1. Journal of Cell and Tissue Research Vol. 11(2): 2731-2735 (2011)

Identification and differential distribution of the prostatic interstitial cells in the mouse prostate: Implications for prostate-specific diseases

Brody, K.M., Ziogas, J. and Kusljic, S.

Department of Nursing, University of Melbourne, Carlton, Victoria, Australia. E mail: skusljic@unimelb.edu.au

Abstract: Smooth muscle tone of the prostate is dependent on the pacemaker activity arising from prostatic interstitial cells (PICs) and sympathetic nerves releasing noradrenaline which acts upon α-adrenergic receptors. PICs have been identified in the guinea-pig, human and rat prostate gland, and have been suggested as a potential new therapeutic target. The activity of PICs appears to be dependent on the signal transduction through c-Kit which is now recognized as a PICs specific marker. The aim of the present study was to identify PICs in the mouse prostate and investigate their relationship with sympathetic nerves. Male C57BL/6J mice were sacrificed by cervical dislocation and the prostate was excised through an abdominal incision. The individual lobes were separated and processed for c-Kit and tyrosine hydroxylase immunohistochemistry. Confocal microscopy of the mouse prostatic lobes showed c Kit immunofluorescence indicating that these c-Kit positive cells had a stellate-shaped aspect in the dorsal and anterior lobes of the mouse prostate. By contrast, the ventral lobe did not contain PICs. Labelling of prostate tissue with tyrosine hydroxylase revealed that the tyrosine hydroxylase immunoreactive fibres coarsed through the smooth muscle layer of the prostate gland, but they don’t seem to be associated with the PICs. We postulate that the PICs communicate with noradrenergic nerves via intercellular communication proteins, specifically the connexin 43. Future experiments are needed to fully characterize the nature of this connection by identifying intercellular communication proteins involved in the PICs communication with each other, smooth muscle cells and nerves.

2. Journal of Cell and Tissue Research Vol. 11(2): 2737-2740 (2011)

Non alcohalic fatty liver disease (NAFLD) aggravates inflammation in metabolic syndrome

Haque, S.F.,  Ahmed, A., Zafar, K.S.  and Khan A.S.

Department of Medicine, Aligarh Muslim University, Aligarh 202 002, E.mail: sfhalig@yahoo.co.in

Abstract: Non-Alcoholic Fatty Liver Disease (NAFLD) is a common comorbid cardio vascular risk in patients with metabolic syndrome. Sub-clinical inflammation, endothelial dysfunction and lipid abnormalities being the potential underlying mechanism.The aim of present study was to evaluate correlation, between cardiovascular risk factors and high-sensitvity-CRP and TNF-α in patients with metabolic syndrome having NAFLD. The metabolic syndrome was diagnosed as per NCEP-ATP III-criteria. Inflammatory markers were estimated by ELISA. All statistical data were analyzed by using SPSS version 12.0. Scheffe’s post-hoc analysis with ANOVA was used for comparing data of different groups. Value of p<0.05 at 95% confidence interval was considered significant. Significant correlation of CRP and TNF-α was found with smoking, hypertension, diabetes, dyslipidemia, obesity and metabolic syndrome, with increasing grades of non-alcohalic fatty liver disease. Metabolic syndrome and its components have been consistently associated with the presence of low grade systemic inflammation (LGSI). This study validates the relationship between inflammatory markers and metabolic syndrome in patients with Non-alchlic fatty liver disease.

3. Journal of Cell and Tissue Research Vol. 11(2): 2741-2750 (2011)

In vitro differentiation of primordial germ cells from mouse embryonic stem cell in presence of different concentrations of bone morphogenetic protein 4

Ghasemi, H.H., Pasbakhsh, P., Amidi, F., Soleimani, M., Frozandeh, M., Sobhani, A. and Sobhani, A.

Department of Anatomy, Faculty of Medicine, Tehran University of Medical Sciences, Poursina Street, Tehran, Iran E. mail: sobhania@sina.tums.ac.i

Abstract: Purpose of this study was to determine optimal dose of BMP4 and its time period on differentiation Primordial Germ Cells (PGCs) from mouse Embryonic Stem Cells (mESCs) in vitro. To differentiate PGCs, Embryoid Bodies (EBs) from mouse ES cells were cultured in concentrations 0, 10, 50 and 100 ng/ml BMP4 at different time intervals. Viability of PGCs was assayed by MTT. Also, germ cell markers were analyzed by flow cytometry, immunocytochemistry and Real Time PCR. There were significant differences between BMP4 at 10 ng/ml and 100 ng/ml concentrations on the fourth day. Data of flow cytometry demonstrated that most of Mvh-positive cells were observed in D4B10. Result of Real time PCR illustrated that expression of gene Oct-4 in the control groups was high and it gradually decreased with adding concentrations 10 and 50 ng/ml of BMP4. Expression of Stella with BMP4 concentration of 100 ng/ml was considerably decreased. Expression of Mvh with BMP4 concentrations of 0 and 100 ng/ml was none. However, the maximum expression of Mvh was observed in D4B10 and this increase was not significant. Results indicate low concentration of BMP4 (10 ng/ml) at day 4th had the optimal effects on differentiation of PGCs from mESC.

 4. Journal of Cell and Tissue Research Vol. 11(2): 2751-2758 (2011)

Protective efficacy of andrographolide on 7,12-dimethylbenz(a)anthracene induced genotoxicity in bone marrow cells of golden Syrian hamsters

Manoharan, S., Singh, A.K., Suresh, K., Silvan, S., Baskaran, N., Manoj Prabhakar, M. and Karthikeyan, S.

Department of Biochemistry and Biotechnology, Annamalai University, Annamalainagar 608 002, Tamil Nadu.
E. mail: rasmanshisak@gmail.com

Abstract: Aim of the present study was to investigate the anticlastogenic and antigenotoxic effect of andrographolide in the bone marrow cells of golden Syrian hamsters treated with 7,12-dimethylbenz(a)anthracene (DMBA). The frequency of bone marrow micronucleated polychromatic erythrocytes (MnPCEs) was used as cytogenetic end point to assess the anticlastogenic effect of andrographolide in hamsters treated with DMBA. The antigenotoxic potential of andrographolide was assessed by analyzing the DNA damage in hamsters treated with DMBA using Single cell gel electrophoresis (comet assay). The status of phase I (cytochromes P450 and b5) and phase II [glutathione-S-transferase (GST), glutathione reductase (GR) and reduced glutathione (GSH)] detoxification agents, lipid peroxidation by-products (thiobarbituric acid reactive substance [TBARS]) and antioxidants [glutathione peroxidase (GPx), superoxide dismutase (SOD), catalase (CAT) and GSH] were used as supportive biochemical end points to assess the antigenotoxic and anticlastogenic effect of andrographolide in hamsters treated with DMBA. We noticed elevated MnPCEs frequency, increased DNA damage (bone marrow) and an increase in the status of liver phase I and phase II detoxification agents, and plasma lipid peroxidation by-products and antioxidants in hamsters treated with DMBA alone. Oral pretreatment of andrographolide for five days to hamsters treated with DMBA significantly protected DMBA induced clastogenesis and genotoxicity as well as above said biochemical abnormalities. The present study thus demonstrated the anticlastogenic and antigenotoxic potential of andrographolide in the bone marrow cells of hamsters treated with DMBA.

5. Journal of Cell and Tissue Research Vol. 11(2): 2759-2764 (2011)

Geraniol protects cell surface glycoconjugates abnormalities during 7, 12-dimethylbenz (a) anthracene induced oral carcinogenesis

Vinothkumar, V., Manoharan, S., Palanimuthu, D., Rajasekaran, D., Srinivasan, R. and Afaq Wani, S.

Department of Biochemistry and Biotechnology, Faculty of Science, Annamalai University,
Annamalainagar-608 002, Tamil Nadu. E. mail: rasmanshisak@gmail.com

Abstract: The study reveals the protective effect of geraniol on 7, 12-dimethylbenz(a)anthracene (DMBA) induced abnormalities in cell surface glycoconjugates (protein bound hexose, hexosamine, total sialic acid and fucose) in the plasma and buccal mucosa of golden Syrian hamsters. Oral squamous cell carcinoma was developed in the buccal pouch of male golden Syrian hamsters by painting with 0.5% DMBA in liquid paraffin three times a week for 14 weeks. Glycoconjugates status was assessed both biochemically and histologically in the buccal mucosa of DMBA painted hamsters. Plasma and buccal mucosa glycoconjugates were increased in hamsters painted with DMBA alone. Oral administration of geraniol at a dose of 250 mg/kg bw to DMBA painted hamsters for 14 weeks brought back the status of glycoconjugates to near normal range. Our results thus suggest that geraniol has the ability to protect cell surface glycoconjugates abnormalities during DMBA induced hamster buccal pouch carcinogenesis.

6. Journal of Cell and Tissue Research Vol. 11(2): 2765-2770 (2011)

Peroxisome proliferator–activated gamma (PPAR–γ) agonists improves metabolic parameters and vascular inflammation in patients with chronic kidney disease

Khan, R., Khan, A.S., Haque, S.F., Quaiser, S. and Kumar, A.

Department of Medicine, Jawaharlal Nehru Medical College, Aligarh Muslim University, Aligarh.
E. mail: sfhalig@yahoo.co.in Phones: 09412517518

Abstract: Peroxisome proliferator-activated gamma (PPAR-γ) receptors are expressed in many tissues of the body including the kidneys. PPAR-γ agonists like glitazones (e.g. Pioglitazone and Rosiglitazone) and angiotensin-1 receptor blockers (e.g. Telmisartan), exert several beneficial effects on kidney in different pathological conditions such as diabetic nephropathy, non diabetic renal diseases and uremia related cardiovascular risk factors in patients with chronic kidney disease (CKD). The study was an attempt to evaluate certain metabolic, vascular and inflammatory parameters in CKD and to compare the effects of different PPAR-γ agonists like pioglitazone and telmisartan on these parameters. In this prospective (case control), parallel group, observational study, 40 patients of CKD, irrespective of the etiology were recruited and followed up for three months. The data was analysed and the results were given in percentage in accordance with previous related studies. The majority of metabolic, vascular and inflammatory markers taken in our study (Blood urea, nitrogen, serum creatinine, serum triglycerides, serum HDL, urinary albumin, carotid intimal media thickness, CRP and TNF-α) depicted statistically significant difference in pre- and post-treatment values with treatment with either telmisartan or pioglitazone or with both telmisartan and pioglitazone over control group. The study showed that both telmisartan (partial PPAR-γ agonist) and pioglitazone (full PPAR-γ agonist) exert significant beneficial effects in CKD patients, through alteration of various inflammatory, metabolic and vascular markers.

7. Journal of Cell and Tissue Research Vol. 11(2): 2771-2774 (2011)

Reduced media salt concentration improves in vitro rooting in Indian Pennywort, Centella asiatica (l) Urban

Hanumantharaya, B.G., Sathyanarayana, B.N. and Waman A.A.

Plant Tissue Culture Laboratory, Department of Horticulture, University of Agricultural Sciences,  GKVK, Bengaluru 560065. E mail: ajit.hort595@yahoo.co.in

Abstract: Micropropagation involves a high establishment and operational cost. In plant species where the market price is comparatively low, it becomes difficult to supply the plants at cheaper rate. Centella asiatica is one of such important threatened medicinal plant species which has good demand but fetches low price in market. Thus, efforts were initiated to test the performance of cultures at reduced media salt concentrations and using market grade sugar as carbon source. Results revealed that, reduced media salts improved the rooting process apart from reducing cost of production. Quarter strength Murashige and Skoog’s media was found to be superior over half strength media for all the rooting parameters such as, number of days for rooting, percent rooting, number and length of primary and secondary roots. Quality of roots produced using quarter strength media was also observed to be better than half strength media.

8. Journal of Cell and Tissue Research Vol. 11(2): 2775-2780 (2011)

Purification and characterization of α-amylase produced by Aspergillus niger using banana peels

Kiran Kumar, V., Ravi Sankar, N.,  Shailaja, R., Saritha, K., Siddhartha, E., Ramya, S., Giridhar, D. and Sahaja, R.V.

Microbiology laboratory, Global Institute of Biotechnology, Hyderabad-500029. E. mail: nrsr2008@yahoo.com

Abstract: The present study was aimed at purification and characterization of α-amylase enzyme from strain of Aspergillus niger isolated from local soil samples. Solid state fermentation (SSF) was carried out to produce α-amylase from A. niger using waste banana peel as substrate. The maximum activity of α-amylase (11926 U/gds) was recorded after 72 hours of fermentation. The extracted enzyme was subjected to purification by ammonium sulphate precipitation, sephadex G-100 gel and through ion exchange chromatography. Through the process 15.3-fold increase in purity with a specific activity of 158.7 U/mg proteins was obtained. The molecular weight of the enzyme determined by SDS-PAGE was found to be 61 kDa. The enzyme was optimally active at pH 5 and 50°C, starch as substrate. This enzyme was almost 60% stable at 600C even after 50 minutes of incubation. It was strongly activated by metal ions such as Mn2+ and Fe2+.

9. Journal of Cell and Tissue Research Vol. 11(2): 2781-2784 (2011)

Anti-inflammatory activity of Ceiba pentandra l. seed extracts

Alagawadi, K.R., and Shah, A.S.

Department of Pharmaceutical Chemistry, KLEU’s College of Pharmacy, Belgaum, Karnataka.
E. mail: amol2473@indiatimes.com.

Abstract: Present study was carried out to evaluate the anti-inflammatory effects of petroleum ether and ethanolic extract of the seeds of Ceiba pentandra (L). Gaertn. (Bombacaceae) commonly known as silk cotton tree. The plant has been used as antibacterial, anti-inflammatory, antipyretic,for rheumatism, diabetes and in headache in Indian system of traditional medicine. Anti-inflammatory activity was assessed by induction of carrageenan in left hind paw. CPE and CPO at both administered doses of 200 mg/kg and 400 mg/kg reduced paw edema volume significantly. These results clearly show anti-inflammatory effect of seed extracts. Further studies are needed to isolate phytoconstituent(s) responsible for anti-inflammatory effect.

10. Journal of Cell and Tissue Research Vol. 11(2): 2785-2788 (2011)

Protective effect of Bauhinia variegata extracts on triton induced hyperlipidemia in rats

Biradar, S.S. and Patil, P.A.

Department of Pharmacology, K. L. E. University’s College of Pharmacy, Belgaum 590 010, Karnataka.
E. mail: ssbiradar81@yahoo.co.in

Abstract: In hypercholesterolemia, the cholesterol content of erythrocytes, platelets polymorphonuclear leucocytes and endothelial cells increases and this leads to activation and enhanced production of oxygen free radicals. In present study, the extracts of barks of Bauhinia variegata were tested for their hypolipidemic effect on Triton induced hyperlipidemia in rats. It showed significant reduction in total cholesterol and LDL (p < 0.01). The HDL level increased but was not significant. However, after 24 hours of Triton injection, the BV aqueous extract 500 mg/kg and petroleum ether extract 500 mg/kg showed significant rise in HDL (p < 0.05 and p < 0.01 respectively). This study demonstrated that extract of BV were effective against Triton induced hyperlipidemia in rats.

11. Journal of Cell and Tissue Research Vol. 11(2): 2789-2794 (2011)

Antioxidant potential of seasonal fruits in fluoride toxicity: An in vitro study

Rupal, A.V. and Narasimhacharya, A.V.R.L

Laboratory for Animal Sciences, Department of Biosciences, Sardar Patel University, Vallabh
Vidyanagar -388 120, Gujarat. E. mail : narasimhacharya@yahoo.com

Abstract: The present study was undertaken to investigate the antioxidant potential of ethanolic extracts of four fruits - Indian gooseberry, mango, star fruit and wood apple on the oxidative changes in hepatic tissue exposed to sodium fluoride in vitro. Liver homogenates were exposed to NaF and subsequently treated with ethanolic extracts of fruits and the levels of antioxidants (ascorbic acid, superoxide dismutase, catalase, glutathione, glutathione peroxidase) and lipid peroxidation (TBARS) were determined. Results indicated that of the four fruits tested, gooseberry exhibited a maximum potency in reducing the lipid peroxidation (56.70%) and increased the antioxidants - TAA, SOD, CAT, GsH and GPX where as wood apple was found to possess a comparatively low antioxidant potential.

12. Journal of Cell and Tissue Research Vol. 11(2): 2795-2801 (2011)
Caudal fin regeneration is regulated by COX-2 induced PGE2 in teleost fish, Poecilia latipinna

Anusree, P., Saradamba, A., Tailor, N., Desai, I. and Suresh, B.

Department of Zoology, Faculty of Science, The Maharaja Sayajirao University of Baroda,
Vadodara- 390 002; E-mail: suved9@hotmail.com,

Abstract: Regeneration is a fundamental attribute of all living things, whether it is simple tissue restoration or the complete replacement of lost body parts, such as limb, tail or even head. Prostaglandin E2 (PGE2) is known to play a cardinal role in wound healing, cell proliferation, cell migration and differentiation. However, insufficient reports are there suggesting the involvement of PGE2 in regeneration. In the present work, role of PGE2 in fin regeneration of teleost fish, Poecilia latipinna, was worked out by inhibiting cyclooxygenase-2 (COX-2), an upstream modulator of PGE2, using a selective COX-2 inhibitor, etoricoxib. The study revealed deranged regenerative process in the PGE2 suppressed animals. Further, an apparent reduction in the nucleic acids and protein content in the regenerates was observed which in turn might have hindered the early regenerative process. PAGE profile revealed alteration in proteins having molecular weights ranging from 42-44KDa. Thus evidently, PGE2 does influence the process of epimorphic regeneration in teleost fish. However, the exact pathway by which PGE2 exerts its mitogenic influence needs to be further evaluated and finer mechanisms, once worked out, might give insight into how an evolutionary conserved regulator of epimorphic regeneration is parted way for the advanced wound healing mechanism developed in recently evolved organisms.

13. Journal of Cell and Tissue Research Vol. 11(2): 2803-2808 (2011)

Adventitious plant regeneration and transient GUS-gene expression in Jjatropha curcas (L.)

Saharan, V.,  Patel, B., Shah, M.A., Ranwah, B.R., Chabra, K.,  Shrimali, D. and Shukla, K.B.

Department of Molecular Biology and Biotechnology, Rajasthan College of Agriculture, Maharana Pratap University of Agriculture and Technology, Udaipur 300 001, Rajasthan. E. mail: vinodsaharan@gmail.com

Abstract: Efficient plant regeneration and transient gus (b-glucuronidase) gene expression was achieved in Jatropha curcas (L) using leaf explants derived from 3.5 years old Jatropha plants. MS basal salts fortified with 1.0 mg/l BAP + 0.5 mg/l Kin promoted 54.4 % adventitious shoot induction. Multiplication and elongation of microshoots were achieved on MS basal salts + 0.5 mg/l BAP + 0.5 mg/l Kin + 0.25 mg/l IAA and 0.25 mg/l GA3. The leaf explants infected with Agrobacterium strain EHA 105, harbouring pBI121 vector was used for standerization of transformation protocol.

14. Journal of Cell and Tissue Research Vol. 11(2): 2809-2814 (2011)

An alternative pathway for plant regeneration via somatic embryogenesis of upland rice (Oryza sativa cvs. Khandagiri and Vanaprava)

Rabha, M. and Rout, G.R.

Department of Agricultural Biotechnology, College of Agriculture, Orissa University of Agriculture
and Technology, Bhubaneswar 751 015. E. mail: grrout@rediffmail.com

Abstract: Rice is an important food crop for human consumption and its production is about 96 million tones every year in India. Now-a-days, the production of rice is hampered due to the climatic changes. Therefore, it is essential to develop variety which is tolerant to drought, salinity and cold. The present investigation was to standardize the regeneration protocol for genetic transformation study. Somatic embryogenesis were achieved from immature zygotic embryos of two emerging Indica rice varieties i.e. “Khandagiri” and “Vanaprava” using MS (Murashige and Skoog) basal medium supplemented with 2 - 3 mg/l 2,4-D (2,4-dichlorophenoxyacetic acid),  0.5 – 1.0 mg/l kinetin and 3% (w/v) sucrose within 6 weeks of culture. The secondary somatic embryogenesis was also developed in subsequent subculture on similar medium. The frequency of embryogenesis was more in subsequent subculture on medium having kinetin and 2,4-D. The inclusion of either L-proline or L- glutamine (500 mg/l) favored the more frequency of  embryogenic calli in both the varieties. The medium having BA (6-benzylaminopurine) did not favour the induction of somatic embryogenesis. The somatic embryos were germinated on half strength basal MS medium supplemented with 2 mg/l kinetin, 0.1 mg/l NAA (1~napthaleneacetic acid) and 2 % (w/v) sucrose. About 90% of somatic embryos were germinated into complete plantlets. The somatic embryo derived plantlets were survived in the greenhouse and grown normally. Further, this protocol will be utilized for the development of stress tolerant rice by using two genes i.e. Ann Bj-2 and Ann Bj-3.

15. Journal of Cell and Tissue Research Vol. 11(2): 2815-2820 (2011)

Alteration in testicular protein profile following administration of embelin and vincristine: Potent reproductive toxicants

 Sonawane, T., Chaudhary, N., Udani, J., Thayyil, L. and Bhiwgade, D.A.

Department of Biotechnology and Bioinformatics, Padmashree Dr. D.Y. Patil University, CBD Belapur,
Navi Mumbai 400 614. tareekasonawane@gmail.com

Abstract: Reproductive organs have been the target for majority of the toxicants including several chemotherapeutic drugs. Embelin, a potential lead chemotherapeutic molecule and vincristine widely used chemotherapeutic agent has shown to cause reproductive toxicity. The present study was aimed to examine the changes in the testicular proteins on embelin and vincristine treatment. The proteins from various cell types of the testis i.e., the germ cells, the Sertoli cells and Leydig cells were found to be affected. There were changes in the pattern of protein bands on 12% SDS PAGE from testicular extract of embelin and vincristine treated rats and in the purified fractions obtained using ion exchange chromatography. This suggests that the compromised male fertility observed in treatment by these drugs is probably because of the alteration in the key proteins involved in the process of spermatogenesis and steroidogenesis.

16. Journal of Cell and Tissue Research Vol. 11(2): 2821-2826 (2011)

Similarties in toxicity of pathogenic fungus (Phoma medicaginis) and chromium (VI)

Gandhi, A.P. and Kale, P.G.

 Depatment of Zoology, Ramniranjan Jhunjhunwala College, Ghatkopar, Mumbai  400 086.
E. mail: amrutagandhi@hotmail.com

Abstract: Phoma medicaginis is a deuteromycete fungus that grows on a variety of plants, a few of which are used in herbal medicines. The current work attempts to establish the possible mechanism of toxicity of this fungus, in case it is consumed by animals as fungus infested food. Chromium (VI) is a known pollutant, the toxicity of which is manifested through the oxidative stress exerted on the animals that happens to consume food contaminated with it. Chromium (VI) is capable of generating the reactive oxygen species (ROS), which in turn, may react with the biomolecules in the tissues leading to degenerative changes; TBARS (thiobarbituric acid reactive substance) and protein carbonyl are routinely used indices of oxidative stress. When Swiss albino rats were fed on food contaminated with chromium (VI) and biomass of Phoma medicaginis it was observed that the TBARS content, which in turn is proportionate to lipid peroxidation of liver, kidney and brain tissues was found to rise above that of control rats. The protein carbonyl content that serves as a measure of degradative changes in proteins by reactive oxygen species was also found to be high in these tissues of rats fed on diet contaminated with chromium (VI) as well as fungal biomass. The weight loss observed in the treated rats could well be ascribed to the decrease in protein as well as carbohydrate contents of the tissues analyzed. A corresponding increase in free amino acid and reducing sugar contents of the respective tissues of treated rats were noticed. Ascorbic acid and α – tocopherol are employed in managing the oxidants and so is the cholesterol that may be utilized for synthesizing regulatory hormones to achieve the same result. The fact that ascorbic acid and α – tocopherol content were low and the total cholesterol content was high in all tissues of rats receiving diet contaminated with chromium (VI) and fungal biomass confirmed that the toxicity of the fungus is manifested through the generation of oxidative stress identical to the one by chromium (VI).

17. Journal of Cell and Tissue Research Vol. 11(2): 2827-2832 (2011)

Statistical media optimization for b--fructofuranosidase production by Saccharomyces cerevisiaeNncim 3287

Vernekar, M. and Supali, P.

Department of Biotechnology and Bioinformatics, Padmashree Dr. D. Y. Patil University, Plot No 50, Sector 15, CBD Belapur Navi Mumbai, 400614. E. mail: madhavi.revankar@gmail.com

Abstract: Saccharomyces cerevisiae NCIM 3287 has been identified as an excellent producer of invertase (b-D- fructofuranoside fructohydrolase, EC 3.2.1.26) which is an important enzyme in confectionary industry. The main aim of the present study was to develop an economically feasible medium for invertase production with appropriately supplemented nutrients. The basal medium gave invertase activity of 250U/L. Screening of different medium components and their effects on invertase production using S. cerevisiae was studied using orthogonal array method. A simple sucrose based medium gave very high enzyme activity of 300U/L. The activity is very high hence this strain was explored for enhancement of invertase of production using different inducers. Addition of 8g/L sucrose enhanced invertase activity to 844 U/L. Invertase activity was further enhanced to 1072 U/L using 0.02 % concentration of K2HPO4 as inducer. Thus, this strain could be an efficient and attractive source for invertase production.

18. Journal of Cell and Tissue Research Vol. 11(2): 2833-2837 (2011)

Acute toxicity of camptothecin and influence of α-tocopherol on hematological and biochemical parameters

Singh, K.C. and Marar, T.

Department of Biotechnology and Bioinformatics, Padmashree Dr. D. Y. Patil University,
CBD Belapur, Navi Mumbai-400 614. E. mail: skanchan19@gmail.com

Abstract: Camptothecin (CPT), a natural alkaloid from Camptothecin acuminate is a potent drug with a broad spectrum antitumor activity. Its use has been limited due to wide ranging toxicities and oxidative stress that havebeen implicated as side effects. The present study is an attempt of modulating CPT induced toxicities by using vitamin E as a nutritional supplement. Hematological parameters such as RBC, WBC and platelet counts that decreased after camptothecin injection were found to be restored in rats which were pretreated with α-tocopherol. Enhanced levels of lipid peroxides in animals administered with CPT showed significant revision after vitamin E administration. Improvement in the alterations of other chemical constituents in blood like glucose, urea, uric acid, hemoglobin, cholesterol and triglycerides were also observed in animals pretreated with vitamin E suggesting a prophylactic role of vitamin E in CPT mediated side effects.

19. Journal of Cell and Tissue Research Vol. 11(2): 2839-2846 (2011)

Antigenotoxic effect of 18ß-glycyrrhetinic acid in 7,12-dimethylbenz(a)anthracene induced genotoxicity in Golden Syrian hamsters

Kowsalya, R., Manoharan, S.,  Silvan, S., Baskaran, N. and Palanimuthu, D.

Department of Biochemistry and Biotechnology, Faculty of Science, Annamalai University, Annamalai nagar
608 002, Tamil Nadu. E. mail: sakshiman@rediffmail.com

Abstract: The aim of this study was to assess the protective effect of glycyrrhetinic acid on 7,12-dimethylbenz(a)anthracene (DMBA) induced micronucleated polychromatic erythrocytes (MnPCEs) frequency, chromosomal aberrations and DNA damage in the bone marrow cells of golden Syrian hamsters. Lipid peroxidation by-products, antioxidants and phase I and phase II detoxification agents were also used as biochemical end points to assess the protective efficacy of glycyrrhetinic acid in DMBA induced genotoxicity. Elevated MnPCEs frequency, marked chromosomal abnormalities, increased DNA damage and an increase in the status of liver phase I and phase II detoxification enzymes, plasma lipid peroxidation by-products and antioxidants were noticed in hamsters treated with DMBA alone. Oral pretreatment of glycyrrhetinic acid at a dose of 45 mg/kg b.w for 5 days to hamsters treated with DMBA significantly decreased the frequency of MnPCEs, chromosomal aberrations and DNA damage as well as restored the status of phase I and phase II detoxification enzymes, lipid peroxidation by-products and antioxidants. The present study thus demonstrated the protective effect of glycyrrhetinic acid on DMBA induced cytogenetic (bone marrow cells) and biochemical (liver, plasma) abnormalities in golden Syrian hamsters.

20. Journal of Cell and Tissue Research Vol. 11(2): 2847-2851 (2011)

Fluctuation of soil bacterial dehydrogenase activity in response to the application of endosulfan and chlorpyrifos

Sumit Kumar

Biotechnology Department, V. V. P. Engineering College, Saurashtra University, Rajkot 360 005.
E. mail: btsumit@gmail.com

Abstract: The effects of two different categories of pesticides viz. chlorpyrifos (an organophosphate) and endosulfan (an organochlorine cyclodiene) were evaluated on soil dehydrogenase activity. The medium and higher dose of endosulfan resulted in the significant decrease in dehydrogenase activity till third and fourth week of treatment, respectively. In case of chlorpyrifos, decrease in dehydrogenase activity was observed till second week at medium dose and till fourth week at higher dose, compared to untreated control. In comparison to control, the activity of dehydrogenase decreased by 16% and increased by 25% in presence of 10 ppm each of endosulfan and chlorpyrifos, respectively, after 14 days. The combined effect of chlorpyrifos and endosulfan showed inhibitory effect at both medium and higher doses, for the treatment duration of four weeks. The maximum inhibition (62%) in soil dehydrogenase activity was noticed in presence of 50 ppm each of chlorpyrifos and endosulfan after first day of treatment.

21. Journal of Cell and Tissue Research Vol. 11(2): 2853-2855 (2011)

Correlation of seminal steroid levels with impaired sperm acrosomal enzyme activity in semen of unexplained infertility cases

Dave, V.S. and Highland, H.N.

Reproductive Biology Laboratory, Department of Zoology, Gujarat University, Ahmedabad 380 009.

E. mail: davevijyalaxmi@yahoo.co.in

Abstract: Alteration in seminal steroid levels is known to induce changes in functional status of sperm, a major cause of unexplained infertility world wide. The present study was carried out on a specific group of fifty males with mean age range 36.5 years, referred with unexplained infertility and poor sperm in vitro fertilizing ability of Ahmedabad region. The study revealed that there was a significant decrease in sperm count and viability in this group as compared to control. The hypo-osmotic swelling test suggested altered membrane permeability. Spermatozoa from the infertile samples showed fewer halos in the gelatin slide test. The activities of acrosomal enzymes indicates prematurely reacted spermatozoa that led to loss of membrane integrity and breakdown of sperm acrosome, resulting in impaired sperm function and fertilizing ability. The results of altered acrosomal activity confirmed low androgen levels further indicating decreased levels of testosterone and elevated levels of estradiol resulting in an imbalance of T/E2 ratio. These findings suggest that the functional status of sperm depends upon testosterone levels.

22. Journal of Cell and Tissue Research Vol. 11(2): 2857-2863 (2011)

Histoarchitectural changes in the adrenal gland of the female bat Taphozous kachhensis (Dobson) during estrus and pregnancy

Chavhan, P.R., Dhamani, A.A. and Misar, S.D.

Department of Zoology, N. H. College, Bramhapuri 441206. E. mail: panksphd@gmail.com,

Abstract: The present study shows the cyclical alternation in the activity of adrenal gland of the bat Taphozous kachhensis during different phases of the reproductive cycle. The adrenal gland is made up of steroidogenic and chromaffin cells. The gland during pregnancy is much larger than that of other phases of reproductive cycle. The cortex is completely encircles the medulla in Taphozous kachhensis. The medulla in this species consists of epitheloid cells arranged in smaller groups surrounded by blood capillaries. The zona glomerulosa consist of numerous bulbous structures which appear as acinus like group of cells. The zona fasciculata occurs in the form of irregular short cords; one or two cells thick arranged radially and merge with each other. These cords consist of large polyhedral vacuolated cells. The zona reticularis is also increase in size and measures. The cells are hypertrophied. Cytoplasm is eosinophilic with darkly stained nucleus and more vacuolated.

23. Journal of Cell and Tissue Research Vol. 11(2): 2865-2871 (2011)

An immunohistochemical study of the interstitial cells of Cajal (ICCS) in the dog intestinal tract

Choudhary, R.K., Das, P. and Ghosh. R.K.

Department of Veterinary Anatomy and Histology, Faculty of Veterinary and Animal Sciences.
West Bengal University of Animal and Fishery Sciences , 37, K. B. Sarani, Kolkata-700037
E. Mail: dr.ratneshchoudhary@gmail.com

Abstract: In the present investigation the distribution pattern of interstitial cells of Cajal (ICCS) population throughout the canine intestinal tract was explored. ICCS were identified by Kit receptor immunohistochemistry. Kit positive ICCS were present all along the segment of intestine but there was a marked difference in their presence, distribution and morphology. The number of ICCS were counted in different parts of intestine and recorded as 102.00 ± 7.74, 93.20 ± 4.17, 104.00 ± 6.60, 74.80 ± 10.02, 134.00 ± 12.42, 96.40 ± 11.97 and 112.00 ± 10.43/mm² in case of duodenum, jejunum, ileum, ileocolic junction, cecocolic junction, caecum and colon respectively. The average number of Kit positive ICCS were significantly (P‹0.05) higher in cecocolic junction and less in the ileocolic junction. According to their occurrence within the intestine ICCS were sub typed into three categories, like Submucosal plexus or Meissner’s plexus ICCS (SMP-ICCS), Intramuscular ICCS (IM-ICCS), Auerbach’s plexus or Myenteric plexus ICCS (AP-ICCS). All the subtypes were not present all along the intestinal tract rather they were region specific. SMP-ICCS were not detected in small intestine. However they were present in the large intestine, ileocolic and cecocolic junctions. At the level of AP, C-Kit positive cells were stellate shaped with long branching process, which formed a network and sometimes extended into circular longitudinal muscle layers. Intramuscular C-Kit positive cells were spindle shaped and ran parallel to the muscle fibre. At the level of SMP C-Kit positive cells were round in shape and formed a network of cells. The IM-ICCS was found in cecum and colon along the outer aspects of the circular muscle bundles.

24. Journal of Cell and Tissue Research Vol. 11(2): 2873-2878 (2011)

Serum cytokine levels of IL-12, IL-4 and TNF-Alpha in malaria: a study from the coastal districts of Odisha

Sarangi, A.,   Mohapatra, P.C. and Dalai, R.K.

Department of Biochemistry, S.C.B. Medical College, Cuttack 753 007, Odisha.
E. mail: anshumanbbsr@rediffmail.com.

Abstract: This study was conducted to assess the levels of IL-12, TNF-alpha and IL-4 in patients affected by Plasmodium falciparum and Plasmodium vivax along with sex and age matched healthy controls. The study included 98 patients of Plasmodium falciparum malaria, 54 patients of Plasmodium vivax malaria and 30 age and sex matched healthy controls. Blood samples were drawn from malaria patients admitted to medicine ward along with healthy controls and transferred to the Biochemistry department for the routine hematological parameters and ELISA for TNF-alpha, IL-12 and IL-4. The levels of TNF-alpha and IL-12 were significantly elevated in falciparum malaria and vivax malaria compared to healthy controls The levels of IL-4 were lower in falciparum malaria and vivax malaria compared to healthy controls. TNF-alpha /IL-12 ratio in falciparum and vivax malaria were significantly elevated compared to healthy controls. IL-4/ IL-12 ratio in falciparum and vivax malaria were significantly reduced as compared to healthy controls. The results indicate that in both falciparum and vivax malaria are characterized by up regulation of TNF-alpha and IL-12 activity contrasting markedly by down regulation of IL-4 in the same patients.

25. Journal of Cell and Tissue Research Vol. 11(2): 2879-2884 (2011)

Evaluation of anticonvulsant activity of Annona squamosa Linn. leaves in mice

Porwal, M. and Sharma, K.

Department of Pharmacology, B. N. Post Graduate College of Pharmacy, Udaipur 313001. 1Vivek College of Technical Education, Bijnor 245 701 (U.P.). E. mail: komalsharmagautam@yahoo.com

Avstract: The aim of the present study was to evaluate the effects of Annona squamosa (AS) leaves on seizures induced by pentylenetetrazol (PTZ) and picrotoxin (PTX). Pretreatment with hydroalcoholic extract of Annona squamosa leaves (125-500mg/kg) induced a dose-dependent decrease in the incidence of both clonic and seizures generalized tonic–clonic seizures (Pd”0.05) following PTZ and PTX administration. Co-administration of a sub-effective dose of AS (125 mg/kg, po) with a sub-protective dose of diazepam (0.5 mg/kg, ip) increased the latency to seizure. The combination significantly enhanced percent protection against PTZ and PTX induced convulsions. The results suggested that the anticonvulsant effect of Annona squamosa leaves against PTZ and PTX induced convulsions may be mediated, at least partly, through GABAA-benzodiazepine receptor complex.