1. Journal of Cell and Tissue Research Vol. 7(2) 1013-1018 (2007)
Functional and receptor modulations of T- lymphocyte and macrophage in acute and subacute dose finding studies of a novel immunotherapeutic molecule T11TS

Sarkar, P., Bhattacharjee, M., Acharya, S., Ghosh, A. and Chaudhuri, S.

Cellular and Molecular Immunology Lab, Department of Physiology, Dr. B.C. Roy Post Graduate Institute of Basic Medical Sciences, Institute of Post Graduate Medical Education and Research. 244B, A.J.C. Bose Road, Kolkata-700020, India. E. mail: swapna_chau@rediffmail.com

Abstract:  T11TS, a membrane glycopeptide of sheep erythrocytes (SRBC) helps to regress successfully the ENU induced rat glioma. T11TS ligands with CD2 molecule on various immunocytes and thereby activate them to enter into the brain parenchyma causing apoptosis of glioma cells. The present study deals with animal toxicological studies and its immunological consequences of intravenous administration of T11TS during acute (four groups) and sub acute (three groups) dose finding studies (n = 6). Splenic T lymphocytes of these animals showed no E-rosette formation with SRBC due to the down regulation of CD2 receptor on T cells. The non-specific cytotoxicity of these cells remained unaltered. Growth arrest stage of the T cells following T11TS administration is indicated by CD25 (IL-2R) down regulation, the activation marker of T lymphocytes. Thus, the anergic state acquired by T cell is characterized by intact functional activity and down regulated growth receptors.  Phagocytic activity of the splenic macrophage of these animals showed no  alteration . However, this cell showed marked suppression in CD11b (CR3) receptor expression in the highest dose selected for acute and subacute dose finding study.

2. Journal of Cell and Tissue Research Vol. 7(2) 1019-1029 (2007)
Electron microscopic radioautographic study on macromolecular synthesis in hepatocyte mitochondria of aging mice

Nagata, T.

Department of Anatomy and Cell Biology, Shinshu University School of Medicine, Matsumoto 390 8621 and Department of Anatomy, Shinshu Institute of Alternative Medicine, Nagano 380 0816, Japan. E-mail: nagatas@po.cnet.ne.jp

Abstract: In order to study the aging changes of intramitochondrial macromolecular synthesis in mouse hepatocytes, 10 groups of aging mice during development, each consisting of 3 individuals, total 30, from fetal day 19 to postnatal month 24,  were injected with 3H-leucine,  a protein precursor, sacrificed 1 hr later and the liver tissues were processed for electron microscopic radioautography. On electron microscopic radioautograms obtained from each animal, the number of mitochondria, the number of labeled mitochondria and the mitochondrial labeling index labeled with silver grains due to 3H-leucine showing protein synthesis in each hepatocyte were counted and the averages in respective aging groups were compared.  From the results, it was demonstrated that the numbers of mitochondria, the numbers of labeled mitochondria and the labeling indices of intramitochondrial protein syntheses in mononucleate hepatocytes of mice at various ages from embryonic day 19 to postnatal month 24 increased and decreased due to aging of animals. 

3. Journal of Cell and Tissue Research Vol. 7(2) 1031-1037 (2007)
Production, purification and characterization of an alkaline proteasefrom an alkaliphilic
Bacillus sp.

Sinha, R.,  Joshi, R. H., Dodia, M. S. and Singh S. P.

Department of Bioscience, Saurashtra University, Rajkot- 360005, India. E. mail: satyapsingh@yahoo.com

Abstract: An extracellular serine like alkaline protease was purified and characterized from alkaliphilic Bacillus sp. The enzyme was purified to the homogeneity by a combination of aqueous two-phase system and affinity chromatographic techniques. The apparent molecular weight was estimated as 26-29 kDa based on SDS-PAGE and gel filtration chromatography and the enzyme appeared to be a monomer. The Km and Vmax for the casein were 4.7mgml-1 and 24.5μgml-1 respectively. The enzyme was active over a wide range of temperature (10-65°C) and pH (8-12.5); the optimum activity being at 35°C and pH 11.0. The purified enzyme was quite stable at 35°C up to 1 h of incubation, while 80 and 50% residual activities were detected at 55 and 65°C respectively after 20 min of incubation. Similarly, enzyme exhibited 100% stability in alkaline pH range, which sharply decreased in acidic range. The enzyme lost its activity in the presence of PMSF, Hg+2 and Cu+2 and remained unaffected with EDTA, Mg+2 and Ba+2.

4. Journal of Cell and Tissue Research Vol. 7(2) 1039-1042 (2007)
Serological levels of zinc and iron among Giardia lamblia infected children under 12 years old

Ajami, A. and Izadi, J.

Department of Immunology, Sari Medical School,  School of Medicine, Mazandaran Medical Science University, Sari, Iran. E Mail: A.Ajami@Mazums.ac.ir

Abstract: Giardiasis is one of the most prevalent human intestinal parasite infections, especially among children aged between 2 to 12 years old.  In this study, the effects of giardiasis on the plasma levels of zinc and iron were determined in infected children. A total of 100 children aged under 12 years who were admitted to the pediatric department of hospitals affiliated to Mazandaran Medical Science University with gastrointestinal complain, and diagnosed as having giardiasis by stool examination (direct and formalin-ether technique and trichrome stain) were selected as the case group.  The control group consisted of 100 age-matched healthy children.  A record of age, physical growth status and clinical symptoms were recorded.  Serological levels of zinc and iron were measured by atomic absorption spectrophotometer in all samples. In this study, serum iron levels were 67±25 μg/dl and 77±23 μg/dl and zinc levels were 64±20 μg/dl and 96±23 μg/dl in case and control groups, respectively. Statistical analysis showed significant differences between the serological levels these elements, which was much higher in serum zinc level (P<0.01). These results revealed that serological levels of zinc and iron decreased during giardiasis due to malabsorption.

5. Journal of Cell and Tissue Research Vol. 7(2) 1043-1046 (2007)
Antihyperammonemic effect of morin: A dose dependent study

Subash, S., Subramanian, P. and Sivaperumal, R.

Department of Biochemistry and Biotechnology, Faculty of Science, Annamalai University,  Annamalainagar 608 002, India. E mail: annamalai_rhythm@yahoo.co.in

Abstract: Ammonia is a neurotoxin that has been strongly implicated in the pathogenesis of hepatic encephalopathy and a major pathogenic factor associated with inborn errors of urea cycle, Reye’s syndrome and organic acidurias. The present work is aimed at evaluating the protective effect of morin (3, 5, 7, 2', 4' – pentahydroxyflavone), a naturally occurring plant component on ammonium chloride (AC) induced hyperammonemia in Wistar rats. Four different doses of morin (10, 20, 30 and 40 mg/kg of body weight) were administered to rats treated with AC for 8 weeks. An influence of morin on AC induced hyperammonemia was evaluated by analyzing the circulatory levels of ammonia, urea, uric acid, non-protein nitrogen and creatinine. The levels of these components were significantly increased in the AC treated rats but were decreased significantly on treatment with morin.  30 mg/kg body weight dose of morin was significantly effective. Further the administration of morin (10, 20, 30 and 40 mg/kg of body weight) to normal rats did not show any noticeable side effects. Thus our results show that morin could act as an effective antihyperammonemic agent without causing side effects and may be a quite suitable for further research to find a natural antihyperammonemic agent.

Therapeutic effect of herbal and inherent antioxidants on hexavalant chromium altered circulating thyroid hormones

Sood, P. P., Misra, S., Lavekar, G. S.  and Chundawat, R. S.

Laboratory of Neurobiology and Toxicology, Department of Biosciences, Saurashtra University, Rajkot 360005, India. E mail: ppsood@yahoo.com

Abstract:  Hexavalent chromium (Cr VI) pretoxicated developing broiler chicks were treated with the innate and herbal antioxidants that showed significant therapeutic efficacy in the restoration of circulating thyroid hormones TSH, T3 and T4 decreased by metal toxicity. Two day old chicks were divided into seven groups and treated with a daily dose of 10 mg/Kg potassium dichromate from 2nd to 8th day of their age. Thereafter, one group was kept without intoxication for another 7 days and sacrificed on 16th  day. Five groups of chromium pre-toxicated animals were exposed to therapeutic agents viz., vitamins (B and E), glutathione, ashwagandha and garlic for another seven days. These animals were sacrificed on 16th  day of their age along with control and TSH, T3 and T4 levels were measured in serum. Study shows that Cr (VI) decreases T3 and T4 more severely than TSH.  Almost all the therapeutics tried proved efficient to raise the hormonal concentration though the recovery was not uniform. Nevertheless, in many cases a control level was achieved. Further, study also indicates that a positive T3/T4 ratio is maintained pointing that chromium administration did not leave any impact on a large scale.

Development of primary cell culture from heart explants of common carp, Cyprinus carpio (linn)

Mastan, S. A., Goswami, S., Kakaria, V. K. and Qureshi, T. A.

Department of Applied Aquaculture, Barkatullah University and Regional Institute of Education (NCERT), Bhopal - 462013, India. E mail: shaikmastan2000@yahoo.com.

Abstract: Primary cell cultures of heart cells were initiated from heart tissue explants of common carp, Cyprinus carpio. The hearts were collected aseptically from 8-10 g (2-3 inch) fingerling after killing them by a blow on head. Before seeding, the tissues were minced in phosphate buffer saline (PBS) with antibiotic/antimycotic mixture. The explants were cultured in minimum essential medium (MEM) with 10% foetal bovine serum (FBS) and antibiotic/antimycotic mixture. Approximately 50% of minced heart tissue was subjected to standard trypsinisation method with 0.25% trypsin – EDTA solution. The cell pellets were suspended in MEM with 10 % FBS. They were incubated at room temperature. After 40-48 h of attachment, several types of cells were found to grow. A good and confluent monolayer found in three weeks time. In present experiment, the explant method was found to be better than trypsinisation for initiation of primary culture. The attachment efficiency and growth of cells were better and fast in explant method than trypsinisation.

Evaluation of analgesic and antiinflammatory activities of thiazolidinone derivatives

Taranalli, A. D., Bhat, A. R., Srinivas, S. and Saravanan, E.

Department of Pharmacology, K. L. E. S’s College of Pharmacy, Belgaum 590 100, India.
E mail: ashok_taranalli@yahoo.co.in

Abstract: A comparative study for pharmacological activity was focused on the two sets of thiazolidinone derivatives. 1). Para amino phenol was made to react with aromatic aldehyde followed by thioglycolic acid to form the respective thiazolidine-4-one derivative which was substituted with aryl grouping at 2-position of thiazolidine-4-one.  2). In another set 7-hydroxy, 4-methyl-coumarin was reacted with ethyl chloroacetate to get the corresponding ester which in turn on reacting with hydrazine hydride yielded the corresponding thiazolidinone derivative. These two sets of thiazolidinone derivatives were studied for anti-inflammatory and analgesic activities. The toxicity studies were performed according to the OECD guidelines and dose was fixed as 50mg/kg b.w. The study was performed using acute models like Carrageenan induced paw edema in rats for anti-inflammatory activity and acetic acid induced writhing in mice for analgesic activity. The compounds C3 and C6 showed significant anti-inflammatory and analgesic activities in comparison to other derivatives. This increased activities can be attributed to the substitution of R-H, R’-OH, and   R”-OCH3 in C3, and R-H, R’-H, and R”-Cl in C6.

Evaluation of certain novel thiazolidinones foranti-inflammatory, analgesic, antipyretic and cyclooxygenase inhibitory activity in animals

Taranalli, A. D., Bhat, A. R., Srinivas, S. and Saravanan, E.

Department of Pharmacology,  K.L.E.S’s College of  Pharmacy, Belgaum 590 100.
E mail: ashok_taranalli@yahoo.co.in

Abstract: Nimesulide was hydrolyzed under alkaline conditions to obtain a free amino compound. The Schiff bases were prepared from these and were converted to the thiazolidine-4-one derivatives which were further converted to the corresponding spiro compounds by reacting with 1,4-dibromobutane. These compounds were evaluated for anti-inflammatory, analgesic, antipyretic and cyclooxygenase enzyme inhibitory activity. All the compounds showed significant anti-inflammatory, analgesic and antipyretic activity at 100 mg/kg b.w. The compounds A3, A4, A5, A6 and A7 showed significant inhibition of COX-2 activity without inhibition the COX-1 activity. The nimesulide and paracetamol were used as standard drugs for comparison. The substitution at R, R1 and R2 with the functional groups Cl, OCH3, NO2, and OH in the aromatic ring resulted in increased activity as compared to unsubstituted thiazolidinones.

Induction of apoptosis by piper betel leaf extract on 7, 12 dimethylbenz [A] anthracene induced hamster buccal pouch carcinogenesis

Selvamuthukumar, S., Pushpalatha, R., Sethupathy, S., Manavalan, R. and Karar, P. K.

Department of Pharmacy, Annamalai University, Annamalai Nagar 608002, India.
E mail: smk1976@gmail.com

Abstract: The induction of apoptosis by Piper betel leaf extracts during 7, 12 dimethylbenz [a] anthracene (DMBA) induced hamster buccal pouch (HBP) carcinoma was studied in male Syrian hamsters using DNA fragmentation and the apoptosis-associated proteins, tissue transglutaminase (tTG) and Bcl-2. Hamsters were divided into four groups of six animals each. Animals in group 1 were painted with a 0.5% solution of DMBA in liquid paraffin on the right buccal pouches three times a week for 14 weeks. Group 2 animals painted with DMBA as in group 1, in addition received 500 mg/kg body weight chloroform extract of Piper betel leaves orally on days alternate to DMBA application. Group 3 animals painted with DMBA as in group 1, in addition received 500 mg/kg body weight methonolic extract of Piper betel leaves orally on days alternate to DMBA application. Group 4 animals received neither DMBA nor betel leaf extracts and served as the control. The experiment was terminated at the end of 14 weeks. Administration of Piper betel leaf extracts (500 mg/kg body weight) to animals painted with DMBA inhibited DMBA-induced oral cancer as revealed by the absence of neoplasm, induction of tTG and inhibition of Bcl-2 expression. The results of the present study suggest that Piper betel leaves may exert its chemopreventive effect by inducing apoptosis.

Chemopreventive effect of Piper betel and Alternanthera amoena leaves extracts on cellular redox status during buccal pouch  carcinogenesis

Selvamuthukumar, S., Pushpalatha, R., Manavalan, R., Sethupathy, S. and Karar, P. K.

Department of Pharmacy, Annamalai University, Annamalai Nagar  608002
E mail: smk1976@gmail.com

Abstract: In present investigation the effect of Piper betel and Alternanthera amoena leaves extracts has been studied on cellular redox status during hamster buccal pouch  carcinogenesis. Animals were divided into 8 groups. In group 1 animals were painted with a 0.5% solution of DMBA in liquid paraffin on the right buccal pouches three times a week for 14 weeks. Group 2 animals painted with DMBA as in group 1, in addition received 500 mg/kg body weight methanolic extract of Piper betel leaves (PE) orally on days alternate to DMBA application. Group 3 animals painted with DMBA as in group 1, in addition received 500 mg/kg body weight methonolic extract of Alternanthera amoena leaves (AE) orally on days alternate to DMBA application. Group 4 animals painted with DMBA, in addition 250 mg/kg body weight of PE and AE were administered orally on days alternate to DMBA application. Animals groups 5 to 7 received PE 500 mg/kg body weight, AE 500 mg/kg body weight and 250 mg/kg body weight of both PE and AE respectively. Group 8 animals received neither DMBA nor herbal extracts (PE and AE) and served as the control. The experiment was terminated at the end of 14 weeks. Combinatorial administration of PE and AE leaf extracts to animals painted with DMBA inhibited DMBA-induced oral cancer as revealed by the absence of neoplasm. The results of the present study suggest that combinatorial administration of PE and AE may potentiate the chemopreventive effect more than that of either agent used alone.

Expression and correlation of cytokeratin 19 mRNA with the progression of colorectal cancer: A preliminary study

Bavarva, J. H. and Shah. N. G.

B.R.D. School of Biosciences, Sardar Patel University, Vallabh Vidyanagar 388120, India
E mail: jasmin.spu@gmail.com

Abstract: Cytokeratin 19 is epithelial cell specific intermediate filament protein, which may help to provide the stability to the cell unlike other Cytokeratin family members. There is no report correlating the expression of CK19 with progression of colorectal cancer. In order to explore such correlation of CK19 mRNA with disease progression we designed this pilot study. Twenty-five tumor samples along with six normal mucosal samples were collected. The total RNA was prepared and subjected to reverse transcriptase polymerase chain reaction and the product was quantified on gel documentation system. Amongst twenty five, fifty six per cent (14/25) showed a raised expression of CK19 mRNA than the average value of normal mucosa (cut-off) in the initial stages of colorectal cancer progression, revealing a significant inverse correlation with lymph node status (r = -0.457, p = <0.05). Rests of the parameters were non-significant. Thus, it was concluded that the CK19 expression is up-regulated in the initial stages of the disease and then down regulated with the progression of the disease increasing the invasive potential of the tumor. In particular, results revealed the association of CK19 expression with progression of colorectal cancer.

Effect of salivariadenectomy on the testicular lactate dehydrogenase-X in mice.

Bodare, R. D. and Pillai, M. M.

Department of Zoology, Shivaji University, Kolhapur   416004.  E mail: rdbodare@rediffmail.com

Abstract : The aim of present study is to investigate the effect of salivariadenectomy on the postnatal development of testicular lactate dehydrogenase-X. In general the salivariadenectomy means either the removal of sublingual or submandibular glands or removal of both the glands. These glands were removed in male mice at their age of 20 days. The controls were sham operated. The operated male mice were sacrificed at the age of 45 days, 60 days and 90 days. The testes of these animals were studied biochemically and by polyacrylamide gel electrophoresis for lactate dehydrogenase activity. The epididymis was used to study the sperm count. In control mice, the lactate dehydrogenase were electrophoretically separated into six bands i.e., I, II, III, IV, V and LDH-X, but the testis specific LDH-X band was completely disappeared in salivariadenectomised mice. The total LDH activity and the sperm count were decreased significantly in the operated mice sacrificed at different age intervals. 

14. Journal of Cell and Tissue Research Vol. 7(2) 1089-1091 (2007)

Evaluation of sustained release capsules of metoprolol in rabbits: I. An in vivo study

Kannan, K., Subburaj, T., Manavalan, R. and Karar, P. K.

Department of Pharmacy, Annamalai University, Annamalai Nagar  608 002,
E mail: egkkannan@yahoo.co.in

Abstract: Pharmacokinetic evaluation of metoprolol sustained release 125mg capsules (wax coated pellets of metoprolol filled in hard gelatin shells) has been studied in this study. A single dose was carried out in six rabbits with two sequences, cross over study. Blood samples were collected at 0.5, 1.0, 2.0, 3.0, 4.0, 6.0, 8.0, 10, 12 and 24 hours. The plasma concentration of metoprolol was estimated by reverse phase HPLC. The pharmacokinetic parameters were calculated from the plasma concentration of metoprolol and time data. The sustained Tmax and lower Cmax, was decreased, absorption rate constant (Ka) and prolonged t ½ indicate a prolonged release of metoprolol from sustained release pellets in comparison with immediate release dosage form. The results indicate that wax coated pellets were provide controlled delivery of metoprolol in desired rate.

15. Journal of Cell and Tissue Research Vol. 7(2) 1093-1096 (2007)

Evaluation of sustained release capsules of metoprolol tartrate: II. An in vitro study

Kannan, K., Subburaj, T., Manavalan, R. and Karar, P. K.

Department of Pharmacy, Annamalai University, Annamalai Nagar  608 002, India.
E mail: egkkannan@yahoo.co.in

Abstract: Formulation development and in vitro evaluation of metoprolol tartrate sustained release capsules were studied in this investigation. Non pareil seeds were prepared and coated with metoprolol tartrate using polyvinyl pyrollidone and iso propyl alcohol. Drug coated pellets were then coated with different proportion of microcrystalline wax and glyceryl distearate using carbon tetrachloride as solvent. The release pattern of drug coated pellets and wax coated pellets were evaluated in vitro by dissolution test apparatus. Theoretical sustained release was compared with formulated sustained release. For successful formulation, 10% of drug coated pellets and 90% of wax coated pellets were blended and filled in hard gelatin capsules. Stability study was carried out and the formulation was found to be stable.

16. Journal of Cell and Tissue Research Vol. 7(2) 1097-1100 (2007)

Assessment of antistreptokinase antibody as a risk factor in predicting acute myocardial infarction

Rezaei, A., and Shemirani, H. 

Immunology Department, Isfahan medical Science, Faculty of Medicine, Isfahan, Iran
E mail: rezaei@mui.ac.ir, rezaei1335@yahoo.com

Abstract: Anti streptokinase antibody (ASK) is an antibody which is produced against streptococcus infection or induced by streptokinase (SK) infusion. This antibody may be considered as an inflammatory marker which increase fibrinogen level and then promote myocardial infarction. 135 (104 male and 31 female) patients with myocardial infarction (MI) who hospitalized in emergency ward, Noor hospital, Isfahan, were studied in an observational and cohort study. ASK and fibrinogen levels were estimated from patient’s blood taken at the time of MI diagnosis and before SK infusion. Electrocardiograph response was used to divide patients in two groups’ viz., Q wave and non Q wave MI. For managing patients, fibrinogen level greater than 500 mg/dl considered as high level. Difference between groups tested by Chi-squire or Fisher exact test if p value became significant. Comparison between two groups with high and normal level of ASK Ab and Q wave MI showed that fibrinogen level was higher in patients with high level of ASK Ab P=0.000. Comparison amongst other groups was not meaningful. Patients with AMI who had not ST segment resolution about 70% after 1-2 hours of SK infusion had higher ASK and fibrinogen level. Therefore it could be interpreted that ASK Ab may considered as a risk factor for AMI.

17. Journal of Cell and Tissue Research Vol. 7(2) 1101-1105 (2007)

Effect of antioxidants on essential trace metals, glutathione, and d-aminolevulinic acid dehydratase levels in rats

Kumar, P., Bhatt, K., Pant, S. C., Satish, H. T. and Flora, S. J. S.  

Division of Pharmacology and Toxicology, Defence Research and Development Establishment, Jhansi Road, Gwalior-474 002, India. E mail: sjsflora@hotmail.com

Abstract: The effect of repeated administration of few selected antioxidants on essential metal status and on the activity of metalloenzymes like d-aminolevulinic acid dehydratase (ALAD) and on the concentration of glutathione (GSH) and thiobarbituric acid reactive substance (TBARS) in blood, liver and kidneys were investigated in male rats. The animals were treated with 0.1 mmol/kg dose of each of the 3 antioxidants, intraperitoneally daily for 10 days. The results suggest that among the three antioxidants only d-lipoic acid slightly but significantly inhibited blood ALAD activity which could be due to moderate alterations in essential metal status. No adverse effect of these antioxidants on GSH and TBARS levels in blood, liver and kidney was noted. While, blood zinc and copper concentration showed significant depletion on d-lipoic acid administration along with depleted liver copper in animals given repeated dose of d-lipoic acid. Kidney magnesium concentration showed marginal increase on melatonin administration while administration of N-acetylcysteine led to a significant loss of blood and liver zinc concentration.  The results suggest that among the three antioxidants, melatonin has least adverse effects on repeated administration based on the parameters studied in the present study.

18. Journal of Cell and Tissue Research Vol. 7(2) 1107-1110 (2007)

A comparative study of blood proteins and some of its enzymes in Oziotelphusa senex senex and in Pila globosa during thermal stress

Sridevi, B. and Naidu, B. P.

Center for Biotechnology, Department of Animal Sciences, University of Hyderabad, Hyderabad  500 046, E. Mail: sri27_bonthu@yahoo.co.in

Abstract: In Oziotelphusa senex senex and in Pila globosa on exposure to cold temperature biosynthetic activity is considerably increased besides lowering of maintenance energy requirements. During warm exposure, a few metabolites are either sharply decline or elevated indicating an active utilization of metabolites. This is in consonance with the assumption that during warm exposure the energy requirement is considerably high. Study shows that during warm exposure, the protein synthetic machinery is disturbed causing a decrease in protein level. The elevation in the free amino acid pool, urea, ammonia aspartate aminotransferase and alanine aminotransferase suggests enhanced proteolytic activity during thermal stress.

19. Journal of Cell and Tissue Research Vol. 7(2) 1111-1114 (2007)
Granulosa cell apoptosis in situ in caprine ovary
Sharma, R. K.  and Bhardwaj, J. K.

Reproductive Physiology Laboratory, Department of Zoology, Kurukshetra University, Kurukshetra 136119. E. mail: bhardwajkj111@yahoo.co.in

Abstract: Apoptotic changes in membrana granulosa of ovaries in goat (Capra hircus) have been studied in situ using light and scanning electron microscopy. Histologically, the degenerating granulosa cells were characterized by condensed cytoplasm, pyknotic nuclei and hazy cytosol. Most of the cytoplasmic components were condensed and were seen clumping against the pyknotic nucleus. The nucleus of degenerating cells stain darkly with giemsa while the cytoplasm were eosinophilic. In a few apoptotic cells heterochromatin bodies were seen adhering the nuclear membrane in the hyaline nucleosol. Under scanning electron microscopy apoptosis was marked by asymmetrical shrinkage and vacuolization of cytoplasm. The cell membrane of degenerating granulosa cells was smooth textured with a number of uneven depression and ruffles. Fragmentation of nucleus and pinching off of membrane bound nuclear fragments were the characteristic features observed in situ.

20. Journal of Cell and Tissue Research Vol. 7(2) 1115-1121 (2007)

Cytokine modulation of icam-1 (CD54) in human conjunctival epithelium: Primary cells and cell line compared and contrasted
Trocme, S. D., Sra, S. K.,  Haq, S.  and  Zhang, M.

Department of Ophthalmology and Visual Sciences , Case Western University, Cleveland Ohio, University of Texas Medical Branch, Galveston,  Texas, USA

Abstract: Adhesion molecules play a key role in the migration of effector cells to sites of inflammation in various allergic conditions. At the ocular surface, the intercellular adhesion molecule-1 (ICAM-1 or CD54) functions as the ligand for leukocyte adhesion to the corneal and conjunctival epithelium. In the present study, we investigated the effects of the proinflamatory cytokines IFN-g and TNF-a on ICAM-1 expression on cultured human conjunctival epithelial cells and the HC0597. Human conjunctival epithelial cells (primary cells or the HC0597 cell line) were grown to confluence in 96-well plates and treated with increasing concentrations of IFN-g, TNF-a, or a combination of IFN-g and TNF-a. ICAM-1 expression was measured using a cell-based ELISA. Exposure to IFN-g or TNF-a induced a time and dose-dependent increase of ICAM-1 expression in the HC0597 cells. IFN-g induced up to an eight-fold increase in ICAM-1 expression at 24 hours, while there was less than a 1-fold increase from any TNF-a treatment. A similar trend was observed in the primary conjunctival epithelial cells. Treatment with IFN-g and TNF-a resulted in an additive, but not a synergistic increase in conjunctival ICAM-1 expression. ICAM-1 expression on conjunctival epithelial cells can be modulated by the proinflammatory cytokines IFN-g or TNF-a in a dose- and time-dependent manner. The up regulation of ICAM-1 on the conjunctival epithelium may contribute to the inflammatory response in ocular allergies by binding and activating leukocytes, which express the ICAM-1 receptor LFA-1. The HC0597 cell line proved to be a good model for the human conjunctival epithelial cell response to proinflammatory cytokines.

21. Journal of Cell and Tissue Research Vol. 7(2) 1123-1129 (2007)

Evaluation of antibacterial property of fresh leaf juice of Tridax procumbens: An in vitro study

Deshmukh, A. A., Gajare, K. A., Bele, M. S., Solanke, M. R.  and Deshmukh, V. P.

Department of Zoology, Government Vidarbha Institute of Science and Humanities, Amravati 444 6064, India. E. mail: ashish_cellbio@rediffmail.com

Abstract: Tridax procumbens a commonly occurring weed in several parts of the world is routinely used for several ailments such as cuts and wounds, blister boils, in diarrhoea and dysentery. The fresh leaf juice is directly applied on the wounds. In the present investigation the antibacterial activity of this plant was studied in two species of Gram-positive bacteria (Staphylococcus aureus and Bacillus subtilis) and one species of Gram-negative bacteria (Escherichia coli). The antibacterial activity was studied by Agar well diffusion method, Disk diffusion method and Turbidometric assay. In Agar well diffusion method S. aureus and B. subtilis exhibited zone of inhibition immediately followed by zone of stimulation around the well containing fresh leaf juice of Tridax procumbens. In disk diffusion method there was zone of enhanced growth of S. aureus and B. subtilis. The S. aureus and B. subtilis from the zone of inhibition, inoculated on the nutrient agar plate exhibited prominent growth after twelve hours of incubation suggesting the bacteriostatic action of Tridax procumbens. There was neither inhibitory nor stimulatory effect of leaf juice on E. coli. In turbidometic assay there was highly significant (p<0.001) decrease in the optical densities in leaf juice treated group than the control group of S. aureus and B. subtilis, while in E. coli there was insignificant difference in the optical densities of the control group and leaf juice treated group. From all these results it seems that leaf juice of Tridax procumbens is bacteriostatic on Gram-positive bacteria and ineffective on Gram-negative bacteria.

22. Journal of Cell and Tissue Research Vol. 7(2) 1131-1136 (2007)

Evaluation of plumbago indica callus for its plumbagin content and antimicrobial activity

Karadi, R. V., Kavatagimath, S. A., Gaviraj, E. N., Sastry, D. N., Chandrashekhara, S. and Rajarajeshwari, N.

Department of Pharmaceutical Biotechnology, KLES College of  Pharmacy, Belgaum 590 010, India.
Email: rvkaradi@yahoo.com

Abstract: Plumbagin is commercially important because of its broad range of pharmacological activities such as antimicrobial, anticancer, antifertile and insecticidal. Production of this metabolite by plant cell culture is required as the native Plumbago species produce only small amount of this compound after 2-6 years of growth. In order to study the biosynthetic potential of in vitro cultures, efforts were made to establish callus cultures from leaves and shoot tips of Plumbago indica on Murashige and Skoog (MS) and Gamborg’s (B5) media containing varying amounts of auxins and cytokinin. Callus initiation was seen in both media but a greater frequency was obtained in MS. Growth was highest in MS media supplemented with 2,4-dichlorophenoxy acetic acid (2 mg/l) and Kinetin (0.5 mg/l) but maximal production of plumbagin was seen in the presence of 1-naphthalein acetic acid (2 mg/l) and kinetin (0.5 mg/l). Calli exhibited an initial lag phase before entering an active growth phase around day 12 and reached stationary phase 24 days after inoculation. The synthesis of plumbagin was found to be non-growth-associated with minimum in early exponential phase (0.31 mg/g dry weight on day 12) and maximum in early stationary phase (2.45 mg/g dry weight on day 24) of culture. Calli were elicited with copper sulphate and silver nitrate but the amount of plumbagin was enhanced by about 2-fold to 7.61 mg/g dry weight by the former at 10 μM. Antimicrobial studies showed a close correlation of activity with the production of plumbagin indicating in vitro cultures are capable of producing bioactive secondary substances.

23. Journal of Cell and Tissue Research Vol. 7(2) 1137-1140 (2007)

Topical combination (azithromycin, fluconazole, metronidazole) and systemic glucantime treatments for cutaneous leishmaniasis

Siadat, A. H., Shirani-Bidabadi, L., Zolfaghari-Baghbaderani, A., Saberi, S., Nilforoshzadeh, M. A., Jooya, A. and  Mahmoudi, M.

Skin Diseases and Leishmaniasis Research Center, Department of Dermatology, Isfahan University of Medical Sciences, Isfahan,  Iran. E. Mail: amirhossein1@yahoo.com

Abstract: Leishmaniasis is a parasitic disease transmitted by sand flies. It is characterized by a spectrum of cutaneous, mucocutaneous and visceral clinical manifestations that depend largely on the species of parasite involved and the host immune response. Unfortunately, still there is no definite treatment for this infection. Pentavalent antimony compounds are considered as first-line agents for the treatment of cutaneous leishmaniasis. Our objective in this study was to compare the efficacy of  topical combination treatment with “3% azithromycin + 3% fluconazole + 3% metronidazole” versus “ intralesional glucantime” in the  treatment of cutaneous leishmaniasis caused by Leishmania. Leishmania major (MRHO/IR/75/ER) strain was used.  6 to 8 week old female BALB/C mice were  infected with 1.6 × 106 viable stationary-phase promastigotes  in the base of tail. 31 BALB/C infected  mice were randomly divided into  in to 3 groups. 12 mice were treated with systemic glucantime, 20mg/kg, once daily for 20 days, 7 mice were treated with combination treatment of  topical, “% 4 azithromycin + 3% fluconazole + 3% metronidazole alcoholic solution” twice daily for 6 weeks and 12 mice mice were left untreated and regarded as controls. The statistical tests including student t-test and Kruskal-Walis were used for analysis. There was no significant difference regarding lesion size between the 3 groups at the starting of the treatment (p value = 0.87). At the end of study, no significant difference regarding progression of the lesions was observed between the topical combination and glucantime treated groups (p value=0.64). Both of these treatments were significantly effective in arresting the progression of the cutaneous leishmaniasis lesions as compared to control mice.

24. Journal of Cell and Tissue Research Vol. 7(2) 1141-1146 (2007)

Biochemical alterations in blood parameters by marble stone dust in craft workers
Upadhyay, R. K. and Jaiswal, G.

Department of  Zoology, Deen  Dayal Upadhyay Gorakhpur University, Gorakhpur 273 009, India. E mail: rkupadhya@ yahoo.com

Abstract: A survey of limestone exposure amongst 783 cases of marble stone craft workers was undertaken in 50 crafting firms at Agra where the industry is providing wages to 5-10% of the population. Important hematological changes such as RBC, Hb, PCV, MCV, MCH, MCHC, DLC and platelets counts were observed in regular craft workers and daily wagers based on the age, exposure time and socioeconomic conditions. The mean blood hemoglobin concentration was found 11.24 ± 0.39 gm/100ml in workers which is also observed in more chronic stage among aged workers. In workers mean RBC counts was significantly decreased 3.22± 0.016 in comparison to non-workers, while WBC counts was found increased (8.66± 0.22). Among WBC mean lymphocyte count was decreased (26.08± 0.45) and monocyte count get increased in the workers (4.51± 0.18) in comparison to non-workers. Similarly platelet count in workers gets induced (317± 0.72) due to dust exposure. The cholinestrase, lactic dehydrogenase and alkaline phosphatase activities were  found higher in comparison to control, which indicate that substantial occupational exposure of solid dust particles were generating negative effects and health hazards for the body physiology and biochemistry of craft workers. Most of the workers were unaware of the risks due to inhalation of carcinogenic stone dust as it has significantly induced sister chromatid exchanges (SCEs) in craft workers (20.17 ± 0.202) in comparison to non-workers (5.52 ± 1.65). It was found more erroneous on the part of industries that they have poor safety measures for the protection of stone craft workers.    

25. Journal of Cell and Tissue Research Vol. 7(2) 1147-1154 (2007)

Primer designing for microbial endo-1, 4-b-xylanase gene
Garg, S., Sohani, N., Pundhir, S. and Anil Kumar

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

Abstract: Primer designing for endo-1,4-b- xylanase gene is done using Primer 3 software. Nineteen sequences for endo-1,4-b- xylanase were retrieved from NCBI’s refseq database through Entrez. On multiple sequence alignment of the sequences, three conserved regions were identified. All the combinations of 18 to 24 nucleotides in these regions were selected as potential primers. The properties of these potential primers were analyzed using Premier Biosoft’s NetPrimer tool. One forward (5’end) and one backward (3’end) primers having 50 to 60% GC contents, 54 to 58oC Tm and absence of secondary structures were finalized. Specificity of the primers was validated by carrying out the local alignment against the NCBI’s nr database through BLAST. All the alignments showed significant alignment to xylanase validating the specificity of the primers.

26. Journal of Cell and Tissue Research Vol. 7(2) 1155-1160 (2007)

Comparative study of arylpropionic acid derivatives ketoprofen and naproxen on acute and chronic inflammatory muscle hyperalgesia

Chopade, A. R., Nalawade, A. Y.,  Pol,  R. P. and Naikwade. N. S.

Department of Pharmacology, Appasaheb Birnale College of Pharmacy,  Sangli  416416, India.
E. mail: chopadearv@gmail.com

Abstract: Pain is a common problem, often associated with inflammatory conditions. Nonsteroidal anti-inflammatory drugs (NSAIDs) are the most commonly used as analgesic agents. These drugs inhibit both cyclooxygenase enzymes (COX-1 and COX-2) and are effective in treatment of pain/inflammatory disorders.  The aim of the present study is to compare the profile of anti-inflammatory/analgesic action of two propionic acid derivatives on inflammatory muscle hyperalgesia. carrageenan, a plant-based polysaccharide, was injected into the gastrocnemius muscle of rats. Three percent Carrageenan-induced hyperalgesia was used as an animal model of acute and chronic inflammatory hyperalgesia in rats. The behavioral data then can be compared to the histological data which provided us with a very good opportunity to correlate behavioral effects (hyperalgesia) with physiological effects (sensitization). Current study revealed that nonselective cyclooxygenase inhibitors moderately reversed the acute and chronic inflammatory hyperalgesia. The behavioral and histological differences emphasize the ability of analgesics on chronic hyperalgesia and sensitization.  The present studies suggest that selective cyclooxygenase-2 inhibition is essential to exert maximum antihyperalgesic effect in chronic inflammatory muscle hyperalgesia.

27. Journal of Cell and Tissue Research Vol. 7(2) 1161-1165 (2007)

Effects of in vitro supplementation of µ tocopherol on structural and functional integrity of human spermatozoa

Theng, A. D., Gajare, K. A. and Deshmukh, A. A.

Department of Zoology, Government Vidarbha Institute of Science and Humanities, Amravati 444 6064, India E. mail: ashish_cellbio@rediffmail.com

Abstract: Spermatozoa possess week antioxidant defense system and are rich in unsaturated fatty acids. Therefore, are highly vulnerable to oxidative damage. The process of fertilization carries several biochemical processes viz., capacitation, acrosome reaction, fusion of the sperm and oocyte plasma membrane which require healthy and intact membrane and functional mitochondria.  In the present investigation effects of in vitro supplementation of µ tocopherol on sperm plasma membrane integrity, acrosomal membrane integrity, mitochondrial activity index were studied along with motility and viability. Twenty-five human semen samples were treated with Ringer Tyrode solution containing 1mM µ tocopherol for one hour. The study showed a significant (p<0.001) increase in the percentage of motile and viable spermatozoa, spermatozoa having functionally active plasma membrane, acrosomal membrane and sperm mitochondrial activity index suggesting the protective role of µ tocopherol on human spermatozoa in vitro condition.

28. Journal of Cell and Tissue Research Vol. 7(2) 1167-1172 (2007)

Neuroprotective effect of Bacopa monniera linn. extract on lipofuscinogenesis and fluorescence product in brain of D-galactose induced ageing accelerated mice

Gajare, K. A., Deshmukh A. A.  and Pillai, M. M

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

Abstract: Bacopa monniera is a medicinal plant used in Ayurveda for improving memory and learning. It is also used as anxiolytic, cognition enhancer, antidepressant, anticancer, immuno-modulatory, and anti-inflammatory agent. It has been proven to posess antioxidant property. In the present investigation effect of ethanolic extract of B. monniera was studied on lipofuscinogenesis and fluorescence product in the brain of female albino mice. For this study ageing was accelerated by the treatment of 0.5ml 5% D-galactose for 15 days. This resulted in increased accumulation of lipofuscin granules and increased fluorescence product. After co-treatment of ethanolic extract of B. monniera (40mg/kg body wt.) there was decrease in accumulation of lipofuscin granules and significantly decreased fluorescence product in cerebral cortex, hippocampus, corpora quadrigemina, cerebellum as compared to ageing accelerated group.

29. Journal of Cell and Tissue Research Vol. 7(2) 1173-1179 (2007)

Protective effect of quercetin on sulphur mustard induced oxidative stress in mice

Kumar, O., Kulkarni, A. S.  Nashikkar, A. B. and Vijayaraghavan, R.

Division of Pharmacology and Toxicology, Defence Research and Development Establishment, Gwalior 474002, India. E-mail: omkumar63@rediffmail.com

Abstract:  Sulphur mustard (SM, 2, 2' dichloro diethyl sulphide) is a potent chemical warfare agent for which there is still no effective antidote. SM is known to cause oxidative stress. Therefore, in the present investigation, we studied the protective effect of quercetin, a bioflavonoid following SM toxicity. 2.0 and 4.0 LD 50 of SM (1 LD 50 = 9.67 mg/kg; 14 days observation for mortality) was administered to Swiss albino female mice through percutaneous route. SM exposed mice were treated with quercetin (100 and 200 mg/kg) three times by intraperitoneal injection, one immediately following SM exposure, then once each day for 2 days after SM treatment. The effect of quercetin on survival, body weight, markers of oxidative damage (in blood, liver and kidney), WBC counts and purine metabolite were investigated 7 day post exposure. Survival time increased significantly following quercetin treatment. The decrease in body weight due to SM was prevented to a significant extent by quercetin. Significant decrease in reduced glutathione and increase in the level of malondialdehyde indicated oxidative damage to hepatic and renal tissues. Quercetin protected hepatic and renal tissues from oxidative damage caused by SM.  Alterations in WBC counts and end product of purine metabolite were also prevented by quercetin. This study shows that quercetin enhanced the survival time, restored the decrease in body weight and protected hepatic and renal tissues from oxidative damage.

30. Journal of Cell and Tissue Research Vol. 7(2) 1181-1188 (2007)

Correlation of cellular distribution of mercury and subsequent impairment of spermatogenesis in rat

Vachhrajani, K.  D.
Division of Toxicology, Department of Zoology, The Maharaja Sayajirao University of Baroda,
Vadodara  390 002, India. E. mail: kdv_zdmsu@yahoo.com

Abstract: Male reproductive toxicity of mercury is now well established. However, occurrence of variations in the lesions at different seminiferous epithelial (SE) stages is still not very clearly understood. In present experiment cellular distribution of mercury is correlated with cellular lesions, biochemical status and spermatogenesis. Adult rats were administered 100 mg mercuric chloride per kg body weight, daily for 60 days. Observations were made on day 60 and day 120 including withdrawal period. Qualitative and quantitative spermatogenic analyses and quantification of mercury localization were made at stages II, VII and XII. Nuclear status in terms of degenerative grades was assessed at stages I-XIV. Histochemical demonstration of acid phosphatase activity was studied at stages I-XIV using fresh frozen serial sections for histochemical localization and nuclear staining by toludine blue for SE stage identification. DNA, RNA and protein were estimated in testis. Study shows decline in macromolecule levels and in the acid phosphatase activity suggested functional impairment of testis. Site-specific enzyme activity decline suggested stage-wise variations in the status of Sertoli cells. Mercury specifically localized in close vicinity of dividing primary spermatocytes (pachytene) and in the dislodged cytoplasm masses. Such masses were highly eosiniphilic as well as had diffused acid phosphatase activity. Such finding correlated pattern of mercury localization and spermatogenic impairment. The findings suggest that stage-specific distribution of mercury grain in Sertoli cells is very closely associated with the site and severity of injury, apoptosis of more mature spermatogenic cells and hence, consequent spermatogenesis.

31. Journal of Cell and Tissue Research Vol. 7(2) 1189-1193 (2007)
An analysis of pharmacokinetic models: A review
Puri, A., Medhi, B. and Kumar, M.

Postgraduate Institute of Medical Education and Research, Chandigarh 160012.
E-mail: avinash_puri@hotmail.com

Abstract: Pharmacokinetic models provide the means to store past experience with the behavior of drugs, and the tool to apply that past experience to the case of future patients. This experience is usually stored in the form of population pharmacokinetic model which is used to design the initial regimen for the next patient who appears to belong to that population. The dosage regimen to achieve the target goal is computed and given. The patient is then monitored both clinically and by measuring concentrations. We can see the computed concentration of drugs in a peripheral compartment or in various compartments. These cannot be seen without such models. It helps in surgical patients by comparing the clinical behavior of the patient with the behavior of the patients model. We can also evaluate the patients clinical sensitivity to the drug that can adjust the target goal appropriately. Thus models are helpful in surgical patients for better improvement of human welfare and mankind.

31. Journal of Cell and Tissue Research Vol. 7(2) 1189-1193 (2007)

An analysis of melatonin induced aggregatory responses of the isolated scale melanophores of an air breathing fish Channa gachua (ham.)

Sheikh, I. A. and Ovais, M.

Department of Biosciences, Barkatullah University, Bhopal. 462026, India.
E. Mail: sheikh_irfan99@yahoo.co.in.

Abstract: Effect of melatonin (MT) was examined on the isolated scale melanophores of a teleostean air breathing fish Channa gachua (Ham). Melanophore size index (MSI) was chosen as the parameter for recording of melanophore responses to MT. Increase or the decrease in the MSI value from the control represents dispersion or aggregation of the melanophores respectively. MT in a wide dose-range induced aggregation in the dermal melanophores of this fish. Denervation of melanophores caused a measurable change in the sensitivity of the melanophores to MT. In the denervated melanophores low doses of MT induced responses were potentiated while the higher doses of MT induced responses were inhibited. The results indicate the presence of two different mechanisms for the low and high dose-range of MT induced responses. Melatonin induced responses were effectively and mostly significantly inhibited by phentolamine, yohimbine, atropine and propranolol. The aggregatory responses of the melanophores were not blocked by verapamil and guanithidine. These results suggest that MT induced aggregatory responses in this fish melanophores are due to activation of specific MT receptors.