1. Journal of Cell and Tissue Research Vol. 8(3): 1491-1498 (2008)
Benzodiazepine  receptor binding and anxiolytic studies on anximin®, a polyherbal formulation

Mishra, S., Khanna, V.K. and Vikas Kumar

Neuropharmacology Laboratory, Department of Pharmaceutics, Institute of Technology,Banaras Hindu
University, Varanasi-221 005, India. Email: neuropharmacologist@rediffmail.com

Abstract: Anximin is a polyherbal formulation containing Bacopa monnierae, Convolvulus pluricaulis, Rauwolfia serpentina, Nardostachys jatamansi and Acorus calamus. As scientific validation of Anximin is lacking, the objective of our present study was to evaluate the putative anxiolytic activity of Anximin, through validated animal models of anxiety viz. elevated plus maze (EPM), elevated zero maze (EZM), open field test (OFT) and social interaction tests (SIT). Anximin was administered orally in two doses (20 and 40 mg/kg) as suspension in 0.3 % carboxy methyl cellulose (CMC) for 7 consecutive days. Control group animals were administered equal volume of 0.3% CMC suspension. Diazepam (3 mg/kg, p.o.) was administered acutely to rats as a standard drug. At both dose levels Anximin demonstrated significant increase in open arm entries and time spent on open arms (p < 0.05 and p < 0.001 respectively). In EZM test, significant augmentation in entries and time spent in open arms (p < 0.01) was observed. In OFT, significant increase in the open field ambulation and rearing (p < 0.01) was observed while fecal droppings were significantly (p < 0.01) reduced by Anximin. In SIT, the rats treated with Anximin spent more time in social interaction in comparison to control rats, and effects of Anximin were found to be significant (p < 0.001). Results of this behavioural study indicate that Anximin possesses promising anxiolytic activity, qualitatively comparable to diazepam. To elucidate the mechanism of action, benzodiazepine receptor binding study was also undertaken. Rats treated with Anximin (40 mg/kg p.o., for 7 days) significantly up-regulated benzodiazepine receptors in frontal cortex. Thus it may be concluded that the observed anxiolytic activity of Anximin is regulated through benzodiazepine receptors.

2. Journal of Cell and Tissue Research Vol. 8(3): 1499-1504(2008)

Influence of different levels of calcium on growth and biochemical attributes in fluoride toxicated rats

Patel, B.G.,  Patel, V.H., Patel, I.N., Dhaduk, J.J., Vyas, S.R., Patel, N.V. and Sail, S. S.

Department of Food Science and Nutrition, ASPEE College of  Home Science, S. D.  Agricultural University, Sardarkrushinagar 385 506; E. mail: bgpatel62@yahoo.co.in

Abstract: Forty female albino rats were exposed to dietary fluoride toxicity (100 ppm NaF) on low  (n=15, LC), normal  (n= 10, NC) and high  (n=15, HC) calcium diet for 45 days (Exp. I).  After 45 days, the five animals from each group were either continued with the same diet or shifted to opposite diet (shifting HC diet to LC diet or vice versa).  except  normal calcium diet for further 45 days (Exp.  II). The toxic effects as reflected through growth attributes and altered biochemical profile in general, were apparent over the dietary calcium levels in fluoride toxicated rats fed for 45 days. The effects of calcium levels in  fluoride toxicated rats of experiment II were significant for the almost same growth attributes that were under experiment I. These growth attributes showed improvement on  shifting HC diet to LC diet, while there was a marked decline in growth attributes when the diet regime changed from LC to HC. However, organo-somatic index for heart, kidney, tibia and femur was maximum for the rats fed HC diet continuously.  The effects of dietary calcium levels in fluoride toxicated rats were significant for liver proteins, liver vitamin C, plasma phosphorus, tibial moisture, tibial and femoral lipid, bone calcium, bone phosphorus, bone fluoride, bone hydroxyproline, bone collagen, bone nitrogen and hexosamine. On the contrary, it is  significant that highest values for plasma phosphorus, tibial moisture, bone ash and bone fluoride were observed under low calcium diet. Similarly, the plasma AST, LDH and femoral lipid, hydroxyproline, collagen, nitrogen, hexosamine of bone tissue were highest for rats  fed HC diet constantly for 90 days while these values were lowest for LCàLC subgroup.  Thus, the present findings revealed that increased intake of dietary calcium can be useful in reducing toxic effects of fluoride while calcium deficiency exaggerated the fluoride toxicity. However, level of dietary calcium may cause weight loss due to hypercalcemia.

3. Journal of Cell and Tissue Research Vol. 8(3): 1505-1510(2008)

Withaferin-a protects biochemical changes in the buccal mucosa of Golden Syrian hamsters during 7,12-dimethylbenz [a]  anthracene (dmba) - induced oral carcinogenesis                                                                                                                            

Panjamurthy, K.,  Balakrishnan, S., Vellaichamy, L., Manoharan, S.  and Linsa Mary, A.

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

Abstract: Our aim was to investigate the protective effect of Withaferin-A on the status of  glycoconjugates, lipids, lipid peroxidation by-products and antioxidants in the buccal mucosa of golden Syrian hamsters during DMBA-induced oral carcinogenesis. Oral squamous cell carcinoma was developed in the buccal mucosa of animals by painting with 0.5% DMBA in liquid paraffin thrice a week for 14 weeks. The status of glycoconjugates, lipids, lipid peroxidation by-products, enzymatic and non-enzymatic antioxidants in buccal mucosa was estimated by specific colorimetric methods. The level of total cholesterol  significantly increased whereas phospholipids and free fatty acids  decreased in buccal mucosa of DMBA alone painted animals as compared to control. Decrease in TBARS level and disturbances in antioxidants status (Vitamin E, GSH and GPx SOD and CAT) were noticed in buccal mucosa of oral cancer bearing hamsters as compared to control. The levels of  glycoconjugates  significantly increased in buccal mucosa of DMBA alone painted animals as compared to control. Oral administration of Withaferin-A at a dose of 20mg/kg body weight to DMBA painted animals on days alternate to DMBA painting brought back the status of above said biochemical parameters to near normal range in the buccal mucosa. The present study thus demonstrated that Withaferin-A has the ability to protect biochemical alterations in hamster’s buccal mucosa during DMBA-induced oral carcinogenesis.

4. Journal of Cell and Tissue Research Vol. 8(3): 1511-1514(2008)

Extraction, purification and characterization of thermostable  alpha- amylase

Gudmalwar, R.M.,  Kulkarni, S.S.,  Shembekar, V.S.  and Khobarahgade, C.N.

Department of Biotechnology Engineering, KIT’s College of Engineering, Kolhapur, 4162342, E. mail: grajesh1@rediffmail.com,

Abstract: The microorganism which grows optimally at 60 °C and above is known as thermophilic organisms. Thermostable enzymes remain folded and functional at high temperatures and show maximum enzymatic activity as compared to their mesophilic analogs (thermal adaptation). Because of these characteristics, thermostable enzymes are remarkable tools for the study of protein stability. The primary aim of the work was to optimize the cultural conditions for the optimal enrichment of the microorganisms. The six potent growers were selected and checked for their ability to produce α- amylase. Further, ability of the isolates for maximum degradation of starch was also screened. The enzyme was purified in two step procedure involving ammonium sulphate precipitation and DEAE cellulose anion exchange chromatography. The purified enzyme was subjected to SDS-PAGE for determining molecular weight. Finally the enzyme kinetics study was carried out. Presently the mechanism of thermostbility of the purified amylase is being analyzed.

5. Journal of Cell and Tissue Research Vol. 8(3): 1515-1518 (2008)
Brahmi improves lysosomal membrane integrity in mouse brain during aging

Pillai, M.M. , Vora, S.R.  and Patil, R.B.

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

Abstract: Cure of age associated nervous disorders like dementia, Alzheimer’s Disease may be years away in the field of medicines. Endless antioxidants have been scrutinized by researchers that can benefit  the  nervous disorders, but the results are inconclusive. Many results suggest that dietary supplementation of antioxidants may reduce the risk of developing these diseases. The present study was undertaken to assess the potential of ethanolic extract of Brahmi (Bacopa monniera Linn.) to maintain the lysosomal membrane integrity in the brain tissues of mice during natural aging. The biochemical studies on three lysosomal enzymes acid phosphatase, non-specific esterase and β-glucuronidase were performed in post lysosomal and lysosomal fractions of brain tissues of male and female mice. The enzyme levels were found to reduce in lysosomal fractions of brain tissues as the age progressed. While, in post lysosomal fractions they were found to increase with the age showing leakage of enzymes from lysosomes. Ethanolic extract of Brahmi repaired this imbalance at 12 months age of mice but not in mice aged above 15 months. Thus, the present study suggests that Brahmi, serves as a stabilizing agent, and maintains the lysosomes membrane integrity in the brain in the aging animals but not in the aged mice.

6. Journal of Cell and Tissue Research Vol. 8(3): 1519-1524(2008)

Spermatogenic activity of dietary antioxidant in oxidatively stressed mice

Patil, R.B., Vora, S.R. and Pillai, M.M.

Department of Zoology, Government Rajaram College, Vidyanagar, Kolhapur, 416004,
E. mail: rb_aging@rediffmail.com

Abstract:The present study was carried out to evaluate antioxidant properties of Petroselinum crispum and Bacopa monniera leaf extracts. Six months old male mice were divided into four groups viz. control group, 5% D-galactose treated group, D-galactose along with P. crispum leaf extract treated group and with D-galactose + B. monniera leaf extract administered group. The sperm count in epididymis and qualitative determination of lactate dehydrogenase from testis and epididymis by Polyacrylamide gel electrophoresis were made. Sperm count was found significantly low in case of D-galactose administered group of mice as compared to control. The number of sperm cells was increased in case of plant extract treated groups even though D-galactose was administered to those groups along with leaf extract. A specific LDH-X isoenzyme, a marker enzyme of active spermatogenesis was seen in control group but it disappeared in D-galactose stressed mice. However, it   reappeared in both plant extract treated groups. Further, it was more prominent in B. monniera received group than other one. Study shows abundant antioxidant potential of leaf extracts of these plants that overcomes the stress induced by D-galactose.

7. Journal of Cell and Tissue Research Vol. 8(3): 1525-1530(2008)

Protective effects of curcumin and ferulic acid on circadian pattern of lipid peroxidation and antioxidants in 7,12-dimethylbenz[a]anthracene induced oral carcinogenesis

Balakrishnan, S., Manoharan, S.,  Linsa Mary, A., Vellaichamy, L. and Menon, V.P.

Department of Biochemistry and Biotechnology, Faculty of Science, Annamalai University,
Annamalainagar 608 002, Tamil Nadu, India. E. Mail: manshisak@yahoo.com

Abstract: The protective effect of curcumin and ferulic acid on circadian pattern of lipid peroxidation and enzymatic antioxidants in 7,12-dimethylbenz[a]anthracene  (DMBA)-induced hamster buccal pouch carcinogenesis has been investigated. We developed oral squamous cell carcinoma in the buccal pouches of golden Syrian hamsters at different time intervals (4.00h, 8.00h, 12.00h, 16.00h, 20.00h and 24.00h) by painting with 0.5% DMBA three times per week for 14 weeks. The status of lipid peroxidation by-products [thiobarbituric acid reactive substances (TBARS)] and enzymatic antioxidants [superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx)] were assayed at 4.00h intervals throughout the 24.00h period using colorimetric methods. Disrupted circadian pattern of lipid peroxidation and antioxidants status was observed in the buccal mucosa of hamsters painted with DMBA alone. Oral administration of curcumin (80mg kg-1 b.w) and ferulic acid (40mg kg-1 b.w) to hamsters painted with DMBA for 14 weeks, on alternate days of DMBA painting, significantly protected the circadian dysregulation in the status of lipid peroxidation and antioxidants in the buccal mucosa. Our study thus demonstrates the protective role of curcumin and ferulic acid on circadian fluctuations of lipid peroxidation and antioxidants during DMBA-induced hamster buccal pouchcarcinogenesis.

8. Journal of Cell and Tissue Research Vol. 8(3): 1531-1536(2008)

Action of acute industrial effluent exposure in prepubertal and adult rat testis during one cycle of seminiferous epithelium

Vachhrajani, K.D. and Pandya, A.P.

Division of Environment and Toxicology, Department of Zoology, Faculty of Science, The Maharaja Sayajirao University of Baroda, Vadodara – 390 002, India.  E-mail: kdv_zdmsu@yahoo.com

Abstract: There are several studies in literature dealing with the toxic effects of single or a group of compounds on reproductive organs but attempts to assess toxic potentials of heterogeneous mixture are scanty.  Present studies evaluated the toxicity of industrial effluent following single oral exposure to rat testes. One cycle of seminiferous epithelium (about 12 days) was studied both in prepubertal (age 20 days) and young adult (age 50 days) rats. Qualitative and quantitative analysis of spermatogenesis at different intervals during one SE cycle indicated an initial toxic lesion development at days 2 post treatment both in pre-pubertal and adult animals. In former early–mid pachytene exposed to industrial effluent could not successfully accomplish all the events of meiotic cell division and degenerated, resulting consequently in the absence of round spermatids.  In adult rats, effluent exposure initiated loss of more mature spermatogenic cell types and delayed the progression of spermatogenesis. The tubules at stage IX – XIV of SE were more sensitive to the effluent exposure.

9. Journal of Cell and Tissue Research Vol. 8(3): 1537-1544(2008)

Physiological functions of the haemocyanins of Oziotelphusa Senex senex and Pila globosa in relation to thermal stress

Sridevi, B. and Naidu, B.P.

Environment Protection Training and Research Institute, Gachibowli, Hyderabad 500 032, 
E. Mail: sri27_bonthu@yahoo.co.in

Abstract: The haemocyanin of Arthropods and Mollusks appears to function in gas transport much like the haemoglobin of the vertebrates, although haemocyanin possesses a much larger Bohr effect. The haemocyanin  of Oziotelphusa senex senex and Pila globosa under physiological conditions possesses a high oxygen affinity. The oxygen equilibrium curve is sigmoid with a fairly high degree of interaction occurring between the O2 combining sites of the haemocyanin molecule. It is shown that the low half saturation value of the haemocyanin confers a stability of loading pressure in the face of changing temperatures. It is speculated that the high O2 affinity may also represent an adaptation tending to counter the effect of the rate at which O2 diffuses through the respiratory surface into the haemocyanin. In both the control and temperature exposed animals the different  spectra showed maximum absorption band of the oxyhaemocyanin at 580 nm for Oziotelphusa senex senex and at 575 nm for Pila globosa. The oxygen equilibrium curves of the blood of the animals exposed to cold temperature shifted to the left with a corresponding decrease in the P50 values showing greater affinity of the haemocyanin for oxygen. In the animals exposed to warm temperature the oxygen equilibrium curves shifted to the right with a corresponding increase in the P50 values showing decreased affinity of the haemocyanin pigment for oxygen. The results obtained are discussed in relation with the physiological characteristics of the haemocyanin in relation to temperature. To understand the physiological functions between arthropod and molluscan haemocyanin we studied the interactions between the oxygen-binding curves. The oxygen equilibrium curves in both phyla are  similar with a significant effect of temperature.

10. Journal of Cell and Tissue Research Vol. 8(3): 1545-1550(2008)

Antimicrobial activities of three Indian medicinal  plants

Salunke, B. K.,  Patil, S. V., Lad, R. Chatterjee, S. and  Maheshwari, V. L.

Department of Biochemistry, School of Life Sciences, North Maharashtra University, P. B. No. 80, Jalgaon- 425 001.  E. mail: vlmaheshwari@hotmail.com

Abstract: The antimicrobial activities of root, leaf and stem extracts of three medicinal plants, Barleria prionitis L. (Acanthaceae), Withania somnifera L. Dunal (Solanaceae) and Helianthus annus L. (Compositae) in water, methanol and dichloromethane were evaluated against few microorganisms (bacteria - Staphylococcus aureus, Bacillus subtilis, Pseudomonas aeruginosa, Escherichia coli and Salmonella typhi and fungi -  Aspergillus niger, Aspergillus flavus, Aspergillus awamori, Fusarium oxysporium and Penicillium notatum). Promising antimicrobial potential was observed for the root and leaf extracts of the plants in methanol and dichloromethane. Barleria prionitis root extract in methanol presented highest activity against Bacillus subtilis among all the tested microorganisms. Water extracts, in general showed less antimicrobial activity as compared to other solvents. The antimicrobial activity of the plant extracts was found to be dose dependant and varied with the type and concentration of the extract as well as type of microbial species.  The results were compared with the activity of some standard antibiotics.

11. Journal of Cell and Tissue Research Vol. 8(3): 1551-1556(2008)

Evaluation of Calotropis gigantea R. Br. flowerextract on alcohol induced hepatotoxicity

Kshirsagar, A. and Purnima, A.

Department of Pharmacology, K.L.E.S’s College of Pharmacy, Bangalore 560010.
E. mail: purnimaal1@yahoo.com

Abstract: The protective effect of Calotropis gigantea flower extract in acute alcohol induced hepatotoxicity in rats was evaluated. A significant increase in the levels of enzyme markers of tissues damage (ALT, AST and ALP), lipid peroxidation (TBARS) and decreased serum vitamin C  levels were observed in alcohol intoxicated rats as compared to control animals. Pretreatment with 250 and 500 mg/kg body weight of extract significantly decreased the levels of enzyme markers, lipid peroxidation and markedly increased serum vitamin C level in a dose-dependent manner. Post-treatment with 500mg/kg body weight of extract significantly enhanced the recovery of animals from hepatic damage compared to untreated control. Lipid peroxidation and depletion of vitamin C due to oxidative stress could be the possible mechanism of alcohol induced toxicity and the protective effect of the extract may be due to its ability to inhibit lipid peroxidation and prevent the depletion of vitamins C.

12. Journal of Cell and Tissue Research Vol. 8(3): 1557-1563(2008)

Influence of administration of FGF-2 and ANTIFGF-2 on the nucleic acids and protein profiles of the regenerating tail of northern house gecko Hemidactylus flaviviridis

Yadav, M., Sharma, P., Desai, I., Pilo, B. and Suresh, B.

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

Abstract: We investigated the role of FGF-2 in lizard caudal regeneration. FGF-2 is known to possess broad mitogenic and cell survival activities and in lizard tail regeneration, FGF-2 acts to augment the mitotic activity as evidenced by increased DNA levels in the regenerates, while at the same time it also significantly increased the levels of RNA. However, the animals treated with FGF-2 showed significantly lower transcriptional and translational activities at the blastema stage, while during the differentiation stage the synthetic activity was found to increase in the FGF-2 treated animals. The current notion was further consolidated by the results of antiFGF-2 treated animals. However, the exact pathway by which FGF-2 exerts its mitogenic influence needs to be further evaluated.

13. Journal of Cell and Tissue Research Vol. 8(3): 1565-1569 (2008)

Absence of chytridiomycosis in the skin of amphibians collected from Vadodara district of Gujarat

George, R.  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,

Abstracts: The reports of life threatening fungal infection chytridiomycosis on amphibian populations have emerged from various parts of the world and the fungus has been considered as major contributor to the decline of amphibian population worldwide. However, a meticulous scan through the literature revealed that this disease is yet to be identified in Asia. Therefore, it was thought pertinent to investigate the prevalence of the cutaneous disease the chytridiomycosis in the amphibian community in Vadodara, Gujarat by analyzing the histologic profile of the amphibian skins. A careful analysis of the histoarchitecture of epidermis revealed no evidence of fungal infection in the samples of the skin examined. The thickness of stratum corneum in all the cases remained within the normal range of 2-5µm. Thus, it could be concluded that the anurans collected from  Vadodara might be free from Batrachochytrium dendrobatidis zoosporangia.

 14. Journal of Cell and Tissue Research Vol. 8(3): 1571-1576(2008)

Antioxidant and hepatoprotective activity of Pimenta dioica leaves extract

Nayak, Y. , Abhilash, D., Vijaynarayana, K. and Fernandes, J.

Department of Pharmacology, Manipal College of Pharmaceutical Sciences, Manipal 576104, Mangalore, 574160.  E. mail: yogendranayak@gmail.com

Abstract: The berries of Pimenta dioica (L) Merril (Family Myrtaceae) are commonly known as allspice in culinary. The natives of Kerala and Mangalore use Allspice leaves as medicine for pain, arthritis, fever and stress. In all these pathological conditions oxidative stress is one of the causes, hence we tried extract the leaves  for in vitro antioxidant activity. 20% yield of alcoholic extract obtained from the dry leaves  showed total phenolic content (TPC) equivalent to 20 ± 1.63 mg/g of the dry leaves powder. The extract showed significant antioxidant activity by reducing power, DPPH radical scavenging and Lipid peroxidation assays. The antioxidant activity was proportional to the total phenolic content in the leaves. The high content of phenolic antioxidants in the extract urged us to screen for hepatoprotective activity in carbon tetrachloride (CCl4) intoxicated Wistar rats. The oral doses (250, 500, 750 mg/kg/day) showed statistically significant protection of liver when investigated by histopathological and biochemical parameters.

15. Journal of Cell and Tissue Research Vol. 8(3): 1577-1580(2008)

Antioxidant activity of Syzygium cumini seeds in aspirin induced peptic ulcer in rats

Udapurkar, P.P.,  Kale, M,K. and Duragkar, N.J.

Sharad Pawar College of Pharmacy, Wanadongari, Hingna Road, Nagpur 441110. E. mail: prachi.udapurkar@gmail.com

Abstract: In the present study a hydroalcoholic extract of Syzygium cumini seeds have been investigated for its antioxidant activity in aspirin induced peptic ulcer in rats with the aim of exploring possible correlation between its antioxidant and antiulcer activities. Oxygen free radicals are considered to be important factors in the pathogenesis of peptic ulcer. The level of lipid peroxide was elevated and the activities of antioxidant enzymes were reduced considerably by oral administration of aspirin. The decreased level of  lipid peroxides and increased activity of antioxidant enzymes were observed with treatment of Syzygium cumini seeds. The results indicated that Syzygium cumini seed may exerts its gastroprotective effect by free radical scavenging action and have considerable therapeutic potential in the treatment of peptic ulcer.

16. Journal of Cell and Tissue Research Vol. 8(3): 1581-1584(2008)

Immunomodulatory activity of aqueous extract of Amorphophallus campanulatus tuber

Tripathi, A.S., Chitra, V.,  Sheikh, N.W., Dewani, S.P., Chakma, C.

SRM College of Pharmacy, Kathankulathur  603203, Tamil Nadu. E. mail: shloksk@rediffmail.com

Abstract: Traditionally Amorphophallus campanulatus (AC) tuber is used for tumor, rheumatoid arthritis, carminative and liver tonic. It is also used in the treatment of  piles and given as the restorative in dyspepsia debility, Anti-inflammatory, antihaemorrhoidal, haemostatic, expectorant, digrstaive and anthelmintic.  In present study effect of aqueous extract of Amorphophallus campanulatus tuber on immunological function in mice was studied. The Aqueous extract (AE) of Amorphophallus campanulatus tuber, given orally at the doses of 250 and 500 mg/kg showed a significant results of test parameters viz., charcoal clearance, spleen index and delayed type hypersensitivity (DTH) response. In mice immunized with sheep RBC, aqueous extract of AC decreases the charcoal clearance rate and the cellular immunity by facilitating the footpad thickness response to sheep RBC in sensitized mice.

17. Journal of Cell and Tissue Research Vol. 8(3): 1585-1588(2008)

Mass propagation of orchids through in vitro seed culture technology

Das, A.K.,  Das,  J., Gogoi, H.K. and Srivastava, R.B.

Defence Research Laboratory, Post Bag No. 2, Tezpur- 784 001, E-mail:  ajit_drl@rediffmail.com

Abstract: Seeds of four orchid species from North-east India namely Aerides odoratum, Cymbidium longifolium, Dendrobium aphyllum and Rhyncostylis retusa were germinated on four basal media, Knudson, Burgeff, Vacin and Went and Phytomax. Best response was exhibited by Knudson medium followed by Burgeff and Phytamax in terms of faster and higher frequency of germination.  Root-shoot initiation was found in Knudson and Burgeff media only and in support with natural extracts like 15 % coconut water and 6 % banana extract. Earliest shoot and root initiation was recorded as 70 days in C. longifolium, D.  aphyllum and R. retusa and 98 days in D. aphyllum and R. retusa respectively. Plantlets with well defined shoot and root system were cultured on half strength Knudson basal medium prior to transfer in to pots. About 170-180 days old in vitro plantlets were transferred into pots. The survival rate of the plantlets was found to be 70-80 %. This technique involves three sequential phases – germination, protocorm formation and seedling development. The nutrients and culture requirement  are different in each phase.

18. Journal of Cell and Tissue Research Vol. 8(3): 1589-1594(2008)

Biochemical response during the germination of Raya and Fenugreek seeds under heavy metal stress  

Nataraj, M,  and Parmar, S.B.

B. R. D. School of Biosciences, Sardar Patel University, Vallabh  Vidyanagar- 388120
E. mail: nattusamy@yahoo.com 

Abstract: Heavy metals are known to induce oxidative stress in germinating seeds and plants which is countered by the antioxidative stress mechanisms of tolerant plants. The effect of heavy metals like Cd, Pb, Ni, Cr, Zn and Cu on germination of Brassica compestris (raya) and Trigonella foenum-graecum (fenugreek) and the induction of antioxidative enzymes has been studied. The highest catalase activity was observed in fenugreek seeds treated with 10 ppm Cd. The latter showed highest activity of peroxidase. The total soluble protein was highest in the seeds of raya and fenugreek treated with 10 ppm of Cd. Highest proline content was recorded in 10 ppm Pb and 10 ppm Zn in fenugreek and raya, respectively.

19. Journal of Cell and Tissue Research Vol. 8(3): 1595-1598(2008)

Evaluation of hepatoprotective efficacy of Lippia nodiflora L. against carbon tetrachloride induced hepatic damage in rats

Sampathkumar, P., Dheeba, B., Vinothkannan, R., Veena, V., Balakumar, S. and Shanmugasundaram, A.

Department of Chemistry and Biosciences, SASTRA University, SRC, Kumbakonam  612 001, Tamil E. mail: pskumar16@yahoo.com,

Abstract: Hepatoprotective activity of ethanolic leaf extract of Lippia nodiflora was evaluated against carbon tetrachloride (CCl4) induced hepatic damage in rats. The extract in doses of 100 and 200 mg/kg body weight were administered orally once daily for fifteen days. Hepatoprotective activity was measured based on biochemical parameters. Elevated levels of total bilirubin, aspartate transaminase (AST), alanine transaminase (ALT) and alkaline phosphatase (ALP) in CCl4 intoxicated rats were restored to normal levels significantly (p<0.05) in the animals treated with the extract in doses of 100 and 200 mg/kg body weight and CCl4. The level of total protein appreciated significantly (p<0.05) in animals treated with different doses of the leaf extract and CCl4. The effects were dose dependent. These results suggest that the leaf extracts of Lippia nodiflora have potential therapeutic and preventive efficacies, probably due to its antioxidant effect.

20. Journal of Cell and Tissue Research Vol. 8(3): 1599-1604(2008)

Antitumour promoting potential of Coccinia indica against benzidine  induced hepatocellular carcinoma in mice

Sampathkumar, P., Prabhakaran, K., Dheeba, B., Vinothkannan, R., Vinothprasanna, G. and Sheikabdulla, S.

Department of Chemistry and Biosciences, Srinivasa Ramanujan Centre, SASTRA University, Kumbakonam 
612 001 Tamilnadu. E. mail: pskumar16@yahoo.com

Abstract: In the present study, anti-tumour activity of ethanolic extract of leaves of Coccinia indica against benzidine induced hepatocellular carcinoma in mice was investigated. Hepatocellular carcinoma (HCC) was induced by treating the mice orally with benzidine (200mg/kg of body weight) for 30 days. HCC was manifested by significantly decreased (p<0.05) levels of serum protein, albumin and liver proteins. Serum globulin and bilirubin levels were significantly increased (p<0.05) on tumour development. Activities of liver marker enzymes (SGOT, SGPT, ALP and γ-GT) were also significantly increased. On treatment with the ethanolic extract of Coccinia indica (250mg kg of body weight) the alterations were reversed. Levels of serum protein, albumin, globulin, liver protein and bilirubin were restored in near normal levels in mice treated with the ethanolic extract of Coccinia indica (CLEt).  Activities of liver marker enzymes were also restored at near normalcy in treatment group. Elevated level of tumour marker Alpha – Feto protein (AFP) was observed in mice treated with benzidine alone, whereas AFP level was markedly reduced on treatment with the ethanolic extract of Coccinia indica. In conclusion, these findings indicate that Coccinia indica has a protective effect against the hepatocellular carcinoma induced by benzidine.

21.  Journal of Cell and Tissue Research Vol. 8(3): 1605-1612(2008)

Evaluation of antimicrobial and antioxidantal activities of lime and ginger

Motwani, S., Parekh, T. , Rema, S. and Patel, V. H.

Food Biotechnology Laboratory, Department of Home Science, Sardar Patel University, Vallabh
Vidyanagar-388120. E mail: tarak577@gmail.com

Abstract: Herbs and spices are an important part of  human diet. They have been used for thousands of years to enhance the flavour, colour and aroma of food. In addition to boosting flavour, herbs and spices are also known for their preservative and medicinal values. The present study was carried out to investigate the antimicrobial and antioxidant activities of lime juice and ginger. Different crude and solvent extracts were tested against selected bacterial and fungal strains. Hexane extract showed  significant zone of inhibition against microorganisms. Antioxidant activity was measured by DPPH and ABTS methods. Methanolic extract of dry ginger showed more antioxidant capacity than fresh ginger. Methanolic extract of dry ginger showed higher phenolic values.

22. Journal of Cell and Tissue Research Vol. 8(3): 1613-1616(2008)

Role of GSTP1 gene polymorphism in the development of breast cancer: A case control study

Surekha, D., Vishnupriya, S.,  Sailaja, K., Nageswararao, D., Padma, T.,  and Raghunadharao, D.

Department of Genetics, Osmania University, Hyderabad 500 007; 1Nizams institute of Medical Sciences, Hyderabad 500 082, India. E. mail: sattivishnupriya@gmail.com

Abstract: The glutathione S transferases (GSTs) are involved in the metabolism of xenobiotics, such as environmental carcinogens, chemotherapeutic agents and free radicals. A group of 250 breast cancer (including 11 male breast cancer cases) patients and 250 controls were analyzed for GSTP1 by PCR-RFLP method. The genotype frequency distribution of GSTP1 did not show any significant difference between breast cancer patients and controls. Ile/Val genotype frequency was elevated in postmenopausal breast cancer patients [49.2%] with high BMI and negative for estrogen receptor [45.9%], progesterone receptor [47.5%] and HER2/neu [55.6%]. GSTP1 Val/Val genotype frequency was elevated in premenopausal breast cancer patients who were positive for estrogen receptor [9.0%], progesterone receptor [9.3%] and with high grade tumors [8.0%]. Our data suggested that GSTP1 Val/Val genotype might confer risk to develop breast cancer depending on the level of exposure to carcinogens.

23. Journal of Cell and Tissue Research Vol. 8(3): 1617-1628(2008)

Electron microscopic radioautographic study on mitochondrial DNA synthesis in adrenocortical cells of mice in aging and senescence

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

Abstract: In order to study the aging changes of intramitochondrial DNA synthesis of mouse adrenal cells, 10 groups of developing, aging and senescent mice, each consisting of 3 individuals, total 30, from fetal day 19 to postnatal newborn at day (1, 3, 9), juvenile at day 14, young adult (at month 1, 2) and senescent adult (at month 6, 12, 24) were injected with 3H-thymidine, 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 per cell, number of labeled mitochondria per cell and the mitochondrial labeling index labeled with 3H-thymidine showing DNA synthesis in each adrenocortical cells, in 3 zones, were counted and the results in respective developing groups were compared with each other. From the results, it was demonstrated that the numbers of mitochondria in 3 zones, the zona glomerulosa, fasciculata and reticularis of respective mice at various ages in 10 groups increased from fetal day 19 to postnatal month 2 reaching the plateau from month 6 to 12 and 24 due to development and aging of animals, respectively, while the numbers of labeled mitochondria and the labeling index of intramitochondrial DNA synthesis incorporating 3H-thymidine increased from fetal day 19 to postnatal month 2, reaching the maxima and  decreased from month 6 to month 12 and 24. It was shown that the activity of intramitochnodrial DNA synthesis in the adrenocortical cells in developing and aging mice changed due to aging.

24. Journal of Cell and Tissue Research Vol. 8(3): 1629-1634(2008)

Effect of fresh wheatgrass juice on plasma antioxidant status of geriatric population

Sharma, B., Parekh, T., Dave, N., Patel, V. and Rema, S.

Laboratory of Food Science and Nutrition, Department of Home Science, Sardar Patel University, Vallabh Vidyanagar-388120, E. mail: tarak577@gmail.com

Abstract: Wheat grass is considered to be a nutritional powerhouse. It acts as a natural cleanser of the body. The present study was undertaken with the broad objective of assessing plasma antioxidant levels of geriatric population after supplementation of wheat grass juice for one month. Fresh wheatgrass was analyzed for total phenol, flavonoids and antioxidant capacity. 18 subjects were selected for the study. All experimental subjects were fed with fresh wheat grass juice daily early in the morning for 30 days. It was observed that 30 days supplementation of juice significantly (Pd<0.01) increased the hemoglobin, glutathione, ascorbic acid and vitamins A, E, TAC (mg trolox equivalent per 100 gm). The phenol content of water and methanolic extract was found to  increase trenomdously. The water extract showed significantly higher (pd<0.01) TAC compared with the methanolic extract. It is concluded from the study that wheat grass juice provides genuine therapeutic advantage in the body and can be used as a nutrional supplement for therapeutic implication in the treatment of various diseases.

25. Journal of Cell and Tissue Research Vol. 8(3): 1635-1640 (2008)

Ultrastructure of the adrenal medulla of emballonurid female bat, Taphozous longimanus (Hardwicke) during the reproductive cycle

Nerkar, A.A.  and Gadegone, M.M.

Cell Biology Section, Department of Zoology, Institute of Science, Nagpur-440 001, India
E mail: archana_nerkar1@yahoo.co.in
Abstract:  Adrenal medullary cells of Taphozous longimanus are characterized by the presence of well developed Golgi apparatus, rough endoplasmic reticulum and large number of mitochondria suggesting that these cells are active in synthesis of components during different stages of reproductive cycle. Adrenal medulla consists of two types of cells depending on the presence of secretory granules. The type I cells are characterized by the presence of secretory granules of extra ordinary dense core which is separated from the surrounding membrane by an electron lucent nimbus like space which correspond to nor-epinephrine cells of other mammals. The type II cells are characterized by the presence of granules with less dense core which lack the large electron lucent space surrounding them as observed in the epinephrine cells of other mammals.

26. Journal of Cell and Tissue Research Vol. 8(3): 1641-1643 (2008)

Pimecrolimus 1% cream in the treatment of oral lesions of pemphigus vulgaris: a double-blind, placebo-controlled clinical trial

Iraji, F, Khoramian Fard, M., Asilian, A. and Siadat, A.H.

Department of Dermatology, Isfahan university of Medical Sciences, Skin Diseases and Leishmaniais
Research Center, Isfahan , Iran. E. mail: amirhossein1@yahoo.com

Abstract: Pemphigus vulgaris (P.V.) is a chronic, bullous disorder that is usually characterized by presence of bulla and erosion on the skin and mucosa. Many researches on P.V. are now, therefore, focusing on the use of topical non-steroid agents. One of these agent is pimecrolimus and its efficacy is established in some inflammatory and autoimmune disorders such as oral and genital lichen planus. In present investigation a double-blind study was undertaken in 30 patients with confirmed diagnosis of  P.V.  Patients under treatment with systemic steroid and Azathioprine who had bilateral symmetrical oral lesions were selected and right or left sided lesions of them were randomized to be treated either by pimecrolimus or placebo. The largest diameter of lesions was measured at the baseline and every 10 days  2 times. Epithelization Index (E.I.) was calculated and data were analyzed with SPSS software. Out of 30 patients, 17 (56.7 %) patients were women and 13 (43.3 %) patients were men. There was also no significant difference regarding mean size of the lesions at the baseline between the drug and placebo groups (p=0.102). However, there was significant difference regarding mean size of lesions at the end of study between the pimecrolimus and placebo groups . In addition, epithelization index (E.I.) was significantly different at the end of study in favor of pimecrolimus group (P < 0.0001). Pimecrolimus can be used  as an effective and safe adjunctive treatment of oral lesions of pemphigus vulgaris.

27. Journal of Cell and Tissue Research Vol. 8(3): 1645-1648 (2008)

The efficacy of topical cream composed of vitamins K, C, E  and Coq10 in  the treatment of infraorbital melanosis and wrinkling: An open label, self-controlled study

Enshaieh, S., Jooya, A.,  Iraji, F.,  Asilian, A., Mazloomi, R.,  Mousavi, S.A., Khorvash, F., Asadi, M., Ayati, R. and Mehrpour, K.

Department of Dermatology, Isfahan University of Medical Sciences, Isfahan, Iran. E. mail: aj351350@yahoo.com

Abstract: In present contribution we evaluated the efficacy of a cream formulation that was composed of Vitamin K, Vitamin C, Vitamin E and Coenzyme Q10 in treatment of periorbital darkening and wrinkles. Twenty-nine adult Iranian volunteers with dark under-eye circles with or without wrinkles were enrolled in this open label study. Informed consent was obtained from all of the patients. Patients who had used eye contour cream in the last 2 months or patients with known allergy to the ingredients of cream were excluded from the study.  Also patients who had any dermatologic lesions around the eyes were also excluded. Periorbital darkening and wrinkles were evaluated by a physician and by the patients themselves, using a digital camera and a visual analogue scale respectively, after 6 weeks of treatment. The patients, feed back regarding improvement rating of infraorbital darkening was  the following: 2 patients (10%) reported mild improvement, 12 patients (60%) reported moderate improvement and 6 patients (30%) reported excellent efficacy. According to physicians rating, 100% of the patients with infraorbital melanosis and 63.15% of the patients with periorbital wrinkling had improved melanosis or wrinkling respectively, although with variable results. According to our results, all of the patients with infraorbital melanosis and roughly half of the patients with periorbital wrinkling had improved melanosis or wrinkling respectively, albeit with variable results.