Anil B. Patel

Erythrocyte Abnormality in Human Myopathy

Erythrocyte ghosts isolated from myopathic patients responded to 10-4 molar ouabain with a dramatic increase in adenosine triphosphatase activity, while identical preparations from normal donors were inhibited by the same drug. These results have been interpreted in terms of a disease-related change in membrane integrity bearing upon function of the transport enzyme.

Gas-liquid chromatography of nucleosides. Effect of silylating reagents and solvents.

The aims of this investigation were to study the completeness of silylation of nucleosides with three different reagents, bis(trimethylsilyl)trifluoroacetamide (BSTFA), bis(trimethylsilyl)acetamide (BSA) and trimethylsilylimidazole (TMSI), and to investigate the effect of different solvents (acetonitrile, pyridine, dimethylformamide, chloroform, methylene chloride, hexane, benzene, and toluene) on quantitation of derivatization. Closed-tube silylations of the nucleosides were performed with BSTFA, BSA, and TMSI, and for the most complete silylation, the optimal time, temperature, and molar excess of reagent were: for BSTFA, 150 degrees-15 min and 225 molar excess; for TMSI, 60 degrees-3 h and 1000 molar excess; and for BSA, 120 degrees-2 h and 250 molar excess. Also, silylations of seven major and minor nucleosides were carried out using a 1000 molar excess of BSTFA, BSA, and TMSI at 25 degrees with 5 min sonication, and at optimal silylation conditions as described above for the three reagents. The silylating strengths were determined by the increase in RWR (= weight response of nucleoside/weight response of pyrene) values, and are summarized for the amino group containing nucleosides silylated at room temperature as BSTFA greater than TMSI greater than BSA, and for silylation under optimal conditions as BSTFA greater than BSA greater than TMSI. The efficiency of silylation for the hydroxyl group-containing nucleosides silylated at room temperature was BSTFA greater than TMSI greater than BSA, and for silylation under optimal conditions BSTFA greater than TMSI = BSA...

Glycoside effect upon membrane enzymes of erythrocytes and muscle in duck myopathy

Skelettmuskelmembrane und Erythrozytenstromata normaler Enten zeigen eine adenosine triphosphatase Aktivität, die durch das herzaktive Glykosid Ouabain gehemmt wird. Das gleiche Material von Enten mit Myopathien zeigte eine durch Ouabain stimulierte ATPase-Aktivität.

Derivatization and chromatography of nucleosides and nucleotides

The aims of this study were to determine the effect of levels of various substances and reaction by-products, which are formed during hydrolysis of nucleic acids, on the derivatization and chromatography of nucleosides; and to investigate the silylation of mono- and dinucleotides. The effect of NaCl, KCl, MgCl2, NH4Cl, and (NH4)2SO4 on silylation and chromatography of nucleosides was studies at various molar excesses of salt. The response values for all nucleosides were studied at various molar excesses of salt. The response values for all nucleosides were significantly affected at molar excess salt present values (MSP) between 1 and 10 for KCl, NaCl, NH4Cl, (NH4)2SO4 and between 0.1 and 1 for MgCl2. It was noted that thymidine was more sensitive than other nucleosides if silylated in presence of these salts. Two chromatographic peaks at retention temperatures (RT) 240 and 251 were obtained for cytidine at MSP values of 10(-3) for NaCl, KCl, and MgCl2, and 10(-4) for NH4Cl and (NH4)2SO4. In a mixture of nucleosides the RT = 251 peak was used for quantitative analysis of cytidine as the RT = 240 peak elutes with guanosine. Thus, these salts have a significant effect on the gas-liquid chromatography of trimethylsilyl (TMS) cytidine in a mixture of nucleosides, especially the RT = 241 peak. The effect of salts on derivatization can be explained in part as follows: (a) reduced derivatization of nucleosides due to a decreased solubility in the solvent system; (b) formation of TMS anion derivatives, e.g. TMS-SO4, TMS-PO4, with a reduced molar excess of BSTFA; (c) metal chelation by Mg ions or other divalent cations with nucleosides or BSTFA; and/or (d) an increased breakdown of TMS derivatives in presence of salt in the sample or on the top 3 in. of the column packing. Also, experiments were made on the effect of other substances such as Tris, phosphate, alkaline phosphatase, and KCl on completeness of silylation. The individual impurities showed no significant effect on the relative weight response (RWR) values of nucleosides; however, when a mixture was used, significantly lower RWR values were observed for all nucleosides except thymidine when using 1000 molar excess of BSTFA greater than 1000 should be used for silylation and chromatography of nucleosides in an RNA hydrolysate. As reported earlier the best derivatization of nucleosides was achieved using closed tube silylation at 150 degrees for 15 min with 225 molar excess BSTFA and chromatography on 4% OV-11 on Supelcoport. In general, the presence of salts and other substances can be significant in quantitative work, thus it is suggested that they be removed using chromatographic cleanup methods. The stability of nucleosides as a function of concentration of HCl, at room temperature was studied and very low RWR values for nucleosides were obtained when stored for 48 h in greater than 0.001 N HCl. Trimethylsilylation of various nucleotides and dinucleotides were made at 15 min as a function of temperature, and at 150 degrees at different times...

Analytical Application of Microcalorimetry

Abstract A “bench top” microcalorimeter has been designed and constructed. This instrument, and a more conventional microcalorimeter previously described, have been applied to several analytical determinations including both inorganic and enzymatic reactions. Because of the stability, yet rapid equilibration time of the bench top calorimeter, multiple analyses may be performed. The ease of operation, nearly universal applicability, and the possibility of obtaining thermodynamic as well as kinetic data simultaneously, make this technique extremely useful.

CO-binding pigment (P-450) and other electron transport components in hepatoma bearing rats

Basierend auf früheren Eiweissuntersuchungen bezüglich einer endoplasmatisch-retikulären Elektronenstromkette ergibt sich: NADPH-Oxydase und Zytochrom-c-Reduktase sind in der normalen Rattenleber wie in der Hepatoma (Typ 7777) tragenden gleich, während die Ferricyanid-Reduktase und P-450 in der Leber der letzteren herabgesetzt waren.

Abstract OT3-3-03: Silent risk? Cardiovascular risk factors in survivors of breast cancer treated with anthracyclines.

Withdrawn by Author Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr OT3-3-03.