RESUMEN
Background Six Sigma is a widely accepted quality management system that provides an objective assessment of analytical methods and instrumentation. Six Sigma scale typically runs from 0 to 6, with sigma value above 6 being considered adequate and 3 sigma being considered the minimal acceptable performance for a process. Methodology Sigma metrics of 10 biochemistry parameters, namely glucose, triglycerides, high-density lipoprotein (HDL), albumin, direct bilirubin, alanine transaminase, aspartate transaminase, urea nitrogen, creatinine and uric acid, and hematology parameters such as hemoglobin (Hb), total leucocyte count (TLC), packed cell volume (PCV), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), and platelet were calculated by analyzing internal quality control (IQC) data of 3 months (June-August 2019). Results Sigma value was found to be > 6 for triglyceride, HDL, Hb, TLC, and MCH, signifying excellent results and no further modification with respect to IQC. Sigma value was between 3 and 6 for glucose, albumin, creatinine, uric acid, PCV, and MCHC, implying the requirement of improvement in quality control (QC) processes. Sigma value of < 3 was seen in AST, ALT, direct bilirubin, urea nitrogen, platelet, and MCV, signifying suboptimal performance. Discussion Six Sigma provides a more quantitative framework for evaluating process performance with evidence for process improvement and describes how many sigmas fit within the tolerance limits. Thus, for parameters with sigma value < 3, duplicate testing of the sample along with three QCs three times a day may be used along with stringent Westgard rules for rejecting a run. Conclusion Sigma metrics help assess analytical methodologies and augment laboratory performance.
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The cytochrome P450 (CYP) enzymes, involved in the metabolism of therapeutic drugs, are the major determinants of drug half-life. From a drug industry perspective, variability in drug response owing to CYP polymorphisms makes CYP profiling a commercially interesting option for diagnosis, prognosis and predicting response to drug treatment. Recent studies highlighting microRNA-mediated regulation of CYP genes represents a major advance in our understanding of variations in individual drug responses. Herein we review new perspectives on the molecular mechanisms of CYP regulation and genotyping technologies. Together, these developments present novel therapeutic opportunities and help to explain the integrated response of cells to xenobiotic drug metabolism.
Asunto(s)
Sistema Enzimático del Citocromo P-450/genética , Sistema Enzimático del Citocromo P-450/metabolismo , Xenobióticos/farmacocinética , Animales , Genotipo , Semivida , Humanos , Inactivación Metabólica , MicroARNs/genética , Polimorfismo GenéticoRESUMEN
SIRT1, human ortholog of yeast SIR2 protein, deacetylates histones and several other transcription factors. Recently, SIRT1 has emerged as a drug target for treating age related diseases, type II diabetes, neurodegeneration, inflammation and cancer. Here, we have optimized production of functionally active wild type full-length SIRT1 protein and its N-terminal deleted mutants. In a comparative study, we found that the region containing 192-208 amino acids towards the N-terminus is critical for right conformational folding of the protein to retain its deacetylase activity. The EC(50) and IC(50) values obtained with standard modulators showed that the SRT(748) & SRT(556) can deacetylate substrate and are activated by resveratrol, whereas, deacetylase activity of all the other deletion mutants (SRT(540), SRT(532), SRT(507) and SRT(503)) was lost. We further report that the peptide substrate K(m) for SRT(748) (70+/-5.2 microM) was comparable to SRT(556) (93+/-5.4 microM). The K(m) for NAD(+) substrate was 176 & 274 microM for SRT(748) and SRT(556), respectively. Similar substrate affinity studies demonstrate that either of the protein (SRT(748) or SRT(556)) can be utilized for screening SIRT1 modulators. We have also examined critical regions in SIRT1 required for deacetylase activity as well as kinetic analyses of SIRT1 proteins.