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Title
DECLARATION
CERTIFICATE
ACKNOWLEDGEMENT
CONTENTS
ABBREVIATIONS
1 Introduction
1.1 PLANTS AND HEALTH CARE
1.2 Cancer
1.2.1 Cancer Treatment Modalities
1.3 Development of a drug for cancer
1.3.1 Development of anticancer drugs from plants
1.3.2 Bioassays of plant products
1.4 Plants used in the treatment of Cancer
1.4.1 Cytotoxic and antitumour principles
1.5 Apoptosis
1.5.1 BCL2 family: Killers and protectors
1.5.1 Therapeutic implications
1.6 Chromosomes
1.7.1 Modulators of the toxicity of the anticancer drugs vincristine and cyclophosphamide
1.8 Infrared Spectroscopy
1.8.1 Spectroscopy of major cellular components
1.8.2 Infrared Absorption Spectra of Protein and Nucleic acids
1.9 Rationale for the selection of the plant of this study
1.9.1 Properties and Uses of B.racemosa seed
1.9.2 Chemistry and Pharmacology
1.I0 Aims and Objectives
2 MATERIALS AND METHODS
2.1 MATERIALS
2.1.1 Plants
2.1.2 Experimental animals
2.1.3 Tumour Cell Lines
2.1.4 Chemicals
2.1.5 Instruments
2.2 METHODS
2.2.1 Methods for separation and purification of phytochemicals
2.2.2 Extraction and Purification of B.racemosa seeds
2.2.2.1 B.racemosa seed extract
2.2.2.2 Purification of the methanol extract of B.racemosa seed extract
2.3 Toxicity studies
2.3.1 Acute toxicity
2.3.2 Short-term toxicity
2.4 Assessment of anticancer activity
2.4.1 Experimental Animal Maintenance
2.4.2 Maintenance of tumour cell lines in mice
2.5 Tissue Culture
2.5.1 Determination of cytotoxicity using cell culture
2.5.2 Short-term Cytotoxicity
2.6 Anti tumour testing using murine models
2.7 Determination of Apoptosis
2.7.1 in vitro study
2.7.2 in vivostudy
2.8 Chromosomal aberration studies
2.9 Modulation of the toxicity induced by the antiturnour drugscyclophosphamide and vincristine
2.9.1 Modulation of Vincristine induced toxicities in normal mice
2.9.2 Modulation of toxicities in tumour bearing mice treatedVincristine
2.9.3 Modulation of cyclophosphamide induced toxicities in normalmice
2.9.4 Modulation of toxicities in tumour bearing mice treated withCyclophosphamide
2.9.5 Haematological parameters
2.9.5.1 Method
2.9.5.2 Red and White cell counting
2.9.5.3 Platelets count and size distribution
2.9.5.4 Measurement of hemoglobin concentration
2.9.5.5 Derivation of parameters
2.9.5.6 Determination of serum alkaline phosphatase
2.9.5.7 Determination of SGPT
2.5.9.8 Determination of SGOT
2.5.9.9 Determination of Urea
2.5.9.10 Quantitative determination of inorganic phosphate in serum
2.5.9.11 Estimation of calcium [0- cresolphthalein Complexone Method]
2.6 IR Spectroscopy
2.6.1 Mice
2.6.2 Preparation of cell lines
2.6.3 Solid tumour growth formation
2.6.4 Histopathological studies
2.7 Statistical analysis
3 CYTOTOXICITY TO TUMOUR CELLS AND CELLS IN CULTURE (IN VITRO)
3.1 introduction
3.2 Materials and Methods
3.2.1 Preparation of seed extract
3.2.2 Purification of the seed extract
3.2.4 Toxicity
3.2.4.1 Acute toxicity
3.2.4.2 Short-term toxicity
3.3 In vitro cytotoxicity
3.3.1 Ascitic tumour cells
3.3.2 In vitro cell culture
3.5 Results
3.5.1 Toxicity
3.6 Discussion
4 ANTI -TUMOUR ACTNITY STUDIES
4.1 INTRODUCTION
4.2 Materials and Methods
4.2.1 Antiturnour testing using rnurine models
4.2.2 Effect of methanol extract of B.racemosa seed on the growth of ascitic tumour in mice
4.2.3 Treatment Schedule
4.2.4 Effect of purified fractions F5 (7: 3) and F8 (2: 8) of B.racemosa seed extract on the growth of ascitic tumours in mice.
4.2.5 Effect of the drug on solid tumour in mice
4.3 Results
4.3.1 Effect of the drug (s) on the growth of solid tumour
4.4 Discussion
5 THE EFFECT OF THE DRUG ON APOPTOSIS
5.1 INTRODUCTION
5.2 Biochemistry of apoptosis
5.3 Apoptosis induced by Medicinal plants
5.4 Materials and Methods
5.4.1 Mice:
5.4.2 DLA Cell-line:
5.4.3 Purification of B.racemosa seed extract:
5.4.4 lnvitro study:
5.4.5 Invivo study:
5.4.6 Morphologic determination of apoptosis
5.5 Result
5.6 Discussion
6 ANTI -CLASTOGENIC ACTIVITY OFBARRINGTONIA RACEMOSA SEED EXTRACT
6.1 INTRODUCTION
6.2 Materials and Methods
6.3 Slide preparation and chromosome aberration scoring
6.4 Results
6.5 Discussion
7 MODULATION OF TOXICITY OF THE ANTI-CANCER DRUGS, VINCRISTINEAND CYCLOPHOSPHMIDE
7.1 INTRODUCTION
7.2 Materials and Methods
7.2.1 Mice
7.2.2 Drugs
7.2.3 Cyclophosphamide treated normal mice
7.2.4 Cyclophosphamide treated tumour bearing mice
7.2.5 Vincristine treated normal mice
7.2.6 Vincristine treated tumour bearing mice
7.2.7 Haematological and biochemical parameters
7.3 Result
7.3.1 Cyclophosphamide treated normal mice
7.3.2 CYP treated tumour bearing mice
7.3.3 Vincristine treated normal mice
7.3.4 Vincristine treated tumour bearing mice
7.4 Discussion
8 INFRARED SPECTROSCOPIC STUDIES OF NORMAL DLA INDUCED TUMOUR AND DRUG TREATED TUMOUR
8.1 INTRODUCTION
8.2 Materials and Methods
8.2.1 Mice:
8.2.2 Preparation of cell lines
8.2.3 Solid tumour growth formation
8.2.4 Infrared Spectroscopic Analysis of DLA induced tumour tissue, drug treated tumour and control (normal thy muscle)
8.2.5 Histopathological studies
8.3 Results
8.3.1 Inhibition of solid tumour growth
8.3.2 IR spectroscopic studies
8.3.3 Histopathological studies
8.4 Discussion
9 GENERAL DISCUSSION
9.1 GENERAL DISCUSSION
SUMMARY AND CONCLUSIONS
REFERENCES
LIST OF PUBLICATIONS
ABSTRACT