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1.0 Introduction

1.1 Medical treatment

Modern medical treatments use the latest research to combat disease.

Medicine is the applied science or practice of the diagnosis, treatment,

and prevention of disease [1]. It encompasses a variety of health care

practices evolved to maintain and restore health by the prevention and

treatment of illness in human beings.

Contemporary medicine applies health science, biomedical research,

and medical technology to diagnose, treat injury, and disease,

typically through medication or surgery, and through therapies as

diverse as psychotherapy, external splints, and traction, prostheses,

biologics, ionizing radiation, and others. The word medicine is derived

from the Latin airs medicine, meaning the art of healing [2, 3].

1.2 Classification of Medical Treatment

Medical treatment may be classified in two categories.

1.2.1 Allopathic medicine

German physician Samuel Hahnemann (1755-1843) invented the term

“allopathy”. “Allopathy” is derived from the Greek words “allo,”

(meaning “other”), and “pathos” (meaning “suffering”). A system of

medical practice that aims to combat disease by use of remedies (as

drugs or surgery) producing effects different from or incompatible with

those produced by the disease being treated by homeopathy. Samuel

Hahemann, the founder of homeopathy, coined the term “allopathy.” It

is not a good way of treatment for the diseases because any disease

cured by allopathic way of medication may also have some side effects

of that cure or any other symptoms. Because allopathic medication

does not remove the disease or does not kill the germs, it pushes them

inside the body so patient feels that it is covered or removed from the

body but in real it become stronger and attacks again after a time

interval with high proportion. Antibiotics and different type of agents

are used to cure the diseases. These antibiotics produce effects

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different from the disease that is being treated. Allopathic medication

killing people day by day because of different type of side effects, and

it creating other diseases that do not exist yet. It is not a permanent

way of medication to treat the diseases or to remove them from the

body of suffering patient.

Though, the allopathic medicine is working in a wide range for the

treatment of different kind of diseases because it is faster way of

treatment than other alternative medicine but it has many side effects

[4].

1.2.2 Alternative Medicine

The major schools of alternative medicine in India are Ayurveda,

Unani, Naturopathy, Homeopathy, Yoga, and Meditation. Ayurveda

traces its roots to 5,000 years back and is now fast catching up as the

medical treatment of choice with urban people, who have not found

satisfaction with allopathic treatments.

Naturopathy refers to the treatment of various mental and physical

disorders using natural methods, like magnet therapy, mud therapy,

osteopathy etc. Naturopathy stresses healthy dietary and living habits.

Yoga and meditation developed in India thousands of years ago. One

of the most widely practiced of alternative medicine in India; Yoga has

gained popularity in the West due to the extremely effective therapies

and its offers to problems ranging from weight gain to cardiac

disorders [5, 6].

Alternative medicine may be classified in two categories.

I. Homeopathy

II. Unani

III. Herbal

1.2.2.1 Homeopathy Treatment

Homeopathy, like Naturopathy, is another system of alternative

medicine in India that has its roots in the West. The pioneer in the

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field of homeopathy is Samuel Hahnemann, who postulated in 1796

that "like cures like", known as the principle of similar.

Like other systems of alternative medicine in India, homeopathy is a

system of holistic medicine. Homeopathy treatments include a single

medicine to treat all disorders of the body. Since, it is believed that

entire body functions as a single entity, and a disorder in one system

of the body causes a disturbance in the other system. Homeopathy

practitioners believe that mere symptomatic treatment will not cure

the ailment. Instead, the whole body is treated to bring back the

harmony of the various systems.

The medicines used in homeopathy are sourced from natural sources

like animals, plants, minerals etc. The substances are diluted and

then various doses are prescribed based on the practitioner's

diagnosis. Homeopathy is said to have cured diseases like asthma,

measles, benign nodules, and cysts. India presently has many

registered practitioners of homeopathy. The Central Council for

Research in Homeopathy in New Delhi, India, and the Foundation for

Homeopathic Research in Mumbai are some of the premier institutes

offering homeopathy treatments in India [7].

1.2.2.2 Unani Treatment

The Unani system of medicine traces its origins to ancient Greece.

However, it was the Muslims "Hakims" of India, who developed it

further into an effective system of medicine. The Hakim Ibn Sina is

considered the founder of Unani medicine.

One of the most well developed fields of alternative medicine in India;

the Unani offers cures from cardiac problems to problems like hair

loss, weight gain, and gum bleeding. India has over 100 hospitals

specializing in the system of Unani medicine. The country also has

30,000 registered practitioners running private Unani clinics. The

Tibbiya College at Aligarh and Ahmedabad are considered centres of

excellence for Unani medicine.

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The Unani therapies are based on the use of various herbs, and

medicinal plants that have been used for centuries in traditional

medicine in India. The basic principles of Unani medicine are similar

to those of Ayurveda, and state that the well being of the body

depends on the harmony between the Humors [8-10].

1.2.2.3 Herbal Treatment

Ayurveda: Theory for herbal medicine

Definition

Ayurveda originated in India several thousand years ago. The term

“Ayurveda” combines the Sanskrit words “ayur” (life), and “veda”

(science or knowledge). Thus, Ayurveda means “the science of life.”

According to Charaka, ayurveda is knowledge, which seeks to weigh of

life in the scales of wholesomeness and happiness against their

opposites. Its main themes of health, disease, and recovery of health

from disease take the stage against an inspiring background of

intuitive philosophy, lofty idealism, and vivid compassion, which are

the hallmarks of India’s cultural inheritance [11, 12].

Ayurveda is a type of healing craft practiced in India from the ancient

era. Ayurvedic practitioners rely on plant extracts, both “pure” single-

plant preparations, and mixed formulations. The preparations have

lyrical names, such as Ashwagandha (Withania somnifera root),

Cauvery 100 (a mixture), and Livo-vet. These preparations are used to

treat animals as well as humans. In addition to their antimicrobial

activities, they have been found to have ant diarrheal,

immunomodulatory, anticancer, and psychotropic properties [13-18].

In-vivo studies of abana, an ayurvedic formulation, found a slight

reduction in experimentally induced cardiac arrhythmias in dogs [19].

Two microorganisms against, which ayurvedic preparations have

activity are aspergillus spp. [20], and propionibacterium acnes [21].

The toxicity of ayurvedic preparations has been the subject of some

speculation since, some of them include metals. Prpic-Majic et al. [22]

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identified high levels of lead in the blood of adult volunteers, who had

self-medicated with ayurvedic medicines [23].

Propolis is a crude extract of the balsam of various trees; it is often

called bee glue since, honeybees gather it from the trees. Its chemical

composition is very complex; like the latexes described above,

terpenoids are present as well as flavonoids, benzoic acids, esters,

substituted phenolic acids, and their derivatives [24]. Synthetic

cinnamic acids, identical to those from propolis, are found to inhibit

hem agglutination activity of influenza virus [25].

1.2.2.3.1 History of medicine

All human societies have medical beliefs that provide explanations for

birth, death, and disease. Throughout history, illness has been

attributed to witchcraft, demons, astral influence or the will of the

Gods. These ideas still retain some power with faith healing, and

shrines still used in some places although the rise of scientific

medicine over the past millennium has altered or replaced mysticism

in most cases [26].

The ancient Egyptians had a system of medicine that was very

advanced for its time, and influenced later medical traditions. The

Egyptians and Babylonians both introduced the concepts of diagnosis,

prognosis, and medical examination. The Hippocratic Oath, still taken

by doctors today, was written in Greece in the 5th century BCE. In the

medieval era, surgical practices inherited from the ancient masters

were improved, and then systematized in Rogerius's “the practice of

surgery”. During the Renaissance, understanding of anatomy

improved, and the invention of the microscope would later lead to the

germ theory of disease. These advancements, along with developments

in chemistry, genetics, and lab technology (such as the x-ray) led to

modern medicine [27].

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1.2.2.3.2 Introduction of herbal medicines

Herbal medicines are as old as mankind. Since, ancient time’s man

has adopted remedies from nature. During 19th century a number of

natural products were isolated, and subjected to detailed investigation

of their structure, and pharmacological action. A review of ancient

literature reveals that many drugs of the vegetable, mineral, and

animal origin are being traditionally used for the treatment for the

various diseases [28-32]. This involved the application of chemistry to

isolate the active constituents in medicinal plants, resulting in next

few centuries in the discovery of quinine, caffeine, morphine etc.

Herbalism is a traditional medicinal or folk medicine practice based on

the use of plants, and plant extracts. Herbalism is also known as

botanical medicine, medical herbalism, herbal medicine, herbology,

and phytotherapy.

Herb plants produce, and contain a variety of chemical compounds

that act upon the body, and are used to prevent or treat disease or

promote health, and well being. A herbalist is a

professional trained in herbalism, the use of

herbs (also called botanical or crude medicine) to

treat others. The first Chinese herbal book, the

Shennong Bencao Jing, compiled during the Han

Dynasty but dating back to a much earlier date,

possibly 2700 B.C., lists 365 medicinal plants,

and their uses-including Ma-Huang, the shrub that introduced the

drug ephedrine to modern medicine. Sometimes, the scope of herbal

medicine is extended to include fungi, and bee products as well as

minerals, shells, and certain animal parts [33, 34]. People on all

continents have used hundreds to thousands of indigenous plants for

treatment of ailments since prehistoric times.

From the middle ages on, many practitioners have tried to classify

herbal remedies by observation of their effects. This is closer to the

modern scientific approach of gathering evidence. Eastern herbal

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medicine still adheres to the mystical approach in its theories whilst

Western herbalists tend to use herbs for the ingredients they contain;

mixing, and matching them in the way that conventional medicine

does with modern drugs.

The study of herbs dates back over 5,000 years to the Sumerians, who

described well-established medicinal uses for such plants as laurel,

caraway, and thyme. Ancient Egyptian medicine of 1000 B.C. are

known to have used garlic, opium, castor oil, coriander, mint, indigo,

and other herbs for medicine and the Old Testament also mentions

herb use, and cultivation including mandrake, vetch, caraway, wheat,

barley, and rye.

The Greek physician compiled the first European treatise on the

properties, and uses of medicinal plants, De Materia Medica. In the

first century AD, Dioscorides wrote a compendium of more than 500

plants that remained an authoritative reference into the 17th century.

The ancient Greeks and Romans made medicinal use of plants. Greek

and Roman medicinal practices, as preserved in the writings of

Hippocrates. Similarly, important for herbalists and botanists of later

centuries was the Greek book that founded the science of botany,

Theophrastus’ Historian Planetarium written in the fourth century

B.C.

In some cases, herbal medicines offer an inexpensive and safe

alternative to pharmaceuticals. In the U.S., which has just 4% of the

world's population, 106,000 patients died from, and 2.2 million were

seriously injured by adverse effects of pharmaceuticals in 1994, as

reported in Journal of the American Medical Association.

Many of the pharmaceuticals currently available to physicians have a

long history of use as herbal remedies, including opium, aspirin,

digitalis, and quinine. The World Health Organization (WHO)

estimates that 80 percent of the world's population presently uses

herbal medicine for some aspect of primary health care. However,

most herbalists concede that pharmaceuticals are more effective in

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emergency situations where time is of the essence [35, 36]. Within the

past decade, Americans have consumed ever-increasing amounts of

traditional Chinese herbs, and formulas. Some of these consumers are

under the care, and guidance of practitioners, who have received

specific training in the use of Chinese herbal preparations. However,

many receive haphazard advice from both health practitioners, and

lay people, who have no experience or training.

In the case of Chinese herbal knowledge, its use by people unfamiliar

with its rules, and protocols invariably leads to mishaps: either the

herbs or formulas fail to work as expected or worse side effects may

result whenever herbs are used in contraindicated conditions. Whilst

there is undoubtedly merit in testing plants for beneficial compounds

they may contain, it is through a truly scientific approach that these

benefits will be realized.

Herbal medicine is the oldest form of health care known to mankind.

Herbs had been used by all cultures throughout history. It was an

integral part of the development of modern civilization. Primitive man

observed and appreciated the great diversity of plants available to

him. The plants provided food, clothing, shelter, and medicine. Much

of the medicinal use of plants seems to have been developed through

observations of wild animals, and by trial and error. As time went on,

each tribe added the medicinal power of herbs in their area to its

knowledgebase [37, 38]. They methodically collected information on

herbs and developed well-defined herbal pharmacopoeias. Indeed, well

into the 20th century much of the pharmacopoeia of scientific

medicine was derived from the herbal lore of native peoples. Many

drugs commonly used today are of herbal origin. Indeed, about 25

percent of the prescription drugs dispensed in the United States

contain at least one active ingredient derived from plant material.

Some are made from plant extracts; others are synthesized to mimic

natural plant compounds [39]. Substances derived from the plants

remain the basis for a large proportion of the commercial medications

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used today for the treatment of heart disease, high blood pressure,

pain, asthma, and other problems. For example, ephedra is herb used

in Traditional Chinese Medicine for more than two thousand years to

treat asthma and other respiratory problems. Ephedrine, the active

ingredient in ephedra, is used in the commercial pharmaceutical

preparations for the relief of asthma symptoms and other respiratory

problems. It helps the patient to breathe more easily. Another example

of the use of herbal preparation in modern medicine is the foxglove

plant. This herb had been in use since 1775. At present, the powdered

leaf of this plant is known as the cardiac stimulant digitalis to the

millions of heart patients it keeps alive worldwide.

Herbal Medicine can be broadly classified into various basic systems:

Traditional Chinese Herbalism, which is part of Traditional Oriental

Medicine, Ayurvedic Herbalism, which is derived from Ayurveda, and

Western Herbalism, which is originally came from Greece and Rome to

Europe, and then spread to North and South America.

Chinese and Ayurvedic Herbalism have developed into highly

sophisticated systems of diagnosis, and treatment over the centuries.

Western Herbalism is today primarily a system of folk medicine.

Interest in the United States had been growing in the recent years

from the reported success stories from the use of herbs. For example,

St. John's Wort is widely used in the treatment of mild depression

without the need for Prozac. St. John's Wort does not have the side

effects such as that of Prozac. There are some Ayurvedic herbs that

are very useful for reducing cholesterol, diabetes etc. Similarly, the

popularity of Ginseng and Ginkgo biloba (ginkgo) is rising due to its

beneficial effects.

1.2.2.3.3 Timeline of Herbal Medicine

No one knows, for sure, when humans began using herbs for

medicinal purposes. The first written record of herbal medicine use

showed up in 2800 B.C. in China. Since, then the use of herbs has

gained, and fallen out of, favor many times in the medical field. The

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timeline that follows shows some of the key dates and major points in

the history of herbal medicine.

2800 B.C.- The first written record of herbal medicine use showed up.

(Titled the Pen Ts'ao by Shen Nung)

400 B.C.- The Greeks joined the herbal medicine game. Hippocrates

stressed the ideas that diet, exercise and overall happiness formed the

foundation of wellness.

50 A.D.- The Roman Empire spread herbal medicine around the

Empire with the commerce of cultivating herbs.

200 A.D.- The first classification system that paired common illnesses

with their herbal remedy appeared. This was prepared by the herbal

practitioner Galen.

800 A.D.- Monks took over the herbal field with herbal gardens at

most monasteries and infirmaries for the sick and injured.

1100 A.D.- The Arab world became a center of medicinal influence.

Physician Avicenna wrote the Canon of Medicine, which gave mention

to herbal medicines.

1200 A.D.- Black Death spread across Europe, and herbal medicines

were used alongside “modern” methods such as bleeding, purging,

arsenic, and mercury with equal or better results.

1500 A.D.- Herbal medicine, and herbalists were promoted, and

supported by Henry VII, and the Parliament due to the large number

of untrained apothecaries giving substandard care.

1600 A.D.- Herbs were used in treating the poor, while extracts of

plant, minerals, and animals (the “drugs”), were used for the rich. The

English Physician, an herbal explaining the practice of herbal

medicine was written during this time.

1700 A.D.- Herbal medicine got another high profile endorsement

from Preacher Charles Wesley. He advocated for sensible eating, good

hygiene, and herbal treatments for healthy living.

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1800 A.D.- Pharmaceuticals began to hit the scene, and herbal

treatments took a back seat. As side effects from the drugs began to

be documented, herbal remedies came into favor again. The National

Association of Medical Herbalists was formed, and later renamed the

National Institute of Medical Herbalists (NIMH.)

1900 A.D.- Lack of availability of drugs during World War I increased

the use of herbal medicines again. After the war pharmaceutical

production increased and penicillin was discovered. Herbal

practitioners had their rights to dispense their medications taken

away, and then reinstated. The British Herbal Medicine Association

was founded, and produced the British Herbal Pharmacopoeia. People

began to express the concern over the large number of side effects and

environmental impact of the drugs of the 1950s.

2000 A.D.- EU took action on regulation, and testing of herbal

medicines similar to those used for pharmaceuticals. Herbal

medicines have been documented for almost 4000 years. These

medicines have survived real world testing, and thousands of years of

human testing. Some medicines have been discontinued due to their

toxicity, while others have been modified or combined with additional

herbs to offset sideeffects. Many herbs have undergone changes in

their uses. Studies conducted on the herbs and their effects keep

changing their potential uses.

1.2.2.3.4 Plant derived drugs

Medicinal plants play an important role in the development of potent

therapeutic agents [40]. In the USA deserpidine, reseinnamine,

reserpine, vinblastine, and vincristine in all over the world, ectoposide,

eguggulsterone, teniposide, nabilone, plaunotol, zguggulsterone,

lectinan, artemisinin and ginkgolides, paciltaxel, toptecan, gomishin,

and irinotecan are the drugs derived from higher plants. Plant based

drugs provide outstanding contribution to modern therapeutics; for

example; serpentine isolated from the root of Indian plant Rauwolfia

serpentina in 1953, was a revolutionary event in the treatment of

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hypertension, and lowering of blood pressure. Vinblastine isolated

from the Catharanthus rosesus is used for the treatment of hodgkins,

choriocarcinoma, non-hodgkins lymphomas, leukemia in children,

testicular, and neck cancer [41]. Vincristine is recommended for acute

lymphocytic leukemia in childhood advanced stages of hodgkins,

lymophosarcoma, cervical, and breast cancer. Phophyllotoxin is a

constituent of Phodophyllum emodi currently used against testicular,

small cell lung cancer, and lymphomas. Plant derived drugs are used

to cure mental illness, skin diseases, tuberculosis, diabetes, jaundice,

hypertension, and cancer.

In the Western world, as the people are becoming aware of the

potency, and side effect of synthetic drugs, there is an increasing

interest in the natural product remedies with a basic approach

towards the nature. Throughout the history of mankind, many

infectious diseases have been treated with herbs. A number of

scientific investigations have highlighted the importance, and the

contribution of many plant families i.e., Asteraceae, Liliaceae,

Apocynaceae, Solanaceae, Caesalpinaceae, Rutaceae, Piperaceae,

Sapotaceae used as medicinal plants. The bioactive extract should be

standardized on the basis of active compound. The bioactive extract

should undergo safety studies. Almost, 70% modern medicines in

India are derived from natural products [42, 43].

1.2.2.3.5 Herbal Medicine Today

Herbal medicines are still in use today. In some respects they have

gained a new momentum in the medical field, as many people seek

alternative treatments, and begin to check out traditional, and

Eastern, medicine, herbs are becoming more popular. As physicians

seek new treatments for many common illnesses they are beginning to

revisit the traditional remedies, using herbal medicines.

Pharmaceutical medications, with their potential for harmful side

effects, and addiction are becoming less popular. People are seeking

alternatives to the modern medical interventions. Improving, and

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maintaining, health naturally is a very popular approach to overall

wellness [44-52].

The herbs used today are generally cultivated for those purposes. Very

few herbs are harvested from the wild with the exception of a few still

found in the rainforests, and higher elevations. The cultivation of

herbs for medicinal uses is a large field, and more people are

beginning to plant their own herb gardens. Many monasteries

continue to grow large herbal gardens within their walls.

Elderly people also metabolize medications differently, and generally

are on more medications, and therefore, must also exercise caution

when trying new herbal treatments. Underlying ailments that may

affect the body’s ability to process or absorb medications are also an

issue.

Herbal medicine has enjoyed a long and colorful history. From the

early Chinese Empires to modern physicians’ offices, herbal medicines

have continued to be a part of the medical field. Herbal treatments

have matured throughout history, along with the methods of

delivering them. In the beginning, the herbs were used in a hit or miss

method and required major events to change their use. Research, and

clinical trials have helped to shape the field of herbal medicine, and

the future for herbal medicine looks bright [53-58].

Plants have been used in treating human diseases for thousands of

years. Some 60,000 years ago, it appears that Neanderthal man

valued herbs as medicinal agents; this conclusion is based on a grave

in Iran in which pollen grains of eight medicinal plants were found

[59], which was used by Iranian.

Since, prehistoric time’s shamans or medicine men, and women of

Eurasia, and Americas acquired a tremendous knowledge of medicinal

plants. All of the native plant species discussed in detail elsewhere

was used by native people in traditional medicine. The fact that

hundreds of additional species were also used by First Nations

Canadians [60], suggests that many of these also have important

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pharmacological constituents that could be valuable in modern

medicine.

Until the 18th century, the professions of doctor, and botanist were

closely linked. Indeed, the first modern botanic gardens, which were

founded in 16th century in Pisa, Padova, and Florence (Italy) were

medicinal plant gardens attached to medical faculties or schools.

The use of medicinal plants is not just a custom of the distant past.

Perhaps 90% of the world's population still relies completely on raw

herbs, and unrefined extracts as medicines [61]. A 1997 survey

showed that 23% of Canadians have used herbal medicines. In

addition, as much as 25% of modern pharmaceutical drugs contain

plant ingredients [62].

India is one of the 12 mega biodiversity centers having 45,000 plant

species; its diversity is unmatched due to the 16 different

agroclaimatic zones, 10 vegetative zones, and 15 biotic provinces. The

country has a rich floral diversity as represented in Table 1.1.

Number species

15,000-18,000 Flowering plants

23,000 Fungi

25,000 Algae

1,600 Lichens

1,800 Bryophytes

30 millions Microorganism

Traditional medicine is the synthesis of therapeutic experience of

generations of practicing physicians of indigenous systems of

medicine. Traditional preparation comprises medicinal plants,

minerals, and organic matters etc. Herbal drug constitutes only those

traditional medicines that primarily use medicinal plant preparations

for therapy. The ancient record is evidencing their use by Indian,

Table 1.1 Floral diversity in India

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Chinese, Egyptian, Greek, Roman, and Syrian dates back to about

5000 years. Indian subcontinent is a vast repository of medicinal

plants that are used in traditional medical treatments [63], which also

forms a rich source of knowledge [64, 65]. The various indigenous

systems such as Siddha, Ayurveda, Unani, and Allopathy use several

plant species to treat [66]. In India around 20,000 medicinal plant

species have been recorded recently [67]. More than 500 traditional

communities use about 800 plant species for curing different diseases

[68]. Currently 80 % of the world population depends on plant-derived

medicine for the first line of primary health care for human alleviation

because it has no side effects [69]. Plants are important sources of

medicines, and presently about 25% of pharmaceutical prescriptions

in the United States contain at least one plant-derived ingredient. In

the last century, roughly 121 pharmaceutical products were

formulated based on the traditional knowledge obtained from various

sources [70].

1.3 Cassia genus and its importance

Cassia species: The genus Cassia is one of the most important

genuses of sub family Caesalpinaceae (Family: Fabaceae), most

distributed in tropics and sub tropics. It comprised of about 600

species, most of them are having medicinal values, and used to cure a

broad range of diseases. The available literature on in vitro

micropropagation of Cassia species described the differential response

to cytokinin, and auxin treatment in the media for effective shoot

regeneration, and plantlet propagation. It was also found that the

explants type played a specific role in morphogenesis and

multiplication.

General botanical description and properties of Cassia species

Cassia species belong to the family Caesalpiniaceae. Caesalpiniaceae

is often treated as a sub family, Caesalpinioideae, of the large family

Leguminosae. It is closely related to Mimosaceae, and Papilionaceae,

but can be distinguished by few stamens, and five free petals.

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Caesalpinioideae consist of trees, shrubs, and a few woody herbs

found in the tropics. Economically, woody Caesalpiniaceae is

important for its timber. Cassia and Tamarindus species are used for

medicinal purposes [71-74].

Examples of Cassia species are C. rotundifolia Pers., C. jaegeri Keay in

Bull., C. nigricans Vahl, C. kirkii Oliv., C. mimosoides Linn., C. mannii

Oliv., C. aubreviIlei Pellegr in Bull., C. sieberiana DC., C. arereh Del.,

C. alata Linn., C. podocarpa Guill. and Perr., C. siamea Lam., C. italic

(Mill.) Lam., C. senna Linn., C. absus Linn, C. singueana Del., C.

laevigata Willd., C. bicapsularis Linn., C. tora Linn., C. sophera Linn.,

C. occidentalis Linn., and C. hirsuta Linn. Others are C. auriculata

Linn., C. fistula Linn., C. fruticosya Mill., C. ligustrina Linn., C.

multijuga Linn., C. javanica Linn., C. nodosa Buch.-Ham. ex Roxb., C.

roxburghii DC, and C. surattensis Burm. f.

Cassia species have been of keen interest in phytochemicals, and

pharmacological research due to their excellent medicinal values.

They are well known in folk medicine for their laxative, and purgative

uses [75-77]. Besides, they have been found to exhibit anti-

inflammatory [78], antioxidant [79, 80], hypoglycaemic [81, 82],

antiplasmodial [83], larvicidal [84], antimutagenic [85, 86], and

anticancer activities [87]. They are also widely used for the treatment

of wounds [88], skin diseases such as ringworm, scabies and eczema,

gastro-intestinal disorders like ulcers [89], and jaundice [90]. The

phytochemical studies of the medicinal plants have provided some

biochemical basis for their ethno pharmacological uses in the

treatment, and prevention of various diseases, and disorders [91-93].

The medicinal properties of Cassia species are due to their contents of

hydroxyanthraquinone, alkaloids, flavonoids, phenolic acids,

carbohydrates, glycosides, protein, amino acids, saponins, and

triterpenoids derivatives. Looking to the importance two plants are

selected for present study.

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Cassia siamea (Lam.) (syn: Senna siamea (Lam.) is a very widespread

medicinal and food plant cultivated in Southeast Asia, and sub-

Saharan Africa [94]. Its stem bark is traditionally used against

constipation, malaria, and associated diseases such as fevers and

jaundice [95-97]. A decoction of bark is given to diabetic patients,

while a paste is used as dressing for ringworm, and chilblains. The

roots are used as antipyretic and leaves for constipation,

hypertension, insomnia and asthma. The ethnobotanical survey

among the tradi practitioners of Brazzaville, and Dolisie (Congo)

confirms these uses, with the addition of antinociceptive, and antiviral

activities. Previous studies have confirmed some of these traditional

uses: antiplasmodial activity [98, 99], antioxidant, and

antihypertensive activity, laxative activity, sedative activity [100-101]

have been validated by pharmacological studies. However, there are as

yet no published studies concerning the analgesic activity of Cassia

siamea, though it has been noted that the plant, thanks to barakol

has anxiolytic and CNS inhibitory effects.

Cassia javanica (Linn.) is a beautiful garden tree that belongs to

family Leguminosae. It is cultivated throughout in India for beautiful

pink blossoms. Previous literature provides meagre information about

therapeutic uses of this plant. Bark of C. javanica is used as one of

the ingredients in antidiabetic ayurvedic formulation. Leaves are

proved to be active against Herpes simplex infection [102]. Leaves are

reported to contain variety of secondary metabolites, such as flavones,

sterols, several hydrocarbons, anthraquinones, glycosides etc. Among

these flavones, glycosides and sterols are considered to be antidiabetic

compounds [103]. The presence of these antidiabetic phytochemicals

of C. javanica leaves may give desired pharmacological action. As

there are no scientific data available regarding antidiabetic effects of

leaves, it felt relevant to assess bioactivity of leaves of C. javanica

[104, 105].

Chapter 1

18

1.4 Antimicrobial Survey

What are Antimicrobials?

A wide variety of active phytochemicals include the flavones,

flavonoids, flavonols, phenolics, polyphenolics, quinones, tannins,

coumarins, terpenoids, alkanoids, lectins, and polypeptides are

belived to the present in plant parts, and possesses antimicrobial

properties. Some volatile essential oils of commonly used culinary

herbs, spices, and herbal teas have also exhibited a high level of

antimicrobial activity. From the literature and other survey, we can

tell that flavonoids are naturally occurring substances possessing

several biological properties [106]. The antimicrobial effects of some

flavonoids, and phenolics are studied by several researchers [107].

Antimicrobial chemotherapy has been an important medical treatment

since, the first investigations of antibacterial dyes by Ehrlich in the

beginning of the twentieth century. However, by the late 1940s

bacteria resistant to antimicrobials were soon recognized as a serious

problem in clinical environments such as hospitals, and care facilities

[108]. Bacterial resistance forces the research community to develop

methods of altering structures of antimicrobial compounds to avoid

their inactivation yet structural modifications alone are not enough to

avert bacterial resistance. The increasing use of household

antibacterial products and agricultural antimicrobials fosters

resistance to drugs specific for human therapy, and may have huge

consequences particularly for children, and elderly persons [109,

110]. Antimicrobials contained in manure, and bio solids may

enhance selection of resistant bacteria by entering the aquatic

environment through pathways of diffuse pollution [111]. Surface

water, and shallow groundwater are commonly used for drinking

water, and antimicrobials are now found to pollute many aquatic

sources [112, 113]. Antimicrobials are used worldwide in human

medicine, food, agriculture, livestock and household products. In

many cases, the use of antibiotics is unnecessary or questionable. An

Chapter 1

19

overview of antimicrobial pathway used in human medicine is shown

in Figure 1.1.

Consumption of antibiotics is linked to bacterial resistance. In

hospitals, most common resistant bacteria include methicillin-

resistant, Staphylococcus aureus, vancomycin-resistant enterococci,

and gram-negative rods, including the Enterobacteriaceae, and

Pseudomonas aeruginosa [114]. Many medicinal plants are considered

to be potential antimicrobial crude drugs as well as a source for novel

compounds with antimicrobial activity, with possibly new modes of

action.

This expectation that some naturally occurring plant compounds can

kill antibiotic-resistant strains of bacteria such as Bacillus cereus,

Escherichia coli, Micrococcus luteus, and Staphylococcus aureus [115].

In the past few decades, the search for new anti infective agents has

occupied many research groups in the field of ethnopharmacology. A

pub med search for the antimicrobial activity of medicinal plants

produced 115 articles from the period between 1966 and 1994.

However, in the following decade between 1995 and 2004; this

number increase more than doubled to 307. In these studies one finds

a wide range of criteria related to the discovery of antimicrobial

Figure 1.1 Antimicrobials used in human cell and its biochemical

pathway

Chapter 1

20

compounds in plants. Many focus on determining the antimicrobial

activity of plant extracts found in folk medicine, essential oils or

isolated compounds such as alkaloids, flavonoids, sesquiterpene

lactones, diterpenes, triterpenes or naphthoquinones. After detection

of antimicrobial activity in the plant extract, some of these compounds

were isolated or obtained by bioassay-guided isolation. A second block

of studies focuses on the random screening of natural flora of a

specific region or country, and the third relevant group of papers is

made up of in-depth studies of the activity of a plant or plant

compound against a specific pathological microorganism [116].

(semi)synthetic compounds natural compounds

cocaine morphine

metformin galegine

nabilone Δ9-tetrahydrocannabinol

oxycodon (and other narcotic analgesics) morphine

taxotere taxol

teniposide podophyllotoxin

verapamil khellin

amiodarone khellin

The goals of using plants as sources of therapeutic agents are a) to

isolate bioactive compounds for direct use as drug, e.g., atropine,

scopolamine, digoxin, digitoxin, morphine, reserpine, taxol,

vinblastine, and vincristine; (b) to produce bioactive compounds from

novel or known structures, using them as lead compounds for

(semi)synthesis of novel patentable entities with better activity, and/or

lower toxicity (examples are shown in Table 1.2 ); (c) to use natural

products as pharmacological tools, e.g., lysergic acid diethylamide,

Table 1.2 Some (semi)synthetic bioactive compounds derived

from natural compounds, which demonstrate better

activity and/or lower toxicity.

Chapter 1

21

mescaline, strychnine, yohimbine; and (d) to use the whole plant or

part of it as a herbal remedy, e.g., Cranberry, Echinacea, Feverfew,

garlic, Ginkgo biloba, St. John’s wort, and saw palmetto. The number

of higher plant species (angiosperms and gymnosperms) is estimated

between 215,000, and 500,000 species.

1.5 Antioxidants

1.5.1 Free radical generation: Role of oxidation process in body

To help our body, protect itself from the rigors of oxidation, Mother

Nature provides thousands of different antioxidants in various

amounts in fruits, vegetables, whole grains, cereals, millets, minor

millets, nuts, and legumes. When our body need to put up its best

defense, especially true in today's environment, antioxidants are

crucial to our health. As oxygen interacts with cells of any type an

apple slice or in our body, the cells lining our lungs or in a cut on our

skin oxidation occurs. This produces some type of change in those

cells. They may die, such as with rotting fruit. In the case of cut skin,

dead cells are replaced in time by fresh new cells, resulting in a healed

cut. This birth and death of cells in the body goes on continuously, 24

hours a day. A process is necessary to keep the body healthy.

Oxidation is a very natural process that happens during normal

cellular functions. Yet, there is a downside, while the body

metabolizes oxygen very efficiently, 1% or 2% of cells will get damaged

in the process, and turn into free radicals.

"Free radicals" is a term often used to describe damaged cells that

can be problematic. They are "free" because they are missing a critical

molecule, which sends them on a rampage to pair with another

molecule. These molecules will rob any molecule to quench that need.

1.5.2 Types of free radicals

Most free radicals are coming from oxygen atoms, and are called

Reactive Oxygen Species (ROS), such as superoxide ion, hydroxyl

radical, hydrogen peroxide and singlet oxygen.

Chapter 1

22

Superoxide ion (or reactive oxygen species) is an oxygen molecule

with an extra electron. This free radical can cause damage to

mitochondria, deoxyribonucleic acid (DNA), and other biomolecules.

Superoxide radical is produced in-vivo, and the superoxide dismutase

enzymes are an important protective antioxidant defense [117,118].

Scheme 1.1 shows the free radical mechanism of oxygen. Dismutation

occurs spontaneously at a rate that decreases with a rise in pH, but

the rate can be accelerated enormously by addition of superoxide

dismutase (SOD) enzymes, which are present in essentially all body

tissues, and in most uncooked foods [119]. Fortunately, neither •O2-

nor H2O2 is very reactive. H2O2 can cross membranes readily, which

•O2- is unable to do; the charged natured of •O2- renders it membrane

impermeable unless there is a trans-membrane channel through,

which it can move, such as the anion channel in erythrocytes [120].

Much of the damage that can be done by •O2- , and H2O2 in vivo is

thought to be due to their conversion into more reactive species [121],

probably the most important of which is hydroxyl radical (•OH).

Hydroxyl radical is formed by the reduction of an oxygen molecule in

the electron transport chain as shown in Figure 1.2.

Scheme 1.1 Free radical mechanism of oxygen

Chapter 1

23

It is a neutral (not charged) form of the hydroxide ion. Hydroxyl

radicals are highly reactive and form an important part of radical

biochemistry [122]. Unlike superoxide, the hydroxyl radical cannot be

eliminated by an enzymatic reaction. It has a very short half-life, and

will only react with molecules within its vicinity. Because of its high

reactivity it will damage most organic molecules such as

carbohydrates, DNA, lipids, and proteins. Singlet oxygen is formed by

immune system. Singlet oxygen causes oxidation of Low-density

lipoprotein (LDL) cholesterol [123-125].

1.6 Flavonoids

The flavonoids, derivatives of 1, 3-diphenylpropane, are a large group

of natural products, which are widespread in higher plants but also

found in some lower plants, including algae. Most flavonoids are

yellow compounds, and contribute to the yellow colour of the flowers,

and fruits, where they are usually present as glycosides. Flavonoids

are the largest group of phenols in the plant kingdom, and are

metabolites with a benzopyran nucleus having an aromatic

substituent at carbon number 2 (C-2) of the C ring [126, 127]. Figure

1.3 depicted here shows the basic structure of flavonoids ring.

Figure 1.2 Reduction of an oxygen molecule in the electron

transport chain

Chapter 1

24

The most common types of flavonoids are flavanones, flavonols,

flavones, and flavans. As shown in Figure 1.3, characteristic features

of these flavonoids are a carbonyl at C-4 (flavanones), carbonyl at C-4,

double bond between C-2, and C-3, and hydroxyl at C-3 (flavonols),

carbonyl at C-4, and a double bond between C-2 ,and C-3 (flavones),

and no carbonyl at C-4 but with a hydroxyl at C-3 for the flavans.

Among the types of flavonoids that exist in nature, flavans are the

major group found in sorghum [128], and the major flavans are

leucoanthocyanidins (hydroxyl at C-3 and C-4), (+)-catechin (hydroxyl

at C-3), and anthocyanidin (hydroxyl at C-3, double bond between C-3

and C-4). Anthocyanidins such as luteolinidin, cyaniding, and

apigeninidin have been reported in sorghum. Awika, Rooney, and

Waniska identified apigeninidin and luteolinidin as the major 3-

deoxyanthocyanidins (anthocyanidins that lack a hydroxyl group at

the C-3 position) in black and brown sorghums [129]. Blessin, Van

Etten, and Dimler found an anthocyanidin, fisetidin, in sorghum, and

Yasumatsu, Nakayama, and Chichester reported pelargonidin.

Strumeyer, and Malin however did not find any anthocyanidins in

Leoti and Georgia 615 sorghums [130-132]. Some most common

flavonoids are shown in Figure 1.4.

Figure 1.3 Basic skeletons of flavonoid

Chapter 1

25

1.7 Phenolic acids

Phenolic compounds are substances that possess an aromatic ring

bearing one or more hydroxyl (-OH) substituent [133]. As mentioned

earlier, sorghum contains phenolic compounds that have antioxidant

properties, and provide a protective role to the sorghum plant [134-

138]. These phenolic compounds in grain may be classified very

broadly, into phenolic acids, flavonoids, and condensed tannins.

Figure 1.5 Basic skeletons of benzoic acid

Figure 1.4 Basic structures of common flavonoids

Chapter 1

26

Name of acid Functional group position

R1 R2 R3

Benzoic acid H H H

Gallic acid H -OH -OH

Protocatechuic acid H -OH -OH

p-hydroxy benzoic acid H H -OH

Salicylic acid -OH H H

Basic skeleton of benzoic acid is depicted in the Figure 1.5 and basic

skeleton of cinnamic acid is given in Figure 1.6. Tables 1.3 and 1.4

are given for the convenience to identify the derivatives of benzoic and

cinnamic acid respectively, according to the position of functional

group attached with basic skeleton. Phenolic acids are the simplest

phenolic compounds, and are derivatives of benzoic or cinnamic acids.

They contain hydroxyl or methoxyl groups substituted at various

positions on the aromatic ring. Hahn et al., [139] separated and

identified, by reverse phase high performance liquid chromatography

(HPLC), eight main phenolic acids in sorghum grain extracts in both

white and brown sorghum (gallic, protocatechuic, p-hydroxybenzoic,

vanillic, caffeic, p-coumaric, ferulic and cinnamic acid). These

phenolic acids are found in both the free and bound form, with brown

sorghum containing the highest amount of free phenolic acids. Bound

phenolic acids are those that are complexes or form conjugates with

sugars, proteins or cell wall polysaccharides [140, 141].

Table 1.3 Position of functional groups with respect to R1,

R2 and R3 in benzoic acid

Chapter 1

27

Name of acid Functional group position

R1 R2 R3

Caffeic -OH -OH H

Ferulic -OCH3 -OH H

p-coumaric H -OH H

Sinapic -OCH3 -OH -OCH3

1.8 Anti-inflammatory

1.8.1 Inflammation

Inflammation (Latin, inflamatio, to set on fire) is the complex biological

response of vascular tissues to harmful stimuli such as pathogens,

damaged cells or irritants [142]. It is a protective attempt by the

organism to remove the injurious stimuli as well as initiate the healing

process for the tissue. Inflammation is not a synonym for infection.

Even in cases where inflammation is caused by infection, the two are

not synonymous: infection is caused by an exogenous pathogen, while

inflammation is the response of the organism to the pathogen.

In the absence of inflammation, wounds and infections would never

heal and progressive destruction of the tissue would compromise the

survival of the organism. However, an inflammation that runs

Figure 1.6 Basic skeletons of cinnamic acid

Table 1.4 Position of functional groups with respect to R1, R2

and R3 in cinnamic acid

Chapter 1

28

unchecked can also lead to a host of diseases such as hay fever,

atherosclerosis, and rheumatoid arthritis. It is for that reason that

inflammation is normally closely regulated by the body.

Inflammation can be classified as either acute or chronic. Acute

inflammation is the initial response of the body to harmful stimuli and

is achieved by the increased movement of plasma, and leukocytes

from the blood into the injured tissues. A cascade of biochemical

events propagates and matures the inflammatory response, involving

the local vascular system, the immune system, and various cells

within the injured tissue. Prolonged inflammation, known as chronic

inflammation, leads to a progressive shift in the type of cells, which

are present at the site of inflammation and is characterized by

simultaneous destruction, and healing of the tissue from the

inflammatory process.

1.8.2 Causes of Inflammation

Burns, Chemical irritants, Frostbite, and Toxins, Infection by

pathogens, Physical injury (blunt or penetrating), Immune reactions

due to hypersensitivity, Ionizing radiation, foreign bodies, including

splinters, and dirt, are main cause of inflammation. Compression

between acute and chronic inflammation is given in the Table 1.5.

Acute

Chronic

Causative

agent

Pathogens,

injured tissues

Persistent acute inflammation due to

non – degradable pathogens,

persistent foreign bodies, or

autoimmune reactions

Major cells

involved

Vasoactive

amines,

Mononuclear cells (monocytes,

macrophages,

Table 1.5 Comparison between acute and chronic inflammation

Chapter 1

29

eicosanoids

lymphocytes, plasma cells), fibroblasts

Primary

mediators

Immediate

IFN-γ and other cytokines, growth

factors, reactive oxygen species,

hydrolytic enzymes

Onset

Few days

Delayed

1.8.3 In vitro methods for anti-inflammatory activity [143]

An array of physiological substances, sometimes called autacoids, are

involved in the process of inflammation and repair. These include

histamine, serotonin, bradykinin, substance P, and the group of

eicosanoids (prostaglandins, thromboxanes and leucotrienes), the

platelet- activating factor (PAF) as well as cytokines and lymphokines.

Their discovery makes the use of in vitro studies possible. The

influence of non-steroidal anti-inflammatory agents (NSAID) on the

eicosanoid pathway gave rise to numerous studies. These includes

3H-Bradykinin receptor binding, Substance P and the tachykinin

family, 3H-Substance P receptor binding, Neurokinin receptor

binding. Assay of polymorphonuclear leukocyte chemotaxis, in vitro,

Polymorphonuclear leukocytes aggregation induced by FMLP,

Constitutive and inducible cellular arachidonic acid metabolism in

vitro, Formation of leukotriene B4 in human white blood cells in vitro,

Formation of lipoxygenase products from 14C-arachidonic acid in

human polymorphonuclear neutrophils (PMN), in vitro formation of

eicosanoids from 14C-arachidonic acid in human platelets, in vitro,

stimulation of inducible prostaglandin pathway in human PMNL,

COX-1 and COX-2 inhibition, Induced release of cytokines

(Interleukin- 1alpha, IL-1beta, IL-6, IL-8 and TNF-alpha) from human

white blood cells. In vitro flow cytometric analysis of intracellular

Chapter 1

30

cytokines, TNF-alpha antagonism, binding to interferon receptors,

Screening for interleukin-1 antagonists, Inhibition of interleukin-1β

converting enzyme (ICE) are main causes of inflammation.

1.8.4 In vivo methods for anti-inflammatory activity

Tissue injury or trauma initiates a cascade of reactions, which results

in the proteolytic generation of Bradykinin and kallidin from high-

molecular-weight precursors, kininogens found in blood, and tissue.

The rapid enzymatic cleavage of kininogens is accomplished by the

kallikreins, a group of proteolytic enzymes, which are present in most

tissues and body fluids. Bradykinin produces pain by stimulating A

and C fibers in the peripheral nerves, participates in the inflammatory

reaction and lowers blood pressure by vasodilatation. Since, its

breakdown occurs via the same enzyme responsible for converting

angiotensin I into angiotensin II some of the effects of converting

enzyme inhibitors may be due to presence of Bradykinin. The 3H-

Bradykinin receptorbinding is used to detect compounds that inhibit

binding of 3H-Bradykinin in membrane preparations obtained from

guinea-pig ileum. Two types of Bradykinin receptors (BK1 and BK2

receptors) are known. The existence of a pulmonary BK3 receptor has

been proposed by Farmer et al. in 1989. Evidence was obtained for the

existence of three subtypes of B2 receptors, B2a, B2b, and B2c

1.8.5 Methods for testing acute and sub acute inflammation

Carrageen induced Paw edema in rats, Ultraviolet erythema in guinea

pigs, vascular permeability, Inhibition of leukocyte adhesion to rat

mesenteric venules. In-vivo Oxazolone-induced ear edema in mice,

Croton-oil ear edema in rats, and mice, Pleurisy test, Granuloma

pouch technique, Urate-induced synovitis, are some most common

methods to induce inflammation in animal study.

Chapter 1

31

1.8.5.1 Non-steroidal anti-inflammatory drugs (NSAIDS)

Non-steroidal anti-inflammatory drugs, usually abbreviated as

NSAIDs or NAIDs, are drugs with analgesic, antipyretic (lowering an

elevated body temperature and relieving pain without impairing

consciousness), and in higher doses, with anti-inflammatory effects

(reducing inflammation).

The term “non-steroidal” is used to distinguish these drugs from

steroids, which (among a broad range of other effects) have a similar

eicosanoid-depressing, anti-inflammatory action. As analgesics,

NSAIDs are unusual in that they are non-narcotic. Figure 1.7 depicted

here indicates mechanism of action for NSAID.

NSAIDs are sometimes also referred to as non-steroidal anti-

inflammatory agents / analgesics (NSAIAs) or non-steroidal anti-

inflammatory medicines (NSAIMs). The most prominent members of

this group of drugs are aspirin, ibuprofen, and diclofenac because

they are available over-the-counter in many areas. They are classified

as under [144,145]:

Figure 1.7 Mechanism(s) of Action. All of the non-steroidal anti-

inflammatory drugs

Chapter 1

32

Nonselective COX inhibitors (conventional NSAIDs)

1. Irreversible COX inhibitors Aspirin

2. Reversible COX inhibitors

a. Pyrazolone derivatives - Phenylbutazone, Oxyphenbutazone

b. Indole derivatives - Indomethacin

c. Propionic acid derivatives - Ibuprofen, Naproxen, Ketoprofen,

Flurbiprofen

d. Anthralinic acid derivative - Mephenamic acid

e. Aryl-acetic acid derivatives - Diclofenac, Aceclofenac

f. Oxicam derivatives - Piroxicam, Tenoxicam

g. Pyrrolo-pyrrole derivative - Ketorolac

B. Selective COX-2 inhibitors

1. Preferential COX-2 inhibitors - Nimesulide, Meloxicam,

Nabumetone, Etodolac

2. Highly selective COX-2 inhibitors - Celecoxib, Etoricoxib,

Parecoxib, Lumiracoxib C. Analgesics-antipyretics with poor anti-

inflammatory action.

1. COX-3 inhibitors - Paracetamol (Acetaminophen)

2. Do not inhibit prostaglandin synthesis – Nefopam

These drugs are not derived from opium. They do not interact with

opioid receptors but act primarily on peripheral pain mechanisms.

They may also act on CNS to raise the pain threshold. The Greek

physicians Galen, and Hippocrates described the analgesic effects of

willow bark from which aspirin was synthesized subsequently. They

are considered as non narcotic analgesics [146]. They are also referred

to as non-steroidal anti-inflammatory drugs (NSAIDs). These drugs are

chemically diverse, but most are organic acids. During 1999 to 2000,

per member per year costs for these drugs grew by 37.9%, the highest

growth rate among the top 25 therapeutic categories, as measured by

‘Express Scripts Drug Trend Report’. Contributing to this growth was

Chapter 1

33

the introduction of the newer, more expensive cyclo-oxygenase-2

(COX- 2) specific inhibitors.

The first COX-2 inhibitor approved in the United States was celecoxib

in December 1998, and the second, rofecoxib, entered the market in

May 1999. Valdecoxib was approved by the US-FDA in November

2001(Cox et al., 2003).

Inflammation and oxidative stress plays an important role in various

diseases. Inflammation is an immunological defense mechanism

elicited in response to mechanical injuries, burns, microbial

infections, allergens, and other noxious stimulus [147-149]].

Use of anti-inflammatory agents may therefore, be helpful in the

treatment of inflammatory disorders [150]. Non-steroidal or steroidal

anti-inflammatory drugs are commonly used to treat different

inflammatory diseases [151, 152]. The adverse effects of the currently

available anti-inflammatory drugs, however, pose a major problem in

their clinical use [153]. Therefore, naturally originated agents with

very little side effects are desirable to substitute chemical therapeutics

[154, 155].

1.9 Aim and Objectives of present thesis

Looking to the importance of both plants of Cassia species, the

present research programme is designed accordingly.

The main aim of this research programme is to find out therapeutic

use of phytochemicals present in the aerial parts of two plants Cassia

siamea and Cassia javanica.

The present work will be also focused on separation, isolation,

characterization of various phytochemicals present in the aerial parts

of two above-mentioned plants.

The study also emphasized possible applications of these

phytochemicals in medicinal treatments of various diseases.

To achieve the aim of this research programme following points are

considered.

Chapter 1

34

i. It is assumed that phenolic acids and flavonoids are playing

important role in various biological activities of plants and

therefore, total phenolics and total flavonoids will be

investigated.

ii. Thus, the present investigation is aimed to evaluate the

antimicrobial activity of eight extracts of aerial parts of Cassia

siamea and Cassia javanica

iii. In-vitro antioxidant activity of three extracts of aerial parts of

Cassia siamea and Cassia javanica will also be established

using standard protocols.

iv. In-vivo anti-inflammatory activity of ethanolic extract of aerial

parts of Cassia siamea and Cassia javanica will be evaluated

using paw edema method on Swiss Wistar albino rats.

Chapter 1

35

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