same level of sesame oil (sesame oil diet). Plasma lipids and atherosclerotic lesions were quantified after 3 months of feeding. Sesame oil-containing diet significantly reduced the atherosclerotic lesion formation and plasma cholesterol, triglyceride, and LDL cholesterol levels in LDLR/mice. Finally, they suggested on the basis of finding that sesame oil could inhibit atherosclerosis lesion formation effectively, due to synergistic response of fatty acid and nonsaponifiable components.36

Effect on Serum Cholesterol and in Hypercholesterolemia During a study, all the volunteers taking part are advised to take normal regular food before 2 week from the commencement of the experiment. During experimental period of 4 week, they have to take 40 g roasted sesame followed by normal regular diet for 4 weeks. At the 0th, 4th, and 8th week, weight and fasting blood was examined. They found a significant decrease in the level of total cholesterol and LDL after consuming the roasted sesame. Time for hemolysis of erythrocyte and lag phase of LDL oxidation were increased significantly after adding sesame in the diet. Also, sesame diet decreased the levels of thiobarbituric acid reactive substances in LDLs. On return to normal regular diet, the valuable effects of sesame were disappeared. Finally, they found that sesame exerts valuable effects on serum lipid profile and improves antioxidant ability in hypercholesterolemic patients.37

II. Role of Seeds in Nutrition and Antioxidant Activities

196

15. Sesame Seed in Controlling Human Health and Nutrition

Effects on Plasma Tocopherol Levels The tocopherols, the chief vitamers of vitamin E, are supposed to play a part in the prophylaxis of human aging-linked diseases such as cancer and heart disease, yet little is identified regarding determinants of their plasma concentrations. Confirmation from animal studies proposes that the nutritive source of g-tocopherol can significantly change plasma levels of this tocopherol as well as its functional vitamin E bustle. For studying whether plasma levels of tocopherols in humans are correspondingly altered, a study in which subjects (n ¼ 9) were fed muffins comprising equivalent quantities of g-tocopherol from sesame seeds was carry out. They found that ingestion of 5 mg of g-tocopherol per day over a 3 day period from sesame seeds significantly elevated serum g-tocopherol levels (19.1% increase) and decreased plasma b-tocopherol (34% decrease). No significant alterations in baseline or post-intervention plasma levels of carotenoids, cholesterol, and triglycerides were perceived for any of the intrusion groups. All subjects ingesting sesame seedcontaining muffins had noticeable levels of the sesame lignan (sesamolin) in their plasma. Consumption of adequate amounts of sesame seeds seems to significantly raise plasma g-tocopherol and modify plasma tocopherol proportions in humans and is stable with the effects of dietary sesame seeds experiential in rats leading to raised plasma g tocopherol and elevated vitamin E bioactivity.38

Antioxidant and Anticancer Effect of Sesamol Sesamol has been displayed earlier to produce anti-ageing activity and reactive oxygenmediated antimutagenic activity. Furthermore, it has been established to exert chemopreventive effect as maximum of the antioxidants act by their property to auto-oxidize and the antioxidant activity would rest on the concentration of the chemical used and the free radical source. For evaluations, six dilutions in concentration range of 5e1000 nmoles of sesamol were utilized. Further, the antioxidant activity was compared with ascorbic acid (a standard water-soluble antioxidant). They reported comprehensively (both in terms of the number of doses and also a variety of systems being utilized) on the antioxidant activity of sesamol. Finally, they found that sesamol is an efficient scavenger of the entire range of reactive oxygen species in several test systems directing toward the potential of sesamol to be developed as a possible therapeutic agent.39 Sesamol (from S. indicum) is a dietary compound, which is soluble in aqueous and lipid phases. Its free radical scavenging reactions have been studied using a nanosecond pulse radiolysis technique. Sesamol efficiently scavenges hydroxyl, one-electron oxidizing, organo-haloperoxyl, lipid peroxyl, and tryptophanyl radicals. Cyclic voltammetry was utilized for the evaluation of its antioxidant activity. In biochemical studies, it has been found to inhibit DNA cleavage, lipid peroxidation, and hydroxyl radical-induced deoxyribose degradation.40 Sesamol produces radioprotective effect on g-radiation-induced DNA damage, lipid peroxidation, and antioxidants levels in cultured human lymphocytes.41 Sesamol inhibits UVB-induced ROS generation and subsequent oxidative damage in cultured human skin dermal fibroblasts.42 Sesamol also inhibits doxorubicin-induced oxidative impairment and harmfulness on H9c2 cardiomyoblasts.43

II. Role of Seeds in Nutrition and Antioxidant Activities

Pharmacological Applications

197

The beneficial potential of sesamol was examined intensively, and there is persuasive evidence that sesamol acts as a metabolic controller that possesses antioxidant, antihepatotoxic, antimutagenic, anti-aging, anti-inflammatory, and chemopreventive properties. Numerous studies have described that sesamol exerts potent anticancer effects. The protective role that sesamol exhibits against oxidative stress over its radical scavenging ability and lipid peroxidation lowering potential is analyzed. The capability of sesamol to control apoptosis and numerous stages of the cell cycle is also drawn. Moreover, the signaling paths that sesamol appears to target to implement its antioxidant, anti-inflammatory, and pro-apoptotic/antiproliferative roles were presented. The signaling paths that sesamol targets include the p53, JNK, MAPK, TNFa, PI3K/AKT, NF-kB, caspase-3, Nrf2, eNOS, PPARg, and LOX pathways. The mechanisms of action that sesamol performs to bring its anticancer effects are delineated. Therefore, there is ample evidence signifying that sesamol owns potent anticancer properties in vitro and in vivo.44 Other detailed study on the cancer is depicted in Table 15.6.

Anticancer Effect of Sesamin It is found that a lignan of sesame, that is, sesamin, downregulates cyclin D1 protein expression during studies in human tumor cells.45

Antihypertensive Effect of Sesamin Sesamin, a major lignan in sesame seeds and oil, has been known to lower blood pressure in several types of experimental hypertensive animals. A recent study demonstrated that sesamin metabolites had in vitro radical scavenging activities. They determined the antioxidative effect of sesamin metabolites that modulates the vascular tone and contributes to the in vivo antihypertensive effect of sesamin. Scientist utilized four demethylated sesamin metabolites: SC-1m (piperitol), SC-1 (demethylpiperitol), SC-2m [(1R,2S,5R,6S)-6-(4-hydroxy-3methoxyphenyl)-2-(3,4-dihydroxyphenyl)-3,7-dioxabicyclo[3,3,0]octane], and SC-2 [(1R,2S,5R, 6S)-2,6-bis(3,4-dihydroxyphenyl)-3,7-dioxabicyclo-[3,3,0]octane]. SC-1, SC-2m, and SC-2, but not SC-1m, exhibited potent radical scavenging activities against the xanthine/xanthine oxidase-induced superoxide production. On the other hand, SC-1m, SC-1, and SC-2m produced endothelium-dependent vasorelaxation in phenylephrineprecontracted rat aortic rings, whereas SC-2 had no effect. The SC-1m- and SC-1-induced vasorelaxations were markedly attenuated by pretreatment with a nitric oxide synthase (NOS) inhibitor, NG-nitro-L-arginine (NOARG), or a soluble guanylate cyclase inhibitor, 1H1,2,4oxadiazolo-[4,3-a]quinoxalin-1-one. Neither SC-1m nor SC-1 changed the expression level of endothelial NOS protein in aortic tissues. The antihypertensive effects of sesamin feeding were not observed in chronically NOARG-treated rats or in deoxycorticosterone acetate salt-treated endothelial NOS-deficient mice. These findings suggest that the enhancement of endothelium-dependent vasorelaxation induced by sesamin metabolites is one of the important mechanisms of the in vivo antihypertensive effect of sesamin46. Sesamin shields deoxycorticosterone acetate salt-induced cardiovascular hypertrophy and hypertension.47 Sesamin defends salt-loaded hypertensive than unloaded stroke-prone spontaneously hypertensive rats.48 Sesamin obtained from sesamin seed also protects against twokidney, one-clip renal hypertension and cardiovascular hypertrophy.49 A group of scientist

II. Role of Seeds in Nutrition and Antioxidant Activities

198 TABLE 15.6

15. Sesame Seed in Controlling Human Health and Nutrition

A Detailed Summary of the Main Findings Related to the Anticancer Activities of Sesamol, the Dosages, the Dosage Regimens, the Experimental Models, and Whether the Reported Effects are Dose-dependent And/Or Time-dependent.

Experimental Model

Effect

Dose

Dosedependent And/Or TimeDosage Regimen dependent

References

e

62

1e10 mg/mL Incubation (30 min)

Dosedependent

41

pBR322

50 mM e62 mM

Incubation (1 h)

Dosedependent

63

Sesamol provides protection against g-irradiation by increasing antioxidant profile and reducing the expression of apoptotic proteins

C57BL/6 mice (males)

100 mg/kg, i.p.

Intraperitoneal administration (30 min prior to g-irradiation)

e

64

Sesamol inhibits lipid peroxidation and plasmid DNA degradation

Incubation (1 h) Brain (albino Charles 0.1e6010 6 mol dm3 Foster rats)

Dosedependent

40

Sesamol protects against C57BL/6 mice g-irradiation-induced DNA (males) damage by reducing TCA and micronuclei in bone marrow cells

10 and 20 mg/kg, i.p.

Sesamol protects against Human blood g-irradiation by increasing lymphocytes activities of antioxidant enzymes Sesamol serves a protective role against g-irradiation by scavenging radicals

Intraperitoneal administration (30 min prior to g-irradiation)

Sesamol attenuates DNA Peripheral blood damage and radical formation in lymphocytes (swiss g-irradiated mice albino mice)

100 mg/kg Intraperitoneal 10e50 mg/mL administration Incubation (60 min)

Dosedependent

65

Sesamol inhibits UVB-induced cytotoxicity, lipid peroxidation, reactive oxygen species production, and DNA oxidative damage

8e160 mM

Incubation (30 min)

Dosedependent

42

Sesamol offers a photoprotective Human blood role against UVB-induced lipid lymphocytes peroxidation in lymphocytes

1e10 mg/mL Incubation (30 min)

Dosedependent

66

Sesamol has antimutagenic SK-MEL-2 properties and exerts antioxidant effect by scavenging radicals and lowering lipid peroxidation

100 mL and 1 Incubation (48 h) Dosee4 mM dependent

67

Sesamol inhibits lipid peroxidation and attenuates

10 mg/kg, s.c.

HDFa

Liver (male Wistar rats)

Subcutaneous administration

II. Role of Seeds in Nutrition and Antioxidant Activities

e

68

199

Pharmacological Applications

TABLE 15.6

A Detailed Summary of the Main Findings Related to the Anticancer Activities of Sesamol, the Dosages, the Dosage Regimens, the Experimental Models, and Whether the Reported Effects are Dose-dependent And/Or Time-dependent.dcont’d

Experimental Model

Effect

Dose

Dosedependent And/Or TimeDosage Regimen dependent

levels of hydroxyl radical and superoxide anion Sesamol decreases levels of Plasma, liver, kidney, 50e200 mg/ lipid peroxidation and increases and heart (male kg, p.o. levels of enzymatic antioxidants albino Wistar rats)

(6 42 h, every 6 h) Postoral administration (6 weeks)

Sesamol reverses the effects of diabetes on levels of inflammatory cytokines, antioxidant enzymes, and reactive oxygen species

Plasma (male Wistar 2e8 mg/kg, mice) p.o.

Sesamol competitively inhibits lipoxygenase, resulting in the production of reactive oxygen species and free radicals

Soy LOX-1 (enzyme model)

References

e

69

Oral administration (4 weeks)

Dosedependent

70

20e60 mM

Incubation (3 min)

Dosedependent

71

Antimutagenicity of sesamol is TA100 and TA102 directly linked to its antioxidant properties in radical scavenging in TA100 and TA102 strains

1e10 mmol/ plate

Incubation (20 min)

Dosedependent

72

Sesamol has the highest rate constant for the disappearance of DPPH compared to other natural compounds in sesame cake extract

40e320 mM

e

e

73

Intraperitoneal administration (5 days)

e

74

Freshly prepared solution of DPPH*, methanol, and sesamol

Lens and capsule 50 mg/kg, Sesamol protects against i.p. cataract-induced oxidative stress homogenate by reducing MDA level and TOS (sprague-dawley rats) and elevating GSH level and TAS Sesamol inhibits LOX-1-mediated Plasma (syrian hamsters) HAECs pro-apoptotic p38 MAPK/ caspase pathway and induces anti-apoptotic PI3K/AKT/eNOS signaling

50 or 100 mg/kg, p.o. 0.3e3 mM

Oral administration (16 weeks) Incubation (24 h)

Dosedependent and Timedependent

75

Sesamol protects against doxinduced oxidative damage by increasing levels of antioxidant enzymes and attenuating apoptotic cells

12.5e50 mM

Incubation (30 min)

Dosedependent

43

H9C2

(Continued)

II. Role of Seeds in Nutrition and Antioxidant Activities

200 TABLE 15.6

15. Sesame Seed in Controlling Human Health and Nutrition

A Detailed Summary of the Main Findings Related to the Anticancer Activities of Sesamol, the Dosages, the Dosage Regimens, the Experimental Models, and Whether the Reported Effects are Dose-dependent And/Or Time-dependent.dcont’d

Experimental Model

Effect

Dose

Dosedependent And/Or TimeDosage Regimen dependent Intraperitoneal administration (7 days)

e

References

Sesamol protects the Heart (albino Wistar myocardium through its rats) antioxidant and lipid-stabilizing activity from doxorubicininduced cardiotoxicity

50 mg/kg, i.p.

76

Sesamol inhibits amylase, lipase, AR42J and lipid peroxidation in cerulein-induced pancreatic acinar cancer cells and increases cell survival by resisting oxidative stress

10e1000 mg/ Incubation (24 h) DosemL dependent

77

Sesamol attenuates ferric nitrilotriacetate-induced oxidative stress due to its antioxidant activity

Kidney and serum (male Wistar rats)

2e8 mg/kg, p.o.

Incubation (30 min)

Dosedependent

78

Sesamol inhibits hydroxyl radical SPF BALB/c mice generation and superoxide anion levels in Fe-NTA intoxication

10e100 mg/ kg, s.c.

Subcutaneous administration (3 h)

e

57

Sesamol inhibits lipid peroxidation and scavenges radicals due to its antioxidant property

Brain and liver (rats) 5 e e1000 nmoles

Dosedependent

39

Sesamol protects against oxidative damage induced by cyclophosphamide through its antioxidant and antiinflammatory properties

Serum, liver, kidney, 50 mg/kg, and spleen (Wistar p.o. rats)

Oral administration (6 days)

Dosedependent

79

Sesamol inhibits lipid peroxidation and reduces hydroxyl radical and hydrogen peroxide levels

Liver (albino Wistar rats)

1e30 mg/kg, Intraperitoneal i.p. administration (24 h)

Dosedependent

80

Solid-lipid nanoparticles are used for effective application of sesamol for treating skin cancer

HL-60 and Molt-4

10e100 mg

Incubation (1 3 h) Dosedependent

81

Sesamol decreases proinflammatory cytokines’ levels

Serum (male ICR mice) RAW 264.7

10 mg/kg, s.c. 100e300 mM

Subcutaneous Doseadministration dependent Incubation (24 h)

82

II. Role of Seeds in Nutrition and Antioxidant Activities

201

Pharmacological Applications

TABLE 15.6

A Detailed Summary of the Main Findings Related to the Anticancer Activities of Sesamol, the Dosages, the Dosage Regimens, the Experimental Models, and Whether the Reported Effects are Dose-dependent And/Or Time-dependent.dcont’d

Dose

Dosedependent And/Or TimeDosage Regimen dependent

References

3e100 mM

Incubation (1 h)

Dosedependent

83

Sesamol inhibits MMP-1, MMP- SW1353 9, and MMP-13 expression by inhibiting NF-kB and p38 MAPK

5e20 mM

Pre-incubation Dose(24 h) and dependent incubation (24 h)

84

Sesamol inhibits MMP-1 and MMP-9 expression

30 mg/kg, p.o.

Oral administration (2 weeks)

Effect

Experimental Model

by inhibiting NF-kB translocation to the nucleus Sesamol attenuates proRAW 264.7 inflammatory cytokines by inhibiting NF-kB pathway

e

84

Sesamol attenuates inflammatory Lung (male sprague- 0.3e3 mg/kg, Subcutaneous dawley rats) s.c. administration response by inhibiting NF-kB activity in alveolar macrophages (4 h)

Dosedependent

77

Sesamol attenuates proinflammatory cytokines by inhibiting NF-kB pathway

Dosedependent

85

Wistar rats (males)

Gastric mucosa (male 0.1e1 mg/kg, Oral Wistar rats) p.o. administration (30 min prior WIR treatment)

Sesamol inhibits platelet Human platelet activation by decreasing IKKb suspension phosphorylation and inactivating NF-kB pathway

2.5e25 mM

Pre-incubation (3 min)

Dosedependent and timedependent

86

Sesamol decreases proinflammatory cytokines by inhibiting 5-LOX expression

10 mg/kg, p.o.

Oral administration (15 days)

e

87

10e100 mg/ kg, i.p.

Intraperitoneal administration (30 min prior to g-irradiation)

Dosedependent and timedependent

88

Oral administration (4 weeks)

Dosedependent

89

Oral administration (8 days)

e

90

e

91

Serum and liver (male Wistar rats)

Sesamol attenuates radiationLiver (swiss albino mice) induced increase in inflammatory cells and increases levels of antioxidant enzymes Sesamol inhibits lipid peroxidation and increases antioxidant enzyme levels

Kidney (male Wistar 2e8 mg/kg, rats) p.o.

Sesamol attenuates inflammatory Colon (male albino responses by decreasing nitrite Wistar rats) and neutrophil levels

100 mg/kg, p.o.

(Continued)

II. Role of Seeds in Nutrition and Antioxidant Activities

202 TABLE 15.6

15. Sesame Seed in Controlling Human Health and Nutrition

A Detailed Summary of the Main Findings Related to the Anticancer Activities of Sesamol, the Dosages, the Dosage Regimens, the Experimental Models, and Whether the Reported Effects are Dose-dependent And/Or Time-dependent.dcont’d

Experimental Model

Effect

Dose

Sesamol attenuates inflammatory Colon (female albino 100 mg/kg, responses by decreasing nitrite Wistar rats) p.o. and neutrophil levels

Dosedependent And/Or TimeDosage Regimen dependent

References

Oral administration (3 days)

Serum and Sesamol protects against systemic inflammatory response leukocytes (male through its effect on Wistar rats) inflammatory cytokines and PPARg activation

3 mg/kg, s.c. Subcutaneous administration (1 h after LPS administration)

e

92

Sesamol can be effectively delivered by encapsulating it in phosphatidylcholine micelles

RAW 264.7

0e60 mM of PCS

Incubation with LOX (3 min)

e

93

Sesamol induces either necrosis or apoptosis due to its antiproliferative activity

SK-LU-1

0.05e10 mM

Incubation (48 h) Dosedependent and timedependent

94

Sesamol induces apoptosis by increasing reactive oxygen species level

HCT116

0.05e10 mM

Incubation (48 h) Dosedependent

95

Sesamol induces apoptosis by inhibiting autophagy

HepG2

0.25e1 mM

Incubation (24 h) Dosedependent and Timedependent

96

Sesamol induces slows cell growth and induces cell cycle arrest at S phase

MCF-7 and T47D

50 mM-1 mM Incubation (72 h) Dosedependent

97

Sesamol induces apoptosis by activating caspase pathway

MA-10

0.1e1 mM

Incubation (12 and 24 h)

Dosedependent and Timedependent

98

Sesamol induces apoptosis by HepG2 translocating to the nucleus and binding to DNA via minor groove-binding mode

10e1000 mM

Incubation (24 h) Timedependent

99

Sesamol suppresses colon carcinogenesis by attenuating cyclooxygenase-2 gene expression

12.5e100 mM Incubation (48 h) 500 ppm Oral administration (8 weeks)

DLD-1/COX-2-B2bGal-BSD small intestine (Min mice, Apc-deficient mice)

Dosedependent Timedependent

100

101

II. Role of Seeds in Nutrition and Antioxidant Activities

203

Pharmacological Applications

TABLE 15.6

A Detailed Summary of the Main Findings Related to the Anticancer Activities of Sesamol, the Dosages, the Dosage Regimens, the Experimental Models, and Whether the Reported Effects are Dose-dependent And/Or Time-dependent.dcont’d

Experimental Model

Effect

Dose

Dosedependent And/Or TimeDosage Regimen dependent

Sesamol suppresses DLD-1/COX-2-B2cyclooxygenase-2 by attenuating bGal-BSD small NADPH oxidase 1 mRNA levels intestine (Min mice, Apc-deficient mice)

50e100 mM 500 ppm

Sesamol induces apoptosis through the production of reactive oxygen species

Human platelets

0.25e1.0 mM Incubation (60 min)

Sesamol inhibits tyrosinase activity and induces apoptosis

B16F10

Incubation (48 h) Oral administration (8 weeks)

References

Dosedependent Timedependent Dosedependent

102

25e100 mg/ mL

Incubation (72 h) Dosedependent

103

Sesamol possesses estrogenic and T47D-KBluc anti-estrogenic properties depending on E2 levels

100e500 mM

Incubation (24 h) Dosedependent

104

Sesamol induces apoptosis in tumor cells by reducing expression of Bcl2 and Bax proteins

BALB/c mice

4 mg/kg

Topical administration (5 days/week, 18 weeks)

e

105

Sesamol induces apoptosis and inhibits cell growth

Thymocytes (Wistar rats) and K562

10e30 mM

Incubation (24 h and 72 h)

Dosedependent

106

Sesamol induces carcinogenicity F344 rats and B6C3F1 2% (of diet) in rat forestomach epithelium mice

Oral administration (104 and 96 weeks, respectively)

e

107

Sesamol possesses proliferative activity in rat forestomach epithelium

Oral administration (104 and 96 weeks, respectively)

Dosedependent

108

Oral administration (28 weeks)

e

109

F344/DuCrj rats and 2% (of diet) B6C3F1 mice

F344 rats (males) Sesamol exhibits potential carcinogenicity by increasing rat forestomach papillomas

0.4% and 0.08% (of diet)

With Copyright Permission Majdalawieh AF, Mansour ZR. Sesamol, a major lignan in sesame seeds (Sesamum indicum): anti-cancer properties and mechanisms of action. European Journal of Pharmacology. 2019; 855:75e89.

found the evidence on inhibition of vascular superoxide production by sesamin50. Sesamin feeding-induced improvement of endothelial dysfunction seems to be the result of antioxidative and antihypertensive effects.51 Sesamin produces the effect on altered vascular reactivity in aortic rings of deoxycorticosterone acetate salt-induced hypertensive rat.52 A team of

II. Role of Seeds in Nutrition and Antioxidant Activities

204

15. Sesame Seed in Controlling Human Health and Nutrition

scientist found that dietary sesamin suppresses aortic NADPH oxidase in DOCA salt-hypertensive rats.53

Antihypertensive Effect of Sesame Oil Sesame oil as edible oil lowered blood pressure, decreased lipid peroxidation, and increased antioxidant status in hypertensive patients.54

Metabolism of Sesamin Sesamin is a major lignan found in sesame and is known to have various biological effects. Some of these biological effects occur due to its metabolic conversion to corresponding catechols. The metabolism of sesamin by drug-metabolizing enzymes in rat and/or human liver, such as cytochrome P450 and UDP-glucuronosyltransferase, was found. They dealt with the inhibition of enzymes by sesamin including drug-metabolizing enzymes and other physiologically important enzymes. A remarkable species-based difference was found in sesamin metabolism between humans and other animals; thus, it is very important that precautions are taken when predicting the physiological effects in humans from animal data. A mechanism-based inhibition of human CYP2C9 by sesamin was recently discovered, suggesting that it is important to evaluate the interaction between sesamin and drugs that are mainly metabolized by CYP2C9.55

Effect of Sesame Oil during Endotoxemia Lipopolysaccharide is the main component of outer layer of the cell wall of the gramnegative bacteria, and it triggers multiple organ failure and leads to the death. This model is utilized for the study of effects of sesame oil. The liver and kidney function tests were carried out for the assessment of the mechanism of multiple organ failure. Also, effect on serum uric acid and xanthine oxidase (XO) was examined. Sesame oil corrected hepatic and renal injury in a dose-dependent way and improved animal survival in lipopolysaccharide-treated rats. Sesame oil reduced lipid peroxide level in serum but not in kidney and liver. Serum nitrite fabrication was untouched by sesame oil consumption. Also, the action of xanthine oxidase was decreased in lipopolysaccharide-challenged rats by sesame oil. Finally, it was found that sesame oil decreased multiple organ failure and mortality via inhibition of XO in lipopolysaccharide-dosed rats. XO plays an important role in sesame oil-associated organ fortification during endotoxemia in rats.56

Sesame Oil Protects Against Lead-Plus-Lipopolysaccharide-Induced Acute Hepatic Injury Sesame oil significantly decreased serum aspartate aminotransferase and alanine aminotransferase levels in Pb þ LPS-stimulated mice in Pb-plus-LPS (Pb þ LPS)-induced acute liver damage in mice. Sesame oil reduced Pb þ LPS-induced tumor necrosis factor-alpha, interleukin-1beta, and nitric oxide production in serum and liver tissue. Also, sesame oil

II. Role of Seeds in Nutrition and Antioxidant Activities

Sesame Oil Protects Against Lead-Plus-Lipopolysaccharide-Induced Acute Hepatic Injury

205

decreased inducible NOS expression in leukocytes and liver tissue in Pb þ LPS-treated mice. We hypothesize that the inhibition of proinflammatory cytokines and nitric oxide might be involved in sesame oil-associated protection against Pb þ LPS-induced acute hepatic injury in mice.57

Effect of Sesame Oil on Human Colon Cancer Sesame oil have in vitro antineoplastic properties in human colon cancer58.

Antiaging Effect of Sesamol Sesamol, a highly acclaimed antioxidant, produces anti-aging results on the basis of biochemical and histopathological investigations that the sesamol formulation is effective in preventing photodamage (lesions, ulcers, and changes in skin integrity) due to chronic UV exposure.59

Modulating Effect of Sesamin Sesamin, a major lignan in sesame seeds, has multiple functions such as cholesterollowering and antihypertensive activities through lipid and alcohol metabolizing enzyme’s transcription levels in rat liver. Sesamin significantly augmented the expression of b-oxidation-linked enzymes in peroxisomes and supplementary enzymes prerequisite for degradation of unsaturated fatty acids in mitochondria, via b-oxidation. The consumption of sesamin also resulted in a rise in the gene expression of acyl-CoA thioesterase participated in acyl-CoA hydrolase and very long-chain acyl-CoA thioesterase. Extraordinarily, it encouraged the expression of the gene of an alcohol-metabolizing enzyme (aldehyde dehydrogenase). These results suggested that sesamin controls the metabolism of xenobiotics, lipids, and alcohol at messenger ribonucleic acid level.60

Allergy to Sesame in Humans Immunoglobulin E (IgE)-mediated reactions are supposed to be accountable for utmost food-induced allergic reactions of the immediate hypersensitivity type (type 1), and the identification relies on biological- and scientific-specific features. The supreme common allergic reactions are intermediated by IgE, which happens in the serum and starts activation of effector cells, mainly basophils and mast cells, resulting in an inflammatory response and precise clinical manifestations. When antigens, such as certain foods or proteins from pollen, bind to a definite preformed IgE antibodies linked to the surface of basophils (a part of blood) or mast cells in tissues, then antigen-IgE interaction turns the discharge of mediators such as leukotrienes, histamine, cytokines and prostaglandins, thereby creating an acute inflammatory reaction. In a study, it is found that the reactivity of the 14 kDa protein with maximum of the sera designates that this is the main sesame allergen, later identified as 2S albumin precursor, and its peptide which reacted positively in the dot blot test evidently contains an epitope(s). Some minor sesame allergens, of higher molecular weight, were also revealed.61

II. Role of Seeds in Nutrition and Antioxidant Activities

206

15. Sesame Seed in Controlling Human Health and Nutrition

Conclusions The sesame seed and its constituents are used in the treatment of diseases. It contains a number of constituents, but one of its constituents, sesamol, has effect on various signaling pathways regulating carcinogenesis, making it a valuable and promising bioactive compound. In addition to its anticancer activity, its use as an antihypertensive agent, however, can have major relevance in cardiology. Although, literature convincingly proves the role of sesame seeds and its constituents in therapeutics of different diseases; however, its relevance in clinical surroundings entails further investigation.

References 1. Hegde DM. 17 - sesame. In: Peter KV, ed. Handbook of Herbs and Spices. Woodhead Publishing; 2004:256e289. 2. Visavadiya NP, Narasimhacharya AV. Sesame as a hypocholesteraemic and antioxidant dietary component. Food and Chemical Toxicology. 2008;46(6):1889e1895. 3. Moazzami A, Kamal-Eldin A. Sesame seed oil. In: Moreau RA, Kamal-Eldin A, eds. Gourmet and HealthPromoting Specialty Oils. AOCS Press; 2009:267e282. 4. Namiki M. Nutraceutical functions of sesame: a review. Critical Reviews in Food Science and Nutrition. 2007;47(7):651e673. 5. Shah NC. Sesamum indicum (sesame or til): seeds & oil-an historical and scientific evaluation from Indian perspective. Indian Journal of History of Science. 2013;48(2):151e174. 6. Lyon CK. Sesame: current knowledge of composition and use. Journal of the American Oil Chemists’ Society. 1972;49(4):245e249. 7. Kinman ML, Stark SM. Yield and chemical composition of sesame, Sesamum indicum L., as affected by variety and location grown. Journal of the American Oil Chemists’ Society. 1954;31(3):104e108. 8. Elleuch M, et al. Quality characteristics of sesame seeds and by-products. Food Chemistry. 2007;103(2):641e650. 9. Bailey AE, Swern D, Formo MW. Bailey’s Industrial Oil and Fat Products. New York: Wiley; 1979. Volume 1. Vol. 1. 10. Williamson KS, et al. A survey of sesamin and composition of tocopherol variability from seeds of eleven diverse sesame (Sesamum indicum L.) genotypes using HPLC-PAD-ECD. Phytochemical Analysis. 2008;19(4):311e322. 11. Liu Z, Saarinen NM, Thompson LU. Sesamin is one of the major precursors of mammalian lignans in sesame seed (Sesamum indicum) as observed in vitro and in rats. Journal of Nutrition. 2006;136(4):906e912. 12. Smith KJ. Nutritional framework of oilseed proteins. Journal of the American Oil Chemists’ Society. 1971;48(10):625e628. 13. Maiti S, Hegde MR, Chattopadhyay SB. Handbook of Annual Oilseed Crops. New Delhi: Oxford & IBH; 1988. 14. Budowski P. Sesame oil. III. Antioxidant properties of sesamol. Journal of the American Oil Chemists Society. 1950;27(7):264e267. 15. Budowski P, O’connor RT, Field ET. Sesame oil. VI. Determination of sesamin. Journal of the American Oil Chemists’ Society. 1951;28(2):51e54. 16. Beroza M, Kinman ML. Sesamin, sesamolin, and sesamol content of the oil of sesame seed as affected by strain, location grown, ageing, and frost damage. Journal of the American Oil Chemists’ Society. 1955;32(6):348e350. 17. Budowski P. Recent research on sesamin, sesamolin, and related compounds. Journal of the American Oil Chemists’ Society. 1964;41(4):280e285. 18. Budowski P, Markley KS. The chemical and physiological properties of sesame oil. Chemical Reviews. 1951;48(1):125e151. 19. Shahidi F, et al. Endogenous antioxidants and stability of sesame oil as affected by processing and storage. Journal of the American Oil Chemists’ Society. 1997;74(2):143e148. 20. Rivas RN, Dench JE, Caygill JC. Nitrogen extractability of sesame (Sesamum indicum L.) seed and the preparation of two protein isolates. Journal of the Science of Food and Agriculture. 1981;32(6):565e571.

II. Role of Seeds in Nutrition and Antioxidant Activities

References

207

21. Taha FS, Fahmy M, Sadek MA. Low-phytate protein concentrate and isolate from sesame seed. Journal of Agricultural and Food Chemistry. 1987;35(3):289e292. 22. Wankhede DB, Tharanathan RN. Sesame (Sesamum indicum) carbohydrates. Journal of Agricultural and Food Chemistry. 1976;24(3):655e659. 23. Deosthale YG. Trace element composition of common oilseeds. Journal of the American Oil Chemists Society. 1981;58(11):988e990. 24. Bressani R, Elias LG, Legume Foods V-. In: Altschul AM, ed. New Protein Foods. Academic Press; 1974:230e297. 25. Poneros-Schneier AG, Erdman Jr JW. Bioavailability of calcium from sesame seeds, almond powder, whole wheat bread, spinach and nonfat dry milk in rats. Journal of Food Science. 1989;54(1):150e153. 26. Kiran K, Asad M. Wound healing activity of Sesamum indicum L seed and oil in rats. Indian Journal of Experimental Biology. 2008;46(11):777e782. 27. Hosseini MJ, et al. Protective effects of Sesamum indicum extract against oxidative stress induced by vanadium on isolated rat hepatocytes. Environmental Toxicology. 2016;31(8):979e985. 28. Potipiranun T, Worawalai W, Phuwapraisirisan P, Lamesticumin G. A new a-glucosidase inhibitor from the fruit peels of Lansium parasiticum. Natural Product Research. 2018;32(16):1881e1886. 29. Yu J, et al. The effects of pinoresinol on cholinergic dysfunction-induced memory impairments and synaptic plasticity in mice. Food and Chemical Toxicology. 2019;125:376e382. 30. Bannerman DM, et al. Hippocampal synaptic plasticity, spatial memory and anxiety. Nature Reviews Neuroscience. 2014;15:181e192. 31. Chandrasekaran VR, Hsu DZ, Liu MY. Beneficial effect of sesame oil on heavy metal toxicity. JPEN - Journal of Parenteral and Enteral Nutrition. 2014;38(2):179e185. 32. Basso AS, et al. Reversal of axonal loss and disability in a mouse model of progressive multiple sclerosis. Journal of Clinical Investigation. 2008;118(4):1532e1543. 33. Ghazavi A, Mosayebi G. The mechanism of sesame oil in ameliorating experimental autoimmune encephalomyelitis in C57BL/6 mice. Phytotherapy Research. 2012;26(1):34e38. 34. Mosayebi G, et al. Effect of sesame oil on the inhibition of experimental autoimmune encephalomyelitis in C57BL/6 mice. Pakistan Journal of Biological Sciences. 2007;10(11):1790e1796. 35. Javan M, et al. Cytokine modulatory effects of sesamum indicum seeds oil ameliorate mice with experimental autoimmune Encephalomyelitis. Archives of Asthma, Allergy and Immunology. 2017;1(1):86e93. 36. Bhaskaran S, et al. Inhibition of atherosclerosis in low-density lipoprotein receptor-negative mice by sesame oil. Journal of Medicinal Food. 2006;9(4):487e490. 37. Chen PR, et al. Dietary sesame reduces serum cholesterol and enhances antioxidant capacity in hypercholesterolemia. Nutrition Research. 2005;25(6):559e567. 38. Cooney RV, et al. Effects of dietary sesame seeds on plasma tocopherol levels. Nutrition and Cancer. 2001;39(1):66e71. 39. Geetha T, Rohit B, Pal KI. Sesamol: an efficient antioxidant with potential therapeutic benefits. Medicinal Chemistry. 2009;5(4):367e371. 40. Joshi R, et al. Free radical reactions and antioxidant activities of Sesamol: pulse radiolytic and biochemical studies. Journal of Agricultural and Food Chemistry. 2005;53(7):2696e2703. 41. Prasad NR, et al. Radioprotective effect of sesamol on g-radiation induced DNA damage, lipid peroxidation and antioxidants levels in cultured human lymphocytes. Toxicology. 2005;209(3):225e235. 42. Ramachandran S, Rajendra Prasad N, Karthikeyan S. Sesamol inhibits UVB-induced ROS generation and subsequent oxidative damage in cultured human skin dermal fibroblasts. Archives of Dermatological Research. 2010;302(10):733e744. 43. Nayak PG, et al. Sesamol prevents doxorubicin-induced oxidative damage and toxicity on H9c2 cardiomyoblasts. Journal of Pharmacy and Pharmacology. 2013;65(7):1083e1093. 44. Majdalawieh AF, Mansour ZR. Sesamol, a major lignan in sesame seeds (Sesamum indicum): anti-cancer properties and mechanisms of action. European Journal of Pharmacology. 2019;855:75e89. 45. Yokota T, et al. Sesamin, a lignan of sesame, down-regulates cyclin D1 protein expression in human tumor cells. Cancer Science. 2007;98(9):1447e1453. 46. Nakano D, et al. Sesamin metabolites induce an endothelial nitric oxide-dependent vasorelaxation through their antioxidative property-independent mechanisms: possible involvement of the metabolites in the antihypertensive effect of sesamin. Journal of Pharmacology and Experimental Therapeutics. 2006;318(1):328.

II. Role of Seeds in Nutrition and Antioxidant Activities

208

15. Sesame Seed in Controlling Human Health and Nutrition

47. Matsumura Y, et al. Antihypertensive effect of sesamin. I. Protection against deoxycorticosterone acetate-saltinduced hypertension and cardiovascular hypertrophy. Biological and Pharmaceutical Bulletin. 1995;18(7):1016e1019. 48. Matsumura Y, et al. Antihypertensive effect of sesamin. III. Protection against development and maintenance of hypertension in stroke-prone spontaneously hypertensive rats. Biological and Pharmaceutical Bulletin. 1998;21(5):469e473. 49. Kita S, et al. Antihypertensive effect of sesamin. II. Protection against two-kidney, one-clip renal hypertension and cardiovascular hypertrophy. Biological and Pharmaceutical Bulletin. 1995;18(9):1283e1285. 50. Nakano D, et al. Antihypertensive effect of sesamin. IV. Inhibition of vascular superoxide production by sesamin. Biological and Pharmaceutical Bulletin. 2002;25(9):1247e1249. 51. Nakano D, et al. Effects of sesamin on aortic oxidative stress and endothelial dysfunction in deoxycorticosterone acetate-salt hypertensive rats. Biological and Pharmaceutical Bulletin. 2003;26(12):1701e1705. 52. Matsumura Y, et al. Effects of sesamin on altered vascular reactivity in aortic rings of deoxycorticosterone acetate-salt-induced hypertensive rat. Biological and Pharmaceutical Bulletin. 2000;23(9):1041e1045. 53. Nakano D, et al. Dietary sesamin suppresses aortic NADPH oxidase in DOCA salt hypertensive rats. Clinical and Experimental Pharmacology and Physiology. 2008;35(3):324e326. 54. Sankar D, et al. Effect of sesame oil on diuretics or Beta-blockers in the modulation of blood pressure, anthropometry, lipid profile, and redox status. Yale Journal of Biology and Medicine. 2006;79(1):19e26. 55. Yasuda K, Sakaki T. How is sesamin metabolised in the human liver to show its biological effects? Expert Opinion on Drug Metabolism and Toxicology. 2012;8(1):93e102. 56. Hsu DZ, Liu MY. Sesame oil attenuates multiple organ failure and increases survival rate during endotoxemia in rats. Critical Care Medicine. 2002;30(8):1859e1862. 57. Hsu DZ, et al. Sesame oil protects against lead-plus-lipopolysaccharide-induced acute hepatic injury. Shock. 2007;27(3):334e337. 58. Salerno JW, Smith DE. The use of sesame oil and other vegetable oils in the inhibition of human colon cancer growth in vitro. Anticancer Research. 1991;11(1):209e215. 59. Sharma S, Kaur IP. Development and evaluation of sesamol as an antiaging agent. International Journal of Dermatology. 2006;45(3):200e208. 60. Tsuruoka N, et al. Modulating effect of sesamin, a functional lignan in sesame seeds, on the transcription levels of lipid- and alcohol-metabolizing enzymes in rat liver: a DNA microarray study. Bioscience Biotechnology & Biochemistry. 2005;69(1):179e188. 61. Wolff N, et al. Allergy to sesame in humans is associated primarily with IgE antibody to a 14 kDa 2S albumin precursor. Food and Chemical Toxicology. 2003;41(8):1165e1174. 62. Kumar A, et al. Sesamol attenuates genotoxicity in bone marrow cells of whole-body gamma-irradiated mice. Mutagenesis. 2015;30(5):651e661. 63. Mishra K, Srivastava PS, Chaudhury NK. Sesamol as a potential radioprotective agent: in vitro studies. Radiation Research. 2011;176(5):613e623. 64. Khan S, et al. Protective effect of sesamol against (6)(0)Co gamma-ray-induced hematopoietic and gastrointestinal injury in C57BL/6 male mice. Free Radical Research. 2015;49(11):1344e1361. 65. Kanimozhi P, Prasad NR. Antioxidant potential of sesamol and its role on radiation-induced DNA damage in whole-body irradiated Swiss albino mice. Environmental Toxicology and Pharmacology. 2009;28(2):192e197. 66. Prasad NR, et al. Photoprotective effect of sesamol on UVB-radiation induced oxidative stress in human blood lymphocytes in vitro. Environmental Toxicology and Pharmacology. 2005;20(1):1e5. 67. Srisayam M, et al. Antioxidant, antimelanogenic, and skin-protective effect of sesamol. Journal of Cosmetic Science. 2014;65(2):69e79. 68. Hsu DZ, et al. Sesamol delays mortality and attenuates hepatic injury after cecal ligation and puncture in rats: role of oxidative stress. Shock. 2006;25(5):528e532. 69. Hemalatha G, Pugalendi KV, Saravanan R. Modulatory effect of sesamol on DOCA-salt-induced oxidative stress in uninephrectomized hypertensive rats. Molecular and Cellular Biochemistry. 2013;379(1e2):255e265. 70. Chopra K, et al. Sesamol suppresses neuro-inflammatory cascade in experimental model of diabetic neuropathy. The Journal of Pain. 2010;11(10):950e957. 71. Yashaswini PS, Rao AG, Singh SA. Inhibition of lipoxygenase by sesamol corroborates its potential antiinflammatory activity. International Journal of Biological Macromolecules. 2017;94(Pt B):781e787.

II. Role of Seeds in Nutrition and Antioxidant Activities

References

209

72. Kaur IP, Saini A. Sesamol exhibits antimutagenic activity against oxygen species mediated mutagenicity. Mutation Research. 2000;470(1):71e76. 73. Suja KP, Jayalekshmy A, Arumughan C. Free radical scavenging behavior of antioxidant compounds of sesame (sesamum indicum L.) in DPPH(*) system. Journal of Agricultural and Food Chemistry. 2004;52(4):912e915. 74. Turgut B, Ergen I, Ilhan N. The protective effect of sesamol in the selenite-induced experimental cataract model. Turkish Journal of Ophthalmology. 2017;47(6):309e314. 75. Chen WY, et al. Sesamol reduces the atherogenicity of electronegative L5 LDL in vivo and in vitro. Journal of Natural Products. 2015;78(2):225e233. 76. Chennuru A, Saleem MT. Antioxidant, lipid lowering, and membrane stabilization effect of sesamol against doxorubicin-induced cardiomyopathy in experimental rats. BioMed Research International. 2013;2013:934239. 77. Chu PY, et al. Protective effect of sesamol on the pulmonary inflammatory response and lung injury in endotoxemic rats. Food and Chemical Toxicology. 2010;48(7):1821e1826. 78. Gupta A, et al. Renoprotective effects of sesamol in ferric nitrilotriacetate-induced oxidative renal injury in rats. Basic and Clinical Pharmacology and Toxicology. 2009;104(4):316e321. 79. Jnaneshwari S, et al. Sesamol ameliorates cyclophosphamide-induced hepatotoxicity by modulating oxidative stress and inflammatory mediators. Anti-Cancer Agents in Medicinal Chemistry. 2014;14(7):975e983. 80. Chandrasekaran VR, Hsu DZ, Liu MY. The protective effect of sesamol against mitochondrial oxidative stress and hepatic injury in acetaminophen-overdosed rats. Shock. 2009;32(1):89e93. 81. Geetha T, et al. Sesamol-loaded solid lipid nanoparticles for treatment of skin cancer. Journal of Drug Targeting. 2015;23(2):159e169. 82. Chu PY, et al. Sesamol down-regulates the lipopolysaccharide-induced inflammatory response by inhibiting nuclear factor-kappa B activation. Innate Immunity. 2010;16(5):333e339. 83. Wu XL, et al. Sesamol suppresses the inflammatory response by inhibiting NF-kappaB/MAPK activation and upregulating AMP kinase signaling in RAW 264.7 macrophages. Inflammation Research. 2015;64(8):577e588. 84. Lu YC, et al. Chondroprotective role of sesamol by inhibiting MMPs expression via retaining NF-kappaB signaling in activated SW1353 cells. Journal of Agricultural and Food Chemistry. 2011;59(9):4969e4978. 85. Hsu DZ, et al. Protective effect of 3,4-methylenedioxyphenol (sesamol) on stress-related mucosal disease in rats. BioMed Research International. 2013;2013:481827. 86. Chang CC, et al. A novel role of sesamol in inhibiting NF-kappaB-mediated signaling in platelet activation. Journal of Biomedical Science. 2011;18:93. 87. Yashaswini PS, et al. In vivo modulation of LPS induced leukotrienes generation and oxidative stress by sesame lignans. The Journal of Nutritional Biochemistry. 2017;41:151e157. 88. Parihar VK, et al. Effect of sesamol on radiation-induced cytotoxicity in Swiss albino mice. Mutation Research. 2006;611(1e2):9e16. 89. Kuhad A, Sachdeva AK, Chopra K. Attenuation of renoinflammatory cascade in experimental model of diabetic nephropathy by sesamol. Journal of Agricultural and Food Chemistry. 2009;57(14):6123e6128. 90. Kondamudi PK, et al. Modulatory effects of sesamol in dinitrochlorobenzene-induced inflammatory bowel disorder in albino rats. Pharmacological Reports. 2013;65(3):658e665. 91. Kondamudi PK, et al. Investigation of sesamol on myeloperoxidase and colon morphology in acetic acidinduced inflammatory bowel disorder in albino rats. ScientificWorldJournal. 2014;2014:802701. 92. Periasamy S, et al. Sesamol ameliorates hypotension by modulating cytokines and PPAR-gamma in systemic inflammatory response. EXCLI J. 2015;14:948e957. 93. Yashaswini PS, Kurrey NK, Singh SA. Encapsulation of sesamol in phosphatidyl choline micelles: enhanced bioavailability and anti-inflammatory activity. Food Chemistry. 2017;228:330e337. 94. Siriwarin B, Weerapreeyakul N. Sesamol induced apoptotic effect in lung adenocarcinoma cells through both intrinsic and extrinsic pathways. Chemico-Biological Interactions. 2016;254:109e116. 95. Khamphio M, Barusrux S, Weerapreeyakul N. Sesamol induces mitochondrial apoptosis pathway in HCT116 human colon cancer cells via pro-oxidant effect. Life Sciences. 2016;158:46e56. 96. Liu Z, et al. Sesamol induces human hepatocellular carcinoma cells apoptosis by impairing mitochondrial function and suppressing autophagy. Scientific Reports. 2017;7:45728. 97. Jacklin A, et al. The sesame seed oil constituent, sesamol, induces growth arrest and apoptosis of cancer and cardiovascular cells. Annals of the New York Academy of Sciences. 2003;1010:374e380.

II. Role of Seeds in Nutrition and Antioxidant Activities

210

15. Sesame Seed in Controlling Human Health and Nutrition

98. Chen YH, et al. Effects of sesamol on apoptosis and steroidogenesis in MA-10 mouse Leydig tumor cells. Journal of Agricultural and Food Chemistry. 2011;59(18):9885e9891. 99. Liu Z, et al. The interaction of sesamol with DNA and cytotoxicity, apoptosis, and localization in HepG2 cells. Food Chemistry. 2013;141(1):289e296. 100. Shimizu S, et al. Sesamol suppresses cyclooxygenase-2 transcriptional activity in colon cancer cells and modifies intestinal polyp development in Apc (Min/þ) mice. Journal of Clinical Biochemistry & Nutrition. 2014;54(2):95e101. 101. Shimizu S, et al. Involvement of NADPH oxidases in suppression of cyclooxygenase-2 promoter-dependent transcriptional activities by sesamol. Journal of Clinical Biochemistry & Nutrition. 2015;56(2):118e122. 102. Thushara RM, et al. Sesamol induces apoptosis in human platelets via reactive oxygen species-mediated mitochondrial damage. Biochimie. 2013;95(11):2060e2068. 103. Kumar CM, et al. Interaction of sesamol (3,4-methylenedioxyphenol) with tyrosinase and its effect on melanin synthesis. Biochimie. 2011;93(3):562e569. 104. Pianjing P, et al. Estrogenic activities of sesame lignans and their metabolites on human breast cancer cells. Journal of Agricultural and Food Chemistry. 2011;59(1):212e221. 105. Bhardwaj R, et al. Sesamol induces apoptosis by altering expression of bcl-2 and bax proteins and modifies skin tumor development in balb/c mice. Anti-Cancer Agents in Medicinal Chemistry. 2017;17(5):726e733. 106. Fujimoto A, et al. Apoptosis-inducing action of two products from oxidation of sesamol, an antioxidative constituent of sesame oil: a possible cytotoxicity of oxidized antioxidant. Toxicology in Vitro. 2010;24(6):1720e1726. 107. Hirose M, et al. Stomach carcinogenicity of caffeic acid, sesamol and catechol in rats and mice. Japanese Journal of Cancer Research. 1990;81(3):207e212. 108. Tamano S, et al. Forestomach neoplasm induction in F344/DuCrj rats and B6C3F1 mice exposed to sesamol. Japanese Journal of Cancer Research. 1992;83(12):1279e1285. 109. Hirose M, et al. Carcinogenicity of antioxidants BHA, caffeic acid, sesamol, 4-methoxyphenol and catechol at low doses, either alone or in combination, and modulation of their effects in a rat medium-term multi-organ carcinogenesis model. Carcinogenesis. 1998;19(1):207e212.

II. Role of Seeds in Nutrition and Antioxidant Activities

C H A P T E R

16

Kancolla Seeds: High Nutritional Foods With Nutraceutical Properties Irene Dini Pharmacy Department, “Federico II” University, Naples, Italy

List of Abbreviations AACC American Association of Cereal Chemists ADI Acceptable daily intake CE Catechin GAE Gallic acid L-AA Ascorbic acid LDL Low-density lipoprotein NNMT Acid-N-methyltransferase PGI2 Endothelial prostacyclin 2 RDA Recommended daily allowance

Introduction Kancolla is a sweet variety of Chenopodium quinoa (quinoa) (Chenopodiaceae family) selected in 1950 in Perú. Quinoa is a pseudocereal, a dicotyledonous crop that produces small grain-like seeds. Consequently, their seed nutritional quality is like those of other dicotyledonous plants rather than related to gramineae seeds that are monocotyledonous crops. The main edible parts are seeds that can be eaten in soup, milled into flour, or fermented to make beer, or a traditional ceremonial alcoholic beverage from South America called “chicha.” The seeds of most quinoa varieties contain saponins, located in the outer layers of the seed coat, which deter birds and insects from eating them. These bitter-tasting constituents are removed before consumption by washing or milling to remove the seed coat. Unfortunately, polished quinoa has half the fiber of washed quinoa, along with lower protein, vitamin, and mineral levels. The increased

Nuts and Seeds in Health and Disease Prevention, Second Edition https://doi.org/10.1016/B978-0-12-818553-7.00016-4

211

Copyright © 2020 Elsevier Inc. All rights reserved.

212

16. Kancolla Seeds: High Nutritional Foods With Nutraceutical Properties

demand for quinoa has led researchers to produce several cultivars selected and bred for their tolerance to heat and cold, resistance to disease, and for sweet tastes; among these the oldest and most widespread variety is kancolla.

Nutritional Values of Kancolla Seeds Proximate Composition The quality of kancolla seeds was determined by measuring proximate composition (carbohydrate, lipid, protein, fiber, essential amino acids, and minerals). The protein level in kancolla is 13.4%, which is slightly lower than in other quinoa varieties (13.8%), lower than barley and buckwheat, higher than in wheat, but much higher than corn, millet (pearl), oat, rice, and rye. The amount of fiber present in quinoa seeds and the other grains is less than 4.2%, while in kancolla seeds it is about 3 times higher except buckwheat for which it is 18.2%. The total lipid content in kancolla seeds was found to be 5.9%, which is much higher than barley, rice, rye, and wheat and slightly lower than in other quinoa varieties, buckwheat, corn, millet, and oat (Table 16.1).1

Amino Acid Composition The quality of kancolla seed proteins is noteworthy. In general, animal proteins (meat, fish, poultry, milk, cheese, and eggs) are considered good sources of complete proteins because they contain ample amounts of all essential amino acids, whereas plant proteins are often called incomplete proteins because they generally lack in one or more essential amino acids. Despite this, kancolla seed proteins contain all the essential amino acids in the right dosage TABLE 16.1 Crop

Comparisons of the nutritional quality (% dry weight) of kancolla seeds with various grains and other quinoa varieties. Water

Crude Protein

Fat

Carbohydrates

—————————————— % —————————————— Kancolla

16.6

13.4

5.9

51.7

Quinoa

12.6

13.8

5.0

59.7

Barley

9.0

14.7

1.1

67.8

Buckwheat

10.7

18.5

4.9

43.5

Corn

13.5

8.7

3.9

70.9

Millet (pearl)

11.0

11.9

4.0

68.6

Oat

13.5

11.1

4.6

57.6

Rice

11.0

7.3

0.4

80.4

Rye

13.5

11.5

1.2

69.6

Wheat

10.9

13.0

1.6

70.0

II. Role of Seeds in Nutrition and Antioxidant Activities

213

Nutritional Values of Kancolla Seeds

for human needs. Their concentration is better than in other quinoa varieties and meets or exceeds nutritional FAO standards requirements for all essential amino acids. Moreover, grain proteins have high contents of lysine and sulfur amino acids, which are the limiting amino acids in other grains. Tyrosine and methionine are present in lower amounts, 4.1 and 2.2 g amino acid/100 g sample compared to the other amino acids. Tyrosine is the limiting amino acid. All the essential amino acids have a high chemical score value which implied that the essential amino acids have a high biological value (Table 16.2).1 Moreover, free amino acids were found in highest amounts in kancolla seeds (Table 16.3).1 Free amino acids play an important role in the formation of color and aroma during roasting; they are able to impart bitter, sour, and sweet tastes to foods. The free amino acids present in kancolla seeds contained large amounts of arginine (60 mg/100 g) and glutamic acid (58.0 mg/100 g) along with lesser amounts of aspartic acid (30.7 mg/100 g) and alanine (10.2 mg/100 g). As reported by Schiffman et al.,2 arginine is considered bite and pungent, and most subjects reported salty and sour qualities, glutamic acid has a concentrated taste which is strong, constant, and unpleasant, aspartic acid is considered thick, bite, and pungent, and finally, alanine is a good, sweet, and flavor compound.

Dietary Fiber Analysis Dietary fiber is a broad term defined by the American Association of Cereal Chemists (AACC) in 2000 as the edible parts of plant or analogous carbohydrates that are resistant to digestion and absorption in the human small intestine with complete or partial fermentation in the large intestine. Different classifications are utilized to describe dietary fibers,

TABLE 16.2

Amino Acid

Essential amino acid pattern of kancolla seeds compared to wheat, soy, skim milk, other quinoa varieties and the FAO reference pattern (1990) for evaluating proteins.

Amino Acid Content (g/100g Protein) Kancolla Quinoa Wheat

Soy

Skim Milk

FAO

----------------------- % ----------------------Isoleucine

5.6 (C. I.140)

4.0

3.8

4.7

5.6

2.8

Leucine

9.4 (C. I.130)

6.8

6.6

7.0

9.8

6.6

7.8 (C. I.141) Phenylalanine 6.2

5.1 4.6

2.5

6.3

8.2

5.8

4.5

4.6

4.8

(C. I.172) 3.8

3.0

3.6

5.0

6.3

2.4

2.2

1.4

0.9

2.5

2.2

1.7

1.4

2.6

Lysine

Tyrosine Cystine

4.1 2.7

(C. I.140)

Methionine

2.2

Threonine

6.2 (C. I.155)

3.7

2.9

3.9

4.6

3.4

Valine

6.1 (C. I.122)

4.8

4.7

4.9

6.9

3.5

II. Role of Seeds in Nutrition and Antioxidant Activities

214

16. Kancolla Seeds: High Nutritional Foods With Nutraceutical Properties

TABLE 16.3

Free amino acid pattern of kancolla seeds.

Free amino acid

Kancolla (mg/100g protein)

Isoleucine

4.6

Leucine

7.7

Lysine

5.8

Phenylalanine

6.9

Tyrosine

5.3

Cystine

missing

Methionine

0.9

Threonine

4.4

Aspartic acid

30.7

Valine

6.7

Asparagine

4.1

Glutammic acid

58.0

Serine

4.8

Glycine

3.0

Glutamine

12.5

Hystidine

24.3

Arginine

60.0

Alanine

10.2

Proline

5.8

including origin, chemical composition, and physicochemical properties. The most widely accepted classification differentiated dietary components into two categories: waterinsoluble (cellulose, hemicellulose, and lignin) and the water-soluble (pectin, gums, and mucilage). The first are generally poorly fermented by gut microbes in the distal colon where transit time is slower, and bacterial densities are higher, enhance gut transit rate, and reduce the amount of time available for colonic bacterial fermentation of nondigested foodstuff. Instead, the second, generally known as prebiotics, have high solubility and viscosity, are highly fermentable, and are metabolized by bacteria in the ileum and ascending colon,3 increased colonic fermentation/short-chain fatty acid production, and have a positive modulation of colonic microflora. Noteworthy is dietary fiber content (12.3%) (Table 16.4).1 Dietary fiber (indigestible carbohydrate) is not a nutrient, but it still plays a very important role in maintaining good health. Diets rich in dietary fiber have been associated with reduced risk of all-cause mortality and death from coronary and vascular diseases (dietary fiber reduced total and/or LDL serum cholesterol levels and reduced blood pressure), colon

II. Role of Seeds in Nutrition and Antioxidant Activities

215

Nutritional Values of Kancolla Seeds

TABLE 16.4

Comparisons of fiber and ash content (% dry weight) of kancolla seeds, various grains and other quinoa varieties.

Crop

Fiber

Ash

—————————————————— % —————————————————— Kancolla

12.3

1.7

Quinoa

4.1

3.4

Barley

2.0

5.5

Buckwheat

18.2

4.2

Corn

1.7

1.2

Millet (pearl)

2.0

2.0

Oat

0.3

2.9

Rice

0.4

0.5

Rye

2.6

1.5

Wheat

2.7

1.8

cancer, diabetes (dietary fiber attenuate postprandial glycemia/insulinemia), respiratory disease, and infections and are sometimes considered as useful in the prevention of obesity because they decrease the energy density of foods, reduce postprandial blood glucose and insulin responses, and increase fecal bulk/laxation and promoting satiety.4

Total Carotenoid Content Carotenoids are essential components of human diet mainly because some are precursors of vitamin A. They are plant pigments which act as antioxidants, hormone precursors, colorants, and essential components of the photosynthetic apparatus. Total carotenoid contents of kancolla seeds are determined at precooking seeds and after boiling seeds. Precooking kancolla seeds exhibited a carotenoid content (0.4
0.05 mg/10 g; P < .004) higher than that of precooking bitter quinoa seeds (0.08
0.04 mg/10 g; P < .006) and common cereals, such as einkorn (0.1 mg/10 g) and corn (0.07 mg/10 g). After boiling, data revealed a significant decrease of carotenoid (0.03
0.02 mg/10 g; P < .01) in seeds, but an aliquot was found in cooking water (0.1
0.02 mg/10 g; P < .006). Nevertheless, some carotenoids are broken during the boiling process because the sum of carotenoid content in the postcooking seeds and in the cooking water significantly differed from that of precooking seeds.5

Total Ascorbic Acid Content The biological role of reduced ascorbic acid (L-AA) is that of an enzyme cofactor, radical scavenger, and donor/acceptor in electron transport systems. In humans, the oxidized form (dehydro-AA, DHA) has vitamin character. Within erythrocytes, a reduction of DHA to

II. Role of Seeds in Nutrition and Antioxidant Activities

216

16. Kancolla Seeds: High Nutritional Foods With Nutraceutical Properties

L-ascorbic

acid is reported by a glutathione-mediated pathway. The RDA value (recommended daily allowance) of vitamin C is ranged between 66 and 79 mg/day. Precooking seeds contained the highest value of vitamin C (1.3
0.2 mg/10 g; P < .002), then boiling seed (0.1
0.02 mg/10 g; P < .002), loss due to the passage of the vitamin in water, as shown by higher vitamin C levels detected in cooking waters (0.9
0.3 mg/10 g; P < .01).5

Anion Analysis The anion content was reported in table (Table 16.5).1

Oxalates Oxalates, a ubiquitous constituent of plants, are responsible for a significant number of harmful effects in humans and animals. High dietary oxalate intake plays a key role in secondary hyperoxaluria, determines acute renal failure, reduces the intestinal absorption of calcium and magnesium, determines gastrointestinal irritation, muscle contraction, or tetany accompanied by other nervous symptoms, and decreases ability to clot blood and possible injury to excretory organs, among others, due to deposition of cell substance with high concentrations of crystalline calcium oxalate. Oxalates in kancolla seeds are present in lower amounts, 154.299 mg/100g compared to other quinoa varieties (380 mg/100g).6

Nitrates Nitrates (NO 3 ) are an essential source of nitrogen for plant normal growth. In the human body, nitrates interfere with vitamin A metabolism and in the functions of the thyroid gland. The nitrates reduced to nitrites, collaborate to the formation of metmyoglobin or react with secondary and tertiary amines, forming potentially carcinogenic N-nitrous. The levels of nitrates found in kancolla seeds are lower than other quinoa varieties. Regarding the potential long-term health risks of nitrate levels encountered in the diet, the maximum allowable nitrate levels in kancolla seeds not exceed acceptable daily intake (ADI) of nitrate ions recommended by WHO (222 mg/day for a 60-kg adult); therefore, kancolla seeds do not endanger consumer’s safety. TABLE 16.5

Anions content of kancolla seeds.

Anions

mg/100g

Cl-

47.423

F-

3.470

NO-3

9.857

63

PO34

509.935

383.7

SO24

76.293

C2O24

154.299

II. Role of Seeds in Nutrition and Antioxidant Activities

mg/100g

380

217

Nutritional Values of Kancolla Seeds

Phosphates Phosphorus is an essential nutrient occurring in most foods, both as a natural component and as an approved ingredient added during food processing. Phosphorus from plant-based sources remains less bioavailable than animal sources and animal sources less bioavailable compared to inorganic phosphate additives. Phosphorus is a component of cell membranes (i.e., phospholipid bilayer), nucleic acid (i.e., sugar phosphate), and bones and teeth (i.e., hydroxyapatite). In addition, phosphorus plays important roles in energy metabolism (e.g., in ATP, GTP, ADP, GDP), in acid/base balance, and in an intracellular cell signaling. The most recent USA dietary reference intakes are 700 mg/day. Phosphorus deficiencies are rare. Unfortunately, adverse effects of high dietary phosphorus have been observed on bone health (prevalent with low calcium intakes)7 and on blood pressure regulation through its role in the plasma membrane structure (phospholipids), energy production and storage (adenosine triphosphate, creatine phosphate, and other phosphorylated compounds), enzyme activation, cellular messengers such as G proteins, and acid-base regulation. High phosphorus/mineral (calcium and magnesium) intake is important to lower blood pressure.8 The levels of phosphates found in kancolla seeds are higher than other quinoa varieties; nevertheless, intake of phosphorus relative to calcium is nearly estimated optimal Ca/P intake ratios. In balance studies in human adults, Ca/P molar ratios ranging from 0.07:1 to 2.40:1 (a 30-fold range) had no effect on either calcium balance or calcium absorption.9 These minerals interact in the gastrointestinal tract to limit absorption of the other and are intimately related in tissue (e.g., hydroxyapatite) and in their hormonal regulation.

Mineral Analysis Minerals are important constituents of bones, teeth, soft tissues, hemoglobin, muscle, blood, and nerve cells and are vital to overall mental and physical well-being. Kancolla’s mineral content presents higher levels of calcium (0.36%), potassium (0.96%), iron (107.5 ppm), and zinc (162.5 ppm) and lower level in sodium (107.5 ppm) than wheat, barley, corn, and the other quinoa varieties (Table 16.6). Calcium, iron, and zinc bioavailabilities are low due to high fiber content and low mineral/potassium ratio. A high TABLE 16.6 Crop

Comparison of the mineral content of kancolla seeds with barley, yellow corn, wheat and other quinoa varieties (quinoa data are based on the average of 15 cultivars). Ca

P

Mg

K

———————— % ———————

Na

Fe

Cu

Mn

Zn

—————————— ppm ——————————

Kancolla

0.36

N.D.

0.19

0.96

107.5

107.5

6.5

N.D.

162.5

Quinoa

0.19

0.47

0.26

0.87

115

205

67

128

50

Barley

0.08

0.42

0.12

0.56

200

50

8

16

15

Corn

0.07

0.36

0.14

0.39

900

21

–

–

–

Wheat

0.05

0.36

0.16

0.52

900

50

7

–

14

II. Role of Seeds in Nutrition and Antioxidant Activities

218

16. Kancolla Seeds: High Nutritional Foods With Nutraceutical Properties

potassium/sodium ratio makes kancolla interesting for diets with a defined electrolytic balance. The high content of potassium can be utilized beneficially in the diets of people who take diuretics to control hypertension and suffer from excessive excretion of potassium through body fluid.1 Kancolla seeds have a low sodium/potassium ratio (0.01ppm). The WHO guidelines recommend free-living individuals to satisfy dietary Na/K ratio of AFB2 > AFG2.13

Nuts and Seeds in Health and Disease Prevention, Second Edition https://doi.org/10.1016/B978-0-12-818553-7.00019-X

255

Copyright © 2020 Elsevier Inc. All rights reserved.

256

19. Mycotoxins in Nuts and Seeds

FIGURE 19.1 Chemical structures of aflatoxins B1, B2, G1, G2, M1, M2, and ochratoxin A. Adapted from the Council for Agricultural Science and Technology - CAST 2003, with permission.

AFM1 and AFM2, the hydroxylated derivatives of AFB1 and AFB2, are detected in the milk of dairy cattle or lactating mothers exposed to aflatoxins. AFM1 is less mutagenic and carcinogenic than AFB1, but shows acute toxicity similar to other aflatoxins. The IARC13 classified it as a Group 2B carcinogen (possibly carcinogenic to humans). OTA is a dihydrocoumarin linked to the amino acid phenylalanine (Fig. 19.1).1 In tropical regions, OTA contamination is associated with Aspergillus sp. belonging to the Circumdati (Aspergillus westerdijkiae, Aspergillus ochraceus, A. steynii) and Nigri sections (Aspergillus

III. Fungal Infections on Seeds and Nuts and Health

Natural Occurrence

257

carbonarius, A. welwitschiae, Aspergillus niger).14e16 In cool and temperate regions, OTA contamination occurs mainly by Penicillium verrucosum.17 OTA has been shown to cause nephrotoxic, embryotoxic, cytotoxic, teratogenic, genotoxic,18 and potentially carcinogenic effects. OTA has been classified as possibly carcinogenic to humans (Group 2B), based on evidence for carcinogenicity in animal studies, but there is insufficient evidence in humans.13 The human health hazard associated with the natural occurrence of harmful mycotoxins has compelled several countries to establish regulatory guidelines, but the maximum tolerated levels for mycotoxins vary widely between countries.19 They tend to be higher in tropical and subtropical producing countries and lower in importing countries with temperate climate. Among aflatoxins, AFB1 has triggered the most research efforts because of its high toxicity and worldwide occurrence in staple foods and feeds. At least 119 countries involved in international trade have established regulatory limits for aflatoxins.19 Table 19.1 shows the maximum tolerated levels for aflatoxins20 and OTA21 in foodstuffs by the European Union. For example, the maximum tolerated levels for total aflatoxins (sum of AFB1, AFB2, AFG1, and AFG2) are set at 15 mg/kg in groundnuts (peanuts), hazelnuts, Brazil nuts, almonds, pistachios, and apricot kernels to be subjected to sorting, or other physical treatment, before human consumption or use as an ingredient in foodstuff. The European Union Commission has established maximum levels for OTA in foodstuffs such as cereals (5 mg/kg), dried vine fruit (10 mg/kg), coffee (5 mg/kg), and some spices (15 mg/kg)21 for several years but since OTA has been detected in some other foods the European Union has proposed the expansion of the group of products subject to a maximum level for OTA. For sunflower and pumpkin seeds, pistachios, hazelnuts, or all tree nuts, the maximum level of 5 mg/kg has been proposed for further discussion and, after agreement has been reached, will be included in the guidelines for food contaminants. Considering the great impact on consumer health because of their toxicity and occurrence, this chapter provides an overview of the natural occurrence, effect of heat processing, and toxicological effects of aflatoxins and OTA in nuts and seeds.

Natural Occurrence Fungal growth and mycotoxin production depend on biological (susceptible crop) and environmental factors, with emphasis on regional climatic conditions during plant development and crop harvest. Mycotoxin contamination is also favored by stress or damage to the crop due to drought before harvest, insect infestation, and inadequate drying of the crop before storage.1 Aflatoxins and OTA occur worldwide8,9,22e26 and contaminate a wide range of food commodities, including cereals and cereal products, nuts, spices, and dried fruits. It has been shown that water availability (water activity, Aw) and temperature play an important role in determining the extent of aflatoxin and OTA production. For example, the ideal temperature for growth and mycotoxin production ranges from 25 to 35
C for A. flavus strains. Aw values for aflatoxin production by A. flavus range from 0.95 to 0.99, with a minimum Aw value of 0.82.27 The temperature and Aw values for maximum growth

III. Fungal Infections on Seeds and Nuts and Health

258

19. Mycotoxins in Nuts and Seeds

TABLE 19.1

Maximum Tolerated Levels for Aflatoxins and Ochratoxin A in Some Foodstuffs. Maximum Levels (mg/kg) AFB1 AFB1 D AFB2 D AFG1 D AFG2

Foodstuffs a

Aflatoxins

Groundnuts (peanuts), hazelnuts, and Brazil nuts to be subjected to sorting, or other physical treatment, before human consumption or use as an ingredient in foodstuff

8.0

15.0

Almonds, pistachios, and apricot kernels to be subjected to sorting, or other physical treatment, before human consumption or use as an ingredient in foodstuffs

12.0

15.0

Tree nuts, other than almonds, pistachios, apricot kernels, hazelnuts, and 5.0 Brazil nuts, to be subjected to sorting, or other physical treatment, before human consumption or use as an ingredient in foodstuffs

10.0

Groundnuts (peanuts) and other oilseeds and processed products thereof 2.0 intended for direct human consumption or use as an ingredient in foodstuffs

4.0

Almonds, pistachios, and apricot kernels intended for direct human consumption or use as an ingredient in foodstuffs

8.0

10.0

Hazelnuts and Brazil nuts intended for direct human consumption or use as an ingredient in foodstuffs

5.0

10.0

Tree nuts, other than the almonds, pistachios, apricot kernels, hazelnuts and Brazil nuts, and processed products thereof, intended for direct human consumption or use as an ingredient in foodstuffs

2.0

4.0

Ochratoxin Ab Unprocessed cereals

5.0

All products derived from unprocessed cereals, including processed cereal products and cereals intended for direct human consumption

3.0

a

Commission Regulation (EU) No 165/2010. Commission Regulation (EC) No 1881/2006. Adapted from http://eur-lex.europa.eu, © European Union, 1998e2019 (with permission).

b

for A. carbonarius are 30
C and 0.965 Aw, while for A. niger they are 35
C and 0.98 Aw. The optimum Aw for OTA production is 0.95e0.98 for A. carbonarius and 0.95 for A. niger.28 A. ochraceus shows optimal growth and OTA production at 0.99 Aw and 30
C.29 A number of surveys and monitoring programs on aflatoxins and OTA have been carried out in several countries. Table 19.2 shows selected examples of natural occurrence and co-occurrence of aflatoxins and OTA in nuts and seeds. Sangare-Tigori et al.11 evaluated the co-occurrence of mycotoxins in peanut samples from the Ivory Coast and detected contamination by aflatoxin B1 (range 1.5e10 mg/kg) and OTA (range nondetectede0.64 mg/kg) in 100 and 60% of samples, respectively. AFB1 levels were above the European Union regulatory limits in 40% of samples.

III. Fungal Infections on Seeds and Nuts and Health

TABLE 19.2 Selected Examples of Natural Occurrence and Co-occurrence of Aflatoxins and Ochratoxin A/Other Mycotoxins in Nuts and Seeds. Samples Analyzed Mycotoxins

Positive LOD/LOQ Samples (mg/kg)

Peanuts

Ivory Coast

10

Aflatoxin B1 Fumonisin B1 Ochratoxin A Zearalenone

10 7 6 10

1.5/e 300/e e 25/e

Chestnuts

Italy

14

Aflatoxin B1 Aflatoxin B2 Aflatoxin G1 Aflatoxin G2 Ochratoxin A

3 1 2 1 14

0.04/0.10 (AFB1) 80% is high;