A specific and sensitive enzyme activity assay for aldehyde dehydrogenase 1A1 (ALDH1A1)

Shoeb, M., Xiao, T.L., Hodge, R.P. and Ansari, N.H.

Department of Biochemistry and Molecular Biology, 1Department of Pharmacology and Toxicology,  University of Texas Medical Branch, Galveston, Texas 77555-0647, USA

Abstract: Oxidative stress is known to induce age-related cataract, the leading cause of blindness worldwide. 4-Hydroxynonenal (HNE) is a cytotoxic lipid peroxidation product generated in abundance   in the lens epithelium under conditions of oxidative stress and if not detoxified, can result in the loss of lens transparency and promote cataractogenesis. Aldehyde dehydrogenase 1A1 (ALDH1A1) is an NAD-dependent ALDH isozyme that oxidizes HNE to form 4-hydroxynonenoic acid (HNA). Previous studies have demonstrated the essential role of ALDH1A1 in the maintenance of lens clarity as well as viability of human lens epithelial cells (HLEC) under conditions of oxidative stress. To further investigate the physiological role of this enzyme a convenient enzyme activity assay is necessary.  Due to the overlapping substrate specificity of ALDH1A1 with the other ALDH isozymes, it is difficult to determine the enzyme activity of ALDH1A1 in the tissues without its purification.  Here we standardized a specific and sensitive fluorescent method to assay ALDH1A1 in ocular tissues using a synthetic substrate, 7-methoxy-1-napthaldehyde (MONAL-71). The ALDH1A1 activity determined by this method was approximately 10 times higher than with propionaldehyde as the substrate measured spectrophotometrically. Moreover, compared to the spectrophotometric assay, the fluorometric assay detected higher specific activity of ALDH1A1 in bovine and rat lens epithelium as compared to other ocular tissues consistent to the presence of ALDH1A1 as the main ALDH isozyme in the lens epithelium, shown earlier. These results suggest that fluorometric method using MONAL-71 is specific and sensitive for the determination of ALDH1A1 activity and can be used as an effective tool for further investigating the role of ALDH1A1 in the lens and other tissues.

2. Journal of Cell and Tissue Research Vol. 8(1) 1211-1224 (2008)
Differential functional modulation of the death  proteins in in vivo and in vitro microenvironments in experimental brain tumors (glioma)

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

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

Abstract: Experiments in field of oncology mostly require an in vitro experimental setup. However, the in vitro tumor environment may not always simulate the in vivo milieu. The basic aim was thus to find out the differential modulation of the same proteins in the two environments. For the purpose, the most extensively studied proteins- “the death proteins” were chosen of which Fas/FasL belonged to the surface receptor group, while the cytosolic proteins of choice were the intrinsic death regulators- Bcl-2, Bax, p53, caspase-8, caspase-9 and caspase-3. A distinct disparity in orientation of the surface receptors was observed with that of the cytosolic proteins when the two microenvironments were compared. The death receptor (in this case Fas/CD95) of freshly isolated (in vivo) glioma cells helps the cells to escape immune surveillance while the same receptors in the culture microenvironment primes it towards an apoptotic death. In contrast, the cytosolic death regulators (p53, Bcl-2, Bax ) of cultured glioma cells as compared to freshly isolated  cells strongly stabilizes the cultured cells in favor of survival. The caspases also behaved differentially in the two microenvironment. The study clearly points out that the same proteins orients themselves in different functional patterns in two microenvironments even though the cells belonged to the same tumor type. Though it is known that the culture microenvironment does not always suffice the in vivo condition, the study is the first of its kind in analyzing, documenting and elucidating the functional disparity of the same proteins in two aforesaid microenvironments.

3. Journal of Cell and Tissue Research Vol. 8(1) 1225-1232 (2008)
Preliminary studies on comparison between β-carotene of  prokaryotic and eukaryotic algae: Spirulina platensis sm-pc1 and Dunaliella salina sm-bc1

Joshi, V. S., Desai, P. B.  and Mishra, S.

Central Salt and Marine Chemicals Research Institute, Bhavnagar-364 002. E. mail: smishra@csmcri.org

Abstract: The present study deals with comparison of β-carotene from prokaryotic & eukaryotic algae i.e. Spirulina platensis & Dunaliella salina (rich source of β-carotene) isolated from fresh water and marine habitat respectively. The pigment composition of the prokaryote is reported to differ from eukaryote. An attempt to have a preliminary data on the variation existing between the two natural & synthetic β–carotene is made. All the other analyses such as UV-Vis, FT-IR, MS, and HPLC were done for the determination of β-carotene from both Dunaliella as well as Spirulina spp. The results obtained through FT-IR shows the presence of sharp peak indicating cis isomer(741 cm-1) in Dunaliella salina while in Spirulina platensis the peaks of both cis (739 cm-1) and trans (965 cm-1) isomers were present. HPLC analysis using C18 column successfully eluted out β-carotene approximately after 60 min. The molecular weights of the β-carotene from all the three sources were found to be 537.6 by Mass spectrometry.

4. Journal of Cell and Tissue Research Vol. 8(1) 1233-1238 (2008)
Role of endostatin gene polymorphism in the development of high myopia

Sandhya, A., Vishnupriya, S., Hema Bindu, Ch., and Prasad Reddy, K.

Department of Genetics, Osmania University, Hyderabad 500 007, E. mail: rsattivishnupriya@yahoo.com

Abstract: Myopia is a common ocular disorder characterized by refractive error affecting 30% of general population. High myopia (>6D) is caused by excessive axial elongation that primarily involves the ora-equatorial area and the posterior pole. Pathological myopia often accompanied by glaucoma, cataracts, macular degeneration, and retinal detachment, gradually leading to vision loss. Recent population and family based studies provided evidence for a genetic-component to pathologic myopia. Study on potential candidate genes for myopia through the genomic study of pathological myopia can help in identification of myopia genes. During myopia, retinal defocus effects mechanosensitivity of the eye causing excessive axial elongation, which is associated with altered scleral collagen architecture and defective mechanotransduction. Ocular signal transduction from retina-RPE complexes is mainly effected during scleral remodeling. The most important effectors in scleral coats are collagens, proteoglycans, Matrix metalloproteinases (MMP’s) and tissue inhibitors of matrix proteinases (TiMP’s). The difference of SNPs in different genes might contribute the formation of scleral thinning during myopia development. The endostatin (20 KDa C-terminal fragment of collagen XVIII) has an essential role in ocular development and the maintenance of visual function. In Age-dependent loss of vision the endostatin mutant is associated with pathological accumulation of deposits under the retinal pigment epithelium. Endostatin RNA also blocks the axis duplication induced by beta catenin, partially inhibits Wnt-dependent transcription, and stimulates degradation of beta catenin during the lens development of eye.  Hence, the present study on high myopia (n=152) has been planned to analyze collagen XVIII/endostatin gene polymorphism (D104N) and to understand its functional significance in ocular growth. The association of endostatin gene was analyzed through PCR-RFLP method which resulted in 3 genotypes (DD, DN, and NN). The comparison of gene and genotype frequencies calculated revealed increase in DN genotype frequency in females, non-familial cases, in cases with age at onset group >20yrs and refractive error above >10D. The results indicate that there is risk for myopia progression in individuals with this genotype. Further studies on endostatin can throw light on better understanding of its role in ocular development and help in management of myopia.

5. Journal of Cell and Tissue Research Vol. 8(1) 1239-1242 (2008)
Immuno-cytological localization of progesterone in corpus luteum of goat

Batra, S. and Sharma, R.K.

Reproductive Biology Laboratory, Department of Zoology, Kurukshetra University, Kurukshetra – 136119, India. E. mail: soniabatra2dec@rediffmail.com

Abstract: Immunocytochemistry was used to localize progesterone in the steroidogenic cells of corpus luteum of goat. Variations in the relative abundance of immunologically labelled gold particles within the steroidogenic cells were analysed in corpus luteum of small (1-5 days),  regressing (16-21 days) and that of pregnancy (< 30 days). In corpora lutea of small category labelled gold particles in granulosa luteal cells were 3.00/cm2 and in theca luteal cells were 2.45/cm2 of cytoplasm at 19000X. These values were more than those observed in steroidogenic cells of corpora lutea of regressing category. The number of gold particles of both the theca and granulosa luteal cells were maximum during pregnancy. The variations in immuno-labelled particles shall be discussed in relation to their physiological obligations at specific stage of reproductive cycle.

6. Journal of Cell and Tissue Research Vol. 8(1) 1243-1248 (2008)

Purification and characterization of abrin toxin from white Abrus  precatorius seeds

Kumar, O.,  Kannoji, A.,  Jayaraj,  R. and Vijayaraghavan, R.

Division of Pharmacology and Toxicology, Defence Research and Development Establishment,

Gwalior  474 002 (India).  E. mail: omkumar63@rediffmail.com

Abstract: In nature, evolution has created proteins that are toxic to mammalian cells. Well-studied classes of such cytotoxic proteins are plant and bacterial toxins.  Among plant toxins, abrin and ricin are extremely toxic; a single molecule of these toxins is sufficient to kill a cell.  In the present study we have purified and characterized  abrin toxin from white Abrus precatorius seeds. The toxin was extracted from white abrus seeds following 30% and 90% ammonium sulfate saturations. After dialysis crude abrin was purified by ion-exchange chromatography on DEAE-cellulose column. DEAE purified toxin was further purified by size exclusion chromatography using Bio-gel A-0.5. The purity of toxin was checked by polyacrylamide gel electrophoresis (PAGE). The abrin toxin gives a single band under non reduced condition and two bands under reduced condition. No other band was observed. The purity of abrin was also confirmed  by Western blot. Haemagglutination activity was  studied and found to be 1:2. The molecular weight of abrin molecule was found to be 65,000 approximately. The abrin after treatment with 1% b-mercaptoethanol for 3 minutes converted into two peptides of molecular weight 35,000 and 30,000. The LD50 value was found to be 7.1 (1.5 to 33.9, 95 % confidence limit) mg/kg for male mice through i.p. route for a 7 day observation period. The percent yield of abrin toxin was approximately 2.4 percent of whole abrus seeds.

7. Journal of Cell and Tissue Research Vol. 8(1) 1249-1252 (2008)
Genome size determination of Zanthoxylum oxyphyllum and Meyna spinosa by flow cytometry: A preliminary study

Buragohain, J. and Konwar, B. K.

Department of Botany, Namrup College, P.O. Parbatpur-786 623, Depatment of Molecular Biology and Biotechnology, Tezpur University, Napaam-784028, India. E-mail: jitu_buragohain@yahoo.co.uk

Abstract: An attempt was made to determine the genome size of Zanthoxylum oxyphyllum and Meyna spinosa by flow cytometry using Pisum sativum as reference standard. A one step DNA flow cytometry procedure with slight modifications was followed for isolation and estimation of nuclear DNA content. The 2C nuclear DNA content of Z. oxyphyllum and M. spinosa were found to be 7.590 and 7.862 pg respectively. Knowing of genome size and DNA content of these two plants would provide a useful tool for future molecular biological investigations.

8. Journal of Cell and Tissue Research Vol. 8(1) 1253-1256 (2008)
Comparison between the efficacy of a herbal drug and minoxidil in the treatment of the telogen effluvium

Enshaieh, S., Jooya, A., Rashnoo, H., Siadat, A.H., Sadeghinia, A., Asilian, A. and  Djahehd Nojoucambary, R.

Dermatology Department, Isfahan Medical School, Isfahan University of Medical Sciences and Health Services, Isfahan, Iran. E. Mail: amirhossein1@yahoo.com

Abstract:  Telogen effluvium (TE) is a self-limited, non-scarring disease that shows itself as a diffuse hair loss of the scalp that usually occurs 3 months after an acute illness. Theorically , Minoxidil solution, has been suggested as a treatment for this condition. In this study, we compared the efficacy of the minoxidil and a herbal drug containing Urtica diocia, Chamomilla, Thymus vulgaris, Equisetum avenues and Foeniculum vulgare in the telogen effluvium. It was a double – blind prospective study in that 24 patients with telogen effluvium were allocated randomly in two groups and were treated with minoxidil and the herbal drug. Data were analyzed statistically. The mean duration of telogen effluvium in the minoxidil group was 17 weeks and in the herbal drug group was 7 weeks. This difference was statistically significant. Our finding confirms the good efficacy of this herbal solution in treatment of TE.

9. Journal of Cell and Tissue Research Vol. 8(1) 1257-1259 (2008)
Pro and antioxidant systems in juvenile, rheumatoid  and osteo  arthritis

Padmini, A., Venkateshwari, A., Ravi Babu and Pratibha, N.

Department of Genetics, Osmania University, Hyderabad  500 007.  E. mail: pratinallari@yahoo.com

Abstract: Arthritis is a chronic inflammatory disease with an interaction of various genetic and environmental factors. It is a heterogenous group of disorder with different etiological and pathogenetic mechanisms. Lipid peroxidation is the prevailing mechanism in various inflammatory disorders. Hence the present study is aimed at evaluating the status of pro and antioxidants in the etiology of arthritis. Blood samples from 291 arthritis and 125 healthy controls matched for age and sex have been collected from Orthopedic Unit of Vaidya Vidhan Parishad, King koti , Hyderabad. Levels of malondialdehyde, ceruloplasmin and nitric oxide were analysed in serum and plasma samples by following appropriate methods. Mean levels of malondialdehyde was found to be significantly elevated in rheumatoid and osteoarthritis compared to controls where as a slight decrease was observed in juvenile arthritis compared to juvenile controls.  Ceruloplasmin was also found to be elevated in juvenile arthritis compared to controls indicating the role of CP as an antioxidant in the etiology of Juvenile arthritis. Nitric oxide levels were significantly elevated in rheumatoid arthritis compared to control subjects. The findings can be correlated to the homeostatic role of pro/anti oxidant mechanisms resulting in delineating the subset of arthritis with juvenile arthritis being purely genetic and autoimmune rheumatoid arthritis being inflammatory change with oxidative  stress and osteoarthritis resulting due to the disturbance in homeostasis of pro and antioxidants.

10. Journal of Cell and Tissue Research Vol. 8(1) 1261-1263 (2008)
Role of chymotrypsin alpha  and beta  isozymes  in peptic ulcers

Sulekha, S., Venkateshwari, A., Pratap, B. and Pratibha, N.

Department of Genetics, Osmania University, Hyderabad  500 007. E. mail: pratinallari@yahoo.com

Abstract: Peptic ulcer is a multifactorial disorder with different etiological and pathogenetic mechanisms.  The present study is aimed at identifying the role of chymotrypsin in the etiology of peptic ulcers.  Endoscopically confirmed 210 duodenal ulcers, 49 gastric ulcers and 50 gastric cancers and 185 age and sex matched  control subjects were considered for the  study .  The findings highlighted the association of chymotrypsin  a and b  isozymes with the disease group compared to controls.  This implicates the association of a and b complex with increased proteolytic activity and tissue damage associated with these inflammatory disorders. Since H. pylori is one of the risk factor in the etiology of peptic ulcer the levels were compared in H. pylori positive and negative cases and found a significant increase there by indicating the  host pathogen interactions at the physiological level.

11. Journal of Cell and Tissue Research Vol. 8(1) 1265-1269 (2008)
Assessment of antimicrobial activity for the smaller chain dipeptides and tripeptides
Nagarajan, K., Senthamarai, R., Devi, K., Deepa Shalini, S., Anandh, N., Krishnaveni, P., Mazumder, A., Ghosh, L.K. and Umadevi, G.

Department of  Pharmaceutical Chemistry, Periyar College of  Pharmaceutical Sciences for Girls, Trichy 620 021, E. mail: nagarajan_mph@yahoo.co.in

Abstract: Disc diffusion method was employed to determine the antimicrobial effect of test compounds I, II and III (Trp-Phe-Asn, Gly-Pro, Gly-Asn) against eight microbial species viz., Staphylococcus aureus, Staphylococcus albus, Streptococcus fecalis, Escherichia coli, Proteus vulgaris, Pseudomonoas aeruginosa, Klebsiella aerogenes and Candida albicans. The disc was saturated with 100 ml of compound I and 200 ml of compound II and III, allowed to dry and introduced on the upper layer of seeded agar plate. The plates were incubated over night at 37oC. Microbial growth was determined by measuring the zonal inhibition diameters. Compound I showed maximum potency against gram positive S. aureus (28 mm) in comparison with standard Ciprofloxacin (40 mm). Compound I was found to be highly sensitive against S. albus (20 mm), S. fecalis (16 mm) and E. Coli (15 mm). Compound II shows good potency against S. aureus (13 mm) and S. albus (10 mm). Compound III was found to be moderately sensitive against  S. aureus (8 mm) and Proteus vulgaris (10 mm). Among all the compounds tested, none of them was found to be effective against the fungi Candida albicans.

12. Journal of Cell and Tissue Research Vol. 8(1) 1271-1274 (2008)
Larvicidal activity of indigenous entomopathogenic soil bacteria from Northeastern region of India

Baruah, I., Laishram, S.K.S., Das, S.C. and Kalita, J.

Defence Research Laboratory (DRDO), P.B No. 2, Tezpur  784001, India.  E. mail: indrabaruah@yahoo.co.in

Abstract: During a search for entomopathogenic bacteria, 1154 bacterial isolates were obtained from 246 soil samples of different mosquito breeding habitats of Sonitpur district of Assam, India. Seven bacterial isolates (0.6%) that showed mosquito larvicidal activity was identified as Bacillus sphaericus GC subgroup IV (4), Bacillus brevis (2) and Bacillus circulans (1). LC50  and LC90 values of B. sphaericus GC subgroup IV against Culex quinquefasiatus, Anopheles stephensi and Aedes albopictus were 0.02mg/l and 0.05mg/l, 0.03 mg/l and 0.06 mg/l and 1.0 mg/l and 1.8 mg/l respectively. While LC50 and LC90 values of B. brevis and B. circulans against Cx. quinquefasiatus was recorded as 0.2 mg/l and 0.4 mg/l and 1.8 and 3.0 mg/l respectively. B. sphaericus GC subgroup IV was found more potential when compared to the commercial formulations Spherix with LC50 and LC90 values, as 0.025 mg/l and 0.06 mg/l while that of BG6262 were 0.03 mg/land 0.075 mg/l respectively. It was revealed that B.  phaericus GC subgroup IV was highly toxic, B. brevis moderately toxic while B. circulans was poorly toxic against the tested mosquito larvae.

13. Journal of Cell and Tissue Research Vol. 8(1) 1275-1280 (2008)
Efficacy of herbal and inherent antioxidants on hexavalent chromium induced lipid peroxidation: A comparative study

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

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

Abstract: Hexavalent chromium exerts its effect on the cellular system by generating free radicals and increasing lipid peroxidation. Therefore, any agent that reverse these phenomena would certainly prove to be an ideal therapeutic agent.  An experiment was set in laboratory taking developing chick (broiler) as an experimental model. The animals were intoxicated with Cr(VI) with a daily dose of 10mg/Kg/day for 7 days and thereafter, administered with physiological  (vitamin B complex, vitamin E, glutathione) and herbal products (ashwagandha and garlic) for another 7 days and screened for GSH, sulfhydral groups,  glutathione peroxidase and malondialdehyde levels in liver, kidney and serum. Study shows decrease of first three components and increase in malondialdehyde content indicating the increase of lipid peroxidation. Application of therapeutics reversed all the parameters which reveal decreased lipid peroxidation. Further, the study also shows that vitamins and glutathione show better results than herbal products suggesting long term therapy is require in later case. 

14.  Journal of Cell and Tissue Research Vol. 8(1) 1281-1284 (2008)
A case of 18q- deletion with balanced translocation t(15;18)(p13;q21.3) in the mother

Chandel, D.

Division of Human Genetics, Department of Zoology, Gujarat University, Ahmedabad   380 009, India. E. mail: divya_chandel@yahoo.com

Abstract: The 18q deletion syndrome can be caused by several terminal and interstitial deletions of which terminal deletions of the distal part of 18q are known as the De Grouchy syndrome. A female infant with distal 18q deletion confirmed by G-banding is reported, who was referred for delayed development. Subsequent karyotyping in parents revealed that the deletion was inherited from the mother who carried a balanced reciprocal translocation between chromosome 15 and 18 - 46,XX,t(15;18)(p13;q21.3).

 15. Journal of Cell and Tissue Research Vol. 8(1) 1285-1287 (2008)
Evaluation of some medicinal plants of North-Eastern India for antibacterial activity:
A preliminary study

Laishram, S.K.S., Das, N.G., Bailung, B., Dube, S.N. and   Srivastava, R.B. Defence Research Laboratory (DRDO), Post bag no. 2, Tezpur 784001, India. E. mail: shantiks@rediffmail.com

Abstract: The antibacterial activity of methanolic and ethanolic extracts of four plants from Sonitpur district of Assam was evaluated against five common and clinically significant bacterial pathogens viz.  Staphylococcus aureus (MTCC-96), Escherichia coli (MTCC-739), Klebsiella pneumoniae (MTCC-3040), Pseudomonas aeruginosa (MTCC-2582) and Shigella flexneri (MTCC-1457) by Agar well diffusion method. S. aureus was found to be the most sensitive while K. pneumoniae was the least sensitive among the tested pathogens. The ethanolic extract of Piper thomsoni exhibits the maximum inhibition against S. aureus.

16. Journal of Cell and Tissue Research Vol. 8(1) 1289-1292 (2008)
Comparative antihyperglycemic activity of alcoholic leaf and bark extract of Wrightia tomentosa in streptozotocin induced diabetic rats

Nagarajan, K., Mazumder, A.  and Ghosh, L.K.

Department of Pharmaceutical Chemistry, Periyar College of  Pharmaceutical Sciences for Girls, Trichy  620 021, India. E. mail: nagarajan_mph @ yahoo.co.in

Abstract: Ethanolic bark and leaf extract of  Wrightia tomentosa (200 and 400 mg/Kg) given 48 h after streptozotocin treatment in rats decreased the blood glucose level significantly to greater extent (88.5 ± 10.13 mg/dl  for 400 mg/Kg of  bark extract ) after 10 days indicating the antihyperglycemic activity of the plant product.  

17. Journal of Cell and Tissue Research Vol. 8(1) 1293-1296 (2008)
An outbreak of bacterial gill disease in Indian major carps, Cirrhinus mrigala and Labeo rohita

Mastan, S. A.

Post Graduate Department of Biotechnology, D. N. R. College, Bhimavaram 534202, India.
E. mail: shaikmastan2000@yahoo.com

Abstract: An outbreak of bacterial gill disease (BGD) was reported during the months of April, May and June, 2004 from lower Lake of Bhopal. A total of 1,058 fishes were screened. Except two species namely, Cirrhinus mrigala and Labeo rohita, no other fish species were found suffering from this disease. The infected fishes exhibited the symptoms such as abnormal swimming, lethargic, anorexic and some times, stayed near the surface of water. Their respiratory rate was increased and comparatively high mucus secretion was observed on the gills. The bases of gills become swollen and in severe cases, the gills become rotten and reddish in appearance. Bacteriological examination indicated that, the bacterium involved resembled most closely to the genus Cytophaga. The experimental infection trials indicated that this bacterium is the etiological agent of the disease and is capable of producing disease symptoms.

18. Journal of Cell and Tissue Research Vol. 8(1) 1297-1302  (2008)
Isolation, purification and characterization of venom toxins from indian red scorpion, Mesobuthus tamulus

Upadhyay, R. K. and Ahmad, S.

Department of  Zoology, Deen Dayal Upadhyay Gorakhpur University, Gorakhpur 273009, India.
E. mail: rkupadhaya@yahoo.com

Abstract: Venom proteins from the scorpion Mesobuthus tamulus were extracted. Three major toxin/protein peaks were resolved in continuous fractions at 280 nm, which were also confirmed in the buffer at 640 nm. The molecular weights of three different toxins/proteins were estimated by plotting elution volumes against standard known proteins. From gel filtration chromatogram three proteins were obtained each one of them having molecular weights 64kD (Sco I), 42kD (Sco II) and 36kD (Sco III) respectively. The LD50 of the whole venom and purified toxins was determined in insect and in mice. Toxins were also tested for hemotoxicity in mice and human in vitro, which gave an EC50 value of 2.16 µg/ml of blood for mice and 19.4 µg/ml for human blood. Purified toxins were injected into mice after mixing with Freund’s adjuvant for production of antiserum. After 7 days animals were bled to obtain antiserum. Anti-serum was partially purified by octanoic acid precipitation and antibody specificity was tested by performing immuno-double diffusion test. A visible bow shaped band of precipitin complex of antigen-antibody was obtained. Serotherapy was also performed to neutralize the effect of venom toxin in albino mice.

19. Journal of Cell and Tissue Research Vol. 8(1) 1303-1302 (2008)
Electron microscopic radioautographic study on mitochondrial DNA synthesis in adrenal cortical cells of developing mice
Nagata, T.

Department of Anatomy and Cell Biology, Shinshu University School of Medicine, Matsumoto 390 8621 and Department of Anatomy, Shinshu Institute of Alternative Medicine, Nagano 380 0816, Japan
E mail: nagatas@po.cnet.ne.jp
Abstract: In order to study the aging changes of intramitochondrial DNA synthesis of mouse adreno-cortical cells, 5 groups of developing mice, each consisting of 3 individuals, total 15, from fetal day 19 to postnatal day 1, 3, 9 and 14 were injected with 3H-thymidine, sacrificed 1 hr later and the adrenal tissues were processed for electron microscopic radioautography. On EM radioautograms obtained from each animal, number of mitochondria and the mitochondrial labeling index labeled with 3H-thymidine showing DNA synthesis in each adreno-cortical cells, in 3 zones, were counted and the results in respective developing groups were compared. From the results, it was demonstrated that the numbers of mitochondria and the labeling indices of intramitochondrial DNA syntheses in zona glomerulosa, fasciculata and reticularis of mice at various ages increased from fetal day 19 to postnatal day 14 due to development of animals.

20. Journal of Cell and Tissue Research Vol. 8(1) 1313-1316 (2008)

Influence of the fruit and leaf extract of Psidium guajava linn. on wound healing in wistar rats

Mudliar, V. S., Patil, P. A., Torgal, S. S., Malur, P. R., Mittal, R.

Department of Pharmacology  and Pharmacotherapeutic,  J. N. Medical College, Belgaum 590010, India. E. mail: drpapatil@yahoo.co.in

Abstract: Leaf extract of guava (Psidium guajava Linn.) has been reported to promote wound healing in traditional medicine but there is paucity of information in literature regarding similar activity of fruit extract. Therefore, the present study was planned to investigate the influence of leaf and fruit extract of Psidium guajava Linn. on re-sutured incision, excision, and dead space wounds in male Wistar rats. Wounds were inflicted under light ether anesthesia aseptically. Control animals received vehicle and other groups received aqueous extract of either leaf or fruit orally in the dose of 500mg/kg/day for a period of 10 days in the incision and dead space wounds, whereas treatment continued till complete wound closure in excision wound model. On the 10th day after estimation of breaking strength of the resutured incision wounds, animals were sacrificed and granulation tissue from dead space wounds were used to estimate the breaking strength and hydroxyproline content. Quantification of granulation tissue and histopathological slides were also carried out. Wound closure rate, epithelization time and scar features were studied in the excision wounds from the day of extract administration till complete closure of wounds. Only aqueous fruit extract significantly (P<0.01) promoted the healing process in all three wound models studied. Histopathological slides revealed increased collagen content and granulation tissue in fruit extract treated group as compared to that of control. In contrast, the leaf extract delayed excision wound healing and had no effect on incision and dead space wounds. These findings merit, clinical evaluation of Psidium gaujava fruit.

21. Journal of Cell and Tissue Research Vol. 8(1) 1317-1322 (2008)

Development of latex agglutination test for the rapid detection of abrin toxin

Kumar, O.,  Shakya, B. and Vijayaraghavan, R.

Division of Pharmacology and Toxicology, Defence Research and Development Establishment,

Gwalior  474 002 (India).  E. mail: omkumar63@rediffmail.com

Abstract : Abrin is a plant toxin from Abrus precatorious seeds and extremely toxic in nature. Abrin inhibits protein synthesis by inactivating ribosomes in an irreversible manner. Plant toxins (abrin and ricin) are to be considered as potentials bio-terrorism  or biological weapons. The development of a fast and sensitive method for the detection of biological agents is an important tool to prevent or deal with the consequences of intoxication due to these toxins. In the present study abrin was purified and polyclonal antibodies were raised in rabbits. Latex  particles were sensitized by coating purified IgG antibodies. The optimum binding of antibodies to latex particles was 97 percent.  Latex agglutination test was standardized for the detection of abrin. The 95 to 97 percent saturated latex particles can detect abrin up to 62.5 mg/ml.   The above-developed reagent (sensitized latex particles) was stable for a period of four months without loss in its sensitivity at 4oC and room temperature. The developed field based test system is sensitive, rapid and does not require trained person and instrumentation.

22.  Journal of Cell and Tissue Research Vol. 8(1) 1323-1326 (2008)

Polymorphisms of glutathione-s-transferase genes GSTM1 and GSTT1 and risk for breast cancer

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

Department of Genetics, Osmania University, Hyderabad 500 007, India.
Email:  drsattivishnupriya@yahoo.com

Abstract: The glutathione-S-transferases (GSTs) are involved in the metabolism of xenobiotics, including an array of environmental carcinogens, chemotherapeutic agents and also free radicals generated during oxidative stress. A group of 250 breast cancer patients including 9 male breast cancer cases and controls were analyzed for GSTM1 and GSTT1 polymorphisms by using multiplex PCR method. The frequency of GSTM1 null genotype (M0), GSTT1 null genotype (T0) and GST double null (T0M0) did not show any significant association with breast cancer. M0 (38.4%, p=0.03) genotype frequency was significantly elevated in postmenopausal patients when compared to premenopausal patients. T0 (13.6%, 15.6%) genotype frequency was also increased in overweight and obese women with breast cancer. When the occupation of patients was considered M0 (48.1%) genotype frequency was elevated in women who are exposed to agricultural pesticides. T0 (15.4%) frequency was elevated among patients with receptor status positive for HER2/neu. Double null (T0M0) genotype was found to be elevated among breast cancer patients who were positive for progesterone and HER2/neu receptor status, negative nodal status and with obesity.  In conclusion, our data suggested that GSTM1 and GSTT1 genes might be acting independently in the development and the clinical outcome of the of breast cancer.

23.  Journal of Cell and Tissue Research Vol. 8(1) 1327-1332 (2008)

Effect of Piper betel and Argyreia speciosa extracts on brain acetylcholinesterase and monoamines concentrations in mice

Bodhankar, S. L. and Vyawahare, N.S.

Bharati Vidyapeet University, Poona College of Pharmacy, Erandwane, Pune 411038,  1AISSMS College of Pharmacy, Kennedy Road, Near R.T.O, Pune-4110001.E. mail: neerajsv@rediffmail.com

Abstract: In present investigation effects of ten days administration of  two doses (200 mg and 400 mg/kg)of hydroalcoholic extracts of leaves of Piper betel (PB) and roots of Argyreia speciosa (AS), on brain acetylcholinesterase, dopamine, serotonin (5-HT)  and noradrenaline levels was explored in mice. Both these plants are traditionally claimed as nootropic plants and have effect on learning and memory. Present study showed that the brain AChE level was unaltered with both the extracts. The brain dopamine concentration was found to be reduced significantly with both the doses of AS and higher dose of PB. The lower dose of PB increased level of dopamine in brain. Serotonin level was significantly increased with PB  and decreased with AS, both with lower and higher doses. Contrary to these, noradrenaline was decreased in all the cases. The significance of these alterations has been discussed in the contribution.

Assessment of antinutritional factors and protein content in the seeds of  chickpea cultivars and their divergence

Bhagyawant,  S. S. and Srivastava, N.

School of Studies in Biotechnology, Jwajji  University, Gwalior  474 011, India.
E. mail: bhagyawant123@rediffmail.com

Abstract: Legume grains constitute the primary source of diet protein for the population of many countries including India. However, these legume seeds are also known to contain factors like protease and amylase inhibitors as well as polyphenols etc. which are nutritionally antagonistic. These may reduce the availability of otherwise good proteins in the diet and cause diseases originating from malnutrition. However, presently protein antimetabolites are getting much attention due to their role in defense against insects or microbes. Biochemical characterization of these antinutritional factors in chickpea seed cultivars is attempted in the present investigation.

25.  Journal of Cell and Tissue Research Vol. 8(1) 1339-1345 (2008)

Sub-acute toxicity of fluoxetine hydrochloride on testis of reserpine-depressed Wistar rats

Unnikrishnan, G., Shivabalan, R. and  John, L.

Department of Zoology, Birla College, Kalyan,  421304, E. mail: eeta_ukrishnan@hotmail.com

Abstract: The present study was designed to investigate the effect of a commonly used antidepressant drug fluoxetrine hydrochloride (FH) on male reproduction. Reserpine depressed   Wistar male rats were exposed to a sub- acute toxicity analysis using a low (20mg/kg) and a high (80mg/kg) doses of FH.  The treatment with FH enhanced the food and water intake in low doses and induced hypoglycemic condition irrespective of the dose levels. Exposure of depressed  rats to higher doses of  FH also induced hypercholestremia and decreased the serum testosterone, testicular proteins and cholesterol, reduced the number of epididymal sperms and altered the histo-architecture of testis  suggesting its adverse effect on reproduction. Thus, FH in lower doses can well be tolerated by the animals but higher doses may affect the reproductive system and may interfere with fertility.  The data suggest that higher doses for prolonged  duration depressed individuals should not be recommended.