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Dhananjaya B.L.,University of Mysore | Dhananjaya B.L.,Adichunchanagiri Biotechnology and Cancer Research Institute | Dhananjaya B.L.,Central University of Kerala | Gowda T.V.,University of Mysore | J.M. D'Souza C.,University of Mysore
Cell Biochemistry and Function | Year: 2010

Pharmacologically active 5' nucleotidase is a ubiquitously distributed enzyme in snake venoms. In this study the effect of concanavalin A (Con-A) on different snake venoms 5' nucleotidase activity is tested in order to know the protein nature which will ultimately help in purification of the enzyme with high yield. Con-A inhibited Naja naja, Naja kauthia, Naja melanoleuca, Naja naja sputatrix, Agistrodon halys blomhoffii, Bothrops asper and Oxyranus scutellas venom 5' nucleotidase activity at different concentrations. This indicates the presence of glycopart in the protein, thus glycoprotein in nature. Vipera russellii, Vipera plaestenae, Agistrodon contratrix, Bitis orientis, Echis carinatus and Trimeresures malabaricus was not inhibited by Con-A, indicating absence of glycopart in the protein. This study for the first time shows existence of 5' nucleotidase in multimeric forms. © 2010 John Wiley & Sons, Ltd.

Srinivas L.,Adichunchanagiri Biotechnology and Cancer Research Institute
Journal of Biomedical Science | Year: 2014

Background: Snake venoms are a complex mixture of active principles mainly peptides and proteins also including amino acids, nucleotides, free lipids, carbohydrates and metallic elements bound to proteins that interfere in several biological systems. In this study, we aimed to understand the mode of action of the apoptosis inducing ability of Naja naja venom phospholipase A2 (NV-PLA2) using isolated human peripheral lymphocytes. Results: Human peripheral lymphocytes when incubated with Naja naja venom phospholipase A 2 (NV-PLA2) induced up to 68% DNA fragmentation. The dialysed conditioned media obtained by incubating lymphocytes with NV-PLA 2 at 15th min induced 44% DNA fragmentation, referred to as cmlp-active. Cmlp-active showed 20.5% increased protein concentration than the corresponding control condition media cmlp-c-15. Test for creatine kinase activity in cmlp-active proved negative and negligible amount of lactate dehydrogenase did not show significant DNA fragmentation. Fractionation of cmlp-active on Sephadex G-25 showed two peaks, major peak induced 38% DNA fragmentation, which was further rechromatographed on Sephadex G-25. The single peak obtained was named PID15 (Phospholipase A 2 Induced DNA fragmentation factor secreted at 15th min). Q-Tof MS/MS analysis of PID-15 showed it is a 6 kDa peptide. PID15 sequence analysis gave 40 amino acids in the following order, msilpcknvs iwvikdtaas dkevvlgsdr aikflylatg. The homology search for the sequence revealed it to be an Apoptosis Inducing Factor (AIF). Conclusion: Results indicate that the secretion of PID15 is dependent on concentration of NV-PLA2 treatment, incubation time and also on temperature and the probable membrane origin of PID15 and not of cytosolic origin with apoptosis inducing ability. © 2014 CHETHANKUMAR and SRINIVAS; licensee BioMed Central.

Sivapriya M.,Aurigene Discovery Technologies | Gowda S.S.T.,Adichunchanagiri Biotechnology and Cancer Research Institute | Srinivas L.,Adichunchanagiri Biotechnology and Cancer Research Institute
Journal of Membrane Biology | Year: 2015

Lipid peroxidation by ROS at the membrane level disturbs the inherit integrity of components activating subsequent alterations in the function. In this study, the protective effect of purified Sundakai (Solanum torvum) seed protein (SP) was tested against oxidative membrane damage in erythrocyte membrane. SP prevented oxidative RBC lysis induced by pro-oxidants; Fe:As (2:20 μmol), periodate (0.4 mM), and t-BOOH (1 mM) up to 86, 81, and 86 %, respectively. Further, SP prevented the Fe:As-induced K+ leakage up to the tune of 95 %. The inhibition offered by SP on K+ leakage was comparable to inhibition offered by quinine sulfate, a known K+ channel blocker. SP dose dependently restored Na+K+ ATPase and Ca2+Mg2+ ATPase activities in erythrocyte membrane. The restoration of ATPase activity by SP was two times more than standard antioxidants BHA and α-tocopherol. Besides, SP at 1.6 μmol restored the membrane proteins over Fe:As induction when analyzed by SDS-PAGE, which was comparable to protection offered by BHA. In conclusion, SP is an effective antioxidant in preventing oxidative membrane damage and associated functions mediated by ROS. As SP is non-toxic, it can be used as an effective bioprotective antioxidant agent to cellular components. © 2015, Springer Science+Business Media New York.

Ramadas D.,Adichunchanagiri Biotechnology and Cancer Research Institute | Srinivas A.,Adichunchanagiri Biotechnology and Cancer Research Institute
Asian Journal of Pharmaceutical and Clinical Research | Year: 2011

A 28 kDaglyco protein isolated, purified and characterized from boiling water extract of Turmeric (Curcuma longa L) and named as BGS-Haridrin. The protein showed sharp single peak in RP-HPLC with a retention time of 32.6 min. The apparent molecular weight of BGS-Haridrin was w28 kDa showed in SDS PAGE and MALDI/MS analysis. BGS-Haridrin scavenged hydroxyl, DPPH radicals, superoxide radicals 76-82 % about and inhibited lipid peroxidation about 78% at a maximum dosage of 0.9 n M concentration when compared to BHA, Curcumin (400 μM) and α-tocopherol (400μM). BGS Haridrin effectively protects H2O2 (100μM) induced cell death in human peripheral lymphocytes. Further BGS-Haridrin prevents H2O2 (1mM) caused calf thymus DNA damage as evidenced by agarose gel electrophoresis. In summary, the results represent in vitroantioxidant effects of BGS-Haridrin antioxidant protein from Turmeric (Curcuma longa L).

Itgappa M.,S. S. I. M. S. and R. C. | SubhasChandrappa M.,S. S. I. M. S. and R. C. | VidyaNadigere M.,S. S. I. M. S. and R. C. | Raju G.M.,JJM Medical College | And 2 more authors.
International Journal of Pharmaceutical and Clinical Research | Year: 2011

The present study aim is to determine the most common blood groups for purpose of emergency blood transfusion and to know the high incidence of certain disease in some blood group carriers. The observational cross sectional study was done at department of Bio-chemistry (central Lab) and Pathology at SSI Medical Science &RC, Davanagere, Karnataka of South Indiaduring August 2009 to December 2009. The sample blood was collected by finger prick method. A drop of monoclonal reagent anti A, anti B, and Anti D were added to a drop of blood on clean and fresh glass slides and mixed well with glass rod. Results of agglutination were recorded immediately. During the observation,total number of subjects studied were 600, out of 312 is males and 288 are females. Study showed Blood group-B is more predominant in males. Females O-blood group is more predominant. The Males have Rh positive is 94.57% and 5.13% negative and female 94.44% is Rh positive and 5.66% negative. From the above study we came to a conclusion that, group-B is slightly higher (0.64%) than the group-O in males. But group-O is predominant in Females than group-B by 4.5%. The patterns of Rh factors are similar in both males and females.

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