1. Journal of Cell and Tissue Research Vol. 10(2): 2185-2190(2010)

Berberine protects cellular integrity during 7, 12- dimethylbenz[a] anthracene-induced oral carcinogenesis in golden syrian hamsters

Sindhu, G. and Manoharan, S.

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

Abstract: The protective effect of berberine on cellular integrity was assessed by measuring the status of glycoconjugates (protein bound hexose, hexosamine, total sialic acid and fucose) and lipids (cholesterol and phospholipids) in circulation and buccal mucosa of golden Syrian hamsters in 7,12-dimethylbenz[a]anthracene (DMBA) induced hamster buccal pouch carcinogenesis. Oral squamous cell carcinoma was developed in the buccal pouch of male Syrian golden hamsters by painting with 0.5% DMBA in liquid paraffin three times a week for 14 weeks. Glycoconjugates and lipids status in circulation and buccal mucosa were determined by specific colorimetric methods. Increase in plasma and tumor tissue glycoconjugates were noticed in tumor bearing animals. The molar ratio of cholesterol-phospholipids was increased in plasma and buccal mucosa of tumor bearing animals. Oral administration of berberine (75mg/kg b.w., po.) to DMBA painted animals on days alternate to DMBA painting for 14 weeks brought back the status of glycoconjugates and lipids to near normal range. The present study suggests that berberine has significant protective effect on cellular integrity during DMBA-induced hamster buccal pouch carcinogenesis.

2. Journal of Cell and Tissue Research Vol. 10(2): 2191-2199(2010)

Biochemical investigations on the cancer therapy potential of antioxidant vitamins A,C,E and their analogues on p-dimethylaminoazobenzene-induced hepatoma in rats

Velanganni, A.A.J. and Balasundaram, C.

Department of Biochemistry, J. J. College of Arts and Science, Pudukkottai-622 422, Tamilnadu,
E. mail: ajvelanganni@gmail.com

Abstract: The effect of antioxidant vitamins A, C, E and their analogues on chemically-induced hepatoma by p-dimethylaminoazobenzene (DAB) in male albino rats has been studied. In this experiment 275 rats divided into 3 sets were used. First set served as control; second set received 0.06% DAB and third set received 0.06 % DAB along with analogues of vitamins A, C and E. Rats fed with 0.06% DAB were provided water ad libitum for a period of 4 months, followed by a normal (basal) diet for further 2 months. Vitamins A (10,000-50,000 IU), C (75-1000 mg) and E (50-500 mg) and their analogues were given (per kg body wt) to the third set of rats by gavages route once in a week for a period of 6 months. Supplementation of antioxidant vitamins has a protective effect on the incidence of liver tumour monitored on the basis on liver weight, and biochemical analysis. Individual supplementation of high doses of vitamin A, L-ascorbic acid and vitamin E succinate significantly reduced the content of DNA, RNA, AFP and ã-GT in DAB-induced hepatoma of rats. However, administration of mixture of these vitamins at low doses enables restoration of ã-GT, a marker of hepatoma and the chosen biochemical levels to normal which may due to their synergistic effect.

3. Journal of Cell and Tissue Research Vol. 10(2): 2201-2205(2010)

Association of CYP2D6*4 polymorphism with acute leukemia

Nageswara Rao, D., Manjula, G., Sailaja, K., Surekha, D., Raghunadharao, D., Senthil, R. and Vishnupriya, S.

Department of Genetics, Osmania University, Hyderabad 500007, E. mail: sattivishnupriya@gmail.com

Abstract: Human Cytochrome P450 2D6 (CYP2D6) (Debrisoquine hydroxylase) plays a pivotal role in drug metabolism, metabolizing over 25% of the most commonly prescribed drugs. The CYP2D6 gene is highly polymorphic, leading to wide-inter individual ethnic differences in CYP2D6-mediated drug metabolism. The aim of the study was to find the association between CYP2D6*4 polymorphism G1934A and acute leukemia. The present study includes 305 acute leukemia cases comprising of 158 acute lymphocytic leukemia (ALL), 147 acute myeloid leukemia (AML) and 252 control samples for analysis of CYP2D6*4 polymorphism using PCR-RFLP method. The CYP2D6*4 polymorphism G1934A was significantly associated with acute leukemia development. With respect to clinical variables, the Mean WBC, Blast %, LDH levels were increased in both ALL and AML patients having poor metabolizer genotype (PM). 52.6% (9 out of 10) of the poor metabolizers failed to achieve complete remission (CR) in AML group. However, this trend could not be observed in ALL patients as CR data was available only for few ALL patients with PM genotype. In conclusion, CYP2D6*4 PM genotype might confer risk to develop ALL or AML. The PM genotype was also associated with clinical variables such as elevated WBC, blast %, LDH and failure to CR thus contributing to poorer survival.

4. Journal of Cell and Tissue Research Vol. 10(2): 2207-2211(2010)

Sialoadenectomy effect on skeletal muscles of female mice (Mus musculus)

Walvekar, M.V., Bhopale, L.P. and Sarvalkar, P.P.

Department of Zoology, Shivaji University, Kolhapur 416004, Maharashtra,
E-mail: madhuri_walvekar@rediffmail.com

Abstract: Body organs are known to be influenced by growth factors secreted by salivary gland. Objective of present study was to assess whether the polypeptides secreted by submandibular gland affect the physiology of skeletal muscles or not. For this purpose female mice at the age of 20 days, when submandibular gland started the secretion of polypeptides, were sialoadenectomised (removal of submandibular glands). The operated females were maintained under normal conditions in the departmental animal house up to the age of two months. Thereafter, protein content and LDH activity was determined from gastrocnemius, soleus and rectus abdominis muscles of both sialoadenectomised and control animals. Study shows significant increase of proteins content and LDH level in all the three muscles of sialoadenectomised mice as compared to the control. The electrophoretic separation of LDH also demonstrated the increase in the intensity of LDH bands in experimental animals.

5. Journal of Cell and Tissue Research Vol. 10(2): 2213-2222(2010)

Electron microscopic radioautographic study on mitochondrial RNA synhesis in adrenal medullary cells of aging and senescent mice

Nagata , T.

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

Abstract: In order to study the aging changes of intramitochondrial RNA synthesis of mouse adrenal cells, 10 groups of developing and aging ddY mice, each consisting of 3 individuals, total 30, from fetal day 19 to postnatal newborn at day 1, 3, 9, 14, adult at month 1, 2, 6, 12 and 24 were injected with 3H-uridine, sacrificed 1 hr later and the adrenal tissues were fixed and processed for electron microscopic radioautography. On electron microscopic radioautograms obtained from each animal, number of mitochondria and the mitochondrial labeling index labeled with 3H-uridine showing RNA synthesis in each adrenalin cell and noradrenalin cell in the adreno-medullary tissues, were counted and the results in respective aging groups were compared respectively. From the results, it was demonstrated that the numbers of mitochondria in both adrenalin and noradrenalin cells in the adrenal medullae of mice at various ages increased from postnatal day 1 to month 1, reaching the maximum and keeping the plateau to month 2, 6, 12 and 24, respectively. The number of labeled mitochondria and the labeling indices of intramitochondrial RNA syntheses incorporating 3H-uridine also increased from postnatal day 1 to month 1, 2, 6, 12 and 24 due to aging. It was shown that the activity of intramitochnodrial RNA synthesis in the adreno-medullary cells in developing and aging mice increased due to aging of animals.

6. Journal of Cell and Tissue Research Vol. 10(2): 2223-2227(2010)

Radiologically guided fine needle aspiration cytology (FNAC) of body organs

Dhruva Gauravi, A., Agravat Amit, H. and Parmar Satyam, A.

Department of Pathology, P.D.U. Medical, College, Rajkot 360 005. E. mail: gauravidhruva@yahoo.co.in

Abstract: A study on “ Radiologically Guided FNAC of Body Organs” consisting of 150 cases was carried out at Pathology Department, P.D.U. Medical College, Rajkot, between June 2007 to August 2009. The study was carried out in relation to various body organs, mentioning age incidence (with maximum of 38 cases, in 40 to 50 years of age group), sex ratio ( with M : F ratio 2.5:1), organ involvement (with predominently involving lung 90 cases out of 150), efficacy of the procedure (94.7%), type and behaviour of tumour (with malignant cases comprising 103 cases) was observed.

7. Journal of Cell and Tissue Research Vol. 10(2): 2229-2234 (2010)

Evaluation of pesticide induced developmental immunotoxicity in RIR chicks

Gowri, U.K., Patel, P.V. and Suresh, B.

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

Abstract: The present study is an attempt to evaluate immunotoxicologic potential of pesticides in the developing RIR chicks. Two commercial insecticide formulations, Anaconda 505 (Combination of 50% Chlorpyrifos and 5% Cypermethrin) and Tracer (Spinosad 45% SC) were injected separately into fertilized RIR eggs. Seven days after hatching, the chicks were tested for nonspecific immune response and phagocytic ability by conducting the bacterial clearance test and NBT salt reduction test. The chicks subjected to combination insecticide showed an apparent reduction in bacterial clearance as evident by high number of bacterial colonies in the plates inoculated with blood, liver and spleen homogenate compared to that of the controls. However, spinosad evoked a lag in bacterial clearance only at the highest tested dose. The phagocytic activity of splenic macrophages too was found reduced in chicks exposed to in ovo combination insecticide, while spinosad intoxicated chicks showed no significant change in the said phagocytic activity. Moreover, the combination insecticide treated chicks showed a reduction in the absolute body weight and relative weight of thymus and spleen at the highest dose tested. However, spinosad treated chicks showed no significant variations in the body as well as the lymphoid organ weights. Therefore, from the present study it could be construed that the combination insecticide induces far more immunotoxic afflictions to the developing embryo than the naturalyte pesticide spinosad.

8. Journal of Cell and Tissue Research Vol. 10(2): 2235-2242 (2010)

Optimization of culture conditions for poly β -hydroxybutyrate production from
isolated Bacillus species

Flora, G. D., Bhatt, K., and Tuteja, U.

Amity Institute of Biotechnology, Amity University, Sector 125, Noida - 201 301, U.P
E. mail: gaganflora@gmail.com

Abstract: The capacity for poly β-hydroxy bytyrate (PHB), a biodegradable polymer, synthesis was tested for strain isolated from root nodules of Cicer arietinum plant. Out of 30 different strains, one strain was selected for further screening for PHB production in different conditions like media, temperature, pH, and agitation speed. The MTCC culture of Azotobacter beijerinckii (MTCC-2641) was used as a control for PHB production. When growth conditions were optimized in different media, the highest growth of isolate was observed in PHB glucose medium followed by yeast extract mannitol (YEM) agar. Other observed optimum growth conditions in PHB medium like temperature, pH, shaking were 30-37 0C, 6.5, 200 rpm respectively while in YEM medium all optimized conditions are same as in PHB glucose medium except agitation speed that was 150 rpm. After optimization of different culture conditions, isolated strain was characterized biochemically. Based upon biochemical studies, isolated strain was identified as Bacillus species. The present study provided the useful data about the optimized conditions for PHB production that can be utilized in industrial production of PHB, a fast emerging alternative of nonbiodegradable plastic.

9. Journal of Cell and Tissue Research Vol. 10(2): 2243-2250 (2010)

Evaluations of antimicrobial activity of some pharmacological important dihydropyrimidines compounds

Joshi, K. K., Akbari, J. and Joshi, H. S.

Department of Biotechnology, V.V. P. Engineering College, Rajkot 360 005; E. mail: krishnaonline06@gmail.com

Abstract: Antibiotics are the products of microbes that in dilute solution inhibit or kill other organisms. Antimicrobial agents include antibiotics produced by microbes and similar synthetic compounds that have the same effect. Naturally occurring antibiotics may be modified to give semi synthetic derivatives. In present study the effectiveness of a new synthetic compounds of dihydropyrimidines against a panel of pathogenic bacteria including Gram-positive and Gram-negative species, aerobes, facultative anaerobes, and obligate anaerobes have been analyzed. Pathogenic strains were isolated or obtained from the local pathology laboratory. Compounds were also subjected to find out its potency using broth dilution assay in combination with antibiotics. Study shows several of the synthetic compounds of dihydropyrimidines show antimicrobial activity and worth material for further investigation.

10. Journal of Cell and Tissue Research Vol. 10(2): 2251-2256 (2010)

Efficacy of Bacillus sphaericus strain isolated from north east region of India as potential mosquito larvicide

Yadav, K., Baruah, I. and Goswami, D.

Defence Research Laboratory (DRDO), P.B. NO.2, Tezpur, Assam 784001
E. mail: kavitanami@gmail.com

Abstract: Efficacy of Bacillus sphaericus strain GC subgroup IV (Bs) isolated from Assam, India as water dispersible powder formulation has been evaluated in laboratory and field conditions. Laboratory evaluation revealed that Bs, Spherix, ABG 6262 and Bti 164 (serotype H- 14) produced 90% mortality in Cx. quinquefasciatus @ 0.5, 0.6, 0.75, and 0.7 ppm respectively. In field trial Bs dosage @ 0.1 g/m2 in fresh water ditches resulted in 92.7 % reduction in An. crowfordi and An. annularis larvae whereas, Spherix and ABG 6262 resulted in 82.2 and 78.67 % reduction respectively at same concentration. The application of 0.2 g/m2 dosage in fresh water small ponds Bs, Spherix, ABG 6262 produced 97.43, 85.57, 81.33% reduction respectively in An. vagus and Cx. vishnui larvae. In polluted drains @ 0.5 g/m2 dosage Bs, Spherix, ABG 6262 resulted in 95.57, 86.93, 87.25 % reduction respectively in Cx. quinquefasciatus larvae. The shelf life of Bs was found more and its larvicidal efficacy persisted up to 30 months at room temperature. The study showed that Bs is an effective biolarvicide that can be used in different types of mosquito breeding habitats against different mosquito larvae.

11. Journal of Cell and Tissue Research Vol. 10(2): 2257-2262 (2010)

Optimization and production of alkaline protease from Bacillus thuringiensis cc7

Chudasama, C.J., Jani, S.A., Jajda, H.M. and Patel, H.N.

V.P. & R.P.T.P. Science College, Microbiology Department, Sardar Patel University, Vallabh Vidyanagar, Gujarat. E-mail: hnpvpscience@gmail.com

Abstract: A protease producing bacteria were isolated from soil collected around detergent industry and identified as Bacillus species. They could grow within a broad temperature range between 20 °C & 60 °C and pH range of 6.5 to 10.5. The optimum condition for protease production was 29 °C at pH 8.5 with 3 % inoculum in the medium containing Mg2+ and Ca2+ ions supplementation. Maximum growth and maximum enzyme production was observed at 48 h when grown in 100 ml medium (pH 8.5) under shaking condition (~150 rpm) at 29 °C. The best carbon and organic nitrogen sources for this bacterium were glucose and casein, respectively, while the most effective inorganic nitrogen sources were urea and (NH4)2SO4. It is envisaged that the isolate can be a potential source of alkaline protease for use as additive in industrial applications like detergent industry.

12. Journal of Cell and Tissue Research Vol. 10(2): 2263-2267 (2010)

Distribution of endophytic fungi in lemon grass (Cymbopogon citratus (DC.) Stapf.)

Deshmukh, S.K., Kolet, M.J. and Verekar, S.A.

Department of Natural Products, Piramal Life Sciences Limited, 1, Nirlon Complex, Off Western Express Highway, Near NSE Complex, Goregaon (East), Mumbai 400 063;
E. mail: sunil.deshmukh@piramal.com

Abstract: Fresh leaves and rhizome pieces of Cymbopogon citratus were used for the isolation of fungal endophytes using standard methods. Cymbopogon citratus is a grass with medicinal value found all over the different parts of India. Three hundred and twenty five endophytic fungi belonging to ascomycetes (5.23%), coelomycetes (16.30%), hyphomycetes (51.69%) and sterile mycelium (26.76%) were isolated from 800 segments of Cymbopogon citratus collected from Thane and Mulund areas. Twenty four different fungal species belonging to 21 genera viz. Alternaria alternata, Arthrinium phaeospermum, Aspergillus sp., Chaetomium globosum, Chaetomium bostrychodes, Cladosporium cladosporioides, Colletotricum gloeosporioides, Colletotricum dematum, Drechslera sp., Emericella nidulans, Fusarium sp., Nigrospora sphaerica, Nigrospora oryzae, Periconia atropurpurea, Pestalotiopsis sp., Phomopsis sp., Phyllosticta sp., and seven different sterile forms were isolated. The most frequently isolatedendophytes were Cladosporium cladosporioides (22.46%), Drechslera sp. (6.76%), Colletotrichum gloeosporioides(6.76%)and Phyllosticta sp (5.53%). Leaf samples contained more endophytes than rhizome samples.

13. Journal of Cell and Tissue Research Vol. 10(2): 2269-2280 (2010)

Evaluation of anti-inflammatory and anti-arthritic activities of ethanolic extract of Vernonia anthelmintica seeds

Otari, K.V., Shete, R.V., Upasani, C.D., Adak, V.S., Bagade, M.Y. and Harpalani, A.N.

Department of Pharmacology, Rajgad Dyanpeeth’s College of Pharmacy, Bhor, Dist. Pune 412 206.E. mail: kvotari76@rediffmail.com

Abstract: The present study was undertaken to investigate the anti-inflammatory activity and antiarthritic activity of ethanolic extract of seeds of Vernonia anthelmintica (EVA) in different experimental paradigms. The seeds were extracted by soxhlet method using ethanol (99.9%) and subjected to the preliminary phytochemical and acute toxicity studies. The effect of EVA was evaluated for acute inflammation in carrageenan induced rat paw edema and xylene induced ear edema in mice and for chronic inflammation in complete Freud’s adjuvant (CFA) induced arthritis in rats. Further, the biochemical, histopathological and radiographic evaluation was performed. Additionally, ulcerogenic potential of EVA in rats was evaluated.The phytochemical evaluation revealed the presence of alkaloids, flavonoids, steroids, triterpenes and polyphenols in EVA. In acute toxicity test, no signs of toxicity and mortality were observed on oral administration of EVA in mice up to a dose of 5000 mg/kg. In acute study, pretreatment with EVA (250, 500 & 750 mg/kg, po) was significantly inhibited the carrageenan induced rat paw edema and xylene-induced ear edema in mice. In chronic study decreased paw volume and paw thickness; increased liver weight and decreased spleen weight; decreased serum SGOT and increased serum TP level and inhibition of histopathological changes and soft tissue swelling destruction of the knee joints in radiographic examination were evident after treatment with EVA (250 & 500 mg/kg, po). Further, in the animals treated with EVA (500 mg/kg, po) ulceration was not evident. The results of present study were reveled the effectiveness of EVA in acute as well as chronic inflammatory conditions without ulcerogenic potential. The EVA may possibly act by preventing production of nitric oxide from nitric oxide synthase or by preventing neutrophilic infiltration thereby decreasing the generation or release of chemotactic factors and inflammatory T cell mediators mediators such as IL, TNF-á and LTs. These effects may be attributed to phytochemicals present in EVA.

14. Journal of Cell and Tissue Research Vol. 10(2): 2281-2286 (2010)

Adaptogenic activity of milk and aqueous decoction of Asparagus racemosus willd. in acute and chronic stress paradigms in mice

Garg, R. and Gupta, V.B.

B. R. Nahata College of Pharmacy, Mandsaur 458 001  (M.P.)
E. mail: rajeshgarg@brncop.com

Abstract: Asparagus racemosus is classified in Ayurveda, the ancient Hindu system of medicine, as a rasayana, a group of plant-derived drugs reputed to promote physical and mental health, augment resistance of the body against disease and diverse adverse environmental factors, revitalise the body in debilitated conditions and increase longevity. It has been reported to have potent adaptogenic activity.  A traditional belief regarding mode of administration of Asparagus racemosus is still unrevealed. According to this conviction Ksheerpaka (milk decoction) of Shatavari (Asparagus racemosus) is therapeutically more potent than any other form of dosage. In present study Adaptogenic activity was tested to validate cited traditional claim regarding potentiality of milk decoction of A. racemosus by determining anti stress effect using forced swimming stress model and chronic fatigue stress model in rodents. Various behavioral parameters (anxiety level, memory, muscle relaxation and locomotor activity) were also assessed in mice 24 hours after the last chronic forced swim test. The results indicate that milk decoction of A. racemosus has significant antistress adaptogenic activity, confirming the validity of traditional claim.

15. Journal of Cell and Tissue Research Vol. 10(2): 2287-2292 (2010)

Histopathology study of human’s soft tissue tumours and tumour like lesions

Agravat, A.H., Dhruva, G.A. and Parmar, S.A.

Department of Pathology, P.D.U. Medical College, Rajkot 360 005.  
E. mail: agravatah@gmail.com

Abstract: Histopathology of soft tissue tumours and tumour like lesions of human consisting of 100 cases was carried out at Pathology Department, P.D.U. Medical College, Rajkot, between September 2006 to August 2008. The tissues were the biopsy of patients submitted to the Pathology department for investigations.  The study was carried out in relation to various soft tissue lesions, mentioning histological typing of tumours with maximum of adipocytic tumours (33%), site involved with lower extremities comprising 43% cases, behavior of lesions with maximum of benign cases (86%),   age incidence ( with maximum of 13 cases, in 20 to 30 years of age group), sex ratio ( with Male : Female ratio= 1.1:1.4) have been discussed.

16. Journal of Cell and Tissue Research Vol. 10(2): 2293-2297 (2010)

Protective effect of berberine on immunoexpression pattern of P53 and BCL-2 in 7,12dimethylbenz [A] anthracene-induced skin carcinogenesis in  Swiss albino mice

Manoj Prabhakar, M., Madavan Nirmal, R.,  Silvan, S., Baskaran, N., Muneeswaran, M. and Manoharan, S.

Department of Biochemistry & Biotechnology, Faculty of Science, Annamalai University,
Annamalainagar-608 002.Tamilnadu E. Mail: sakshiman@rediffmail.com

Abstract: The investigation deals with the effects of berberine on immunoexpression pattern of p53 and bcl-2 in skin tissues of 7,12-dimethylbenz(a)anthracene treated Swiss albino mice. DMBA treatment for 8 weeks (25 ìg in 0.1 ml acetone/mouse) on the depilated back of mice developed squamous cell carcinoma in the skin tissues. Overexpression of p53 and bcl-2 was noticed in the skin tissues of mice treated with DMBA alone. Oral administration of berberine at a dose of 75 mg/kg body weight to DMBA treated mice significantly suppressed the over expression of p53 and bcl-2. Our results suggest that berberine might have stimulated apoptosis in skin tissues by down regulating the expression of p53 and bcl-2 during DMBA-induced skin carcinogenesis.

17. Journal of Cell and Tissue Research Vol. 10(2): 2299-2305 (2010)

Natural fluoro power

Gupta, P.D.

Adjunct Professor, Manipal University, Manipal, Karnataka, India. E mail pdg2000@hotmail.com

Abstract: Fuorophore, a compound of protein with the chromophore 4-(p-hydroxybenzylidene)-5-imidazolidinone, is found natually in many hydrozoans. The fluorescent proteins have been exploited in biochemistry and cell biology because of the incalculable value of fluorescent proteins for the study of cells, tissues, and even whole animals at level of detail and subtlety that would otherwise be experimentally inaccessible. The power of fluorescence is not used only in microscopy but it can be used—from measuring gene expression to using light to manipulate cells themselves.