Final approved nature cures book draft Sept 7th 2022

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Swati Save Nature Cures Trusting the biomolecules

© 2022 Europe Books| London www.europebooks.co.uk | [email protected] ISBN XXX-XX-XXX-XXX-X First edition: XXXX 2022 Printed for Italy by Rotomail Italia S.p.A. – Vignate (MI) Stampato presso Rotomail Italia S.p.A. – Vignate (MI)





POETRY I believe I’m here for the results and the opportunity of living in pure & honest intentions. I believe I’m the possibility of being a river of dreams and miracles through commitment. I believe I am here to have my life be on the hot seat, to be willing to go through breakdowns or whatever else happens to get to the breakthroughs that give me my life. I believe I’m here to invent new ways of being -based on nothing – to show up in my life and in the world. I believe I am here because in my intuitive heart I know that my life on earth revolves around bringing the new model of conversations into the world, especially with innovators and for world peace. I believe I’m here to give my love to others, because I can and because it is the same love that they have for me. I believe I’m here to rediscover the hidden – the motives, the rewards, the intentions, the castles in the sand, the moon and the planets, wishes, dreams and fantasies and the unknown world of other people. Dr. Swati Editor in Chief



To all Women Scientists who strive to make a difference in their communities, & in the world... Epigraph Honorable Her Excellency Nobelist Sirleaf Madam, “Generation of citizens in African countries particularly in rural communities have known and experienced the value of life-saving trees, fruits and plants. I applaud the 12 women scientists who have undertaken the research and produced for the first time the book \"Nature Cures\" that will validate this truth and open the doors for corporation to support the research for this natural, cost saving way, of saving and prolonging life”. Ellen Johnson Sirleaf

Capparis spinosa, Nubra Ladakh, Science Citation & Photo Courtesy: Surinder Kitchlu

Hypericum perforatum (Basant) from Kashmir, Science Citation: Surinder Kitchlu



Prologue Food IS Medicine Dr. Anne-Marie Clement Hippocrates Health Institute, Florida, USA From the beginning of time, we homo sapiens utilized food and plants medicinally. It was not until 1905 that Wil- liam Fletcher; a British physician isolated and identified the first nutrient, Thiamine (B-1). In 1928, the science of nutri- tion was born at Cornell University in the United States. Now, for a century, scientists internationally have explored, discovered, and validated the power locked into the cells of botanicals. Arguably the smartest man alive, in the twentieth century, Linus Pauling, Double Noble Laureate, stated that every disease and all aging is in some way linked to nutri- tional deficiencies. He created the field of what is now known as “orthomolecular medicine”. Most relevant to this conversation is the 65 year old institute that I have directed for the last 40 years. We practice and utilize what is today called lifestyle medicine. Where nutrition is at the core of helping people prevent and reverse disease and prolong life. Clinically, we have worked with hundreds of thousands, ob- serving the profound and successful impact that plant based nutrition and targeted food based supplements have on their recoveries. Thousands of people who have learned and ap- plied this dietary lifestyle have been medically diagnosed free of everything from Type-2 diabetes to advance cancers. What is surprising is how little time and effort has been placed on the application of nutritional science in main stream healthcare. Physicians, at best receive a handful of hours of cursory information on the subject. This lost oppor- tunity has seeded a healthy skepticism in the main stream public’s eyes about allopathic medicine. Doctors would be 13

more than happy to have a solid and viable education weaved into the pharmaceutically driven curriculum that they under- take. With such emphasis placed on writing prescriptions and magic bullets, there is no time for them to benefit from this basic and simple knowledge. Ironically, every medical physi- cian takes “the Hippocratic Oath”, where quite possibly his foremost statement was “Let food be thy medicine, and med- icine be thy food”. Today the schools that foster the knowledge for these young, enthusiastic professionals have hijacked the foundation of healthcare and would like the graduates to take an oath that sounds more like this “Let drugs be the answer in every case every time, with no excep- tion”. Rational people, which there seem to be more of these days, realize that the choices you make, will determine your health outcomes. There are certain disorders, such as Type 2 Diabetes, that are 100% diet and exercise related. Cardiovas- cular disease comes in a close second. Even America’s CDC states that 80% of all chronic disease is lifestyle related. This is all easy to correct, but will require a commitment from the powers that be (pharmaceutical Industry) to relinquish their bloodletting profits. Your future and that of humanity’s should be based upon wholesome and healthy living, and healthcare should be rele- gated to an important place when all that you do is not quite enough. Unnecessary trillions of Euros/Dollars and endless pain and suffering is endured because of our personal incom- petence and that of the professions that use no sense in their approach to wellbeing. All doctors know that the body inher- ently has the strength to heal itself. At best we can guide in- dividuals to support their immunity and physiology, even if it requires temporary medicine. Ultimately, it is not the medi- cine that heals; it is the body that finds homeostasis. All of us have had cuts or wounds, in some cases there has been pro- 14

fuse bleeding, and as you see, your cells bond together to stop the injury. Similarly, when a person is in the quest of re- versing a cancerous tumor, they support their immune sys- tem, so its numbers and strength increase ultimately aggres- sively eating up the mutagenic growth. After the disease is placed in remission, balance reoccurs and healthy cells re- place those that were killers. Excitingly, when studying the human anatomy, biology, and biochemistry, it is an enriching and awe filled endeavor. Your hundred trillion cells create every part of your being. Symbiotically working together in total harmony, allowing all systems to function and flourish as they should. What is even more remarkable is how this body is not static, and con- stantly is renewed when cells die and new stem cells replace them creating bone, organs, and all. As an example, if we test the structure of your heart and then 30 days revisit it, 100% of the cells have changed so you actually have an anatomi- cally brand new heart. This profoundly magical process re- quires nutrition, hydration, and electrical charge, which is of- ten sparked from mineralized electrolytes. There is little dif- ference between building a dwelling and that of an organ. Imagine if I decided to put weak bricks into a structure and not cement them. Before long, the wall would collapse. Do you not think the same would occur with a cell when it is not supported with the proper material? We have to rethink fu- ture medicine and humbly sit at a round table authentically expressing our real contribution. If we just allow the driving factor to be what is commonly called the “Band-Aid” ap- proach, we can expect to suffer the continuation of perpetual failure. When more than half of our youth has a chronic dis- order, whereas a quarter of a century ago it was 12%, the time has come for change. 15

Two thirds of the people enduring what was once called adult onset diabetes, are young people. It used to take 50 years or more of poor diet and living to acquire this problem, today it is happening to 10 year olds. Deaths from heart at- tacks and strokes were once expected in maturity, today it is common for youngsters to suffer such fate. Cancer will touch nearly 60% of us at some point in our life. Whereas this was at best a minor concern in 1900. Autism is predicted to infect 50% of the newborns by the early 2030’s, we can go on and on, yet the message is clear. We are out of control, our habits are killing us and mainstream medicine is not addressing the cause. Hippocrates Health Institute has had the opportunity to work with open minded participants over the decades, ob- serving the dynamic and dramatic improvements in their blood profiles and outcomes. Limitless numbers of physi- cians who have been attending these patients have called us, inquiring; most often asking, “What medicine was utilized to get this result?” Surprisingly to them, the answer is a change in habits. I dream of a day where routine modifications and healthy lifestyles are foremost in the curriculum of every cli- nician. Food is medicine, is fundamental in healing. Ac- knowledging and practicing this as a healthcare professional should not be optional. All indigenous and traditional socie- ties had their medicine men, who guided people in such mat- ters. Science has become categorical and specialized. There are different schools of thought that display unique levels of rigidity. Inflexibility arises from these dogmatic, concrete approaches, limiting the ability the caretaker has in compre- hensively helping the patient. Rarely, but on occasion, team approaches from a variety of disciplines are employed to give unlimited assistance in bringing about healing. 16

Hippocrates Institute employs a full spectrum of profes- sionals that utilize their expertise in the process of creating affirmative outcomes. In one day, our participants may be guided and counselled by nutritionists, psychotherapists, ac- upuncturists, exercise physiologists, both medical and natur- opathic doctors and nurses, etc. Food and food based sup- plements are central in all of these healthcare providers minds. It is like taking the person (rough splintery wood) and refining with guidance (sandpaper) the direction, shape, and ultimate result in the totality approach. Future care ethically should be universal and without limits. In the best case sce- narios, it is impossible to keep up with cutting edge science and proven research findings. For this reason, to advantage patients, it should be man- dated that every aspect of what makes a human life healthy and successful should be considered when a protocol is of- fered. Needless to say, what you choose to chew and swal- low, or drink is the ultimate fuel that will either help or hin- der your wellbeing. There is no doubt that physicians taken serious will have to have a full repertoire and use the team approach in caring for their clients. Hospitals in the future should have two departments, an educational department, that insurance will cover, that will teach people how to live healthfully so they will not require crisis care and long term pharmaceutical consumption. The second department should be the critical care area where accidents, necessary surgeries, and medicines when needed are employed. Food is medicine and we all better know it. Intellectually it makes total sense, but until we employ it in our own lives, we will not reach the consensus that those of us have that use plant based fare. All proteins, vitamins, minerals, essential fats, and in addition needed hormones, oxygen, phytochemi- 17

cals and enzymes come from unprocessed, unheated plant food. Every other species and creature on earth in their natu- ral habitat only consumes raw food. One of the missteps that homo sapiens have made over millennia is heating, adding chemicals to, and processing the vary sustenance that we chew and swallow. Modern advanced nutritional science leads us to know that it is on fire vegan choices that fuel the cells that make up our very bodies. We have a long way to go, but the path is well lit and clear and HHI is ready and willing to guide serious professionals via the seven decades we have been in the front line of progressive health. Each and every person has a choice, whether to endure the self- imposed status quo that suggests disease and premature aging is normal or to reject such nonsensical and unsubstantiated ideology and embrace the courageous path to a harmonious and healthy existence. Contact: Dr. Anna Maria Clement [email protected] 18

Viola biflora, Ladakh - Science Citation & Photo Courtesy: Surinder Kitchlu 19

Hypericum Perforation Photo courtesy: Surinder Kitchlu 20

Introduction The intention behind authoring this book was to be the ―change we want to see within ourselves and our communi- ty. A change, which will assure a sustainable future for our future generations; for OUR COMMON FUTURE. Nature has always been the harbinger of good practices, abundant resources, and reliable remedies, which we have been applying in our daily lives for centuries. Nature has al- ways been in balance with the ecosystem and in peaceful co- existence with humans. But the 21st century scenario is alarming. Conservation & nurturing of this ecosystem has been replaced by deforestation & over exploitation ― “Greed has replaced the Nurturing” of nature. Nature has been abused and exploited by the humans, with whom she has peacefully co-existed for centuries. COVID Pandemic is the indirect outcome of this greed and over exploitation. Time has come to reverse this trend, and probably scientist should be in the fore front of this venture. And, that's why this book was written - with the intention to educate people, how different aspects of nature balance the ecosystem & hu- manity. We all know that herbs have been discovered over 5000 years ago by ancient civilizations of the Chinese, Indian, Greek, Roman, and Egyptians, right from using them in daily chores, for curing the diseased and simply for enhancing the knowledge. Today, the most popular ethno medicinal sys- tems that use plant extracts are Chinese herbal medicine (zhongyao and pinyin), Indian (ayurveda and siddha), tradi- tional African medicine, Graeco-Arabic medicine (unani tibb), Native American medicine, classical Greek and Roman 21

herbal medicine, Japanese (kampo) (Paibir, Sheela et al, 2012). The search for new plant - derived cures should be a priority in the 21st century, towards sustainable conservation and rational utilization of biodiversity (Philipson, 1990). In India the knowledge of medicinal plants is very old whose properties are described in Rigveda and in Atharvave- da (3500-1500 B.C.) from which Ayurveda was developed. In Ayurveda, the ancient well-known scripture is the ―Charak Samhita dealing mostly with plants and while the Sushrut Samhita scripture addresses medicine as well as sur- gery. Besides India people of China, Egypt and Mesopotamia use medicinal plants for their daily use. Ancient Egyptians are known for use of plants for medicinal purposes. The Nile Valley is known as the ―Cradle of Medicine. This book therefore has made an attempt of introducing different forms of nature that contribute towards curing the aliments, such as spices, fruits, microbes, endophytes, ocean algae’s, and medicinal plants, with special focus on Himala- yas. It has explained in details the science of how these sys- tems contribute towards curing the aliments such as immune- modulation, inflammation, metabolism, allergies, infections, and energy retention. Medicinals plants have known to contain polyphenols, an- tioxidants, vitamins, phase II inducers, and potent chemopro- tectants that are thought to be major contributors to their healthy effects, and their individual nutrients are also known to modulate gene expression. The purpose of us taking multi vitamins and minerals in our diet either through food or tablets is to build our immune 22

system, which is designed to protect us from invading patho- gens and to eliminate diseases. At the time of infection, our immune system undergoes attacks from a large number of vi- ruses, bacteria and fungi. The immune system is designed to detect the pathogens by using a specific receptor to immedi- ately deliver a response by activation of immune cells, cyto- kines, chemokines and various inflammatory mediators. Functionally, an immune response can be divided into two related activities – recognition and response. Immune recog- nition is remarkable for its specificity. The immune system is able to recognise subtle chemical differences that distinguish foreign pathogens from another. Furthermore, the system is able to discriminate between foreign molecules and the body‘s own cells and proteins. Our body generates two types of immune responses – an innate response and an adaptive response. These responses can be manipulated using drugs and medications. We believe it would be safer to invest in drug molecules that are easily accessible, cheap and give fewer side effects. This is defined thoroughly in the immonomodulation chapters. Immonumodulation is basically a process which can alter the immune system of an organism by interfering with its functions. The interference results in either immunostimula- tion or enhancement of immune reactions or immunosup- pression imply mainly to reduce resistance against infections and stress which may be because of environmental or chemo- therapeutic factors. Plants provide excellent raw material for the discovery and development of novel immunomodulatory compounds. A good number of medicinal plants have stood the test of time, 23

particularly for the treatment of allergic, metabolic and de- generative diseases associated with ageing. Our interest in nature can be traced back thousands of years for their useful- ness of human kind, and this continues to the present day. Compounds and extracts derived from the biosphere have found uses in medicine, agriculture, cosmetics and food in ancient as well as modern societies around the world. There- fore, the ability to access natural products understands their usefulness and derive applications has been a major driving force in the field of natural product research. A large variety of natural bioactive plants mentioned in Ayurveda for their immunomodulating, adaptogenic and re- juvenating properties have been under study. These plants are believed to promote positive health and maintain organic re- sistance against infections by re-establishing body equilibri- um and conditioning the body tissues. Immunomodulation is a strategy for overcoming incurable auto-immune diseases involving Cancer, AIDS, Arthritis, and Allergies. An in-depth study if the immune system is sup- posed to provide both the theoretical and therapeutic back- ground of many chronic disorders. Advancement in the field of surgery has made organ transplantation possible but organ transplantation is endowed with a serious problem of organ rejection which in fact comes under the purview of immune system, proper handling of this situation through modifica- tion of immune response can give new lease of life to pa- tients with organ transplantation. In modern day life, with extreme climate changes, exten- sive exposure to industry based pollutants and xenobiotics; has resulted in emergence of a variety of immune deficien- 24

cies or hypersensitivity situations such as COVID19 Pan- demic, which calls for a changed strategy for handling the severely impacted. This calls for Scientists and Corporations to look differently in the Drug Discovery arena from plants and microbes to safeguard both Nature & Humanity. This book is a call to Science to usher in \"Medicine's of Future\" to address the 21st century challenges and invisible threats and improve our preparedness beyond the visible and fathomable. Rose Geranium; Pelargonium species from Kashmir, Science Citation: Surinder Kitchlu 25

Zanthoxylum Armatum, Udhampur (Jammu), Science Citation: Surinder Kitchlu 26

Chapter 1 Current Perspectives of Fruits as Potent Immunomodulators Dr. Anamika Khajuria Introduction The survival of an organism in this environment, teeming with multitude of potentially harmful micro-organism, de- pends upon it ability to resist widespread infections. There- fore as means of protection from the destructive effects of these organisms, we have developed an elaborate system of lymphoreticular cells, the immune system. Immunomodula- tion is the process of modifying an immune response in a positive or a negative form by administration of a drug or a compound. Recently, progress in clinical immunology has unraveled the development of immune system in the pathogenesis of several diseased conditions. The association of such diseases has a chromic and recurrent viral and bacterial infections (urie amd Duma, 1970; Turk and Bryceson, 1971; Wheelock and toy, 1973), autoimmune diseases (Tan et al., 1973; Zvaifler, 1973), Hypersensitivities and allergies (Ishizaka, et al., 1966; Ellis, 1969; Blaylock, 1976; and Ishizaka, 1978), immunodeficiency diseases (Good et al., 1962; Good and Bergsma, 1968; Hitzig et al., 1978) and cancer (Good and Finstad, 1969; Prehn, 1970; Bortin and Rimm, 1977; Rein- herz, et al., 1979) with the altered functioning of the immune system and aggravation of the defect by conventional therapy (Renoux, 1984), has led to enormous search for development of agents which could act on the immune system for the treatment of the disease of an individual. Many proteins, amino acids and natural compounds have shown significant effect to regulate immune response. 27

A large number of agents capable of modifying the rela- tionship between the disease and host’s response to disease termed as the biological response modifiers (BRM‘s) (Fenichel and Chirigos, 1984) have as a result been devel- oped (Table-I)(Reizenstein and Mathe, 1984). These agents which have also been referred to as immunomodulators, im- munostimulators or immunorestoratives in accordance with their mode of action. Researchers continue to explore the benefits of ―superfruits, a unique group nutrient-rich fruits that contain natural compounds shown to have potential of disease fighting properties in combination with cytotoxic drugs. The literature is replete with instance to their thera- peutic value. Following are the few examples of such ―superfruits dis- cussed in brief of the roles they play in curing aliments: Prunus cerasus Few fruits fall in this category and emerging science shows sour cherries isolated from P. cerasus are one among them. Cherries are rich in anthocyanins with a strong neuro- degenerative activity, and thus they can serve as a good source of bio-functional phytochemicals in our diet, provid- ing health-promising effects in humans. The principal nutri- ents thought to provide the protection afforded by fruits and vegetables are antioxidants such as vitamin C, vitamin E, β- carotene, and flavonoids (including flavones, isoflavones, and anthocyanins). Sour cherries are considered as good sources of both flavonoids and phenolic acids like anthocya- nins, etc. Convincing phytochemical research studies show that sour cherries are one of the few known food sources that are a rich source of powerful antioxidants including melato- 28

nin, quercetin, kaempherol, chlorogenic acid, p-coumaric ac- id, gallic acid, perillyl alcohol and ellagic acid. Melatonin is a powerful antioxidant considered more po- tent than vitamins C, E, and A, because it is soluble both in fat and water. A recent study published in the American Journal of Clinical Nutrition found that sour cherries ranked 14 in the top 50 foods for highest antioxidant content per serving size-surpassing well known leaders such as red wine, prunes, dark chocolate and orange juice. However, the im- munostimulatory potential of P. cerasus on immune system has not yet been explored Plants are being used since antiqui- ty for the treatment of various ailments of body. Ayurveda, one of the ancient systems of medicines, gives more stress on the prevention of the disease rather than cure. A class of herbal drugs, known as immunomodulators, alters the activi- ty of immune system through cellular/humoral or both im- mune responses. For a healthy life and to prevent occurrence of diseases it is important to strengthen the immune system. Immunosuppressants also have their major use in organ transplantation and autoimmune diseases. Prunella vulgaris It was reported that the crude drug of Spica Prunellae consists of the fruiting spikes of the perennial plant Prunella vulgaris L (Labiatae). It is having medicinal activity and used in traditionally Chinese medicine as antipyretic. As well as it was reported that it is widely used for tuberculosis, thyroid gland swell, infectious hepatitis, hypertension, jaundice, can- cer, and others in the clinical treatment. It was observed that the aqueous extract of spica prunellae showing the Inhibition of HIV-1 reverse transcriptase activity and the ethanolic ex- tract is showing immunosuppressive activity due to mixture of triterpenoids, flavonoids, tannins and polysaccharide. In 29

immunosuppressive activity experiment it has been observed that the ethanolic extract of fruit reduces the Con-A and LPS- stimulated splenocyte proliferations. On mitogen-stimulated splenocyte proliferation in-vivo assay the Con A- and OVA- induced splenocyte proliferation was found to be significant- ly and dose dependently suppressed by ethanolic extract. Randia dumetorum Randia dumetorum belongs to family Rubiaceae and also known as Mainphal (Hindi), Emetic nut (Eng). It is a small tree found in the tropical and subtropical region. It was proved by many researchers that the Plant of R. dumetorum is having anthelmentic, insecticidal, antidysentric, diaphoretic, antibacterial, antifungal, ant fertility, analgesic, anti- inflam- matory and antitumor activity and the fruits of it are consid- ered to be tonic, alterative, demulcent, diuretic and restora- tive. The drug is used for the treatment of piles, asthma, jaundice, diarrhoea, and gonorrhoea. The main active com- ponents obtained from the fruit pulp R. dumetorum are Sapo- nins. This was named as Dudumentoronin, Dumetoronin A, B, C, D, E and F etc and some hemolytic triterpenoid sapo- nins that are Randianin, Mannitol and leucoanthocyanidin, glycoside Randioside A, Beta- D-galactopyranosyl (1 - 3)- oleanolic acid. In the experiment to investigate the immuno- modulatory activity of methanol extract of R. dumetorum the humoral immune response was evaluated by humoral anti- body titre. Delayed type of hypersensitivity response was checked by increased footpad thickness. In the experiment it was observed that administration of methanol extract created dose dependent increase in thickness of footpad of mice. 30

Luffa Cylindrica Luffa cylindrica (L.) commonly known as Ghiatarui, sponge gourd, loofa, vegetable sponge, bath sponge or dish cloth gourd, belongs to family cucurbitaceouse. Young fruit can be eaten raw or cooked. The dried fruit fibers are used to remove dead skin and to stimulate the circulation. The major components of fruit are triterpenoid saponins lucyosides A, B, C, D, E, F, G, H, I, J, K, L, M, ginsenosides Re, Rg1 etc. The fruits are reported as anthelmentic, carminative, laxative, depurative, emollient, expectorant, tonic and very much use full in treatment of fever, syphilis, tumours, bronchitis, splenopathy and leprosy. It was studied that the Oral admin- istration of the ethanol extracts of fruits increases the cy- tophagic action and acid phosphatase activity of peritoneal macrophages in mice as well as it is also having the effect on interleukin 1 and 2 and tumour neurosis factor-α. Trapa bispinosa Trapa bispinosa is an aquatic floating herb belongs to the family trapaceae. The whole herb and fruit have traditionally used medicinally as a cure for various diseases. It was report- ed that the whole herb has hepatoprotective, antimicrobial, antibacterial, antitumor, antioxidant and free radical scaveng- ing activity. Further, the fruits were also reported as intestinal astringent, aphrodisiac, anti-inflammatory, in leprosy, urinary discharges, fractures, sore throat, bronchitis and anaemia and the juice of fruit has been used for diarrhoea and dysentery. In the experiment the freshly prepared solution of extract with distilled water was examined as immunomodulatory with sheep red blood cells (SRBC) as antigen. Delayed typed hypersensitivity was measured by footpad thickness, and found that the animal showed significant increase in volume of paw oedema due to production of antibodies in response to 31

the antigen. These were indicated that Trapa bispinosa has stimulatory effect on lymphocytes thus increases cell mediat- ed immunity. The aqueous extract was found to increase the humoral response as it enhanced the production of circulating antibody titre. Haemagglutination antibody titre indicated the enhanced responsiveness of macrophages and T and B lym- phocyte. The carbon clearance test showed the increase in phagocytic index thus support effect on macrophages. It also observed raised in neutrophil index. The experiments proved that T. bispinosa is potent immunostimulant. Citrus aurantifolia Citrus aurantifolia (Lime) belongs to family Rutaceae. It has been traditionally used in treatment of cold, asthma, in- fections, and joint pain. The immunomodulatory effect of concentrated lime juice extract was investigated and it was found that citrus lime juice extract exerts inhibitory effect on the lymphocyte proliferation. The experiment was carried out by production of specific polyclonal antibodies in rabbits and the in vitro immunomodulatory capacity of lime juice extract was tested on human tonsilar mononuclear cells. Phyto he- magglutinin and Staphylococcal protein A were used for ac- tivating T and B lymphocytes respectively in the culture. The experimental data also indicated that anti-citrus lime juice antibody could cancel the inhibitory effect of the extract on the proliferation of Staphylococcal protein A and Phyto he- magglutinin activated mononuclear cells. Morinda citrifolia Morinda citrifolia is a tree in the coffee family, Rubiaceae It is commonly known as great morinda, Indian mulberry, and noni in Hawaii. The scientific evidence of biological ac- tivity of the fruits is very less but on the basis of experiment it was suggested the possibility that morinda citrifolia may 32

suppress tumour growth through activation of the host im- mune system. The fruit juice of Morinda citrifolia (noni) contains a polysaccharide-rich substance (noni-ppt) It was found that Noni-ppt was also capable of stimulating the re- lease of several mediators from murine effector cells, includ- ing tumour necrosis factor-a, interleukin-1b, IL-10, IL-12, interferon-g and nitric oxide, but had no effect on IL-2 and suppressed IL-4 release. The NO production was measured by Griess reagent and Cytokines (interleukins, interferon and tumour necrosis factor) release were measured with ELISA kits. The cytokines and nitric oxide production considered that noni-ppt can induce macrophage activation. The NO, TNF-a, IL-1b and IL-12 are mainly composed of macrophag- es. The immunomodulatory effect of noni-ppt on NO, IL- 1b and TNF-a production was found to be similar to LPS. Piper longum Piper longum Linn. Commonly known as Indian long pepper, is a flowering vine belongs the family Piperaceae. It is used in traditional medicine in Asia and Pacific islands es- pecially in Indian medicine. Piper longum is a major compo- nent of medicines believes to treat gonorrhoea, menstrual pain, tuberculosis, sleeping problems, respiratory tract infec- tions, chronic gut related pain, and arthritic conditions. As well as it is also reported that Piper longum is also having analgesic and diuretic effects, relaxation of muscle tension, and alleviation of anxiety and the first isolated amide of piper species piperine was reported to display central nervous sys- tem depression, antipyretic, and anti-inflammatory activity. In the experiment it was reported that Administration of Piper longum extract and piperine showed increased number of total WBC count thus believed to stimulate the hemopoiet- ic system and increase in bone marrow cellularity and α- 33

esterase positive cells indicating its effect on stem cell prolif- eration. In the investigation of immunomodulatory effect the extract of Piper longum and piperine was found to increase the circulating antibody titer and antibody forming cells indi- cating its stimulatory effect on the humoral immune system. It is also concluded that it may activated the cytotoxic effec- tor cells, such as cytotoxic T lymphocytes, natural killer (NK) lymphocytes, macrophages, and activated neutrophils. Momordica Charantia Momordica charantia Linn often called bitter melon, bit- ter gourd or bitter squash in English, and karela in hindi. It is a tropical and subtropical vine of the family Cucurbitaceae, it widely grown in Asia, Africa, and the Caribbean for its edi- ble fruit, which is among the most bitter of all fruits, Its vari- ety of medicinal activity is reported such as antidiabetic, anti- tumor and immunomodulating properties. In the results ob- tained from this study indicated that 80% ethanolic extract of M. charantia, showed a strong decrease of T-and B-cell pro- liferation and the effect was more potent than that for the wa- ter extracts. The decreased count of lymphocytes might be due to toxicity of the extracts, which was observed both in the presence and absence of mitogen. Punica granatum Punica granatum L. is a fruit-bearing deciduous shrub or small tree belonging the family of Punicaceae. The pome- granate tree typically grows 12-16 feet, has many spiny branches, and can be extremely long lived. Pomegranate (punica grantum) is used in several systems of medicine for a variety of disease. In Indian Ayurvedic medicinal system the pomegranate is considered ―a pharmacy unto itself and rec- ommended for used as an antiparasitic agent, a blood tonic, and to heal aphthae, diarrhoea, and ulcers. Pomegranate also 34

use as a remedy for diabetes in the Unani system of medi- cine. In the experimental study the aqueous suspension of fruit rind powder of punica grantum was found to stimulate the cell-mediated and humoral components of the immune system in rabbits. It showed increase in antibody titer to ty- phoid -H antigen. It was reported that treatment of punica grantum fruit extract increases the percent of T cell count compared to control Group. In delayed type hypersensitivity test the alcoholic extract of fruit was observed to produce significant decrease in paw swelling which suggests cell me- diated immune response. The level of IgG was also observed higher in Punica granatum treated animals. Thus, the punica grantum fruit is examined as potent immunostimulant. Withania somnifera Withania somnifera (L.) Dunal commonly known as Ashwagandha, Indian ginseng, winter cherry, poison goose- berry, or winter cherry, belong from the family solanaceae (nightshade family). It is a most useful medicinal plant which is having many therapeutic properties and use in the different traditional Indian systems of medicines such as Ayurveda, Unani and Siddha. On the basis of experiment results the ethanolic extract of Withania somnifera was found to enhance humoral immune as well as cell mediated immune response. The methanolic extract of the plant was studied on the hematological parame- ters and showed showed an increase in number of RBCs but a decrease in WBCs compared to the control mice. [50,7] Solanum xanthocarpum Solanum Xanthocarpum is one of the members of the dashmula (ten roots) of the Ayurved and commonly known as Kantkari belongs from the family of Solanaceae. It is 35

commonly found throughout India and has been used in many ayurveda formulations. It is believed that the whole plant showing the improvement in some respiratory disease. The fruits of the solanum xanthocarpum exerts their action in immunomodulatory activity as it was found that it shows its methanol extract stimulated the action of immune as well as increase percent neutrophil. Annona squamosa Annona squamosa, commonly known as sugar apple, cus- tard apple, sweetsop is prominent edible fruit but also it is uses in the treatment of anaemia, malignant tumours, burning sensation, cough and for strengthening muscles. The im- munomodulatory activity of Annona squamosa was investi- gated and it was observed that it shows significant increase in B cell count. The action on macrophages, T and B lympho- cytes were also reported after the treatment of extract of an- nona squamosa. The treatment enhances the activity with the response of sheep red blood cells as antigen. On the basis of delayed type hypersensitivity reaction the paw swelling was found to be decreased on the treatment thus suggests cell mediated immune response and the level of IgG increased. Cucumis melo Cucumis melo commonly known as muskmelon, melon. it is recommended as moisturizer for the skin, stomach ache and also used in the first aid treatment for burns. The im- munomodulatory activity was investigated in the experiment on mice paw, and was observed that extract of cucumis melo exerts no increase in paw volume. Thus, significant decrease in paw swelling suggests the cell mediated immune response. The level of IgG was higher in Cucumis melo treated ani- mals. The fruit showed immunomodulating activity in de- layed hypersensitivity reaction the fruit extract does not in- 36

crease after 24 hrs because B cell involved in the reaction and the fruit extract suppress the antigenic cell mediated im- mune response. [8] Emblica Officinalis Emblica officinalis (Amla) belongs to family Euphorbia- ceae and are widely used in the Indian system of medicine. According to believe in ancient Indian mythology, it is the first tree to be created in the universe. It is also known as Amla, Phyllanthus Emblica or Indian gooseberry. It is most reviewed plant and reported as action and treatment on can- cer, diabetes, liver treatment, heart trouble, ulcer, anaemia and various other diseases. Similarly, it also has property as antioxidant, immunomodulatory, antipyretic, analgesic, cyto- protective, antitussive and gastroprotective. As well as it is very useful in memory enhancing, ophthalmic disorders, lowering cholesterol level, neutralizing snake venom and as an antimicrobial. E.officinalis has been reported to inhibit chromium- induced free radical production and also having the inhibitory action on apoptosis and DNA fragmentation induced by chromium. In immunomodulatory activity signif- icant increase in total WBC count, percentage lymphocyte distribution was observed. Emblica officinalis contribute to the regulation of both humoral and cellular immune respons- es. immunostimulatory activity of Emblica officinalis which might be due to its combined action on humoral and cell me- diated immune responses along with macrophage phagocyte. [39, 30, 23] Litchi chinensis Sonn Litchi chinensis Sonn commonly known as litchi is a pop- ular fruit of Southeast Asia, especially China. Its pericarp is about 15% of the total weight of fruit which contains a good quantity of flavonoids. Much of the pharmacological activity 37

of litchi is supposed to be due to its flavonoids contents as flavonoids play important role against cardiovascular diseas- es, cancer, inflammation & allergy. Water soluble ethanolic extract of litchi fruit pericarp reported to have good anti- cancer, activity against human breast cancer & hepatocellular carcinoma in-vitro & in-vivo. The ethyl acetate fraction of li- tchi pericarp mainly contains epicatechin, Proanthocyanidin B2 & Poranthocyanidin B4. The ethyl acetate fraction of the litchi fruit were examined using splenocyte proliferation of mouse splenocyte. The results showed that the ethyl acetate fraction is having good immunostimulatory activity. At the lowest concentration of the study i.e. 12.5 µg/ml, significant- ly (P<0.05) stimulatory effect on splenocyte proliferation was observed. Prunus cerasus Prunus cerasus is a rich source of flavonoids & phenolic acids like anthocyanins. It is a source of powerful antioxidant which includes melatonin, quercetin, kaempherol, chlorgenic acid, p-coumeric acid, galic acid, perillyl alchol, & ellagic acid. Recent study reveals that Prunus cerasus ranked 14 in the top 50 foods for highest antioxidant contents. It is a me- dicinal fruit which is having a number of therapeutic uses in- cluding anti-inflammatory & immunological effects. The in vivo antibody titer studies carried out on the balb-c mice suggests that there is an increase in the anti SRBC antibody on the mice treated orally with ethyl acetate fraction. There is a dose related increase in delayed type hypersensitivity reac- tion after 24 & 48 hr. proliferation of lymphocytes signifi- cantly increased as compared with control. There is an in- crease in the macrophage activity & nitric oxide production expressing the increase in the phagocytic activity of macro- phages treated with the ethyl acetate fraction. Increasing dose of the extract significantly increase the release of TNFα from 38

peritoneal macrophages. There is a dose dependent increase in the TH1 (IFNϒ, TNFα) & TH2 (IL4) cytokines. Conclusion There are numbers of natural agents which was used from the ancient time to treat or cure the pathological and diseased condition some are used as crude drug and many of it are used as various formulations from the plant extract. Fruits are only edible part of the plants from where the therapeutic ef- fect is obtained directly. The immunomodulators reported from the various fruits are very much useful for further study many of the fruits are very much known to the daily life and some of them can be used as different formulations. References: Appendix I - page 179 Contact: Dr. Anamika Khajuria [email protected] 39

Podophyllum Hexandrum from Zanskar, Ladakh, Science Citation: Surinder Kitchlu 40

Ephedra Gerardiana, Nubra (Ladakh) Science Citation: Surinder Kitchlu Artemisia biennis, Leh (Ladakh), Science Citation: Surinder Kitchlu 41

Euphorbia characias, Khardungla: Ladakh; Science Citation: Surinder Kitchlu 42

Chapter 2 Ex Situ Approaches Medicinal Plant Biodiversity Dr. Sushma Koul Introduction: If we try to define Biodiversity, it refers to the variety and variability of living materials and ecological complexes in a given area and comprises species, genetic, and ecosystem di- versity. Diversity can be at level of Molecular sequences, Gene, Individuals, Populations, Species, Higher taxa, Habi- tats, Ecosystems or Biomes. India is one of the twelve mega- diversity countries in the world (namely Mexico, Columbia, Equador, Peru, Brazil, Zaire, Medagascar, China, India, Ma- laysia, Indonesia and Australia). Ecosystem wise, India has 42 vegetation types, 16 major forest types, 10 biogeograph- ical zones and 25 hot spots of endemic centers. India along with being a mega diversity centre is also centre of origin for many crops. It holds 2.4% land area and encompasses 8% world’s biodiversity. Among plant rich countries it stands fourth among Asian countries. In India, Himalaya is a store house of endemics and hot spots of plant biodiversity. Rea- son being: • Compression of thermal zones and fragmentation of land- scape into multitude of microhabitats inhibiting suite of spe- cies. In addition: • Steep terrain and rough climate in combination with severe land use pressure cause high altitude ecosystems to rank among the most endangered ecosystems worldwide. 43

1. MEDICINAL PLANT BIODIVERSITY Medicinal plants have always been a basic resource for human health. Appreciation for the preventative and thera- peutic value of herbal remedies, and the additional benefits of their low cost, wide accessibility, and cultural relevance remains strong in many traditional cultures Out of so far de- scribed 17,000 higher plant species 9000 are medicinal plants and 25-30 % are endemic to Himalayas. Increased consump- tion of medicinal plants, through expansion of local, regional, and global markets, has increased pressure on natural re- source that is largely harvested from depleted wild popula- tions in shrinking wild habitats. About 70% of India‘s popu- lation depends on plants for primary health care and 75% of world‘s raw material (low value) for plant based drugs comes from Indian plants. With the advent of a market-orientated economy, the medicinal plants wealth of the Himalaya has become a high price commodity. Loss of Biodiversity can be attributed to following major causes: • Over Exploitation • Climatic Changes • Increasing population • Habitat destruction • Spread of certain alien weeds affect native flora. • Practice of shifting agriculture by local people. • Selective harvesting of specific groups of plants for economic benefit. • Developmental activities - Tourists influx - Low priority for conservation 2. CONSERVATION OF BIODIVERSITY Interest in and demand for traditional remedies and other plant-based health products (the so-called \"botanicals\") is in- creasing worldwide, particularly in rapidly expanding urban 44

societies. Trade of medicinal plants is expected to rise to US$ 5 trillion by year 2050 (Sharma, 2003). The key question at the forefront is that the number of species threatened with ex- tinction far outstrips the scientific response to the challenge of large-scale extinctions ahead. Conservation of Plant Ge- netic Resource (particularly the plants with narrow genetic base/endemics) can help posterity by preventing the further loss and, at the same time, will help to meet the demands of pharmaceutical companies and herbal healers. This is evident in the vision and mission statement of World Health Organization (WHO) on health improvement and in community-based conservation initiatives by interna- tional organizations, including the World Bank, the Interna- tional Development Research Centre (IDRC) and UNDP. In a country like India both TSM and the folk knowledge sys- tems of medicine are of significance TSM exists in the form of well-known classical traditions of Ayruevda, Unani and Siddha, which are characterized by a large number of practi- tioners trained through formal institutions, a well-codified body of texts either in the form of the scriptures or other written forms and an official recognition as 'Indian Systems of Medicine (Singh, 2004). International organizations like UNDP, IDRC, OXFAM, WHO, Ford Foundation and the World Bank are the leading sources of funding for broad-based programs dealing with medicinal plant conservation and development in India and Asia. At a national level, the Department of the Indian Sys- tems of Medicine (Health) and the Medicinal Plant Boards are the most relevant government agencies that deal directly with the medicinal plant sector and associated local knowledge systems. Further, their direction, design and de- livery have largely been limited to the state level. At the sub - 45

state level (district, block or taluka and village), the pro- grammatic interventions related to medicinal plant and asso- ciated local knowledge systems are organized by local NGOs and community groups. In addition various departments in Universities / organizations like Department of Bio - Technology (DBT) / CSIR / DRDO / Ministry of Environ- ment & Forest (MoEF) have surveyed Himalayan regions for documenting information and development of conservation strategies for these plants. Goals of World Conservation Strategy can be summarized as follows: • Maintenance of essential ecological process and life support systems on which human survival and economic activities depend. • Preservation of species and genetic diversity • Sustainable use of species and ecosystems, which support millions of rural communities as well as major industries. Advances in biotechnology have generated new opportu- nities for genetic resources conservation and utilization (Rao 2004). Techniques like in vitro culture and cryopreservation have made it easy to collect and conserve genetic resources, especially of species that are difficult to conserve as seeds. The conservation of biodiversity can be achieved through an integrated approach balancing in situ and ex situ conserva- tion strategies. Purposes of these strategies are: • Rescue threatened germplasm 46

• Produce material for conservation biology research • Supply material for various purposes to remove or reduce pressure from wild collecting • Grow those species with recalcitrant seeds that cannot be maintained in a seed store • Make available material for conservation studies • Produce material for reintroduction, reinforcement, habitat restoration and management The in-situ preservation of species offers all the ad- vantages of allowing natural selection to act which cannot be recreated ex situ. But habitat destruction is inevitable and important. Endangered species need to be preserved and ex situ conservation provides an opportunity. In Himalayan region a number of protected areas- biosphere reserves, national parks and wildlife sanctuaries are in existence and can be created more. For ex situ conser- vation in Himalayan region, the need of seed, organ, tissue or gene banks is emphasized, although these can be established at minimal cost because of the proximity of glaciers in the region (Khoshoo 1996). 3. MODUS OPERANDI FOR EX SITU CONSERVA- TION 3.1 Planning Planning is the first step in the management process. The scientific and organizational mission should be well defined in terms of goals of the gene bank, i.e. short, medium or long-term. The gene bank should enlist the help of different 47

specialists/advisory committees to deal with specialized sub- jects, such as individual crops, methods of conservation and documentation. Well defined delegation of responsibilities, long range plans and contingencies for emergencies or urgent needs must be prepared in advance. The importance of stand- ard procedures and protocols in this regard should not be overlooked. 3.2. Germplasm Collection Collecting involves gathering samples of a species from populations in the field or natural habitats for conservation and subsequent use. The unit of collection may be botanic seeds or vegetative propagules, depending on the breeding system of the species. Collecting may be easy in species pro- ducing small botanic seeds in abundance. However, it be- comes problematic when seeds are unavailable or non-viable due to damage of plants by grazing or diseases; large and fleshy seeds that are difficult to transport; or where samples are not likely to remain viable during transportation due to remoteness of the collecting site from the gene bank. Ad- vances in biotechnology provide useful solutions for collect- ing such problem species (Karp, 2002). 3.3. Conservation Procedures: Involves conservation outside the native habitat and is generally used to safeguard populations in danger of destruc- tion, replacement or deterioration. Approaches to ex situ con- servation include methods like seed storage, field gene banks and botanical gardens. Vegetative propagules in form of tissue or cell cultures, DNA and pollen storage also contribute indirectly to ex situ conservation. 48

3.3.1 Field gene banks: Field gene banks or living collections are the main con- servation strategy for long-lived perennials, recalcitrant spe- cies and vegetatively propagated species. It is an assemblage of diverse plant species and their range of genetic diversity in an area. The plant materials are conserved and are available for breeding, reintroduction, research and other purposes. This method is useful for long living perennials trees and shrubs. Field gene banks have an important educational value and can easily be accessed for research purposes. Their main lim- itation is that they take a great deal of space and are difficult to maintain and protect from natural disasters. They are sus- ceptible to the spread of diseases and may suffer from ne- glect. Furthermore, out-breeders require controlled pollina- tion for regeneration from seed. In many circumstances they are the only available option for the conservation of im- portant germplasm. 3.3.2. Seed Banking Among the various ex situ conservation methods, seed storage is the most convenient for long-term conservation of plant genetic resources especially for sexually reproducing plants (Phartyal et al 2002). The main advantage of seed banking is that it allows large populations to be preserved and genetic erosion to be minimized by providing optimum conditions and reducing the need for regeneration. The ac- tivities in seed banks should take the following sequence: collection, seed preparation, seed drying, packaging, storage, periodic germination tests, seed regeneration, re-storage and documentation at each stage of activity. The preferred stand- ards for long-term seed storage of orthodox species to dry the seeds to a moisture content of below 7% and seal the dried 49

seeds in a moisture-proof container such as laminated foil bags, aluminum cans or glass jars for storage at a low tem- perature of -18ºC. However, because less is known about wild species, a temperature of -4ºC and a moisture content of 7-8% is advisable to begin with. However, when a natural population still exists, it may be advisable to re-collect rather than regenerate a new supply from the previous collection as damage can occur such as mutations associated with the loss of viability during storage. Clearly this is only applied to true orthodox species because it involves desiccation of seeds to low moisture contents and storage at low temperatures. However, there are a large num- ber of important tropical and sub-tropical tree species which produce recalcitrant seeds that quickly lose viability and do not survive desiccation, hence conventional seed storage strategies are not possible. 3.3.3. DNA Banks DNA banking could constitute a complementary conser- vation strategy for safeguarding the genetic diversity of a crop's gene pool, especially if combined with in vitro conser- vation or cryopreservation (De Vicente et al 2006). It is an- other form of biological resource that offers tremendous op- portunities of practical and academic value is the DNA. In fact, the concept of DNA as a gene bank resource has emerged out of the revolution in genomic information brought about by the analysis of DNA extracted from numer- ous plant species in laboratories across the world. DNA is a highly stable molecule; degradation kinetics models suggest that fully hydrated DNA kept at room temperature takes about 10,000 years to depolymerases into small fragments. However, degradation due to presence of endonucleases and 50