EFFECT OF EFFECT OF α-KETOGLUTARATE ON SOME HEPATIC VARIABLES ALTERED BY CYANIDE IN VIVO AND IN VITRO
TULSAWANI, R.K., HARIHARAKRISHNAN, J., JATAV, P.C. AND BHATTACHARYA, R.

Defence Research and Development Establishment, Jhansi Road, Gwalior, 474 002, (M.P.), India Email: rbhattacharya41@rediffmail.com

Abstract: Dose and time-dependent effects of cyanide on various biochemical variables related to hepatotoxicity in vivo and in vitro, and their response to the treatment of a-ketoglutarate (α-KG) were investigated. Female rats were administered oral doses [0.75 LD50 (10.5 mg/kg) or 2.0 LD50 (28.2 mg/kg)] of potassium cyanide (KCN) with or   without pre-treatment (- 10 min), simultaneous treatment (0 min) or post-treatment (+ 2 to 3 min) of α-KG (2.0 g/kg; oral) and/or sodium thiosulphate (STS, 1.0 g/kg; intraperitoneal: - 15 min, 0 min or + 2 to 3 min). Rats receiving 0.75 LD50 KCN were sacrificed 30 min and 60 min after treatment and those given 2.0 LD50 KCN were killed at the time of onset of signs and symptoms of toxicity (2 to 4 min) or at the time of death (5 to15 min). Cytochrome c oxidase (CYTOX), rhodanese (RHO), malondialdehyde (MDA) and catalase (CAT) were measured in liver and thiocyanate (SCN), alanine aminotransferase (ALAT) and aspartate aminotransferase (ASAT) were measured in serum. No significant change was caused at lower dose of KCN, but 2.0 LD50 KCN significantly reduced the CYTOX activity by 30% and 54% within 2-3 min after treatment and at the time of death, respectively. Also, a 21% decrease in serum ALAT levels was observed at the time of death. Pre-treatment or simultaneous treatment of a-KG alone or a-KG + STS and post-treatment of a-KG + STS significantly restored the changes. All the groups receiving STS showed significantly enhanced levels of SCN. In a separate experiment, primary culture of rat hepatocytes was treated with 5.0 or 10 mM cyanide for 6 h, in the presence or absence of a-KG (5.0 mM). Significant leakage of intracellular lactate dehydrogenase accompanied by loss of mitochondrial membrane potential (rhodamine 123 staining) and nuclear integrity (propidium iodide uptake) indicated cyanide-induced cytotoxicity, and leakage of ALAT and ASAT could be related to cyanide-induced hepatotoxicity in vitro. The ameliorative effect of α-KG was more conspicuous against 5.0 mM cyanide. The study reveals that a significant inhibition of liver CYTOX by a lethal dose of cyanide was insufficient to mediate hepatotoxicity in rats but in vitro results showed changes in parameters associated with cytotoxicity, and more particularly hepatotoxicity. The study also indicates promising role of a-KG as an alternative treatment for cyanide poisoning.

electromorphic variation in complements c3, c4, c8 in dilated cardiomyopathy

USHASREE, B., NARSIMHAN, C., JAIN, R. K.  AND  PRATIBHA, N.

Department of Genetics, Osmania University, Hyderabad 500007, E-mail: prathinallari @ yahoo.com.

Abstract: Cardiomyopathy represents a diverse group of heart muscle disorders, which are classified on the basis of their anatomic and hemodynamic findings. Among which, dilated cardiomyopathy is the most prevalent, but poorly understood group of disorders. Hereditary and environmental factors, in combination with an altered immune response appear to play a causative and pathogenic role in the etiopathology of the disease.  Oxygen derived free radicals and their products play a critical role in the lipid peroxidative damage of membrane lipids and are known to be involved in the pathogenesis of heart failure. The tissue damage thus initiates a complex sequence of events collectively known as inflammatory response through the activation of complement cascade system and contributes further to the susceptibility of the disease. Hence, a study on polymorphic variation of complement proteins in dilated cardiomyopathy may help in understanding their role in the susceptibility to the disease condition. 97 Dilated cardiomyopathy patients between 2 to 65 years of age and 105 healthy volunteers were considered for the study. A predominance of C3 ‘F’ (17.5%), C4 ’F ’(22.6%) and C8 ‘A’ (32.9%) homozygous alleles was observed, with the C4 ‘FF’ phenotypes being significantly associated (c2  8.35) with the disease condition, confirming that, differences in the interactions of C4 with specific allele combinations and with other complement proteins may alter immune complex transport and/or immune complex solubilization functions, opsonisation, etc, that may be genetically related to autoimmunity in dilated cardiomyopathy (DCM). Therefore, it can be concluded that individuals who are homozygous carriers of C3, C4 ‘F’ and C8 ‘A’ alleles are susceptible to exacerbation of the disease condition to muscle degradation/tissue injury which can lead to ischemia/heart failure, a complication associated with DCM. Further, the homozygous phenotype associations confirm the recessive mode of the polymorphisms.

TOXICOLOGICAL STUDIES ON SNAKE VENOM - SPECIFIC ANTIBODIES (IgY) FROM CHICKEN EGG

MANJULA, J., KUSUM, P., SAIRAM, A. K., MURTHY, P. B., AND SUBBARAO, P.V.

Vittal Mallya Scientific Research Foundation, Bangalore 560029. E-mail: prakhya@yahoo.com

Abstract: In recent years, IgY antibodies produced in chicken egg are being explored as an alternative to mammalian antibodies for diagnostic and therapeutic use. Leghorn chickens were injected with a sub-lethal dose of venom of the poisonous snake, Echis carinatus (Krait). The antibodies were purified from the egg yolks of immunized birds by a combination of water dilution method followed by salt fractionation. The efficiency of these antibodies in protecting mice against the lethal effects of the venom has been demonstrated by in vitro neutralization of snake venom in mice. Since there are no studies on the biosafety of IgY antibodies which have great potential for therapeutic use, comprehensive toxicological studies viz. acute oral, intraperitoneal, intramuscular, mucous membrane irritation potential, 28-day repeated intraperitoneal as well as allergenicity and mutagenicity evaluation were carried out. These studies revealed that therapeutic IgY antibodies could be safely used for evaluation in human trials.

FLUCTUATIONS OF γ-GLUTAMYL TRANSPEPTIDASE IN NORMAL AND CATARACTOUS HUMAN EYE

PANDYA, A. V. AND RAWAL, U. M.

C. U. Shah Science College, Ashram Road, Ahmedabad - 380014, India.E mail: ajitpandya@sancharnet.in

Abstract: The most common form of cataract is the “Senile Cataract” occurring in the aged population, which has great socio-medical prevalence for many third world countries, where annually 2-3 million people become blind due to cataract. The cause and phenomenon of cataract development may be same but the changes related with γ- glutamyl transpeptidase (GTP) activity depends on age and types of cataract. Therefore, in present investigation the fluctuation of this enzyme was studied in the lens of patients suffering from senile cataract. The study shows an intensive decrease of GTP   activity, which is 9.595 ± 0.094 n moles/min/mg (mean ± SD) in normal lenses as compared to 3.7 ± 0.216 n moles/min/mg (mean ± SD) in cataractous lens. Since GTP reacts very effectively with glutathione (GSH) amongst all the enzymes involved in the glutathione cycle it appears that GSH is entirely degraded within the lens. This study may help in finding degree, types and progress of cataract in patients.

Effect of sumatriptan on cortical 5-HT1B receptors  in rat brain

SUBHASH, M.N.  AND  RAMAKRISHNA, D.

Department of Neurochemistry, National Institute of Mental Health and Neurosciences, Post  Box  No. 2900
Bangalore 560 029 India. E-mail: subhash@nimhans.kar.nic.in

Abstract: Sumatriptan is a novel and highly effective drug against migraine and cluster headache attacks. The drug is known to exert its effect through the modulation of serotonin (5-HT)-mediated neurotransmission. 5-HT1B receptors in rats and 5-HT1D receptors in humans have been thought to be likely targets of sumatriptan. In the present study the effect of varying dose and duration of sumatriptan on the density of 5-HT1B receptors in the rat cortex were studied to understand its mechanism of action. Sprague Dawley rats were administered with different doses of sumatriptan (0.2-16 mg /kg body wt, i.p.) for seven days and 2, 4 and 8 mg/kg of sumatriptan for 7, 14 and 21 days. The radioligand binding assays were performed in cortical membranes using [3H]5-HT. Treatment with different doses of sumatriptan for seven days showed a significant (p<0.0001) downregulation of 5-HT1B receptors in a dose dependent manner.  A significant decrease in the density of 5-HT1B receptors was observed with 0.2 mg (45%), 0.5 mg (72%) and 1 mg (70%) of sumatriptan treatment, with a significant decrease (p<0.0001) in Kd values.  No further decrease in either the density or in the Kd values was observed with increasing doses of sumatriptan from 2 to 16 mg/kg body wt.  The magnitude of decrease in the receptor density was more significant with a lower dose (2 mg) of sumatriptan for a prolonged period of exposure (21 days). However, such change was not observed with higher doses (4 and 8 mg/kg).  Furthermore, sumatriptan showed a higher affinity for 5-HT1B receptors with a Ki value of 9.4+0.9 nM, when compared to other agonists and antagonists. Taken together, these findings suggest that desensitization of 5-HT1B receptor is dose and time dependent, which may be an important factor underlying the mechanism of action of sumatriptan as an antimigraine drug.

EFFECT OF MELITTIN ON ELECTRICALLY DRIVEN ACTION POTENTIAL OF GUINEA PIG PAPILLARY MUSCLE

RAGHU, K. G.

Pharmacology Division, Central Drug Research Institute, Chatter Manzil Palace, Post Box 173, Lucknow 226 001.E-mail:  raghukgopal@rediffmail.com

Abstract: The effect of various concentrations (1-10µm) of melittin, a major amphiphilic polypeptide components of European honey bee (Apis mellifera) venom, on electrically driven (1Hz) action potential (Ap) of papillary muscle of Guinea pig were studied using classical microelectrode technique. Cumulative application of melittin to perfusion bath caused significant depolarization of resting membrane potential (Em), inhibition of upstroke velocity (Vmax) and reduced action potential duration (APD) at different level of repolarisation. These results suggest the effect of toxin on various ion transport mechanism of cardiac cells such as ion channels, exchangers and pumps.

D-GALACTOSE INDUCED AGEING IN SHORT DURATION: A QUICK MODEL OF ACCELERATED AGEING IN MICE

DESHMUKH, A.A. GAJARE, K.A.  AND PILLAI, M.M.

Department of Zoology, Govt. Vidarbha Institute of Science and Humanities, Amravati 444604.
E- mail: ashish_cellbio@rediffmail.com

Abstract: Literature reveals an increased lipid peroxidation during ageing. The purpose of this study was to develop short term model of ageing in young animals. The animals were grouped into three: control group, experimental group 1 (treated with daily subcutaneous injections of 5% D galactose for fifteen days) and experimental group 2 (treated with daily subcutaneous injections of 5% D galactose for fifty six days).  Study showed that fifteen days subcutaneous injections of 5% D galactose 0.5 ml/day resulted in increased lipid peroxidation and decreased antioxidant enzymes: superoxide dismutase, catalase and glutathione peroxidase in various regions of brain such as cerebral cortex, hippocampus, corpora quadrigemina, cerebellum as well as heart auricle and ventricle. Increasing the duration of treatment for 56, the results were similar to that 15 days treatment as level the fluctuations of various biomolecules were almost similar in both the cases.  The present model minimizes the duration of D galactose treatment from 56 days (as reported in earlier model) to 15 days and hence becomes a quick animal model for researches on ageing.

CORRELATION BETWEEN OXIDATIVE STRESS AND TYPE II DIABETIC PATIENTS

DESHMUKH, A.A., GAJARE, K.A., PULLAJWAR, V.S., RUIKAR, A.A. AND BULE, P.P.

Department of Zoology, Government. Vidarbha Institute of Science and Humanities, Amravati 444 604 India.
E-mail: ashish_cellbio@rediffmail.com

Abstract: Fifty type II diabetic patients (non insulin dependant diabetes mellitus) and 50 healthy individuals (without diabetes) were included in this randomized study. None of the patients studied had any diabetic complications. The levels of serum malondialdehyde (MDA) from type II diabetic patients was 5.33 ± 1.46 nmol/ml as compared to control (2.74 ± 0.4 nmol/ml). There was age-dependent increase in serum MDA in healthy individuals and age-independent increase in MDA levels in type II diabetic patients. There was sufficiently high degree of positive correlation between blood glucose level and the serum MDA level (r=0.78, p<0.001) as well as between the duration of the disease and serum MDA level in TYPE II DIABETICpatients (r=0.82, p< 0.001) indicating that hyperglycemic condition and duration of disease are responsible for increased oxidative stress in these patients.

LEAD INDUCED OXIDATIVE STRESS AND DNA DAMAGEUSING COMET ASSAY IN RAT BLOOD

SAXENA, G., KANNAN, G. M., SAKSENA D. N., TIRPUDE, R. J. AND FLORA, S.J.S.

Division of Pharmacology and Toxicology, Defence Research and Development Establishment, Jhansi Road,
Gwalior. 1School of  Studies in Zoology, Jiwaji University, Gwalior 474 002, India.  Email: sjsflora@hotmail.com

Abstract: Lead is a known environmental pollutant that induces a broad range of physiological, biochemical and behavioral dysfunctions and thus considered to be of serious personal and public health concern. One of the first sites of toxic effect of lead is red blood cells and possible mechanism of lead action on various target tissues has recently been reported to be oxidative stress. The present study was designed to investigate the extent of lead induced oxidative stress and DNA damage following short and long term exposure in male wistar rats. Animals were exposed to 0.2% lead acetate in drinking water for 3 or 30 weeks for determining several blood biochemical variables indicative of oxidative stress, alteration in haem biosynthesis pathway, DNA damage (based on comet assay) besides  lead concentration. The poly acrylamide gel electrophoresis (PAGE) was also carried out to estimate the activity of glucose-6-phosphate dehydrogenase (G6PD) in red blood cells (RBCs). Exposure to lead caused a significant decrease in blood d- aminolevulinic acid dehydratase (d-ALAD) activity and glutathione (GSH) level at both time intervals. There was a significant increase in reactive oxygen species (ROS) accompanied by decreased band intensity of G6PD activity (at 30 weeks) indicating oxidative stress in lead exposed animals. Further, alkali single cell gel electrophoresis assay (comet assay) was adopted for detecting the DNA damage. Comet with a prominent tail was seen in chronic lead exposed animals supporting injury to DNA and oxidative stress. Thus the present study suggests  significant alterations in haem synthesis pathway, DNA injury and oxidative stress following short and long term exposure to lead.

10.  Journal of Cell and Tissue Research Vol. 6(2): 769-774 (2006)

ACUTE TOXICITY OF ALPHA-KETOGLUTARATE AFTER INTRAPERITONEAL ADMINISTRATION IN RATS

BHATTACHARYA, R, TULSAWANI, R. K., KUMAR, D., HARIHARAKRISHNAN, J., SATISH KUMAR, H. T.  AND VIJAYARAGHAVAN, R.

Division of Pharmacology and Toxicology, Defence Research and Development Establishment, Jhansi Road, Gwalior 474002 India. E-mail: rbhattacharya41@yahoo.com

Abstract: Cyanide antagonism by oral and intraperitoneal (ip) administration of 2.0 g/kg alpha-ketoglutarate (A-KG) was reported earlier in rodents. While, oral treatment of 2.0 g/kg A-KG did not produce any toxicity in rats, it was interesting to determine acute effects of ip administration of 2.0 g/ kg A-KG in female rats. Various biochemical, hematological and histological variables were studied 1 h, 24 h and 7 d after A-KG administration. We also studied several physiological parameters in anesthetized rats, after 1.0, 2.0 and 4.0 g/kg A-KG administration. Except for a transient increase in blood lactate dehydrogenase, sugar and creatinine levels, 1 h after treatment, no significant change in biochemistry, hematology or histology of the vital organs was observed. No change was also observed in the animal body weight or organ-body weight index of the vital organs. Except for a decrease in neuromuscular transmission (NMT), none of the physiological parameters were significantly altered by 2.0 g/kg A-KG. However, a significant decrease in mean arterial pressure was observed 210 min after 4.0 g/kg A-KG administration. There was no significant change in the heart rate and rectal temperature, but a significant decrease in respiratory rate (73.1% of control) was observed after 90 min, which further declined (56.8%) by 240 min. Significant decrease in NMT was produced by all the doses of A-KG after 30 min of treatment. The study indicates few deleterious effects of ip administration of A-KG on physiological variables, particularly at the dose of 4.0 g/kg. Although, a reduction in the dose of A-KG by ip route is envisaged, its oral treatment is presumed to be safer and more realistic as compared to ip route.

11.  Journal of Cell and Tissue Research Vol. 6(2): 775-778 (2006)

BENEFICIAL EFFECT OF BENINCASA HISPIDA METHANOLIC EXTRACT AGAINST NEUROLEPTIC DISORDERS

MHETRE, N., AMBAVADE, S., MUTHAL, A., BODHANKAR, S. L. AND THAKURDESAI, P. A.

Poona College of Pharmacy, Bharati Vidyapeeth Deemed University, Erandwane, Pune 411038, India.
E-mail: sbodh@yahoo.com

Abstract: Neuroleptic activity of methanolic extract of Benincasa hispida was assessed using apomorphine induced cage climbing model and catalepsy test in mice. Methanolic extract showed significant reduction in apomorphine induced cage climbing activity at 600 mg/kg and catalepsy in mice .The phytochemical analysis of methanolic extract showed presence of phenolic compounds and triterpene saponins, which may be responsible for the neuroleptic activity.

12. Journal of Cell and Tissue Research Vol. 6(2): 779-782 (2006)

IMMUNOMODULATORY EFFECT OF MANGIFERIN

BALEKAR, N.S., BODHANKAR, S.L. AND JAIN, D.K.

Poona College of Pharmacy, Bharati Vidyapeeth Deemed University, Erandwane, Paud Road, Pune 411 038
1College of Pharmacy, IPS Academy, Indore 452 012. E mail: sbodh@yahoo.com

Abstract : Immunomodulatory activity of the mangiferin, a naturally occurring xanthone-C- glucoside, was evaluated in Swiss albino mice using peritoneal macrophage, reticuloendothelial system, absolute lymphocyte, splenic lymphocyte, hematological parameter and E. coli induced abdominal sepsis. Mangiferin (15 mg/kg) exhibited significant increase in peritoneal macrophage count, phagocytic activity, WBC count, absolute lymphocyte, splenic lymphocyte count and also provided protection against E. coli induced abdominal sepsis. Study shows that mangiferin has stimulatory effect on immune functions.             

13. Journal of Cell and Tissue Research Vol. 6(2): 783-786 (2006)

ELLAGIC ACID - AN IMMUNOMODULATOR!

BALEKAR, N. S., BODHANKAR, S. L.AND MOHAN, V.

Poona College of Pharmacy, Bharati Vidyapeeth Deemed University, Erandwane, Paud Road, Pune411042.
E mail: sbodh@yahoo.com

Abstract: Immunomodulatory activity of ellagic acid was studied in various groups of animals by delayed type of hypersensitivity (DTH) response, plaque forming cell (PFC) assay, haemagglutination (HA) titre and in vitro phagocytosis. Ellagic acid (EA) 500 mg/kg elicited significant increase in DTH. Humoral immunity, as measured by PFC and HA, showed a dose dependent increase and a dose of 500 mg/kg showed a significant effect. EA also elicited a significant increase in phagocytic index and phagocytic capacity of polymorphonuclear (PMN) leucocytes.  EA was found to have stimulatory effect on immune functions in mice.

14. Journal of Cell and Tissue Research Vol. 6(2): 787-791 (2006)

IN VITRO EVALUTION OF ANTIMICROBIAL PROPERTIES OF TWO SPECIES OF GENUS GOMPHOSTEMMA

DEKA, H., GOGOI, D. K., GOGOI, H. K. AND  HANDIQUE, P.J.

Defence Research Laboratory, Post Bag No: 2, Tezpur-784001,  E. mail: hemashree_deka@redifmail.com

Abstract: Two rare species of the genus Gomphostemma (G. crinitum and G. niveum), belonging to family Lamiaceae, available only in few pockets of North East India, were evaluated for their antibacterial and antifungal properties. Leaf extracts, prepared in different organic solvents,  were screened against five human pathogenic microorganisms viz.,  two Gram positive (Bacillus subtilis and Staphylococcus aureus), two Gram negative (Escherichia coli and Klebsiella sp.) and a fungal pathogen (Candida albicans).  Out of five microbial test organisms, S. aureus was found to be the most susceptible. On the other hand, the extract of both the species of Gomphostemma showed weak inhibitory effect against B. subtilis and C. albicans. The MIC of the crude extracts ranged from 180µg/ml to 600µg/ml. Comparative study of the activity of the standard antibiotics and chloroform extracts of both the plants revealed that these extracts have broad spectrum activity than some commercially available antibiotics, therefore,  can be tested further for raising important antimicrobial agent against important pathogen.

15. Journal of Cell and Tissue Research Vol. 6(2): 793-796 (2006)

CHLORPYRIPHOS INDUCED INHIBITION OF AChE IN CYPRINUS CARPIO AND RECOVERY DURING DRUG LEACHING

TEMBHRE, M., AHIRWAR, S., GOUR, S.  AND GAUR, M.

Department of Zoology, Sri Sathya Sai College for Women, Bhopal 246003.  1Department of Zoology,
Government College,  Barelly M.P. India. E-mail: m_tembhre@yahoo.co.in

 Abstract: The objective of present investigation was to screen the affect of  sublethal concentration of widely used pesticide,  chlorpyriphos,  on acetylcholinesterase (AChE) in fish tissues and the recovery of the enzyme during drug  leaching. The AChE activity in brains, heart and gill were evaluated after 24, 48, 72, and 96 hours of pesticide exposures. The chlorpyriphos-induced AChE inhibition was highest in the fore brain (66%) followed by  heart (55%) and gill (44%) respectively. In second experiment fish, exposed to chlorpyriphos for 96 h, were transferred to pesticide free water for 1 week. Study showed about  83 to 90% recovery of the enzyme in  brain, heart and gill on 7th day. Protein contents were also found to be gradually decreased during the chlorpyriphos exposure.

16. Journal of Cell and Tissue Research Vol. 6(2): 797-802 (2006)

BENEFICIAL EFFECTS OF VITAMIN E IN MITIGATING CHROMIUM-NICKEL INDUCED NEPHROTOXICITY IN MICE

PAREKH, S., CHAWLA, S. L. AND RAO, M. V.

Zoology Department, School of Sciences, Gujarat University, Ahmedabad  380 009 India.
E-mail: zooldeptgu@satyam.net.in

Abstract: Nickel chloride (NiCl2;-8, and 16mg/kg body weight) and potassium dichromate (K2Cr2O7 - 5 and 10 mg/kg b wt) administered alone and in combination daily for 30 days were studied for their effects on histological and  biochemical changes in mice. Alongwith this, the effect of vitamin E (2mg/kg body wt) supplementation with metals was also studied. Nickel and chromium treatments to mice revealed a significant decline in the activities of acid phosphatase and alkaline phosphatase as well as in the levels of protein and creatinine in the kidney. These changes appear to be due to the damage in kidney caused by heavy metals. The histological and ultra structural studies supported this fact. Interestingly enough, the supplementation of vitamin E along with nickel and chromium did not show much damage in kidney demonstrating the ameliorative role of vitamin E.

17.  Journal of Cell and Tissue Research Vol. 6(2): 803-806 (2006)

DECLINE IN RESISTANCE PROBLEMS IN SOME MOST FREQUENTLY ISOLATED BACTERIAL PATHOGENS TO ANTIMICROBIALS IN IRAN

MAHDAVI, M.R., EBRAHIMZADEH, M.A., SAEEDI, S.S. AND SHABANIAN, K.H.

Department of Microbiology, Sari School of Paramedicine, Mazandaran University of Medical Sciences, Sari, Iran. E-mail: zadeh20@yahoo.com

Abstract: In Iran, information about bacterial resistance to antimicrobial agents is limited. In present investigation the samples were collected from urine of 14382 patients with UTIs that were refereed to one regional and one governmental diagnostic microbiology laboratories during two-year periods (2004-05). A total of 1731 aerobic cultures were reviewed (12 %). The antibiogram disc agar diffusion technique was used to evaluate the sensitivity of microbial agents to antibiotics. The most common bacterial pathogens were E. coli (36.1%), Staphylococcus sp.. (aureus and coagulase-negative) (36.0%), Streptococcus sp.. (10.7%), Klebsiella (7.7 %), Enterobacter sp. (4.7 %), P. aeruginosa (2.6 %) and Citrobacter sp. (1.4 %). High levels of resistance to nalidixic acid  and co-trimoxazole have be found in nearly all bacterial pathogens, nitrofurantoin in P. aeruginosa (42.2%) and Klebsiella (38.1%)  and penicillin in Streptococcus sp. (35.7%). Levels of resistance to majority of antibiotics were higher than other countries but decreased during last 2 years.

18. Journal of Cell and Tissue Research Vol. 6(2): 807-813 (2006)

PRIMER DESIGNING FOR DREB2A, A DROUGHT RESISTANT GENE IN GLYCINE MAX.

GARG, N.  AND  ANIL KUMAR

School of Biotechnology, Devi Ahilya University, Khandwa Road, Indore 452017 India.
E mail:  ak_sbt@yahoo.com

Abstract: By feeding the cDNA clone sequences of DREB2A gene of Glycine max (soybean), a known drought resistant gene,  in Primer 3 as well as Web Primer software, number of forward and backward primers were designed. Using set theory, permutation and combination, finally eight forward and eight backward primers were selected. The properties of these primers were determined using Gene runner software. Using OLREA software, various restriction endonuclease cutting sites were also determined in finally selected primers. Out of all the primers tested, only F4, F22, 23 and R4 primers were found not to have hairpin structure. Genomic DNA of 20 different varieties of soybean  were isolated, purified and amplified using F4 and R4 primers in PCR. Universal primer for soybean was also amplified for confirming the PCR amplification protocol. Only DNA of  HARDEE, SL 96 and MAUS 71 varieties of soybean got amplified with F4 and R4 primers. These varieties of soybean are known drought resistant varieties. Therefore, F4 and R4 primers may be used for checking drought resistance due to DREB2A  gene in soybean.

19. Journal of Cell and Tissue Research Vol. 6(2): 815-818 (2006)

EFFECT OF BACOPA MONNIERA EXTRACT ON LYSOSOMAL MEMBRANE INTEGRITY IN THE BRAIN AND HEART OF D-GALACTOSE INDUCED OXIDATIVE STRESSED MICE

GAJARE, K. A., DESHMUKH, A. A.  AND PILLAI, M. M.

Department of Zoology, Government Vidarbha Institute of Science and Humanities, Amravati 444604,
E Mail: kavita_kanu@rediffmail.com>

Abstract:  Bacopa monniera is a medicinal plant used as memory tonic in Ayurvedic medicines. In present investigation antioxidant property of Bacopa monniera was studied with respect to protection of the lysosomal membrane from the oxidative damage induced by D galactose.  For this purpose the female albino mice were treated with D-galactose and D-galactose in combination with Bacopa monniera.  The lysosomal enzyme acid phosphatase was measured in both lysosomal and post lysosomal fractions and ratio of lysosomal to post lysosomal acid phosphatase activity was studied. The ratio was more than one in control group and less than one in D-galactose treated group. In Bacopa monniera co-treated group the ratio was also more than one in various brain regions (cerebral cortex, hippocampus, corpora quadrigemina, cerebellum) as well as in  heart (auricle and ventricle). This indicates that Bacopa monniera extract provides protection against oxidative stress and prevents lysosomal membrane leakage, thus acting as a potent membrane stabilizer.

20. Journal of Cell and Tissue Research Vol. 6(2): 819-824 (2006)

EFFECTS OF HIBISCUS SABDARIFFA ON LIPID PEROXIDATION IN HYPERAMMONEMIC RATS

ESSA, M.M. AND SUBRAMANIAN, P., 

Department of Biochemistry, Faculty of Science, Annamalai University, Annamalai Nagar  608002,  India.
E mail: annamalai_rhythm@yahoo.co.in

Abstract: Influence of ethanolic leaf extract of Hibiscus sabdariffa, an indigenous plant used in Ayurvedic Medicine in India, Thailand and China on the temporal patterns of circulatory lipid peroxidation products and antioxidants during ammonium chloride (AC) induced hyperammonemia were investigated in Wistar rats. Experimental rats were divided into control, AC treated, AC + H. sabdariffa leaf extract (HSEt) treated, and HSEt treated. The characteristics of 24 h variations (acrophase, amplitude and mesor) of lipid peroxidation products (thiobarbituric acid reactive substances - TBARS) and antioxidants (reduced glutathione (GSH), glutathione peroxidase (GPx), superoxide dismutase (SOD) and catalase (CAT) were analyzed. Enhanced lipid peroxidation (increased mesor of TBARS) associated with decreased antioxidant activities (decreased mesor of GPx, GSH, SOD and CAT) were found in hyperammonemic rats. Variations in amplitude and ‘r’ values also were found in between the hyperammonemic rats and other groups. These alterations clearly indicate that temporal oxidant-antioxidant status is modulated during hyperammonemia, which may also play a crucial role in disease development.

21. Journal of Cell and Tissue Research Vol. 6(2): 825-836 (2006)

AGE DEPENDENT CHANGES IN STEROID HORMONES LEVEL MODULATE PROGRESSION AND REGRESSION OF BREAST CANCER

GUPTA, P. D. AND PUSHKALA, K.

Atmiya Institute of Gerontology Research, Kalawad Road, Rajkot 360 005, E mail: pdg2000@hotmail.com

Abstract: The mitogenic effect of sex steroid hormones on the breast tissue has been reviewed. Two separate pathways have been hypothesized to operate in the predisposition of breast cancer: 1.Involving the sex hormones and their respective specific receptors inducing the breast cancer specific genes like BRCA-I, BRCA-II and genes like P53 interfering with regular cell cycle check points. 2. Another pathway (estrogen negative breast cancer) by phosphorylation of growth factors such as insulin, insulin–like growth factors, which ultimately interfere with gene transcription to tamper with the regular cell cycle activity. Experimental data are reviewed to substantiate the efficiency of estrogen to induce the secretion of growth factors. The role of steroid hormones on the regression of breast cancer has also been discussed. The risk factors for breast cancer in older women and the efficiency of melatonin to suppress the proliferation of breast cancer cells have been discussed in this review.

22. Journal of Cell and Tissue Research Vol. 6(2): 837-839 (2006)

IN VITRO EVALUATION OF SOME MEDICINAL PLANTS AGAINST CANDIDA ALBICANS

GOSWAMI, S., BORA, L., DAS, J. AND BEGAM, M.

Defence Research Laboratory, Post Bag No 2, Tezpur- 784001 India. E-mail: sewali_drl@rediffmail.com

Abstract: The antifungal activity of methanolic extracts of nine medicinal plants viz., Cinnamomum porrectum, Meyna spinosa, Duranta repens, Sida carpinifolia, Tephrosia purpurea, Cestrum nocturnum, Trachyspermum ammi, Lippia nudiflora and Ludwigia parviflora were studied against clinically important pathogen Candida albicans. The medicinal characteristics of these plants were compared with commercially used antibiotic clotrimazole. The antimicrobial assay was done by agar well diffusion method. The leaf extract of Lippia nudiflora showed the minimum (13.0 mm) activity in comparison to commercial antibiotic. Among various plants used in the present investigation, Trachyspermum ammi and Cestrum nocturnum were found to be more potent with inhibition zone of diameter 38.3 and 31.5 mm respectively, hence can be further screened for potent commercial antifungal agents against Candida albicans.

23. Journal of Cell and Tissue Research Vol. 6(2): 841-844 (2006)

PROTECTIVE EFFECT OF HYDRASTIS CANADENSIS  EXTRACT ON HUMAN LYMPHOCYTES: AN IN VITRO STUDY

KALEENA, P.K.,  DAWOOD SHARIEF, S., VINCENT, I.,  ILANGO, B., AND SUKUMAR, E.

Post Graduate Department of Zoology, Presidency College, Chennai 600 005.  E mail: sdawoodsharief@gmail.com

Abstract: Aerobic organisms are continuously exposed to exogenous and endogenous reactive oxygen species. Oxidatively induced cell damage has been proposed to play an important role in the etiology of numerous pathological conditions. The present study evaluates the antioxidant potential of extract of Hydrastis canadensis (HC) against hydrogen peroxide induced oxidative damage in peripheral blood lymphocytes (PBLs). Incubation of PBLs with 100 µM H2O2  for 2 hours significantly increased lipid peroxidation and decreased the levels of glutathione and the antioxidant enzymes. Pretreatment with HC for 18 h could effectively inhibit lipid peroxidation and enhance the activities of antioxidant enzymes and glutathione content significantly. The observed results indicate that the cytoprotection may be due to its direct action in scavenging free radicals and thereby modulating the antioxidant defense system.

24. Journal of Cell and Tissue Research Vol. 6(2): 845-847 (2006)

ANTI-INFLAMMATORY AND ANALGESIC PROPERTIES OF CASSIA AURICULATA ROOT EXTRACT

SHIRADKAR, M.R., SHIRADKAR, K., LAHOTI, T., SHIVAPRASAD, H.N. AND BAVISKAR, B.

AISSMS  College of Pharmacy, Kennedy Road,  Pune 411 001. E mail: rrshiradkar@rediffmail.com

Abstracts: The methanolic extract of Cassia auriculata (a member of Family Caesalpiniaceae) root was investigated for anti-inflammatory and analgesic activities at a doses of 100, 200 and 300 mg/kg, body weight. The experimental procedures used were carrageenan induced pedal edema for anti-inflammatory activity and hot plate method for analgesic activity. In acute phase inflammation, a maximum inhibition of 62% was noticed at the dose of 300 mg/kg after 3 h of treatment. The extract also produced significant (p<0.01) analgesic activity in hot plate reaction time in mice. This study exhibits that the methanolic extract of roots of Cassia auriculata possess both anti-inflammatory and analgesic activities.

25. Journal of Cell and Tissue Research Vol. 6(2): 849-851 (2006)

IN VITRO ANTIBACTERIAL ACTIVITY OF ESSENTIAL OIL FROM RHIZOME OF HOMALOMENA AROMATICA AGAINST PATHOGENIC BACTERIA

LAISHRAM, S.K.S.,  NATH, D.R., BAILUNG, B. AND BARUAH, I.

Defence Research Laboratory (DRDO), Post bag No. 2, Tezpur, Assam 784001, India.
E-mail: Shantiks@rediffmail.com

Abstract: The antibacterial activity of the essential oil from rhizome of Homalomena aromatica was evaluated against five common and clinically significant bacterial pathogens viz. Staphylococcus aureus (MTCC-96), Escherichia coli (MTCC-739), Klebsiella pneumoniae (MTCC-3040), Pseudomonas aeruginosa (MTCC-2582) and Proteus vulgaris (MTCC-742). The antimicrobial bioassay was evaluated by agar disc diffusion method. The used oil shows a varying degree of potential inhibitory effect against the tested pathogens. S. aureus was found to be the most sensitive while P. aeruginosa was the least sensitive among the tested pathogens.