Development and Validation of New HPLC Method for Simultaneous Estimation of L-Lysine Hydrochloride and L-Carnitine-L-Tartrate in Pharmaceutical Dosage Form.

A new analytical method is developed and validates for simultaneous estimation of L-lysine hydrochloride and L-carnitine-L-tartrate in single pharmaceutical dosage form by means of high resolution HPLC. The chromatographic procedure was carried out using C18-5 µm (4.6 mm × 250 mm), column. Optimally suited mobile phase was made to run in an isocratic elution mode whose composition was 10 mM of potassium dihydrogen phosphate adjusted with triethylamine to a pH of 7.5 and pumped at a flow rate of 0.50 ml/min through chromatographic system. The detector's wavelength was 214 nm. The accuracy of the analytical method was determined keeping into account the recovery of analytes, which in this case remains well within the accepted standards that is 95-105%. Detection limit for L-lysine hydrochloride and L-carnitine-L-tartrate are 1.47 µg/ml and 0.85 µg/ml while quantitation limit for L-lysine hydrochloride and L-carnitine-L-tartrate is 4.41 and 2.55 µg/ml, respectively. All the statistical results were validated performing precision, accuracy, linearity, specificity studies for analytes concentration from 70-130%. The outcome of results confirmed that the method was in consonance with the acceptance criteria.

Stability-indicating High-performance Liquid Chromatography Method for Simultaneous Determination of Aminophylline and Chlorpheniramine Maleate in Pharmaceutical Formulations.

The present work deals with the development and validation of method for simultaneous determination of antihistaminic drugs in pharmaceutical formulations. A precise, specific and accurate reverse phase-high-performance liquid chromatography method for the simultaneous measurement of aminophylline and chlorpheniramine maleate was developed. The separation of drugs was achieved on C-18 (5 µm, 250×4.6 mm) high-performance liquid chromatography column. The runtime for analysis was 10 min. Mobile phase is mixture containing dilute H2SO4:methanol (60:40% v/v) with flow rate adjusted at 1.5 ml/min. The detection of components was performed at a wavelength of 264 nm. Retention times of aminophylline and chlorphinramine maleate were found to be 2.00 and 3.25 min, respectively. Linearity was found in the range of 16-24 µg/ml for chlorpheniramine maleate and 102.4-153.6 µg/ml for aminophylline with a correlation coefficient of 0.9998 and 0.9996, respectively. High peak purity index of 99.99% indicated the complete separation of analytes in the presence of degradation products is justification of method stability. Linearity, accuracy, specificity, precision and robustness studies were performed for method validation.

Macroautophagy inhibition sensitizes tamoxifen-resistant breast cancer cells and enhances mitochondrial depolarization.

Macroautophagy (autophagy), a process for lysosomal degradation of organelles and long-lived proteins, has been linked to various pathologies including cancer and to the cellular response to anticancer therapies. In the human estrogen receptor positive MCF7 breast adenocarcinoma cell line, treatment with the endocrine therapeutic tamoxifen was shown previously to induce cell cycle arrest, cell death, and autophagy. To investigate specifically the role of autophagy in tamoxifen treated breast cancer cell lines, we used a siRNA approach, targeting three different autophagy genes, Atg5, Beclin-1, and Atg7. We found that knockdown of autophagy, in combination with tamoxifen in MCF7 cells, results in decreased cell viability concomitant with increased mitochondrial-mediated apoptosis. The combination of autophagy knockdown and tamoxifen treatment similarly resulted in reduced cell viability in the breast cancer cell lines, estrogen receptor positive T-47D and tamoxifen-resistant MCF7-HER2. Together, these results indicate that autophagy has a primary pro-survival role following tamoxifen treatment, and suggest that autophagy knockdown may be useful in a combination therapy setting to sensitize breast cancer cells, including tamoxifen-resistant breast cancer cells, to tamoxifen therapy.

Postembryonic expression of the myosin heavy chain genes in the limb, tail, and heart muscles of metamorphosing amphibian tadpoles.

Thyroid hormone is presumed to play a role in initiating and/or orchestrating the postembryonic expression of the genes encoding isoforms of the myosin heavy chains (MHCs) that characterize the muscle fibres in an adult organism. The fact that the postembryonic development of a free-living amphibian tadpole takes place during its thyroid hormone-dependent metamorphosis has made the metamorphosing tadpole an ideal system for elucidating the molecular mechanism(s) by which this hormone affects these postembryonic changes. In this review, we summarize the results from recent studies focused on the postembryonic expression of the MHC genes in the skeletal muscles and hearts of metamorphosing anuran (Rana catesbeiana) tadpoles. The demonstration that mRNAs encoding at least five of the MHC isoforms present in the tadpole tail muscles are also present in the adult hind-limb muscles and that an mRNA encoding a cardiac-specific MHC isoform is present in the heart of both the tadpole and adult organism, rules out the possibility that thyroid hormone initiates the expression of these MHC genes. Instead, it seems more likely that this hormone acts by modulating the expression of one or more of the genes encoding these particular MHC isoforms. Whatever the case, the fact that sequence homology suggests that the five distinct skeletal muscle-specific MHCs are all "fast" isoforms raises the question of how these MHCs are distributed among the three different fibre types described for Rana. On the other hand, the possibility exists that the mRNAs for one or more of these fast MHC isoforms encode developmental isoforms that are present but not translated in the muscles of the tadpole and/or adult frog. Finally, an evaluation of the evolutionary relatedness of the R. catesbeiana MHCs to the MHCs in another species of Rana and to the MHCs in other vertebrates discloses, among other things, that the nucleotide sequence in the R. catesbeiana cardiac MHC isoform is more closely related to the chicken ventricular MHC isoform than it is to any of the other MHC isoforms examined.