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Ph.D Thesis Summaries

Titles of the Thesis :


Title: The anatomical and physiological responses of Syzygium cumini (L.) Skeels to coal-smoke pollution with special reference to wood production    

Subject: Botany
Faculty : Science
 

Department:
Chemistry
Name of the student:
Malini Jacob
Name of the supervisor:
Prof. M. Iqbal
Name of the Co-supervisor : Dr. Mahmooduzzafar

Date of Viva Voce:
19 October 2002

Summary 

The present work was carried out to investigate the impact of coal smoke emissions on the overall growth performance cambial activity and the consequent wood production of Syzygium cumini (L.) Skeels growing at various distances (0.5 km, 2 km, 6 km, 9 km and 15 km) from the point source of pollution i.e. the Badarpur Thermal Power Station (BTPS) in Delhi. Growth performance was assessed by analyzing its carbon assimilation capacity and biochemical alterations as evident in the leaf tissue of the stressed trees and by investigating the effect of these metabolic alterations on the meristematic activity of vascular cambium, and the consequent wood development under a gradient of pollution regime. 

The following conclusions can be drawn from the present investigation regarding the changes that S. cumini trees experienced due to coal-smoke pollutants present in the air. 

SO2, NO2 and TSPM values were maximum at the most polluted site (6 km away) whereas at site V the values were below the permissible limit. Visible injury symptoms due to air pollutants were not observed at any of the sites and in any of the seasons. The stomata were smaller at site III (6 km) than at the other sites. Their size was maximum in pre-monsoon and minimum in post-monsoon season. The size of stomatal aperture declined up to a distance of 6 km. Stomatal index (SI) touched the minimum at site III. The SI was positively correlated with nitrogen content and vessel width. The concentration of photosynthetic pigments were lowest at site III and recovered thereafter. Carotenoid content also exhibited a similar trend. The total chlorophyll content was severely affected by TSPM & SO2 concentrations. Chlorophyll content did influence strongly the net photosynthetic rate, NR activity rate, protein content and nitrogen content. The sulphur content and sugar content remained negatively related to each other throughout the year. The sulphur content went parallel to pollution load. The S content was negatively correlated with chlorophyll and net photosynthetic rate in the monsoon months. The span of cambial activity in S. cumini was of about 6½ to 7 months at different sites. However, pollution stress delayed the cambial reactivation for about 2 months at site III. Xylem differentiation preceded phloem production. Curiously, the span of xylem production was longer than phloem under the pollution stress. The annual quantum of wood produced was consequently greater in trees at the polluted sites. Relative proportion of vessels in the wood was relatively low under pollution stress. Width of vessel elements was also smaller at the polluted sites, but their length increased with the degree of pollution. Wood fibres were shorter at the polluted sites, compared to those at the reference site. A high degree of negative correlation emerged between vessel length and fibre length. The photosynthetic rate was inversely proportional to the pollution load. It was positively correlated with nitrogen content. The stomatal conductance dropped to its lowest at the most polluted site. It maintained a positive relationship with sugar content throughout the year. The NR activity rate ran counter to the pollution load. It bore a significant positive correlation with chlorophyll content, sugar content and nitrogen content but a negative correlation with nitrate content. Contrary to NR activity, nitrate content was maximum at site III and minimum at the non-polluted site throughout the year. It was negatively correlated to photosynthetic rate, chlorophyll content and nitrogen content. A linear negative correlation was observed between the amount of total nitrogen and the pollution load. A significant positive correlation existed between sugar content and NR activity rate.

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Summary No. December 2002/01  

The Title : Air pollution effects on the structural, pharmacognostical and phytochemical traits of the barks and leaves of Azadirachta indica A. Juss and Dalbergia sissoo Roxb "  

Subject: Botany
Faculty: Science
Name of Student: AB. Rashid Trag
Supervisor: Dr. T. O. Siddiqui
Co-Supervisor: Dr. Mahmooduzzafar
Date of Viva-voce: 12.12.2002  

Summary

This study was carried out on Neem (Azadirachta indica) and Sheesham (Dalbergia sissoo) trees grown abundantly around the Badarpur Thermal Power Plant where atmosphere is polluted due to coal-smoke pollutants emitted from the stacks of the power plant. The observations were carried out for three consecutive years. Plant materials (mature leaves) collected from five selected sites, located at different distances from the pollution source, were studied and analyzed so as to assess the impact of environmental degradation on anatomical, physiological, biochemical, pharmacognostical and phytochemical traits of the test plants.

The SO2, NO2 and SPM  (Suspended Particulates matter) concentrations were maximum at site III, whereas site V had the minimum stress (comparable with the non-polluted atmosphere) and was, therefore, considered as the reference site.

The soil was slightly alkaline sandy loam at all the sites. Nitrogen content of the soil was minimum at the highly polluted site (site III), and maximum at site V. Soil sulphur was low at sites I and IV. Potassium in soil was lowest at the most polluted site.

The length and width of phloem fibres, seive tube elements, relative proportion of seive tubes and fibres, amount of conducting phloem and the vascular tissues in the petiole of leaf of both species decreased at all sites in all seasons. However, proportion of axial parenchyma and ray parenchyma of conducting phloem in the stem increased.

The net photosynthetic rate and stomatal conductance of leaves declined at the polluted sites but the intercellular CO2 concentration increased.

The nitrate, sugar and sulphur contents of leaf and secondary phloem enhanced at almost all sites, with the maximum enhancement at site III. However, the reduced nitrogen, reducing sugar and sulphate contents in both leaves and bark decreased at all sites.

The pharmacognostic parameters varied with respect to pollution. Loss on drying, per cent ash value content, water-soluble ash, sulphated ash and acid insoluble ash contents were enhanced with respect to pollution load. The various extractive values increased at various distances from the pollution source. The maximum increase was recorded at 8 km from the pollution source. However, the petroleum ether extractive decreased with respect to site V.

Tannin content decreased proportional to the pollution load in case of A. indica.

Reduction in the crude fibre was also observed. The protein content (crude protein) declined under pollution stress in both A. indica and D. sissoo.

Several compounds, mostly flavonoids, terpenoids and fatty acids, were isolated from the barks of A. indica and D. sissoo. Their amounts declined under pollution stress.

Flavonoid content in the leaves of both the trees also reported decreased. 

Few new compounds were isolated through column chromatography in order to mark them as reference or marker compounds for quantitative estimations.

Compounds isolated from the bark of A. indica are; 

RN-I                5-methyl-n-triacotane.

RN-2               n-heptacosan-8-ol

RN-3               n-hexacosanoic acid

            RN-4               2,6,10,14-tetramethyl-pentadeca-8-en-2, 7-diol

RN-5               2,6,10-trimethyl-14-cyclopentanyl-n-tetradecan-6a-ol

RN-6               5,7-dien,3b-ol-b-D-glucopyranoside

RN-7               9,9-dimethyl-decan –7-en-2-ol-11,12-dioic acid 

Compounds isolated from the bark of D. sissoo are; 

RD-I                n-hexacosa-5-ol -yl propionate

RD-2               n-tetracosa-5-ol -yl  propionate

RD-3               6-hydroxy-7-methoxy-4-(2’-hydroxybenzyl)-benzopyron

RD-4               9,11,12-trihydroxy-10-methoxyanthracare-14,17-[2H]-pyran

Plants have a greater capacity to accommodate very wide fluctuations in their environment but the buffering capacity they endow finally fails, resulting in chronic sub-optimal growth. The recorded accumulative toxic effects of pollution in the morphological, physiological, biochemical and phytochemical characteristic are suitable indicators for biological monitoring. The pollution load alter, not only the quantity but also the quality of active compounds of the trees, thereby decreasing the therapeutic value  of a compound or a combination of compounds. The maximum effect was observed during winter, confirming that the test species were more sensitive to pollution during winter.

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Summary No. December 2002/02 

Title of the thesis: "Influence of different diets on bioavailability of conventional cefaclor formulations in healthy human male volunteers"
Name of Candidate: Shahid Karim
Research Supervisor: Prof. P.L.Sharma, Dr. Nilajan Saha, Dr. Tausif Monif
Department:
Pharmaceutical Medicine,

Faculty of Pharmacy,
Date of Viva             :            20.12.2002
 

SUMMARY

This randomized, open-label, balanced, five-treatment, five-period, five-sequence, single-dose and crossover pharmacokinetic study assessed the effect of different types of food on the bioavailability of cefaclor in 18 healthy male volunteers. A single dose of cefaclor, 250-mg capsule was administered at five occasions: after overnight fasting, after two vegetarian (high-fat and low-fat) diets and two non-vegetarian (high-fat and low-fat) diets. Serial blood samples were collected upto 8 h post dose. Serum cefaclor concentrations were determined by a validated HPLC method. AUC values were not significantly affected by food intake, but the Tmax was prolonged and Cmax was decreased, depending on the type of meal. The non-vegetarian diets affected the rate of absorption of cefaclor more than the vegetarian diets. The least decrease in Cmax was produced by low-fat vegetarian diet, while the maximum decrease was produced by high-fat non-vegetarian diet. The results of this study indicate that while the rate of absorption of cefaclor is significantly decreased, the extent of absorption and the rate of elimination is not significantly decreased in the presence of food. As compared to high-fat non-vegetarian diet, the time above MIC50 concentration was significantly increased by low-fat vegetarian diet. The implications of these findings for the large vegetarian Indian population are considerable.

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Summary No. December 2002/03 

Title: "Physio-chemical Responses of Cassia angustifolia Vahl. and Cichorium intybus   L. to Salt Stress"

Subject                        : Botany
Faculty                        : Science  
Name of student     
    : Ms. Anjum Arshi  
Supervisor                
   : Prof. Muhammad Iqbal  
Co-Supervisor          
   : Dr. M. Z. Abdin  
Date of vivo-voce        : 27-12-2002

Summary

A rapidly increasing world population and the fast approaching geographical limitations of the world agriculture system have led to a serious consideration for a possible expansion of agricultural activities on to marginal lands which are unsuitable for plant growth and development. Among the main environmental stresses responsible for low productivity of crop plants on marginal lands are the salt and water stresses. Salinity is characterized by the presence of an abnormally high content of readily soluble salts, primarily chlorides, sulphates and carbonates of Na+, Mg2+, K+ and Ca2+. Consequently, these soils cannot be utilized for cultivating agricultural crops and medicinal herbs. NaCl is usually the dominant salt in the saline soils. Although physiochemical responses of crop plants to environmental stresses have been studied, relatively little is known about the performance of medicinal plants under these hardships.

It was observed in this study that salinity caused by NaCl hampers growth processes of Cassia angustifolia and Cichorium intybus. Treatment consisting of 160 mM NaCl caused maximum growth reductions. Calcium resisted the stressful condition caused by salinity and minimized its deleterious effects on plant performance.   Carbon and nitrogen assimilatory processes, viz. photosynthetic rate, stomatal conductance, contents of chl a, chl b, total chl, soluble protein and nitrogen in leaves, and nitrate reductase activity in the leaves, were also adversely affected by NaCl treatment applied at any phenological stage. Application of CaCl2 with NaCl mitigated the adverse effect of NaCl treatments. The nitrate content, on the other hand, increased in the NaCl-treated plants. The increase caused by NaCl and NaCl + CaCl2 treatments was almost equal but greater than one caused by CaCl2 alone in both the species studied.

Proline content in the leaves increased with age of the plants irrespective of treatments. The increase was eight-fold and seven-fold against NaCl+CaCl2 treatments in C. angustifolia and C. intybus plants, respectively, as against a five time increase with NaCl treatments alone. It was enhanced by three and two times against CaCl2 treatments of the two species respectively. From these results it can be inferred that salinity induces accumulation of proline that might help in osmoregulation of the plant, and that proline in cells works as an osmolyte is proved by its increased accumulation with application of CaCl2 along with NaCl as both together create a more hypertonic soil environment giving way to more accumulation of proline.

The sodium and chloride contents in the roots, stems and leaves increased with increasing NaCl level in the medium. Maximum accumulation occurred in the leaves; followed by stem and roots with each treatment of NaCl. The amount of Na+ taken up by the leaves was greater than the amount of Cl-. CaCl2 treatments reduced the sodium accumulation in different plant parts, but increased Cl- ion. Combined treatments mitigated the increased accumulation of sodium caused by NaCl treatments in both the species. Enormous accumulation of Na+ and Cl- ions in leaves suggests that these plants possibly act as ion accumulators and can be considered for being used to phytoremediate the degraded saline lands. On application of NaCl + CaCl2, the chloride content was higher than one against any of the two salts applied alone.

NaCl stress had an adverse effect on potassium and calcium contents in roots, stems and leaves in both the species. The extent of decline was dose dependent. CaCl2 treatments were beneficial and increased the potassium and calcium contents at any stage of plant development. However, the reduction in the amount of these ions was less against combined treatments than against NaCl treatments. Elevated calcium mitigates the adverse effect of NaCl through inhibition of Na+ uptake and restores the adequate level of K+, and thus enhances the plant growth. 

Sennoside contents were maximum in pods of C. angustifolia followed by those in the immature leaves and mature leaves. The sennoside a, b, c and total sennoside contents in the senna plants decreased under high salinity effect, while supply of calcium promoted production of these substances responsible for the therapeutic property of the plant. Salinity of soil might alter the seed oil composition in chicory plants; the proportion of short carbon chain fatty acids decreased whereas long carbon chain fatty acids increased. This conversion was less with CaCl2 treatments, thus calcium playing a restorative role. Since the relative proportion of the component fatty acids changes in the seed oil, properties of the oil may also change. The efficacy of the given oil may thus be affected by the levels of soil salinity, meaning thereby that salinity-induced alterations in medicinal plants are not only quantitative but these may sometimes be qualitative too, thus affecting the efficacy of the plant drug.

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Summary No. December 2002/04 

Title “Studies on immunomodulatory activity of some herbs used in Indian systems of medicine”

Subject: Toxicology
Name of Student: Bilal Bin hafeez (indianbilal@yahoo.com)  

Department: Medical Elementology & Toxicology, Faculty of Science  
Supervisor: Dr. S. Raisuddin (sraisuddin@hotmail.com)  
Source of funding:Central Council of Research on Unani Medicine (CCRUM) and Council of Scientific
and Industrial Research (CSIR), Govt. of India    
Date of viva voce: December 18, 2002

Summary

Modulation of immune functions by using medicinal plants and their products as a possible therapeutic measure has become fundamental principles of therapeutic approach. Plants and minerals have been used since ancient times for the treatment of many ailments and diseases. It is now being recognized that immunomodulation of immune response could provide an alternative to conventional chemotherapy for a variety of disease conditions, especially when host's defense mechanism have to be activated under the conditions of impaired immune responsiveness or when a selective immunosuppression has to be induced in situation like autoimmune disorders and organ transplantation.

1.         In the present study, aqueous extract of Cassia occidentalis (KASAUNDI), Trigonella foenum-graecum (METHI), Delphinium denudatum (JADWAR) and Anthum graveolens (SOWA) showed no mortality and apparent toxicity signs and symptoms in the treated animals.

2.         Among all the herbs, Cassia occidentalis and Trigonella foenum-graecum showed the stimulatory effect on specific and non-specific immunity. It is also interesting to note that in spite of reduction in spleen weight and its cellularity no suppressive effect was observed in spleen associated functions (antibody production as measured by PFC assay and antibody titre). It is suggested that these plants may be beneficial in ongoing chemotherapy.  However, Delphinium denudatum showed dual effect on immune functions. It suppressed the humoral immunity and stimulated the cell mediated and non-specific immunity. This herb may not be good candidate for immunomodulatory activities. Anthum graveolens depicted no effect on immune functions at any dose used in this study. These reports highlight the needs for further study, in particular to investigate the possible action of C. occidentalis, T. foenum-graecum, D. denudatum extracts and their components in interfering with cytokine production. 

3.         In this study, C. occidentalis T. foenum-graecum and D. denudatum showed antitumor and antibacterial activities against ehrlic’h ascites tumor cell line and S. typhimurium bacteria respectively.

4.         During the functioning of immune system, such as in phagocytosis and respiratory burst activity, reactive oxygen species are generated, if they are left unchecked they may lead to peroxidative damage to other tissue. In this investigation all the herbs showed an inhibition effect on in the lipid peroxidation in both the liver and kidney.

5.            Modulatory role of plant extract on drug metabolizing enzymes and antioxidants was investigated. The extracts of C. occidentalis and D. denudatum showed significant decrease in the level of cytochrome P450 in the both the liver and kidney of mice. However, no effect was observed in case of T. foenum-graecum at any dose level. These observations of C. occidentalis and D. denudatum might be indicative of inhibition of oxidative metabolic processes and thereby a blocking of the activation of xenobiotic compounds including toxic chemicals. There is persuasive evidence to support the induction of glutathione S-transferase and protection against a wide spectrum of cytotoxic, mutagenic and carcinogenic chemicals. All the plant extracts showed significant increase in the activity of glutathione S-transferase in both the liver and kidney.

9.            Modulation of hepatic and nephrotic detoxification enzymes by these herbs plays an important role in the detoxification of drugs and chemicals. It is therefore proposed that use of C. occidentalis, and T. foenum-graecum may be helpful in patients suffering from immunosuppression and they may also provide relief by facilitating fast detoxification of drugs that are known to cause toxicity in the patients undergoing chemotherapy. Anti-mutagenic and hepatoprotective effect of C. occidentalis are well established in animals. It is also an important constituent of Ayurveda and Unani formulations. Further investigation in animals and human are warranted on this plant.

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Summary No. December 2002/05 

Title "Studies On the Neurotoxicology Of Argemone oil"

Subject: Toxicology
Name of Student: ALMAS SIDDIQUI (almassiddiqui@yahoo.com)
Department: Medical Elementology &Toxicology, Faculty of Science
Name of Supervisor: Dr. Fakhrul.Islam (fislam2001@yahoo.com)
Source of funding: Central Council of Research in Unani medicine (CCRUM)
Date of Viva Voce: December 20, 2002


Summary

Chapter 1 describes the in vivo protection of argemone oil induced neurotoxicity by Argemone mexicana aqueous extract. All the rats were sacrificed and their brain were dissected out for the estimations of Na+ K+ ATPase, lipid peroxidation, reduced glutathione content and the activity of glutathione metabolizing enzymes, glutathione peroxidase, glutathione reductase and glutathione-S-transferase and serum was used for the study of the marker enzymes of hepatotoxicity; serum glutamate pyruvate transaminase (SGPT) & serum glutamate oxaloactate transaminase (SGOT). Argemone oil depleted the content and activity of all the above parameters significantly and increase the activity of SGPT & SGOT. The A. mexicana aqueous extract protected the activity of Na+ K+ ATPase, GPx, GR, GST and content of GSH to a significant level and brings down the activity of SGPT & SGOT and the content lipid peroxide in brain when compared to the group treated with argemone oil alone. These results indicate that prophylactic treatment of A. mexicana aqueous extract offered partial protection against argemone oil induced neurotoxicity and hepatotoxicity.

Chapter 2 describes the in vivo protection of argemone oil induced neuro & hepatotoxicity by argemone maxicana aqueous extract for a period of 15days.. On day 16th all the rats were sacrificed and there brain parts and liver were dissected out for the estimations of lipid peroxidation, reduced glutathione and the activity of glutathione metabolizing enzymes, glutathione peroxidase, glutathione reductase and glutathione S-transferase. The Argemone maxicana aqueous extract was found to be helpful in up regulating the enzymes activity to a significant level and lowering down the lipid peroxidation in brain areas as well as in liver when compared with argemone oil treated group. These results indicate that prophylactic treatment of Argemone mexicana aqueous extract offered partial protection against argemone oil induced toxicity in discrete brain areas and liver.

Chapter 3 describes the in vivo effect of acute and sub acute doses of argemone oil on antioxidant status in striatum, hypothalamus, hippocampus & thalamus. Adult male Wistar rats (125±10 g) from Jamia Hamdard animal breeding colony were used in this study and kept on commercial pellet diet and water ad libitum for 12 hr light and dark cycle each. The animals were divided into four groups two control and two experimental each having 8 rats. Group 1 and 2 received intra peritoneal (i.p) injection of saline for 3 and 15 days respectively which served as control. The group 3 and group 4 received argemone oil 1.5 mL/kg b wt & 0.2 mL/kg b wt through i.p. route for3 and 15 days respectively. On day 16th animals were sacrificed and brain regions were dissected out for estimations of lipid peroxidation, glutathione & its metabolizing enzymes; glutathione peroxidase, glutathione reductase and glutathione-S-transferase. These results suggest that a lower dose of argemone oil when given for longer duration is more toxic as compare to the higher dose which was given for short duration.
 
Chapter 4 describes the in vivo effect of argemone oil on antioxidant status, dopamine (DA) and its metabolite dihydroxy phenyl acetic acid (DOPAC) in striatum, hypothalamus, hippocampus & thalamus regions of the brain. The SGPT and SGOT were estimated in the serum of the animals. The male albino rats (125±10g) were divided into 4 groups each having 8 animals. The group I received normal saline i.p. for 14 days. Group II, III & IV received argemone oil 0.05, 0.1 and 0.2 ml/kg b.wt (i.p.) respectively for 14 days. On day 15, all the animals were sacrificed and brain regions were dissected out for estimations of DA, DOPAC, lipid peroxidation, glutathione & its metabolizing enzymes; glutathione peroxidase, glutathione reductase and glutathione-S-transferase. It has elevated the level of DA and DOPAC in all the regions significantly and dose dependently. Argemone oil has depleted the level of reduced glutathione and the activity of its dependent enzymes and increase the content of LPO at the dose of 0.1 & 0.2 ml/kg, but no significant changes were observed with 0.05 ml/kg. The argemone oil (0.1 & 0.2 ml/kg) doses has increased the activity of SGPT and SGOT but no significant changes were observed with 0.05 ml/kg argemone oil. Our results suggest that use of argemone oil at lower doses might be of therapeutic importance in the diseases where there is a potential loss of dopamine take place.

CONCLUSIONS
Argemone oil shows its acute and subacute as well as dose dependent toxicity in whole brain as well as in discrete areas of brain; Striatum, hypothalamus, hippocampus and thalamus..Oral supplementation of aqueous extract of A. mexicana stem & leave shows the protective effect on the brain and liver.At lower doses, argemone oil might have some beneficial effect in neurodegenerative disorders like Parkinson's and Huntingtoncorea where there is a severe loss of Dopamine as argemone oil enhances the level of Dopamine and its metabolite DOPAC without any apparent signs of toxicity

"Nature Has Provided Toxicity and Remedy In the Same Plant"

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Summary No. December 2002/06 

Title: ‘Phytochemical investigation of some plants used in Indian system of medicine.’
Subject:
Chemistry
Department:
Chemistry
Name of the student:
Hinna Hamid (hinna03@rediffmail.com)
Name of the supervisor:
Dr. M.S. Alam
Date of Viva Voce:
31st December 2002

Summary

The present research work involves isolation and characterization of twenty-three compounds, including nineteen new compounds isolated from the extracts of four different plants. Various extracts of these plants have also been screened for their anti-inflammatory, analgesic and antibacterial activities.

Quercus Infectoria Olivier (Fabaceae) is a small tree, which yields galls that find extensive use as antiseptic, against inflammation and eczema, in the treatment of piles, dysentery and diarrhea. The compounds isolated from this plant include.

Ø      A new protocatechuic acid ester, named as infectoyl ester [M+ 196, C10H12O4], characterized as 2,6-dimethylprotocatechuic acid methyl ester.

Ø      A novel phenolic compound, named as quercanaphthalene [M+ 252, C12H12O6] characterized as 2-methyl-3-hydroxymethylene-4,5,6,7,8-pentahydroxy naphthalene.

Ø      A new ellagic acid glycoside, named as quercoside [M+ 772, C32 H36 O22] characterised as ellagic acid-4-O-[b-D-glucopyranosyl]-10-O-[b-D-glucopyranosyl]-(4®1)-b-D-rhamnopyranoside.

Uraria lagopoides D.C. (Fabaceae) is a trailing perennial herb is prescribed as an alcoholic extract in inflammation of chest and remittent fever. It cures asthma, fevers, dysentery, delirium, ulcers and is good for tumors, malarial fevers, eye diseases and fractures of bones. The compounds isolated from this plant include.

Ø      A new hydrocarbon, named as Urariane, [M+ 618, C44 H90], characterized as n-tetratetracontane

Ø      A new molecule, designated as Uraroic acid A [M+ 554, C33 H62O6] characterized as n-7¢-carboxylic-heptacosanyl-glutarate.

Ø      An unreported acidic compound, designated as Uraroic acid B [M+ 498, C30 H58O5] characterized as n-nonacosan-5-ol-1, 13-dioic acid.

Ø      A novel sterol, called urariosterol [M+ 416, C29 H52O], characterized as 8,14-13,17-diseco-stigmast-5, 22 diene-3-a-ol

Ø      An unknown acid, given the name Uraroic acid C [M+ 280, C17H28O3], identified as 4,12-dimethyl-n-tetradeca-6,8,10-trien-1-ol-3-oic acid.

Ø      A known steroid glycoside, characterized as stigmasta-5-en-3-b-D-glucopyranoside [M+ 577, C35 H61O6]

Ø      An unknown triterpene glycoside, given the name Urarialanostenoside [M+ 913, C48 H82O16] characterized as Lanost-9(11)-en-3-O-b-D-glucopyranosyl-(4®1)-D-glucopyranosyl-(4®1)-D-glucopyranoside.

Astragalus hamosus Linn. (Fabaceae) An annual herb, has been reported to be emollient, demulcent, aphrodisiac, diuretic, laxative and good for inflammation, ulcers and leucoderma.

The compounds isolated from this plant include.

Ø      An unknown hydrocarbon, named Hamosane [M+ 310, C22 H46], which was identified as 10-methyl-n-heneicosane.

Ø      A known sterol, namely b-sitosterol [M+ 414, C29 H60O]

Ø      A new phenolic molecule called Hamosol [M+ 164, C11 H16O] characterized as 2,3,4-trimethyl-5-ethyl phenol.

Ø      A novel phenol designated as Astragalone [M+ 424, C26H32O5], characterized as 1-(n-undeca-3-on-yl)-3,4,6-trihydroxy-7-methoxy-antharacene.

Ø      A known sterol glycoside i.e., b-sitosterol glycoside

Ø      Hamosoic acid characterized as Benzene-2-acetaldehyde-1-oic-3-O-b-D-glucopyranoside .

Ø       Hamoside, characterized as 1,6-dihydroxy-4-methoxy-7-O-b-D-glucopyranosyl naphthalene.

Ø      A known disaccharide identified as sucrose

Lavatera kashmeriana J.C (Malvaceae).The compounds isolated from this plant include.

Ø      A new sesterpene, called Lavaterone [M+ 362, C25 H46O] identified as 11-(4,8,10-trimethyl decalinyl)-13,17-dimethyl decan-19-one.

Ø      An unknown homoditerpene designated as Lavaterepene [M+ 306, C22 H42], characterized as 4,8,10-timethyl-9-(13,16-dimethyl heptanyl) decahydro-naphthalene.

Ø      A new molecule named Lavateral, [M+ 230, C13 H11O4], identified as 2¢,3,3¢-trihydroxy-2-formyl biphenyl.

Ø      A novel sterol glycoside named as Lavaterosterol [M+ 578, C35 H62O6] characterized as 8,14-seco-stigmasta-5-en-3b-ol-b-D-glucopyranoside.

Ø      An unreported glucuronic acid derivative named as Lavateronic acid [M+ 274, C12 H18O7] characterised as 2,4,hexadienyl-1-b-D-glucuronic acid.

 

Various plant extracts were also evaluated for their anti-inflammatory, analgesic and antibacterial activity.

Orally administered aqueous extracts (200 mg/Kg body wt.) of U. lagopoides and A. hamosus showed significant inhibition  (62.54, 65.41, respectively) of carageenan induced rat paw edema in comparison to standard, indomethacin (76.68). They also showed marked inhibition (62.16, 60.71, respectively) of acetic acid induced writhing in mice in comparison to standard, acetyl salicylic acid (71.59). Orally administered alcoholic extracts of U. lagopoides, in doses 200 and 100 mg/Kg body wt. showed significant inhibition 70.25 and 60.13, respectively, of rat paw edema induced by carageenan. The orally administered alcoholic extracts of A. hamosus, in same doses inhibited the rat paw edema by 69.93 and 63.02 percent. Orally administered alcoholic extracts of U. lagopoides, in dosages 200 and 100 mg/Kg body wt. showed significant inhibition 65.99 and 59.24 of writhing induced by acetic acid, further alcoholic extracts of A. hamosus inhibited the acetic acid induced writhing in mice by 67.16 and 58.35 percent, respectively.

The aqueous and alcoholic extracts of L. kashmeriana failed to show inhibition in both the tests. 

Petroleum ether, chloroform and alcohol extracts of U. lagopoides, A. hamosus and L. kashmeriana in various concentrations were found to be active against Gram-positive bacteria, while as only the alcohol extracts of these plants showed significant activity against Gram-negative bacteria. Chloroform extract of L. kashmeriana also showed a weak activity against Gram-negative bacteria.

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Summary No. December 2002/07 

Title: Phytochemical and Pharmacogistical studies on Some Indian Medicinal Plants.
Subject:
Chemistry
Department:
Chemistry
Name of the student:
Syed Tarique Abdullah
Name of the supervisor:
Dr. M.S. Alam
Date of Viva Voce:
24th December 2002

Summary

The present research work involves isolation and characterization of twenty-six compounds, including nineteen new compounds isolated from the extracts of three different plants, their standardization and biological evaluation for their anti-inflammatory and antibacterial activities.

Parmelia perlata Ach. A lichen of family Parmaleaceae, known for its emollient, tonic, febrifuge, diuretic, demulcent and sedative properties yielded

Ø       29, 30-nordimethyl-lanost-2-ene, named parmelialanostene

Ø       naphtha-4-(4¢-methylbutan-4¢-olyl)-6-methyl-8,9-diol-1-one, named parmenaphthone A

Ø       3-ethyl-6-methyl-7-carboxy-8-hydroxymethylene-9-hydroxy-naphtha-1-one, named parmenaphthone B

Ø       3-(pentdienyl)-2,5-dimethyl-11-aldehydic-13-hydroxymethylene-10,12-dimethoxy stilbene, named Parmenal

Ø       4¢-hydroxy-5¢-methyl-1¢-pyrogallolyl-(2-formyl-3-hydroxy-4,5-dimethyl) benzoate, named parmelyl ester A

Ø       4¢-methylcarboxy-3¢-hydroxy-2¢,5¢-dimethyl-1¢-phenyl-(3-aldehydic-2,4-dihydroxy-6-methyl) benzoate, named Atranorin.

Ø       4¢-methylcarboxy-3¢, 5¢-dihydroxy-2¢-methyl-1¢-phenyl (3-aldehydic-4-hydroxy-2,6-dimethyl) benzoate, named parmelyl ester B

Ø       Lup-20(29)-en-3a,27-diol, named parmelupenol A

Ø       Lup-20(29)-en-3a,28-diol, named parmelupenol B

Ø       16,18-normethyllabdone-18-(3¢-O-methoxy-benzoyloxy)-O-methoxypyrocatechol, named parmelabdone

Ø       b-3-(deca-11-one-yl)-5-methoxy-pyrogalloyl-(2¢,7¢-dihydroxy-5¢-methoxy) naphthyl ether, named parmenone.

Rubia cordifolia Linn. The plant has antioxidant, antihaemorrhagic, hepatotoxic, antineoplastic, antiallergenic, antibacterial and antiviral properties. Dried roots are considered to be useful in alleviating dropsy, paralysis, jaundice, amenorrhoa and visceral obstruction. It exerts uricolytic activities and thus promotes disintegration as well as elimination of urinary stone. The plant yielded

Ø       1-hydroxy-3-carboxy-6-ethyl-9, 10-anthraquinone, named as Cordifoloic acid

Ø       1,3-dihydroxy-2-methyl-9, 10-anthraquinone, named Rubiadin.

Ø       1,3-dihydroxy-9, 10-anthraquinone, named Xanthopurpurin

Ø       1, 2-dihydroxy-9, 10-anthraquinone, named Alizarin.

Ø       1-methyl-2, 4-dihydroxy-9, 10-anthraquinone named as Rubiol

Ø       Urs- 12- en-3b-ol, 3-epi-a-amyrin

Ø       1-hydroxy-3-ethyl-9, 10-anthraquinone, named as Cordifoliol

Ø       1,8-dihydroxy-11,20 (15-Pentyl- naphthaquinonyl)  phenanthranene, named Cordifodiol.

Rumex maritimus Linn. The plant has refrigerant, antipyretic, cathartic and aphrodisiac activity. It has been used to treat burns and back pain and the alcoholic extract has antifungal activity against Trichophyton mentagrophytes and Micospermum canis.

The plant yielded

Ø       n-dotriacont-3-one, named as Maritimone

Ø       n-heptacos-18,20,22-trien-1-ol, named as Rumexenol,

Ø       22-n- hexyl-12-(a-tetradecahydroanthracenyl)-undecan-3-ol-1-oic acid, named as Rumexoic acid

Ø       quercetin

Ø       sitosterol glycoside

Ø       3,7,11,15-tetramethyl-n-octadac-13-en-3,4,6,7,8,11-hexol-12-one, named as Rumexenone

Ø        8-normethyl-16-(19,22-dimethyl-19,22-dihydroxy-heptanyl)-spongiene-5-en-7-on-8-ol-22-b-glucopyranosyl-b-D-glucopyranoside, named as Maritiaspongiene.

Different extracts of these plants exhibited significant anti-inflammatory and anti- bacterial activity against staphylococcus aureus (G+).

Standardization parameters included morphological and microscopical characters, Physiochemical properties and TLC fingerprints.

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Summary No. December 2002/08

Title: "Role of Nitric oxide in Pressure overload Cardiac Hypertrophy"
Faculty: Center for Biotechnology, Faculty of Science
Name of student: Srinivas M. Tipparaju
Supervisor: Professor S K Jain, Dr M P Dubey
Date of viva-voce: 16-12-2002

Cardiac hypertrophy has reached epidemic proportions worldwide and with no solid evidence of an independent benefit beyond blood pressure control newer therapeutic strategies have to be designed. It has been known since long that hypertension leads to hypertrophy, which disrupts the normal homeostatic mechanisms. We undertook this problem with a view to understand the mechanisms involved in the hypertrophic (two-kidney one-clip; 2K1C) heart subjected to ischemia-reperfusion or hypoxia-reoxygenation challenge.

The study was divided into two parts, the first dealing with the calcium regulatory mechanisms and the second with the Nitric Oxide to assess the performance of hypertrophic hearts during ischemia-reperfusion injury. Evaluation was done for both functional and cellular responses by performing contraction studies and peroxynitrite estimations in whole heart preparations, whereas intracellular calcium, nitrite/nitrate ratio and total ROS estimations were done in myocytes. Assessment was done using various nitric oxide modulators such as L-NAME (NOS inhibitor), SNP (NO donor), L-arginine (NO supplement) and uric acid (ONOO- scavenger). The results suggest that Nitric oxide synthase inhibition by L-NAME offers a strong strategy that gives protection to the hypertrophic heart during ischemia-reperfusion injury, in terms of functional changes and providing protection from cytotoxic effects of peroxynitrite. At cellular level it lowered intracellular calcium transients, attenuated reactive oxygen species generation and maintained a stable mitochondrial membrane potential.

With Ca2+ and NO taking center-stage during ischemia-reperfusion injury, this is the first time that an elaborate cross talk between these two key events has being reported in the hypertrophic rat heart.

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Summary No. January 2003/01 

Title: Studies On Antitumour Activity Of Indole-3-Carbinol In Experimental Animals.   
Subject: Toxicology.   
Name of Student: Ms Bhawna Srivastava   
Department: Medical Elementology & Toxicology, Faculty Of Science.   
Supervisor: Dr Y. Shukla (yogeshwer_shukla@hotmail.com)
Scientist, Industrial Toxicology Research Center, Lucknow.   
Co-Supervisor:  Dr S. Raisuddin (s.raisuddin@hotmail.com)
Date of Viva ATE OF VIVA Voce: January 08, 2003. 

SUMMARY:   

In this dissertation it has been tried to evaluate the antitumour activity of Indole-3-carbinol in the living beings. For this purpose Swiss albino mice were selected. This study was conducted in Industrial toxicology & research center, LUCKNOW. Detailed investigations involving tumour  developing techniques & histological techniques were used to demonstrate I3C action. 

The antimutagenic, anti-tumourogenic & anti-tumour promoting activity were studied using tumourogenic & tumour  promoting chemicals. These chemicals included 7, 12dimethylbenz (a)anthracene(DMBA), Benzo(a) Pyrene(BaP),Cyclophosphamide(CP) & 12-0,tetra decanoylphorbol-13acetate(TPA). Along with this cytotoxity of I3C was observed on Ehrlich ascites cells. BaP & DMBA are widely occurring Polycyclic

aromatic hydrocarbons. CP is prominently used anti-cancer drug showing its effect on cells due to its mutagencity. To show anti-mutagenicity & anti-genotoxicity of I3C CPwas used as a positive control.Here study was conducted on bone marrowcells of mice. CP has been shown to Cross Link with DNA both in vitro & invivo & thus  induces point mutations. CP directly affect cell divison by inhibiting Mitotic index. I3C also mildly shows inhibition of Mitotic Index. But when given with CP shows decrease of cytotoxicity of  CP. 

Swiss albino skin model was used to show antitumourogenic activity of I3C when it was applied topically on skin either one hour prior or one hour after carcinogen administration . I3C showed tumour initiation in both methods but it was more pronounced when it was applied after carcinogen application. Parameters used to show this  were tumor induction time ,average tumour per mouse ,percentage of animals with tumour  and cumulative number of tumours. For  anti tumour promotion activity of I3C  two stage skin model was used . A sub carcinogenic dose of DMBA was applied on skin then I3C was applied  topically prior to application of TPA a well known tumour promoter. Here also the parameters were the same as in previous experiment which demonstrated antitumourogenic activity of I3C. I3C has been shown to increase life span of Ehrlich ascites  tumour bearing mice.It also showed  cytotoxicity  against EA cells.These investigations and experiments showing anticarcinogenic& antimutagenic potential of I3C  involving an array of skin tumour model, transplantable tumour model& mutagenicity test have provided  important information that I3C can be used as adjuvant for cancer chemoprevention.

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Summary No. January 2003/02 

Title: Studies on modulatory role of antioxidants against immunotoxicity of anticancer drugs

Subject: Toxicology
Name of the student: Mr. Rizwanul Haque (haquerizwan@hotmail.com)
Department of Medical Elementology & Toxicology, Faculty of Science
Supervisor: Dr. S. Raisuddin (sraisuddin@hotmail.com)
Co-supervisor: None
Source of funding: CCRUM & CSIR (Govt. of India)
Date of viva voce: January 10, 2003

 Summary

There are many synthetic and natural chemotherapeutic agents available for the treatment of cancer but none of them is effective alone. Some of the most widely used chemotherapeutic agents are doxorubicin, cisplantin, methotrexate, mitomycine and cyclophosphamide. Cyclophosphamide (CP) remains the most widely used alkylating agent used in cancer chemotherapy and still extensively studied both clinically and experimentally. Because of its widespread use side effects with CP treatment are frequently reported. The immunosuppressive effects of CP are the major cause of concern for oncologists. 

It is believed that used of immunomodulating herbs and/ or antioxidants may provide great relief from the side effects of CP in the patients. The present study was undertaken to assess potential of some antioxidant agents in the form of herbal extracts and purified antioxidant compounds in abrogating toxicity of CP in mice. In this study, three herbal extracts viz; Emblica officinalis, Juglans regia and Ginkgo biloba and two well-known antioxidants viz; vitamin C and catechin have been used. 

All the plant extracts alone and in combination with CP-treated animals showed improvement in the cellularity of lymphoid organs. Plant extracts treatment caused a significant improvement in humoral immunity, cell mediated immunity and non-specific immune functions in CP-treated animals.  Animals treated with vitamin C (10 mg/kg body weight) showed no improvement in the relative organ weights. Vitamin C and catechin along with CP restored the specific and non-specific immune response.  

J. regia extract showed appreciable tumor inhibitory activity in Ehrlich ascites tumor challenged animals. It also showed an increase in percentage survival time and showed reduction in the mean body weight of tumor bearing mice.  

A significant reduction was observed in cytochrome P450 content in liver and kidney of E. officinalis and in kidney of J. regia treated animals where as no change was observed in G. bioloba treated animals. Ginkgo and J, regia extract along with CP resulted in a significant improvement in the Cyt. P450 content in both the tissues. Extracts of E. officinalis and J. regia did not show induction in the glutathione S-transferase (GST) activtiy except G. biloba. All the three plants showed restoration in the activities of GST in CP-treated animals in both the tissues. J. regia and G. biloba extracts significantly induced reduced glutathione (GSH) levels in both the tissues (liver and kidney) whereas induction by E. officinalis was not significant. These plants also restored the depleted level of GSH in CP treated animals. Almost all the plant extracts showed significant augmentation in the activities of antioxidant enzymes in both the tissues especially in CP intoxicated mice.  

Vitamin C alone failed to stimulate the drug metabolizing enzymes effectively in kidney but in combination with CP showed an improvement in the cytochrome P450 content in both the tissues. Vitamin C did not induce GST and GPx in control animals but it restored the depleted levels of GST, glutathione peroxidase (GPx), superoxide dismutase (SOD) and catalase (CAT) in CP treated animals. Catechin + CP treatment showed a significant improvement in the content of Cyt. P450 in liver and kidney as compared to CP treated animals. Catechin and catechin + CP groups showed a significant induction in the activity of CAT, SOD, GPx, GST and in the level of GSH in liver and kidney except glutathione peroxidase in the kidney. 

CP-treated animals that were administered plant extracts and antioxidant compounds showed a decreased LPO level in kidney and liver as compared to CP-treated animals.

Large number of natural antioxidants and herbal preparations are available to scavenge the reactive oxygen species and modulate the immune system. It is the need of hour to conduct detailed studies on selected natural antioxidant compounds that have potential to modulate the immune system as well as suppress the toxicity of the synthetic drugs used for the treatment of cancer.  Immunomodulatory therapy could provide an alternative to conventional chemotherapy for the varieties of advantage its offers. The present study conducted on mice offers some weightage to those advantages.

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Summary No : January 2003/03 

Title : Studies on Antimutagenic potential of Black tea and its constituents.
Subject : Toxicology
Faculty/Department: Department of Medical Elementology andToxicology,
Faculty of Science, Jamia Hamdard, New Delhi
Name of the student:
Pankaj Taneja(pankaj1275@yahoo.co.in)
Name of the Supervisor: Dr. Yogeshwer Shukla(yshukla@itrc.res.in).
Name of the Co-Supervisor : Dr. Sarwat Sultana(sarwat786@rediffmail.com)
Address of the Co-Supervisor:
Chemopreventive and Nutrition Toxicology.

Laboratory, Department of Medical Elementology
andToxicology, Faculty of Science, Jamia Hamdard,
Source of Funding : National Tea Research Foudation, Calcutta

Date of viva: 16th January, 2003
 

Summary

Black tea (Camellia sinensis) is one of the most popular and commonly consumed beverage worldwide possessing many chemopreventive properties. In the present set of investigations antimutagenic potential of Aqueous black tea extract (ATE) and Black tea polyphenols (BTP) was evaluated. Antimutagenic activity profile of ATE and BTP was notified towards Benzo(a)pyrene [BaP] and Cyclophosphamide [CP] in Salmonella typhimurium TA 98 and TA 100 tester strains. The induction of his+ revertant colonies by above mutagens was found to be inhibited by black tea. The frequency of chromosomal aberrations, micronucleus formation, and SCE’s induction by BaP/CP was reduced by ATE/ BTP administration in bone marrow cells of Swiss albino mice. The anticytotoxic potential of ATE and BTP was also evidenced by increase in the status of mitotic index. In addition black tea also exhibited protection in germ cell in dominant lethal mutation assay. Decrease in the number of living implants by BaP and CP was found to show increment by ATE and BTP treatment. The post implantation losses induced by BaP/CP were found to be reduced by ATE/BTP. The frequency of dominant lethal mutations induced by BaP/CP in different mating weeks was notified to inhibited by ATE/BTP. Hence the study depicts that black tea has potential in suppressing mutagenicity and cytotoxicity in microbial and mammalian somatic and germinal test systems.

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Summary No : January 2003/04 

Title: Pharmacognostical and Phytochemical Appraisal of Some Herbal Drugs of Indian Medicine.
Subject: Pharmacognosy
Name of the Student: Mr. Parwaiz Akhtar
Departmant/Faculty : Department of Botany, Faculty of Science.
Supervisor : Dr. M. P. Sharma
Co-supervisor : Prof. Mohd. Ali, Faculty of Pharmacy
Source of funding : None
Date of viva voce : January 10, 2003
.

Summary 

The present study pertains to detailed pharmacognostical and phytochemical investigation of four herbal drugs namely: ‘Funduq’ Corylus colurna (Betulaceae), the officinal part of the drug is fruit used in Unani system of medicine as a brain and intestinal tonic, aphrodisiac and expectorant; ‘Kurchi’ Holarrhena antidysenterica (Apocynaceae), the stem bark is used as a principal remedy in cases of various types of diarrhoea and dysentery and this drug is generally adulterated or substituted with the bark of Wrightia tomentosa and W. tinctoria;  ‘Khulanjan’ Alpinia galanga (Zingiberaceae), the rhizome is commonly used for many respiratory ailments in the indigenous system of medicine   and this drug is usually substituted with A. officinarum and  ‘Aatrilal’    Ammi majus (Apiaceae) the fruit is an officinal part used in powdered form in the indigenous systems of medicine for the treatment of leucoderma with promising results and the fruit of this drug is mostly substituted with its allied species A. visnaga .

Macro- and microscopical characters, behaviour of powdered drug on treatment with different chemical reagents, fluorescence analysis,   extractive values, ash values and preliminary phytochemical tests were carried out to study the distinctive features of the drugs. Such  parameters provide basis for standardisation/characterisation of genuine drug. 

Detailed chemical analysis of fruit of ‘Funduq’ resulted in the isolation of four chemical compounds, namely colurnasterol glycoside, b-sitosterol glycoside, campesteryl glycoside and corylusterol glycoside. Two compounds: (i) 20-b-hydroxycampesterol-3b-D-glycopyranoside (colurnasterol glycoside) and (ii) chloest-5-en b, 12, b-triol-b-D-glyopyranoside (corylusterol glycoside) have been reported for the first time from a natural or synthetic source. From the bark of ‘Kurchi’ four compounds were isolated which are holardysenterine A (1), holardysenterine B (2), holardysenterine C (3) and holarkolavene (1). These compounds have close resemblance to conessine, principal alkaloid, but differ in their orientation of C21 –methyl groups. Of these four compounds, one compound holardysenterine B (2) has been reported as new epimer of conessine and established as 3 b-dimethylamino-19b-21a-methylcon-5-ene. Rhizome concentrate of ‘Khulanjan’ on steam distillation yielded 11 volatile constituents. 10 constituents were completely identified. Of the eleven constituents, citronellyl acetate. n-tridecane, geranyl acetate, eincosanol, n-docasane, n-docosan-8-ol have been reported for the first time in the oil. Steam distillation of ‘Aatrilal’ fruit concentrate revealed the presence of 53 compounds, of which 43 compounds have been identified after the analysis of the isolate, comprising 86.8% of total volatilates including 5 partially characterised compounds (ca 4.8%). The most abundant classes of the compounds are 14 monoterpenes (37.3%) and 19 sesquiterpenes (20.2%). Piperitone (10.0%) has earlier been reported from Pakistan sample of A. majus but could not be detected in our sample.

A serious limitation encountered in the use and research of traditional medicine is the lack of standardisation and quality control of raw material forming the drug. The ultimate objective of the pharmcognostic investigation is identification of the genuine crude drug and determining the extent of adulteration/substitution, if any. Advancement in recent years in pharmacognosy, phytochemistry and physicochemical instrumentation techniques can be of immense value in removing this major shortcoming of traditional medicine. These techniques can be utilised for correct botanical identification of plants. The details of organoleptic, macro- and microscopic characters, various evaluative parameters, fluorescence analysis, results of preliminary and detailed phytochemical analysis established in the present study will facilitate in identifying the genuine drug from any substitute or spurious samples and will also be useful in preparation of monographs on these plants.

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Summary No. February 2003/01

Title: “PRODUCTION, CHARACTERIZATION, CHEMOSELECTIVE HYDROLYSIS AND MOLECULAR GENETICS OF EXTRACELLULAR MICROBIAL LIPASE”
Subject: Biotechnology
Name of Student: Mr. Manish Kumar Tripathi (mktcdri@yahoo.com)

Department: Centre For Biotechnology, Faculty Of Science
Supervisor: Dr. P.K.Roy, Deputy  Director, Fermentation Technology Div., Central Drug Research Institute, Lucknow-226001, U.P., India. (prodyutr@yahoo.com)
Co-Supervisor: Prof. S.K.Jain, Centre For Biotechnology, Jamia Hamdard, Hamdard University, New Delhi – 110 062 India. (sk608@rediffmail.com)
Source of Funding: CSIR
Date of viva voce: February 24, 2003

Summary

The importance of thermostable and alkaline lipases is growing rapidly, since these enzymes are relatively stable as well as are capable of carrying out diversified reactions. The inability of the plant and animal lipases to meet current world demands has led to an increased interest in microbial lipases. Microorganisms represent an excellent source of enzymes owing to their broad biochemical diversity and their susceptibility of genetic manipulation. Lipases have a large variety of applications mainly in the detergent, cosmetic, drug, leather, paper, food industry and in several bioremediation processes. So owing to their vast and varied applications newer microbes are to be screened for production of lipases of desirable properties. Cloning and sequences analysis of the lipase gene will help to understand the structure and activity relationship of the enzyme, which will enable researchers to tailor new lipases for biotechnological applications.

Screening of lipase producing microorganism from soil, Optimization of process variables, Physico-chemical  characterization and purification of the enzyme

Lipase producing microorganism, identified by MTCC (IMTECH, Chandigarh) as Streptococcus sp. (N1) was isolated from soil using enrichment technique. To the best of our knowledge, to date there is no report available on lipase from genus Streptococcus, and more than hundred species and subspecies of Streptococcus are listed on Taxonomy browser at NCBI site (Taxonomy ID: 1301). The isolate Streptococcus sp. (N1) has been given Taxonomic ID: 167506 at the Taxonomy browser at NCBI site. A two parameter complete experimental design for optimization of carbon and nitrogen sources was applied. Other requirements such as soyameal, potatoflour, trace elements and parameters such as pH, temperature, inoculum age etc. were optimized to give a maximum production of 3.54 x 103 lipolytic units per ml of fermentation broth in 48 h.

The biochemical properties namely pH optima and stability, temperature optima and thermal stability, organic solvent stability, activation and inactivation by different metal ions, detergents and inhibitors were studied. The enzyme activity was maximum between pH 8.0 and 8.4 and was stable between a pH range of 5.0 – 10.5 upto 96 h. 37oC was the optimum temperature for N1 lipase. The majority of tested metal ions had no inhibitory effect on N1 lipase activity except Hg2+. The activity of the lipase was influenced by DEPC, whereas PMSF had little effect. Enzyme stability was affected by PMSF. No effect of EDTA was seen on enyme activity and stability. Streptococcus sp. N1 lipase retained 100% activity on incubation with Protenaise K for 90 min. No effect on stability was observed on incubation with organic solvents. An electrophorically homogenous lipase having specific activity of 295.3 X 103 U/mg and 361 purification fold has been obtained. The mol. Wt. of purified lipase was deduced to be 44.7 kD.

Probe experiments on biotransformations

Lipase are today enzymes of choice for organic chemists, pharmacists, biophysicists and biochemists. Lipase from Streptococcus sp. N1 has been successfully used for different biotransformation reactions using medicinally important CDRI compounds such as Loganin, Acetyl protected D-ribose and  Spermicidal compound.

Molecular cloning, sequencing and analysis of Lipase gene

A 3489 bp from mesophilic soil isolate Streptococcus sp. (N1) having structural lipase gene have been cloned in vector pBst II KS at EcoRI site (MCS) using genomic library approach. Southern hybridization confirmed the origin of insert from the host. Lipase producing recombinant clone C1N1 conferred a stable lipolytic phenotype. Full length sequencing was performed adopting the strategy of subcloning, end terminal sequencing, designing of primers for over lap and orientation. The 3489 bp fragment is having four significant ORFs. ORF-1 having a size of 1032 bp is giving signatures with alpha/beta hydrolase fold [104-325] and Esterase/lipase/thioesterase family active site [78-168]. ORF-1 is the structural lipase gene and encodes a protein of 343 a.a. with theoretical mass of 38.6 kD, and pI of 9.31. The instability index was computed to be 34.73, which classifies the protein to be stable. Streptococcus sp. N1 lipase contain highly conserved pentapeptide (G-X1-S-X2-G) containing the active site serine (G-H-S-I-G) [151-155]. This has been identified as interfacial lipid recognition site in several lipase sequences from prokaryotic and eukaryotic organisms. It also contain another consensus seuqnece (His-Gly) [84-85] in the N-terminal part of the enzyme. The searches made on databanks showed it to be novel sequence. The predicted amino acid sequence are giving significant homology with cold adapted lipases of antarctic psychrophiles Moraxella sp. TA144 (lip 3) and Psychrobacter immobilis B-10 (lip 1). ORF-2 of 642 bp is giving signatures with GDSL lipolytic enzyme [34-116] and ORF-3 of 729 bp with ABC transporter family [45-232]. The nucleotide sequence has been assigned Genbank Accession No. AF 395190.  

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Summary No. February 2003/02  

Title: “Studies on non-enzymatic antioxidants of fish with special reference to their use as biomarkers of aquatic pollution”
Subject: Toxicology

Name of Student: Mr. Suhel Parvez (parvezsuhel@yahoo.co.in)

Department: Medical Elementology & Toxicology, Faculty of Science

Supervisor: Dr. S. Raisuddin (sraisuddin@hotmail.com)
|
Source of funding: Ministry of Environment & Forests & Indian Council of Agricultural Research (Government of India)
Date of viva voce: February 07, 2003
 

Summary
 

In recent years, the impact of aquatic pollution on human and animal life has become a matter of great concern. Fish responses have been used as biomarkers of aquatic pollution. The use of a suitable biomarker with different degrees of specificity is an important aspect of environmental monitoring based on biomarkers.  In this direction an effort has been made to develop the biomarker responses as an early warning signal in pollution assessment. Studies were conducted on the different sites of river Yamuna for assessing the pollution profile. The oxidative responses, as well as antioxidant potential of fish differed in relation to species, habitat and feeding behaviour. The levels of heavy metals and pesticides were also found to be in varied concentrations at different sites. When comparisons were made between two sites, (Yamunanagar and Mathura), results revealed that Mathura site was more polluted compared to Yamunanagar. The water samples of Yamunanagar did not show any traceable amount of heavy metal and pesticides when compared to that of Mathura sample. The results obtained provide an insight for the assessment of non-enzymatic antioxidants at two different sites for their successful use as biomarkers.

 An attempt was also made to study protein carbonyl as a biomarker of exposure in fish. Use of protein carbonyl has been well documented in humans and rodents. Till date no study has been reported in case of fish. The present study has shown a biomarker approach using the protein carbonyl content in fish. A significant increase in protein carbonyls have been observed in all groups of fish exposed to different pesticides. Assay of carbonyl groups in proteins provides a conventional technique for detecting and quantifying oxidative modification of proteins. A study was undertaken to investigate the modulatory role of copper on non-enzymatic antioxidants viz., protein and non-protein thiols, ascorbic acid and metallothionein. It was also intended to investigate possible regulatory role of copper on iron in fish. The findings of the present investigation provided a new insight into the multifarious role of low-level exposure to copper in fish. It seems to protect the fish from peroxidative damage by inducing both non-enzymatic antioxidants and possibly antioxidant enzymes by the induction of ceruloplasmin and metallothionein. Findings of this study will be helpful in pollution monitoring and identification of pollutant-sensitive and pollutant-resistant Indian fish species. It will enable us to predict the pollution profile of an aquatic habitat viable for fishery activities. It will also contribute to the development of a battery of mechanism-based biochemical assays that can be used to characterize the complex mixtures of chemicals in different potentially toxic environments and thus enhance our ability to assess the long-term risk of environmental contaminants to human health.  

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Summary No. March 2003/01  

Title: “ Characterization and Standardization of Some Traditional Plant Drugs.” 
Subject: Botany
Department: Botany
Name of the student: Sageer Ahmad Khan
Name of the supervisor: Dr. Javed Ahmad
Date of Viva Voce: 27-03-2003

Summary 

In the following paragraphs the anatomical and phytochemical characters of root, leaf and flower drugs of six selected plants species have been described for the purpose of the standardization with their physical constants and pharmacognostical standards. There is resurgence of interest in ethnobotany and plants are reemerging as a significant source of new pharmaceuticals. The present research will certainly lead to systematic documentation of traditional knowledge and other botanical studies. 

Acacia leucophloea (Roxb.) Willd. (Mimosaceae) is a small or medium sized deciduous tree upto 6-8 meter in height. It commonly occurs in west land and open forest. The plant is highly medicinal having astringent, demulcent, aphrodisiac and anti-syphilitic properties.   

Macro and microscopic characters of the flowers of A. leucophloea show that there are numerous, globose, small, pale yellow, sweet scented heads. Solitary rhombohydron crystals are distributed in various regions of the peduncle, mainly in cortical parenchymatous cells and between the vascular bundles. 

Physico-chemical constants include: loss in weight up to 6.05% on drying at 105 °C. The solid content, crude fiber, total ash, water soluble ash, acid insoluble ash, sulphated ash and total alkaloids were reported up to 55.24%, 30.08%, 8.5%, 3.6%, 1%, 0.30% and 0.03% respectively.  

Sucrose (C12 H22 O11) was isolated from the flowers having m. p. 186-188 °C. 

Calliandra haematocephala Hassk. (Mimosaceae) is a bushy shrub upto1.5-4 m in height. It is distributed throughout tropical and sub tropical areas. The decoction product of the flowers is said to be blood purifier and also regarded as tonic. 

Macro and microscopic characters of the flowers of C. haematocephala are studied in detail. Pinkish red and scarlet powder puff like heads are very common. Pericycle contains a continuous and composite ring of 3-5 layers of hexagonal, thick walled sclerenchymatous cells in the peduncle zone.

Physico- chemical constants show loss in weight on drying at 105 °C up to 10.69%. The solid content, crude fiber, total ash, water soluble ash, acid insoluble ash,  sulphated ash, and total alkaloids were found up to 53.70%, 27.72%, 5.56%, 1.57%, 0.82, 0.02% and 0.06% respectively.  

Octacosane (C28H58) isolated from the flowers having m. p 60-620 C. 

Ehretia aspera Willd. (Ethretiaceae) is a shrub upto 3-5 m in height. It also occurs commonly in India. A decoction of the fresh roots is given in venereal diseases. 

Macro and microscopic characters of the leaves of E. aspera have been studied. The leaves are  3-10 x 2-6.5 cm in length, elliptical-oblong in shape, petiole 5-20 mm long. Stomata of anomocytic type, stomatal index (15), vein termination numbers (9-15), vein islet numbers (10-18) and palisade ratio (6-10) have been recorded. 

Physico- chemical constants show loss in weight up to 85.3% on drying at 105 °C. Crude fiber, total ash, water soluble ash, acid insoluble ash, sulphated ash and total alkaloids were measured and reported up to 49.5%, 10.67%, 2.57%, 1.06% 0.51% and 0.06% respectively.

 b-amyrin isolated from the leaves of this plant having m.p 196-1980C. 

Leucaena leucocephala (Lamk.)Wit. (Mimosaceae) is a large erect shrub over small tree upto 1.8-9.0 m in height. It is distributed in tropical regions. The bark of this plant is eaten for internal pain in Assam. The leaves are a good source of protein and carotene. 

Macro and microscopic characters of the leaves are studied. The leaves are bipinnate, 10-25 cm long, cauline and ramal. The taste is bitter and its odour is characteristic, stomatal index (12-20) and vein termination numbers (3-6) are recorded. The vascular bundles are arranged in horseshoe shape manner as shown in the transection of the rachis. 

Physico- chemical constants show loss in weight up to 91.4% on drying at 105 °C. The solid content, crude fiber, total ash, water soluble ash, acid insoluble ash, sulphated ash and total alkaloids were reported 49.9%, 37.39%, 11.27%, 4.87%, 1.02%, 0.53% and 0.05%.

The impure compound from the leaves could not be characterised. 

Trianthema  portulacastrum Linn.(Aizoaceae) is a diffuse, prostrate, branched herb, upto 30-65 cm long. It is found mostly in tropical regions. It is attributed with analgesic, antipyretic, anti-inflammatory, CNS depressant and stomachic properties and used in asthma, bronchitis, jaundice and oedema.s 

Macro and microscopic characters of the root of T. portulacastrum (white variety) are studied. The roots are small, fusiform, ranging between 8-18 cm in length. The Colour is yellowish white, odour is pungent and bitter in taste. Cluster crystals occur in some of the cortical cells of the root. Anomalous structure is also observed.  

Physico-chemical constants show loss in weight up to 5.5% on drying at 105 °C. The solid content, crude fiber, total ash, water soluble ash, acid insoluble ash, sulphated ash and total alkaloids were reported 35.6%, 41.5%, 10.5%, 7.7%, 1%, 0.51% and 0.05%. 

The compounds isolated from the root are b-sitosterol, stearic acid, palmitic acid and potassium nitrate. 

Mallotus philippinensis Muell Arg. (Euphorbiaceae) is an evergreen shrub and found throughout the tropical regions of India. Kamala powder alone is applied over syphilitic ulcer. It is also used as an anthelmintic, vermifuge and purgative medicine. 

Macro and microscopic characters of the commercial sample of the drug under the name kamala show that it is a granular brick-red powder. Microscopic examinations of powder reveal that it is a fine, purplish red or brick red powder consisting of no cellular structure. 

Physico-chemical constants show that the maximum percentage of the extract is found in chloroform (13.7%) and minimum in methanol (3.12%). Loss in weight on drying at 105°C was found up to 4.31%. The solid content, total ash, water soluble ash, acid insoluble ash and Sulphated ash were found to be 90.85%, 53.51%, 0.17%, 45.72%, and 20.55%. 

Rottlerin has been found in dark brown rhombic crystals, m. p.203-205 °C.

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Summary No. March 2003/02  

Title :Studies on the synthesis and characterization of oil epoxy from non conventional seeds oil”  

Subject: Chemistry
Name of Student: Mohd Aslam Khan (staratom@yahoo.co.in)
Department: Department of Chemistry, Faculty of Science
Supervisor: Dr. Najm Zaheer Khan (najmkhan@rediffmail.com)
Co-Supervisor: Dr. Sharif Ahmad (JMI, New Delhi)
Date of Viva-voce: 29th March 2003 

Summary:

During the last two decades, the synthesis of polymers from renewable resources has attracted considerable attention of research workers throughout the world because of the escalating prices of petrochemicals and high rate of depletion of the natural mineral resources. This necessitates a look at the renewable natural resources that can serve as alternative feed stock for monomers of the polymer industry. In this respect the oils of certain seeds, hold considerable promise as a source of unsaturated hydrocarbon, an excellent starting material for epoxidation and subsequent polymer production. The main objectives of the present studies was to epoxidized the unsaturated oil by in situ peracetic acid and cure it with conventional hardener like polyamide which was further modified by blending with natural polymers like starch and alike polymers to develop biomaterials. A detail investigation of the modified materials was carried out by various techniques/methods. The modified materials or blended materials were evaluated for their physico-chemical, mechanical, spectroscopical and thermal behaviour. A detail study of their external stimuli responsive characteristics was also carried out.

Extraction of oil, its characterization, epoxidation and further characterization of the epoxidized oil by physico-chemical analysis, GC, Gel Permeation Chromatography, FT-IR, NMR, DSC & TGA were studied. Molecular weight and polydispersity was determined by GPC. The epoxidized oil were found to be of low molecular weight with a polydispersity of unity showing that ECTO & EJCO had a fairly homogeneous structure and that ECTO is of high molecular weight than EJCO. 

Curing and blending of epoxidized oil: ECTO with polyamide and natural polymer i.e., starch or gum acacia respectively. The cured & blended products have been characterized by spectroscopic techniques; FT-IR and 1H-NMR, DSC, TGA and Mechanical strength as well as various physico-chemical tests were carried out. FT-IR & 1H-NMR spectroscopic techniques confirmed the curing of epoxidized Chukrasia tabularis oil with polyamide. The curing took place at the site of oxygen ring resulting in epoxy ring opening which is accompanied by prominent peak disappearance of 1H-NMR signal at d = 2.9 ppm, the characteristic peak of epoxy ring present in the epoxidized oil of Chukrasia tabularis. Mechanical strength was tested for the blended polymeric films where the EP (epoxy-polyamide) cured films showed very flexible as compared to the starch blended films which showed brittleness in characters. The blended polymeric films developed were studied for its swelling behaviour at different temperatures, pH, ionic strength, types of buffer. It was investigated that all the polymeric films or hydrogels showed maximum swelling in acidic medium. The equilibrium and oscillatory swelling behaviour have also been investigated in response to change in the pH of the swelling media. 

The new low molecular weight soluble polymers were synthesized by reacting Chukrasia tabularis oil with monocarboxylic acid in the presence of tetrabutylammonium halide salt (catalyst). The in vitro biocompatibility based on the epoxy-polyamide/starch blended polymers was investigated. It was observed that for the selected cell line cultured on the polymer do not induced cytotoxicity effects. 

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Summary No. April 2003/01  

Title: Molecular regulation of the 3-hydroxy-3-methylglutaryl Coenzyme A reductase in Artemisia annua.
Subject                        :  Biotechnology
Department/Faculty 
:  Centre for Biotechnology, F/o Science
Name of student      
:  Mr. Mohd. Israr
Supervisor                  :  Dr. M.Z. Abdin
Co-Supervisor            :  Prof. S.K. Jain
Date of viva-voce   
   :  08-04-2003 

Summary 

Malaria is still a major health problem in many developing countries. The parasite responsible for the vast majority of fatal malarial infections is Plasmodium falciparum. The first effective antimalarial drug was quinine, which was isolated from the bark of Cinchona. Since then malaria has been treated with quinoline based drugs such as quinine, chloroquine, mefloquine and primaquine. Unfortunately, many Plasmodium strains have now become resistant to these drugs. Artemisinin, a sesquiterpene-lactone is a novel antimalarial drug isolated from Artemisia annua . It may meet the dual challenges posed by drug resistant parasites and rapid progression of malarial illness. The relatively low yield of artemisinin is, however, a serious limitation to the commercialization of this drug. Hence, optimization of artemisinin yield either in vivo or in vitro is highly desirable. One of the approaches to enhance the production of this compound in vivo is through the regulation of rate limiting enzyme(s) of its biosynthetic pathway employing both exogenous and endogenous factors.

It has recently been shown that isoprenoid compounds are produced via two pathways. Plastid isoprenoids are formed via the novel pathway or Rohmer pathway, while cytosolic isoprenoids like sesquiterpenes, triterpenes, polyterpenes and sterols produced via common biosynthetic pathway i.e. acetate-mevalonate. Henceforth, to confirm the role of alternate pathway in artemisinin biosynthesis, feeding experiments were performed by us with radioactive precursors. The results obtained in these studies strongly suggest that artemisinin is synthesized via acetate/mevalonate rather than Rohmer pathway. Isolation and assay protocols of HMG-CoA reductase, the key enzyme of acetate/mevalonate pathway, for A. annua L. leaves were standardized. The HMG-CoA reductase activity and artemisinin content were monitored in the leaves of A. annua L. at different phenological stages and a strong correlation was observed between the enzyme activity and artemisinin content throughout the growth period of the crop. The combined application of 100 ppm growth hormones (IAA+GA3) at the vegetative stage resulted into optimum activity of HMG-CoA reductase and the higher contents of artemisinin as well as its immediate precursors, viz. artemisinic acid and arteannuin B. The in vivo and in vitro regulations of HMG-CoA reductase activity were studied employing exogenous (light and temperature) and endogenous factors (precursors and products of isoprenoid pathway). Both these factors were significantly modulated the activity of this enzyme both in vivo as well as in vitro conditions. Finally, the enzyme was purified and characterized from the leaves of A. annua L. plants. One major band of molecular weight of 60 Kda was present in 45-60% (NH4 )2 SO4 fraction while absent in other (NH4 )2 SO4 fraction. This fraction also showed the maximum HMG-CoA reductase activity. This band, therefore, may represent the HMG-CoA reductase protein. Six fold purification of the enzyme was achieved by (NH4)2SO4 precipitation. The thiol compounds were found necessary for the long term stability of HMG-CoA reductase enzyme. 

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Summary No. April 2003/02  

Topic: Studies on regeneration in chicory (Cichorium intybus L.) and transformation by Agrobacterium tumefaciens.
Subject: Biotechnology
Name of Student: Reiaz-Ul-Rehman
Name of Supervisor: Dr. M.Z.Abdin
Name of Co-Supervisor: Prof. P.S.Srivastava.
Date of viva-voce  : 16 April, 2003
 

Summary 

There remains a less space for the medicinal plants to be cultivated at large on the existing agricultural land, as it is mainly used for the production of crops to meet the demand for feeding ever-increasing population. Among the options remaining for the cultivation of medicinal plants are the marginal lands, which are infested with various kinds of stresses such as salt and drought. Molecular biology has proved a strong tool and many gene products have been identified from various organisms which confer to them the resistance and /or tolerance against abiotic stresses. These genes provide an opportunity for the biotechnologists to manipulate and explore the possibilities to improve upon the medicinal plants so that they can be profitably grown on the marginal lands. The work carried out by us is an effort in this direction, where we have mobilized tobacco osmotin gene via Agrobacterium tumefaciens mediated genetic transformation in chicory (Cichorium intybus L.). Our results confirm the integration of osmotin gene in the chicory genome and its role as osmoprotectant has been proved.

To achieve micropropagation of chicory various hormonal combinations (Auxin and Cytokinin) with or without growth adjuvent, Casein Hydrolysate (CH) were tried. Based on the percentage callusing, regeneration in cultures , growth and morphology of plantlets the hormonal combination; IAA(2mM) + KN(5mM) + CH (1000mgl-1) was found optimum.The micropropagated plantlets were transferred to pots for acclimatization so that they can sustain and survive in the natural conditions. The in vitro cultures of transformed and non-transformed chicory were found to accumulate the secondary metabolite, esculin at different morphogenetic stages. The concentration of esculin in in vivo grown plants was compared with different developmental stages of in vitro grown cultures. No significant difference in the esculin content was observed, when comparison was made among cultures of transformed and non-transformed chicory. 1.5 fold increase in esculin content was  however, observed in in vitro grown plantlets when compared with in vivo grown plant at flowering stage. The Agrobacterium mediated transformation protocol was optimized considering all the factors for successful transformation. The maximum inhibitory concentration of selectable marker (Kanamycin:150mgl-1) was established. The leaf segments of chicory were co-cultivated with Agrobacterium strain (GV2260) harbouring osmotin gene in the plasmid pBinAR tagged with 35S CaMV promoter. The transformed explants were regenerated on the selection medium optimized for regeneration of chicory. After selection of, the transformants, the putative transgenics were characterized employing molecular biology techniques viz. PCR-utilizing the gene specific primers of osmotin and nptII. These putative transformants were further tested for the presence of osmotin using non-radiolabelled southern blotting of PCR amplified DNA with nptII specific primers, as it was part of the cloning casette of osmotin gene. The southern blotting confirmed the successful integration of the osmotin gene.

To validate the performance of these putative transgenics, physiological and biochemical assessments were made employing leaf disc assay on salt solution. The leaf discs from transgenic chicory plants showed less senescence over the marked period and retained more chlorophyll than those from non-transformed plants. Also proline, the biochemical marker of osmotolerance in plants accumulated upto 90% higher in the transgenic plants as compared to wild type, at 250mM NaCl. These results confirm the over-expression of the osmotin gene in transgenic plants and their improved tolerance when exposed to salt stress. 

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Summary No. May 2003/01  

Title: "Purification and physico-biochemical characterization of sulfite oxidase"

Subject : Biochemistry
Faculty : Science
Name of student : Mr. Ausaf Ahmad
Supervisor : Prof. M. A. Baig
Co-Supervisor : Dr. Zulfiqar Ahmad
Date of viva-voce : 21-05-2003
 

Sulfite oxidase catalyses the physiologically vital oxidation of sulfite to sulfate, the terminal reaction in the degradation of the sulfur containing amino acids, cysteine and methionine.

The physiological importance of an active sulfite oxidase was emphasized by the discovery of a child apparently lacking hepatic sulfite oxidase. Genetic deficiency related to human sulfite oxidase  is associated with severe clinical abnormalities namely mental retardation, seizures, characteristic dysmorphic features and dislocated lenses. The urine of patient contains abnormally large amount of S-sulfocysteine, sulfite and thiosulfate and virtually no inorganic sulfate, making the enzyme of biomedical importance.

The main objectives of the work presented in this thesis were, screening of some plant  leaves and different tissues of goat  for the presence of sulfite oxidase, isolation and purification of the enzyme from the source showing  highest  enzyme activity and physico-biochemical characterization of the enzyme.

Liver was found to have highest activity amongst all goat tissues in which the sulfite oxidase activity was determined. Similarly, leaves of Syzygium cumini showed highest activity amongst all five plant leaves screened for sulfite oxidase activity. Sulfite oxidase from goat liver has been purified to homogeneity by using number of techniques. Ninety- three fold purification was achieved with a yield of 7%.

The absorption spectrum of enzyme indicated the presence of heme in the sulfite oxidase. Circular dichroic  spectra showed the characteristic spectra of proteins in far UV region. Secondary structures of sulfite oxidase  were also calculated to be  45 % a-helix, 9% b-sheet anb 26% b-turn.

The size of sulfite oxidase was found to be 113 kDa and size of subunits of the enzyme was found to be 56 kDa. Sulfite oxidase is composed of two subunits of equal size. The hydrodynamic properties, stokes' radius was determined to be 5.01 nm and frictional ratio, ¦/¦o, was found to be 1.57.

The pH and temperature optima for sulfite oxidase were found to be 8.5 and 25 °C respectively. Sulfite oxidase  showed appreciably less activity in phosphate and carbonate buffers when compared with Tris-Cl buffer under same conditions.

Kinetic parameters, Km and Vmax were found to be 6.98 ´ 10-4 M and 0.5824 nmoles/mL/min respectively. Ferricyanide was 8 times more effective electron acceptor than, physiological electron acceptor, cytochrome c. A number of salts had inhibitory effect on  sulfite oxidase activity and inhibition caused by these salts were mixed non-competitive type.

Atomic absorption spectroscopy  indicated the presence of molybdenum and sodium dithionite reduced sulfite oxidase showed characteristic spectrum of b5 heme, indicating the presence of heme in goat liver sulfite oxidase.

Limited sequence analysis from N-terminal end was done. The  sequence was found to be  trp-glu-pro-ser-gly-ala.

Sulfite oxidase from plant leaves of Syzygium cumini  was partially purified. The absorption spectrum indicated that heme prosthetic group is not present in plant sulfite oxidase. The Km and Vmax for plant sulfite oxidase were found to be  1.38 ´ 10-3 M and 19.23 nmoles/mL/min respectively. Analysis revealed that there is no homology between animal and plant sulfite oxidases.

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Summary No. June 2003/01  

Title: “Plants bearing anti-Salmonellae activity: Molecular and immunological evaluation of their role in protection against typhoid.”

Subject: Biotechnology
Name of the student: Kishwar Hayat Khan
Department/Faculty: Centre for Biotechnology, Faculty of Science
Supervisor: Prof. S.K.Jain
Source of funding: Indian council of Agriculture research (ICAR) and Central Council of Research on Unani Medicine (CCRUM)
Date of Viva Voce: June 20, 2003
 

Typhoid fever is a prolonged febrile illness caused by bacterium Salmonella typhi. It is acute generalized infection of reticuloendothelial system, intestinal lymphoid tissue and gall bladder. Salmonellosis can be experimentally induced in mice by Salmonella typhimurium. The pathogenesis of enteric fever has been studied using S. typhimurium infection in mouse as the animal model and for S. typhi in naturally infected humans and in volunteers. Typhoid fever is characterized with high fever with chills, diarrhoea, inflammatory response in the peyer’s patches, enterocolitis, headache, anorexia, weakness, dizziness, muscle pain, abdominal cramps, liver damage and change in the level of enzymes of oxidative stress. The treatment used against this disease was antibiotic and vaccination which has severe side effect. This lead us to explore natural plant products which is cheap, easily available and has no side effects.

In the present study a number of plants extracts were screened out for their antisalmonellae activity in vitro against S. typhi and S. typhimurium. Six plants exhibit antisalmonella activity showing a very clear zone of inhibition. Extracts obtained from Emblica officinalis and Terminalia chebula exhibit potent antisalmonellae activity as indicated by their zone of inhibition. Minimum inhibitory concentration (MIC) of the above two plants extracts were then determined against S. typhimurium. The result encouraged to perform bactericidal kinetic assay by using the lyophilized juice of Emblica officinalis (EO) and water extract of Terminalia chebula (T) against                  S. typhimurium. EO at a concentration of 5 mg/ml exhibits no effect where as 10 mg/ml of it was bacteriostatic for 10 hour and then it retards the growth of bacteria. The same extract at a dose of 20 mg/ml was bactericidal. T at a dose of 10,12 and 15 mg/ml was bacteriostatic and above 15 mg/ml was highly bactericidal. The above in vitro study claimed the drugs to be used against S. typhimurium infection in vivo.

Animals (mice) pretreated with lyophilized juice of Emblica officinalis at a dose of 500 mg/kg body wt (EO500) for 30 days exhibit full protection against 2x LD50 dose of S. typhimurium where as pretreatment with water extract of Terminalia chebula at a dose of 500 mg/kg body wt (T500) for 30 days exhibit full protection against 1x LD50 dose of S. typhimurium. Studying the clearance of bacteria from reticuloendothelial system and serum enzymes viz, SGPT and SGOT then supported this study.

Animals pretreated with EO500 and T500 for a period of 30 days followed by challenge with sublethal dose of S. typhimurium exhibit an increase in the total leucocytes and lymphocyte count. Delayed type of hypersensitivity is considered as an in vivo manifestation of cell-mediated immune response, and development of a positive DTH reaction is correlated with protective cell-mediated immune response. Treatment with EO500 and T500 exhibit an increase in foot pad swelling significantly (p<0.01) as compared to saline treated control animals at 48 hour followed by a decrease in swelling at 72 hour.

Both the above doses of drugs exhibit an increase in lymphocyte proliferation against     S. typhimurium. Thus both plant exhibit immunomodulatory activity against experimentally induced salmonellosis.

There is imbalance in the level of enzymes of oxidative stress viz lipid peroxidation, xanthine oxidase, catalase, glutathione content, glutathione peroxidase and glutathione reductase when the animals were infected with S. typhimurium. Animals pretreated with EO500 and T500 followed by challenge with S. typhimurium nullifies the effect produced on the enzymes of oxidative stress.

Both EO and T exhibit pharmacological activity viz anti-inflammatory, antipyretic and analgesic activities.

Thus the above plants extract exhibit antisalmonellae activity. Pretreatment of animals with the above drugs prevent the animals from salmonellosis by immunomodulation as well as balancing the level of enzymes of oxidative stress and through various pharmacological activities.

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Summary No. June 2003/02  

Title: “Pathophysiology of Diabetes Mellitus and its Modulation by Drugs

Subject: Pharmacology
Name of the student: (Dr. Manju Sharma)
Lecturer,
Deptt of Pharmacology
Faculty of Pharmacy
Jamia Hamdard
New Delhi - 110062

Supervisor: Prof. K.K.Pillai
Date of Viva Voce: June 23, 2003

Summary:

Over the past decade, there has been increasing scientific and public interest in the so-called antioxidant hypothesis. In general, the reducing environment inside cells helps to prevent oxidative damage. This reducing environment is maintained by oxidative metabolism and by the action of antioxidant enzymes and substances, such as glutathione, vitamins E and C, enzymes such as superoxide dismutase (SOD), catalase etc, which serve to remove the ROS. Current hypothesis favor the idea that lowering oxidative stress can have a clinical benefit. Reports about the status of antioxidants and scavengers as defense mechanisms in diabetic patients are very contradictory. Both increase and decrease antioxidant activity are reported. Studies evaluating the various serum circulating antioxidants in diabetes are still conflicting. It is not known whether oral treatment with antioxidants can reduce oxidative stress in patients with type2 diabetes.

The present study was undertaken to assess the status of lipid peroxidation and to evaluate various antioxidants (Melatonin, Silymarin and a-Lipoic acid) in experimental diabetes mellitus. It appears from the observations of the UKPDS that monotherapy with any oral agent either fails at the outset or will fail overtime. It is often necessary to use multiple drugs for an optimal outcome. We have studied the effects of various antioxidants alone and in combination with gliclazide. Gliclazide was used as a reference drug because it has both insulin releasing and antioxidant property.

Melatonin is mainly used in sleep disorders and jet lags. Frequently melatonin is prescribed for NIDDM patients along with oral antidiabetic drugs. However, the possible outcome of this combination therapy is not reported in the literature. Silymarin is a flavonoid extracted from the milk thistle of Silybum marianum. It is a free radical scavenger and a membrane stabilizer, which prevents LPO and its associated cell damage in some experimental model. Similarly, this concept is further examined in an experimental model of diabetes mellitus, using thioctic acid (a-Lipoic acid) for protection. Potential mechanisms for antioxidant intervention are explored in the above studies.

The ultimate goal of the study was to illustrate the therapeutic potential of antioxidants for treatment of the diabetes mellitus.

The diabetic state, in both humans and experimental animals, is associated with oxidative stress. Diabetes and its complications present a serious medical and socioeconomic problem. The management of diabetes has changed significantly during the past half century. The current strategy to combat diabetes focuses on increasingly stringent control of hyperglycemia to prevent or modify the onset and progression of the disease and its complications. Because of the therapeutic limitations of hypoglycemic therapy in practice, further interventional strategies must be developed. In addition to elevated blood glucose levels, increased production of reactive oxygen species (free radicals), which are known to exhibit direct tissue damaging properties, may contribute to a number of diabetic complications and to the development of insulin resistance itself.  These deleterious species can be neutralized by endogenous and exogenous antioxidant substances such as vitamins A, E, C, flavonoids (Silymarin), polyphenols, carotenoids, alpha lipoic acid and other nutrients present in food and beverages.

Increased oxidative stress in diabetic patients appears to be related to the underlying metabolic abnormalities, and is also an early stage in the disease pathology that may contribute to the development of complications. Therefore, in addition to control of blood sugar, control of oxidative stress offers another avenue for the treatment of the disease. The present research work summarizes the current knowledge of the pathogenic role of oxidative stress in the   onset and progression of diabetes and its complications and presents the results of studies aimed at modulating oxidative stress through the use of anti oxidants in experimental diabetes.

The main finding of the study on Silymarin was that it prevented a rise in both blood glucose and pancreatic lipid peroxidation (LPO), induced by STZ in rats. 

Furthermore, in extension of this work, silymarin showed a dose dependent protective effect on STZ induced diabetic dyslipidemia. Silymarin improved the lipid profile (by decreasing the levels of serum triglycerides, total cholesterol, LDL and VLDL and increasing HDL cholesterol) and restored the liver glycogen content in STZ diabetic rats.  

The effect of B-20 drops, a homoeopathic formulation, on lipid peroxidation, liver glycogen content and microscopic structure of pancreas and liver in STZ diabetes was studied. The study demonstrates the dose-dependent reduction in blood glucose levels and TBARS levels in the pancreas of STZ diabetic rats.

The effect of simultaneous treatment with gliclazide and melatonin in n-STZ diabetic rats was evaluated. Gliclazide alone and also in combination with melatonin were administered to NIDDM rats. The influence of the above treatment on LPO and glucose levels was evaluated. 

The modulatory effect a-LA, a naturally occurring compound and a radical scavenger was studied in vivo in STZ induced oxidative stress and hyperglycemia.

Based on the observations made from these studies, it is concluded that, oxidative stress, antioxidants and the antioxidant network can be relevant to diabetes because diabetes appears to involve oxidative stress. The relevance of oxidative stress mediated mechanism, in this context, may be more appropriate. Antioxidants, vitamins, drugs or diet rich in natural antioxidants would enhance the longevity of life by increasing the threshold, sparing co-enzymes for detoxification pathways during early stages of diabetes, delaying the onset of diabetic complications and may be useful as supportive therapy at later stages of the disease.

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Summary No. July 2003/01  

Title: Chemistry and antihepatotoxic activity of Silybum marianum, its active constituent silymarin and related synthetic analogues containing 1,4 dioxane ring system.
Subject: Pharmaceutical Chemistry

Name of the student: Shah Alam Khan
Dept: Pharmaceutical Chemistry
Supervisor: Dr. Bahar Ahmad
Co- supervisor: Dr. Anees A. Siddiqui
Date of viva: 11th July 2003

Summary :  

Liver disease is a leading cause of death in many countries and the causative factors are alcohol consumption, malnutrition, aneamia, hepatotoxic drugs and infections etc. The liver, a vital organ instrumental in metabolism, detoxification and elimination, is responsible for protection of human body against adverse effects of drugs, chemicals, toxins, bacteria, viruses and parasites etc., but in the process liver itself is under threat and obviously needs protection.

So far no effective measures are available for the treatment of liver diseases. The different medical, surgical and therapeutic methods used at present are inadequate with generally poor results. Also some of the modern drugs which are given to treat liver diseases may themselves cause liver damage.

It is therefore, imperative to search alternative drugs for the treatment of liver diseases to replace the existing currently used drugs of doubtful efficacy and safety.

 

Phytochemical investigation and antihepatotoxic activity of Silybum marianum:

Silybum marianum Linn. (Carduus marianus), family: Asteraceae, a medicinal plant widely used in traditional European medicine for the treatment of various liver ailment, was selected for phytochemical and pharmacological investigation to establish the efficacy of plant extracts, and various isolated constituents as potent antihepatotoxic agent.

50 Kg seeds of S. marianum were dried and crushed to coarse powder which was then exhaustively extracted with ethanol by cold percolation. The crude alcoholic extract was fractionated into Pet. Ether (900 gm), Ethyl acetate (800 gm) and Methanol (300 gm) soluble fraction respectively.

The different compounds like silybin a1, silybina2, silybin b1, silybin b2, and various steroids were isolated and characterized on the basis of IR, UV, NMR and Mass spectral data.

One of the isolated compound, silybin was treated with Boron trifluoride in diethyl ether and change in stereochemistry was studied. The studies suggested that proton of ring C of silybin became Trans from Cis, and orientation of proton at position 8’’ in 1, 4 dioxane ring also changed from b to a.

The various extracts and isolated compounds were evaluated for antihepatotoxic activity in albino rats by using CCl4 as toxicant. The degree of protection was measured by performing histopathological  studies and by estimating various biochemical parameters. Like SGOT, SGPT, ALKP, TP & TA. The methanolic extract showed good antihepatotoxic activity, which was comparable to effects produced by standard drug silybon- 70. The other two extracts, Pet. ether and ethyl acetate also showed moderate protection.

All the isolated compounds showed better protection than standard drug, however among all the isolated compounds, SK –16 and SK-1 were found to be most potent antihepatotoxic compounds.

Synthesis and antihepatotoxic activity of compounds containing 1,4 dioxane ring system:

Silymarin (silybon-70), a hepatoprotective drug is a mixture of three isomers namely silybin, silychristin and silydianin. Among the three isomers silybin is the most potent antihepatotoxic agent. On analyzing the chemical structure of 3 isomers, it was observed that silybin contains 1,4 dioxane ring system in its structure, however this ring is absent in other two isomers. Thus the hepatoprotective activity possessed by silybin can be attributed to the presence of 1,4 dioxane ring system in its strucuture.

We therefore, thought worthwhile that 1,4 dioxane ring system might play an important role in exhibiting antihepatotoxic activity. Thus on the basis of above finding a number of heterocyclic compounds like flavones, xanthones, coumarins and chalcones etc. possessing 1,4  dioxane ring in their molecule were synthesized. The synthesized compounds were screened for antihepatotoxic activity against CCl4 intoxicated albino rats, by estimating biochemical parameters. The synthesized compounds were simple, low molecular weight and could be easily synthesized in lab. For incorporating 1,4 dioxane ring the free ortho dihydroxy compounds were condensed with either ethylene bromide to obtain unsubstituted benzo 1,4 dioxane derivatives or with epichlorohydrin to obtain 2- hydroxymethyl benzo 1,4 dioxane derivatives in presence of aq. Alkali.

Among the synthesized compounds the flavono dioxins were found to be the most potent compounds as they decreased the elevated levels of SGOT, SGPT, ALKP and TA, and increased the decreased level of TP. The effects produced by flavono dioxins were comparable to standard drug silybon 70. The dioxino coumarins showed moderate results followed by dioxino xanthones and chalocone derivatives.

The above studies suggested that both flavone moiety and 1,4 dioxane ring along with 2- hydroxy methyl group in the side chain could exhibit potent antihepatotoxic activity and in turn could be used for treatment of various ailments of liver. The synthesized compounds are low molecular weight and could be easily prepared. On the other hand silybin is a complex molecule and thus can not be prepared easily. Furthermore the synthesized compounds are expected to be easily metabolizable in comparison to silymarin, being simple and low molecular weight.

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Summary No. July 2003/02  

Title: “Evaluation Of Mustard Oil As A Health Oil In Rat Model”

Subject: Toxicology
Faculty/Department: Department of Medical Elementology and Toxicology,
Faculty of Science, Jamia Hamdard, New Delhi
Name of the student: Parul Batra
Name of the Supervisor: Dr. Fakhrul Islam (fislam2001@yahoo.com)Source of Funding: Council of Scientific and Industrial Research (CSIR), Govt. of India  
Date of viva:  July 30, 2003 
 

Summary

The aim to take up this study was the recent controversy on the use of mustard oil in our country. Mustard oil contains high amount of erucic acid and glucosinolates that, according to few reports, are toxic to experimental animals. The use of Indian mustard oil is discouraged in the International market due to its high erucic acid and glucosinolate content. Western countries including U.S. have banned the use of Indian mustard oil thus effecting export of this oil. To solve the controversy this study was conducted to evaluate the toxicity of high dose (50 calorie % in fat free diet) of mustard oil, unboiled and boiled (185-190 ºC for 15 min) with other edible oils (corn oil and sunflower oil) for 3, 6 and 12 months. The lipid profile in serum, heart and aorta of male wistar rats were carried out. Serum was also used for the liver function and kidney function tests. Histopathological responses of twelve month rat heart and aorta tissue were investigated.

No significant alterations were observed in the content of serum cholesterol, triglyceride, low-density lipoprotein-cholesterol, high-density lipoprotein-cholesterol, total cholesterol:HDL-cholesterol ratio, cholesterol:phospholipid ratio, serum glutamate oxaloacetate transamimnase, serum glutamate pyruvate transaminase, bilirubin, creatinine and blood urea nitrogen levels after giving mustard oil (unboiled and boiled), corn oil and sunflower oil in the diet for 3, 6 and 12 months. No toxicity with mustard oil either boiled or unboiled was observed.

Cholesterol, phospholipid, ganglioside and esterified fatty acid levels were analyzed in heart and aorta tissues of experimental animals. The total cholesterol levels were decreased non-significantly with sunflower oil in rat heart. Also there was non-significant decrease in esterified fatty acid levels in rat heart with sunflower oil. Cholesterol-phospholipid ratio in heart remained unchanged with all the edible oils throughout the study. The pathological changes observed in rat cardiac muscle fibres given 50 calorie % unboiled and boiled mustard oil in fat free diets for twelve months revealed majority of cardiac muscle fibres normal except a few muscle fibres with pyknotic nucleus and early degenerative changes. However cardiac pathology was not reflected in serum biochemistry of these rats. No changes in cardiac muscle fibres were observed in corn oil and sunflower oil group.

The total cholesterol levels was decreased non-significantly with all the edible oils after three months but increased after six and twelve months of study in rat aorta. Also there was non-significant depletion in the content of esterified fatty acid in rat aorta with sunflower oil. No significant alterations were observed with all the edible oils in the content of cholesterol, phospholipid, ganglioside and esterified fatty acid in rat aorta. Aortic cholesterol-phospholipid ratio remained unchanged with all the edible oils after three, six and twelve months. Pathological examination of the aorta of rats given 50 calorie % unboiled and boiled mustard oil, corn oil and sunflower oil for twelve months showed a smooth intima with no macroscopic evidence of atheromatous lesions. Our study indicated no toxic symptoms of mustard oil (unboiled or boiled).

No significant changes in these parameters were observed in either of the experimental group of animals. Mustard oil has been found equally as effective as corn oil and sunflower oil in reducing plasma total and LDL Cholesterol levels in rats.

Mustard oil is healthy as it has 30 per cent protein, calcium, phytins, phenolics and natural anti-oxidants. Mustard oil contains high amount of mono-unsaturated fatty acids and a good ratio of polyunsaturated fatty acids, which is good for heart. Mustard oil contains the least amount of saturated fatty acids, making it safe for heart patients.

Glucosinolate, the pungent principle in mustard oil, has anti bacterial, anti fungal and anti-carcinogenic properties, which account for many medicinal utilities of the oil.

The modern concept is to take a mix of polyunsaturated and monounsaturated fat based cooking oils. These include sunflower oil, groundnut oil, corn oil, safflower oil and mustard oil. The relatively high level of oleic acid and the favorable balance between linolenic and linoleic acids is present in mustard oil.

It may be concluded that it is the safest oil and is as good as any other edible oil.

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Summary No. August 2003/01   

Title : Biochmical characterrisation of responses of mustard (Brassica juncea L. Czern and Coss) genotypes to cadmium stress
Department/Faculty : Centre for Biotechnology, Faculty of Science
Name of student : Ms. Shaista Qadir
Supervisor : Dr. M.Z. Abdin
Co-Supervisor : Dr. Saleem Javed
Date of viva-voce : 08-08-2003
Summary

Over the last century human activities have resulted in an unprecedented increase in the level of pollutants, e.g., salinity, ozone levels, pesticides, fertilizers, temperature elevations as well as heavy metals in the environment. Heavy metal pollution of soils is currently a serious environmental problem, because these metals are potential risks to humans. Among the heavy metals, Cd is of increasing scientific interest because of its unique physical and chemical characteristics. Cadmium belongs to group IIB of periodic table, with atomic no 48, atomic weight 112.41, sp. gravity 8.64 g/cm3 and melting point 3210C. Cadmium pigments are found to be unreactive towards chemicals, high temperatures, ultra violent radiation, as well as organic and inorganic solvents. Because of these properties it is widely used in a large number of industrial applications like manufacturing of Cd-Ni batteries, coating of stablizers, paintings, and alloys. This has raised its level in the soil from the normal, 500µg Kg-1 to 500 mg Kg-1. Cadmium is highly toxic to living systems even at very low concentration. It accumulates in the human body with a half-life exceeding more than 10 years. It is responsible for a number of diseases like pulmonary emphysema, demineralization of bones, renal dysfunction, and is categorized as a probable human carcinogen. In plants, it is responsible for a number of physiological breakdowns and even crop failures. Because of these adverse effects of Cd on living system, attempts are being made to minimize its levels in the environment. Traditional methods are expensive, environmentally invasive and labor intensive. So new technology “Phytoremediation” was introduced, which is not only cost effective but also protective to humans as well as to the environment.

Plants used for phytoremediation must tolerate high levels of metals and translocate the metal in the aboveground part of the plant. To understand the mechanism of Cd tolerance, its accumulation and translocation, we undertook this study with ten genotypes of B. juncea, a high biomass accumulating crop plant belonging to the family Brassicaceae. The study was completed by considering and analyzing the effect of Cd on growth, phytochemical and biochemical processes of these genotypes including enzymatic and non-enzymatic antioxidants. Further, SDS-PAGE and TEM have also been used to characterize/distinguish the tolerance mechanism(s) using resistant, moderate and susceptible genotypes. Our study revealed that accumulation of Cd in B. juncea cv. Pusa Jai Kisan was highest among all the genotypes studied. Cadmium had induced oxidative stress in all the genotypes, but with varying magnitudes. Minimum MDA, accumulation was observed in cv. Pusa Jai Kisan and maximum in cv. Vardhan. Minimum reduction in biomass accumulation, plant height, chlorophyll degradation and maximum levels of protein, proline, ATP-sulphurylase activity, non-protein thiols, phytochelatins, glutathione levels and high activities of antioxidant enzymes like SOD, APX. GR were reported in cv. Pusa Jai Kisan. Transmission electron micrograph of chloroplast and mitochondria isolated from this genotype showed minimum damage to these organelles by Cd-stress as compared to other genotypes. Further, over expression of 66 kDa protein band and induction of two new bands viz, 54kDa and 36kDa were observed in this genotypes. The genotype, B. juncea cv. Pusa Jai Kisan, hence, was resistant, while B. juncea cv.Vardhan susceptible to Cd stress among all the genotypes studied. Our results suggest that high tolerance of B. juncea cv. Pusa Jai Kisan to Cd-induced oxidative stress could be due to the increased level of antioxidant system as well as stress induced proteins in this genotype. The findings of our study thus can find application not only in phytoremediation of heavy metals, but also in developing resistance/tolerance in various crops by transferring these biochemical traits using conventional and non-conventional methods.

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Summary No. August 2003/02 

Topic: Studies on Parkinson's Disease: Search for Therapeutic Agents From Herbs
and Herbal Drugs

Department: Toxicology

Student: Muzamil Ahmad (sama_4j@yahoo.co.in)
Supervisor: Dr Fakhrul Islam (fislam2003@yahoo.co.in)
Date of viva voce : 30 August, 2003
 

Parkinson’s disease (PD) is an age-related disorder, more common in senior citizens than in younger ones. The disease is accompanied by the symptoms like rest, tremor, bradykinesia, rigidity, stooped posture and instability. The proper cause of this disease still remains a mystery, despite the role of oxidative stress, free radical formation, genetic susceptibility, programmed cell death and some unknown factor, which may be endogenous or exogenous. The disease progresses slowly and may ultimately produce complete akinesia. The neuropathology of the disease is based on the depigmentation and cell loss in the dopaminergic nigrostriatal tract of the brain with the corresponding decrease in the striatal dopamine concentration and the presence of eosinophilic inclusions called Lewy bodies.

Current pharmacological therapies for the disease are inadequate; these are only able to provide symptomatic relief and after long use produce stern side effects and even worsen the condition.

Herbal medicines are being used since antiquity in folklore and indigenous systems of medicine to treat disease from mild headache to dreaded cancer. They are preferred choice because of their easy accessibility and the notion that they are safer than synthetic drugs, which however, may not be always true. The safety of herbal drugs is only relative but the population feels more assured because of their long and wide spread usage and their familiarity with the plants. Knowledge from the use of medicinal herbs and their active ingredients serve as the foundation for much of the modern pharmacology. In recent times herbal medicines are gaining priority in developed pharmaceutical market because of availability, safety and no major regulatory controls. This study was an attempt towards search for effective and safe alternatives from herbs to protect PD.

Three herbal drugs Ginkgo biloba, Nardostachys jatamansi and Withania somnifera were studied for anti-parkinsonian effects in Wistar rats. These herbs were selected after a extensive survey of the literature for the purpose and advice from the practicing physicians of Unani and Ayurvedic systems of medicine. There are ample reports in the literature for these plants to be used in various neuro-disorders in clinical and experimental studies, beside their use in folklore against various health conditions. The parameters of study included: (a) Behavioural studies (b) Biochemical/enzymatic estimations (c) Dopaminergic D2 receptor binding assays (d) Catecholamine quantifications and (e) Immunohistochemical studies.

Parkinsonism was induced in the rats by injecting 6-OHDA into the right striatum stereotactically. Lesioning with 6-OHDA resulted in significant enhancement of lipid peroxidation (LPO) and reduction in the content of glutathione (GSH) and reduced activities of its dependent enzyme system (i.e. glutathione-s-transferase, glutathione reductase and glutathione peroxidase) along with that of superoxide dismutase and catalase. There had also been a marked up regulation of Dopamine D2 receptor binding and a decrease in the content of dopamine and its metabolites (dihydroxyphenyl acetic acid and homovalinic acid). All the changes were further emphasized by the reduced expression of tyrosine hydroxylase, a marker enzyme for PD, in the ipsilateral striatum of lesioned rats as confirmed by immunohistochemistry. Pre-treatment with test drugs for three weeks, prior to lesioning, markedly reversed all the behavioural, biochemical, neurochemical and immunohistochemical changes.

It is premature to propose the mechanism(s) involved in the observed anti-parkinsonian effects of the herbal drugs studied, and at the same time we do not consider that these effects were elicited through a single mechanism. On the basis of present findings and reported properties of test drugs, it is proposed that these agents improve overall physical and mental performance through antioxidant, anti-stress, immunomodulatory and adaptogenic effects; furthermore, animal model used for the disease do not simulate the precise clinical event as most of the behavioural deficits can never be observed in animal models, but are still relevant for elucidating the mechanisms and testing the efficacy of various therapeutic agents. In spite of these limitations, we consider the findings to be very encouraging with good anti-parkinsonian potential and advocate that these plants for further investigations. Undoubtedly elucidation of proper molecular mechanisms and deciphering appropriate genetic pathways can  go a long way in emphasizing our studies.