Investigation of transferrin polymorphism in Garole sheep.

The aim of this study was to determine the genetics of polymorph systems of Transferrin in Garole sheep breed. The present study was conducted on 95 adult Garole sheep comprising 52 ewes and 43 rams, maintained at Sheep and Goat Breeding Farm of West Bengal University of Animal and Fishery Sciences, West Bengal, during the period from April-September, 2009. The polymorphism of transferrin was determined through SDS-Polyacrylamide gel electrophoresis technique. It was found that the transferrin type was controlled by five codominant alleles (TfA, TfB, TfC TfD and TfE) in Garole sheep. These five alleles, because of co-dominant nature of inheritance, determined the occurrence of nine transferrin genotypes in the analyzed flock. Four (TfAA, TfBB, TfCC and TfDD) of these were homozygous and the remaining five (TfAD, TfBC, TfBD, TfCD and TfDE) heterozygous. It was found that the TfDD genotype (0.263) was predominant while TfDE genotype (0.042) was least common in the analyzed flock. Frequencies of other genotypes were as: TfCD(0.242), TfBD(0.126), TfCC(0.084), TfBB(0.074), TfAA(0.063), TfAD and TfBC (0.053 for each genotype ) in whole population. From the result it was found that in whole population combined, the heterozygotic genotypic frequency (0.516) was more than that of homozygotic genotypic frequency (0.484). Considerable variations were recognized in the frequencies of transferrin alleles. In the whole population frequencies of transferrin alleles were found to be TfA = 0.089, TfB = 0.163, TfC = 0.232, TfD = 0.495 and TfE = 0.021. Transferrin system has shown an absence of genetic equilibrium among the analyzed herd (chi2 value = 51.31). In conclusion, there were polymorphism in Transferrin types and the presence of differences among the frequencies of the five alleles by categories could be a source of genetic variation in Garole sheep.

Influence of microenvironment pH, humidity, and temperature on the stability of polymorphic and amorphous forms of clopidogrel bisulfate.

The effect of microenvironment pH, humidity, and temperature was evaluated on the stability of polymorphic and amorphous forms of clopidogrel bisulfate, when present alone or in combinations. Oxalic acid and sodium carbonate were used as solid stressors to create acidic and alkaline pH, respectively. The samples without and with stressors were subjected for 3 months to (1) 0% RH, 25% RH, 75% RH, and 85% RH at 40 degrees C and also to (2) 60 degrees C, 80 degrees C, and 100 degrees C at 0% RH. In case of solid samples without stressors, the mixture of polymorphic and amorphous forms showed more degradation than the individual forms above critical relative humidity (85% RH). Similar higher degradation was observed between 75% RH and 85% RH in case of acid-stressed samples. In alkaline microenvironment, all the samples showed identical decomposition attributed to conversion of bisulfate salt to free base. Thermal studies indicated that polymorphic forms of clopidogrel bisulfate and also its glassy amorphous form were highly resistant to temperature, whereas the rubbery state of the drug degraded significantly at temperatures of > or =80 degrees C.

Characterization of degradation products of amorphous and polymorphic forms of clopidogrel bisulphate under solid state stress conditions.

The present study deals with the stress degradation studies on amorphous and polymorphic forms of clopidogrel bisulphate. The objective was to characterize the degradation products and postulate mechanism of decomposition of the drug under solid state stress conditions. For that, amorphous form, polymorph I and polymorph II of the drug were exposed to 40 degrees C/75% relative humidity (RH), with and without stressors for 3 months. The samples were analyzed by HPLC, and the relative extent of degradation as well as nature of decomposition was compared among three solid forms. In total, eight degradation products were observed under various stress conditions. The structures of all of them were elucidated using LC-MS/TOF and LC-MS(n) studies. While one matched the known hydrolytic decomposition product of the drug in solution, seven others were new. The postulated degradation pathway and mechanism of decomposition are discussed.

Detection and characterization of synthetic steroidal and non-steroidal anti-inflammatory drugs in Indian ayurvedic/herbal products using LC-MS/TOF.

It is claimed that ayurvedic/herbal healthcare products (AHPs) are safe because of their natural origin. However, several reports exist of adulteration of AHPs with synthetic drugs. In this study, a generalized strategy was developed using LC-MS/TOF for the detection and verification of steroidal and anti-inflammatory drugs in 58 AHPs collected from various parts of India. The strategy involved recording of mass spectral information for standard drugs-including ionization mode (ESI/APCI - ve or + ve), mass spectrum, accurate mass, identification of qualifier fragments (two), extracted ion chromatograms (EICs), isotopic pattern and determination of UV max (nm)-through UV-PDA studies. Adulteration was then detected in AHPs primarily through comparison of EICs at accurate m/z for molecular ion peaks and R(T) matching with the standard. It was confirmed by spiking with the standards, and matching mass spectrum, accurate mass, R(T) of qualifier fragments, isotopic pattern and UV spectrum of the standards with the adulterant peaks in AHPs. Dexamethasone and diclofenac were detected as adulterants in ten AHPs whereas one AHP tested positive for piroxicam and another for dexamethasone. All the adulterated products were sold by the healthcare practitioners, while no product marketed by manufacturers or chemist shops had this problem. The study showed that LC-MS/TOF-based screening could be used as a rapid approach to monitor adulteration of steroids and anti-inflammatory drugs in AHPs.

Mild synthesis of a family of planar triazinium cations via proton-assisted cyclization of pyridyl containing azo compounds and studies on DNA intercalation.

An efficient synthesis of a family of heteroaromatic triazinium compounds, [2a]X-[2g]X (X = Cl, ClO4, NO3, and HSO4), from 2-(arylazo)pyridines via proton-catalyzed heterocyclization is described. Characterization of the compounds is made by different spectroscopic, electrochemical techniques, as well as single-crystal structure determination of the triflate salt of a representative compound, [2a]CF3SO3. The bond parameters indicate that the tricyclo compound, 2a(+), is planar and aromatic with a N-N bond length of 1.275(6) A. These exhibited fluorescence with an emission maximum in the range of 540-535 nm with moderate quantum yields. The triazinium salts can be reduced in two successive one-electron steps as probed by cyclic voltammetry and coulometry. The paramagnetic radical intermediate 2a(*) is distinguished by a sharp and intense EPR spectrum. Fluorescence spectroscopy, circular dichroism, cyclic voltammetry, viscosity measurements, together with DNA melting studies have been used to characterize the binding of 2a(+) with calf thymus DNA. The emission quenching of the compound by [Fe(CN)6](4-) decreased when bound to DNA. As determined by a MTT assay, 2a(+) exhibited significant cytotoxicity at a higher concentration range of 1 mg/mL to 1 microg/mL; however, the % survival ratio increased with dilution. Cellular uptake studies of the referenced compound were followed by FACS analysis.