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Physico-chemical and antigenic characterization of unconventional heavy chain antibodies of Indian desert camel (Camelus dromedarius L.)

2016-07-20T05:53:13Z

Title: Physico-chemical and antigenic characterization of unconventional heavy chain antibodies of Indian desert camel (Camelus dromedarius L.) Authors: Sehrawat, Sharvan; Singh, Ajit Abstract: Heavy chain antibodies (HCAbs) of IgG2 and IgG3 subtypes were purified from the sera of Indian desert camel (Camelus dromedarius L.) by ammonium sulphate precipitation, followed by ion-exchange chromatography on DEAE-cellulose and affinity chromatography on protein A-sepharose and protein G-sepharose, and characterized by SOS-polyacrylamide gel electrophoresis, agar gel immunodiffusion (AGIO), counter-immunoelectrophoresis (CIEP), immunoelectrophoresis (IEP), ELISA and immunoblotting. IgG2 and IgG3 were found to have molecular mass 46.77 kDa and 43.65 kDa, respectively by SOS-PAGE under reducing conditions. They migrated in β-region in IEP and could be detected in CIEP, because of being more negatively charged and smaller size. Anti-camel IgG3 cross-reacted in AGIO, ELISA and immunoblotting with IgGs of pig and ruminants (cattle, buffalo, sheep and goat), but not with immunoglobulins from horse, dog, guinea pigs, mice, fish, poultry and human. The present findings suggest close antigenic relationship of camels with pigs and ruminants. Page(s): 299-304

Role of protein tyrosine kinase inhibitors in cancer therapeutics

2016-07-20T05:52:12Z

Title: Role of protein tyrosine kinase inhibitors in cancer therapeutics Authors: Bhise, S B; Nalawade, Abhijit D; Wadhawa, Hitesh Abstract: Protein tyrosine kinases (PTKs) are critical in regulating cell growth and differentiation and are deeply involved in several cancers. PTK-inhibitors are mainly ATP-site directed and are finding use in the treatment of several cancers, and more than 30 such agents are now in phase I-III clinical trials. The present review focuses mainly on the development of PTK inhibitors in clinical trials, with special emphasis on imatinib mesylate, a rationally designed, potent oral anticancer agent and selective inhibitor for Abl tyrosine kinase, including Bcr-Abl, C-kit and platelet-derived growth factor-receptor tyrosine kinases, which has been implicated in several malignancies, including chronic myeloid leukemia and gastrointestinal stromal tumour. Page(s): 273-280

Inhibition of the hexokinase/hexose transporter region in the glycosomal membrane of bloodstream Trypanosoma brucei by oligomycin and digitonin

2009-04-02T16:30:58Z

Title: Inhibition of the hexokinase/hexose transporter region in the glycosomal membrane of bloodstream Trypanosoma brucei by oligomycin and digitonin Authors: Njogu, Muturi R; Kiaira, Job K Abstract: Glycolysis in bloodstream T. brucei is the sole source of energy and remains a favourable chemotherapeutic target. In furtherance of this, an attempt has been made to understand better the contribution of glucose, fructose, mannose and glycerol to the energy charge of these parasites incubated in the presence of oligomycin, salicyhydroxamic acid (SHAM) and digitonin. Their cellular energy charge, when catabolizing glucose was 0.860, and under inhibition by oligomycin (10 μg), SHAM (2 mM) or oligomycin plus SHAM, 0.800, 0.444 and 0.405, respectively. Oligomycin inhibited the rate of catabolism of glucose, mannose and fructose up to 80%. The inhibition could not be alleviated by uncouplers, such as 2,4-dinitrophenol or permeabilization of the membranes by digitonin. Glucose-6-phosphate and other phosphorylated glycolytic intermediates, such as fructose-6-phosphate were catabolized by the permeabilized parasites in the presence of oligomycin, implying that except hexokinase, all the other glycolytic enzymes were active. Glucose oxidation was stimulated by low concentrations of digitonin (up to 4 μg), but at higher concentrations, it was significantly inhibited (up to 90% inhibition at 10 μg). Apparently, the inhibitory effects of oligomycin and digitonin were confined to glucose uptake and hexokinase catalysis. The above observations suggest that the hexose transporter and the enzyme hexokinase might be functionally-linked in the glycosomal membrane and oligomycin inhibits the linkage, by using a mechanism not linked to the energy charge of the cell. Digitonin at concentrations higher than 4 μg disrupted the membrane, rendering the complex in-operative. A hexokinase/hexose transporter complex in the glycosomal membrane is envisaged. Page(s): 329-332

Determination of serum triglycerides using lipase, glycerol kinase, glycerol-3-phosphate oxidase and peroxidase co-immobilized onto alkylamine glass beads

2009-04-09T16:30:30Z

Title: Determination of serum triglycerides using lipase, glycerol kinase, glycerol-3-phosphate oxidase and peroxidase co-immobilized onto alkylamine glass beads Authors: Kalia, Vandana; Pundir, Chandra S Abstract: A method for determination of serum triglycerides (Tgs) using lipase, glycerol kinase, glycerol-3-phosphate oxidase and peroxidase co-immobilized onto alkylamine glass beads (pore diameter 55 nm) through glutaralde¬hyde coupling was developed and evaluated. The minimum detection limit of the method was 0.54 mM. The analytical recovery of added triolein in the serum was 97.55±1.5% (mean ± S.D.). The mean value of serum Tgs, determined by the present method showed a good correlation (r=0.984) with the Bayer’s kit method, employing free enzymes. The within and between batch coefficients of variation (CV) were <2.25% and <1.35% respectively. No significant loss of activity was observed, when co-immobilized enzymes were reused for about 200 times and stored at 4°C in distilled water. The cost of Tg determination for 200 serum samples was less, as compared with Bayer’s kit method. Page(s): 326-328