Current Issue : January - March Volume : 2013 Issue Number : 1 Articles : 9 Articles
Background: Studies have demonstrated that inflammation has a key role in the pathogenesis of atherosclerosis\r\ndue to the abnormal gene expressions of multiple cytokines. We established an accurate and precise method to\r\nobserve gene expression in whole blood that might provide specific diagnostic information for coronary artery\r\ndisease (CAD) and other related diseases.\r\nMethods: The fifteen selected CAD-related genes (IL1B, IL6, IL8, IFNG, MCP-1, VWF, MTHFR, SELL, TNFalpha,\r\nubiquitin, MCSF, ICAM1, ID2, HMOX1 and LDLR) and two housekeeping genes (ACTB and GK) as internal references\r\nhave been measured simultaneously with a newly developed multiplex polymerase chain reaction (multi-PCR)\r\nmethod. Moreover, the precision was evaluated, and a procedure for distinguishing patients from the normal\r\npopulation has been developed based upon analyses of peripheral blood. A total of 148 subjects were divided into\r\ngroup A (control group without plaques), group B (calcified plaques) and group C (non-calcified plaques, and\r\ncombination group) according dual-source CT criteria. Gene expression in blood was analyzed by multi-PCR, and\r\nlevels of glucose and lipids measured in 50 subjects to explore the relationship among them.\r\nResults: The precision results of the multi-PCR system revealed within-run and between-run CV values of\r\n3.695ââ?¬â??12.537% and 4.405ââ?¬â??13.405%, respectively. The profiles of cytokine gene expression in peripheral blood were\r\nset: a positive correlation between glucose and MCSF, HMOX1 or TNFalpha were found. We also found that\r\ntriglyceride levels were negatively correlated with SELL gene expression in 50 subjects. Compared with controls,\r\ngene expression levels of IL1B, IL6, IL8 and MCP-1 increased significantly in group C.\r\nConclusions: A new multiple gene expression analysis system has been developed. The primary data suggested that\r\ngene expression was related to CAD. This system might be used for risk assessment of CVDs and other related diseases....
Biosensor is an analytical equipment in the form of a small portable unit into which test substance is incorporated directly without pre-processing it. It consists of a biological element, detector element and associated electronics or signal processors that are responsible for display of results. In recent years, immuno-sensors have become important due to increased focus on environmental biohazards. Moreover, biosensors have a vast applicability in the fields of food analysis, bio-molecules interaction study, manufacturing of pharmaceuticals, crime detection, medical diagnosis, industrial process control, detection systems for biological warfare agents and organ replacement. Taking advantage of miniaturization, biosensors have become inexpensive and easy-to-handle. Despite the huge potential its application is restricted which can be improved by sensitivity, response time and lifetime. This review focus and throws light on principle, configuration, applications and recent market available biosensors found useful for mankind....
Bioanalytical methods for assessment of drugs is a quintessential part of pre-clinical as well as clinical studies because of its pivotal role in understanding various parameters about a drug like its safety, toxicity, formulation optimization, pharmacokinetic parameters, etc. Bioanalytical methods are widely used to quantitate drugs and their metabolites in physiological matrices and these methods could be applied in study areas of clinical and nonclinical pharmacology/ toxicology. In recent times, LC-MS/MS has become the most dominant instrument for bioanalytical analysis of drugs due to its various advantages. However, many researchers have also developed bioanalytical method by using HPLC with UV as detection system. These methods have been validated as per the guidelines and hence have the applicability in optimum conditions. This review mainly discusses the sample extraction, method development, process optimization and validation parameters on HPLC with focus on the utility of reversed phase column and UV as detection system in a bioanalytical method development....
A simple, specific, sensitive and rapid reverse phase high performance liquid chromatographic (HPLC) method for the determination of irbesartan (IRB) in small volumes of rat plasma was developed and validated. Biological sample preparation involved simple extraction with organic solvent, followed by dilution with mobile phase to eliminate any chromatographic solvent effects. IRB was quantitated on a C8 column (4.6 mm i.d. × 300 mm length), using a mobile phase composed of methanol: acetonitrile (70:30 %v/v) which was delivered at a flow rate of 1.0 mL/min. The method was proven to be linear over a plasma concentration range of 10 -1000 ng/mL with a mean correlation coefficient of 0.9913. The intra-day and inter-day precision (coefficient of variation) were in the range of 1.9% to 4.3% and 2.7% to 5.4%, respectively. The intra-day accuracy (relative error) was in the range of 2.2% to 5.1% and the inter-day accuracy were in the range of 3.4% to 5.8%. The mean recovery of IRB from rat plasma was found to be 96.93%. The limit of detection (LOD) and the limit of quantification (LOQ) of the developed method were determined to be 1 ng/mL and 10 ng/mL respectively. The developed method was successfully applied to quantitatively assess the pharmacokinetics of IRB in wister rats following a single oral dose (6.75 mg/kg). The developed method was established as a rapid analytical tool in a pharmacokinetic study as it required short retention time, high precision, sensitivity and small volumes of plasma for analysis....
History of using natural products of plant and animal origin for healthcare goes back to thousands of years with the existence of various alternative systems of medicines. India has one of the oldest richest and most diverse cultural traditions which were associated with medicinal plants. The history of India shows wide use of products of animal origin in ayurvedic system of medicine. Cow urine has several potential uses, it is necessary to study it scientifically to prove its therapeutic validation and for its worldwide acceptance. The present study was planned to evaluate cow urine by using modern analytical methods. The parameters used for evaluation were organoleptic characters, physicochemical evaluation, and presence of sodium, potassium and trace elements in cow urine by using Flame photometer and ICPAES. This analytical profile was compared with those of Buffalo and Goat urine, which are also mentioned in ancient texts having medicinal properties....
Panchagavya is a blend of five products obtained from cow and used in ayurvedic medicine. They are milk, curd, ghee, urine and dung. Cow urine plays an important role in Hindu practices and its curative properties are claimed in Charak Samhita. In Sushruta Samhita several medicinal properties of cow’s urine have been mentioned. It is known to cause weight loss, reversal of certain cardiac and kidney problems, indigestion, stomach ache, edema, in the treatment of discharging lymph and infested organs etc. The present study was planned to evaluate cow urine by using modern analytical methods. The parameters used for evaluation were presence of allantoin, uric acid and creatinine in cow urine by HPLC and UV-VIS spectroscopic method. This analytical profile was compared with buffalo and goat urine, which are also mentioned in ancient texts having medicinal properties....
Background/Aims: Infants with diabetes insipidus (DI), especially those with impaired thirst mechanism or\r\nhypothalamic hyperphagia, are prone to severe sodium fluctuations, often requiring hospitalization. We aimed to\r\navoid dangerous fluctuations in serum sodium and improve parental independence.\r\nMethods: A 16-month old girl with central DI, absent thirst mechanism and hyperphagia following surgery for\r\nhypothalamic astrocytoma had erratic absorption of oral DDAVP during chemotherapy cycles. She required\r\nprolonged hospitalizations for hypernatremia and hyponatremic seizure. Intensive monitoring of fluid balance,\r\nweight and clinical assessment of hydration were not helpful in predicting serum sodium. Discharge home was\r\ndeemed unsafe. Oral DDAVP was switched to subcutaneous (twice-daily injections, starting with 0.01mcg/dose,\r\nincreasing to 0.024mcg/dose). The parents adjusted daily fluid allocation by sliding-scale, according to the blood\r\nsodium level (measured by handheld i-STAT analyser, Abbott). We adjusted the DDAVP dose if fluid allocation\r\ndiffered from maintenance requirements for 3 consecutive days.\r\nResults: After 2.5 months, sodium was better controlled, with 84% of levels within reference range (135-145 mmol/L)\r\nvs. only 51% on the old regimen (p= 0.0001). The sodium ranged from 132-154 mmol/L, compared to 120ââ?¬â??156 on the\r\nold regimen. She was discharged home.\r\nConclusion: This practical regimen improved sodium control, parental independence, and allowed discharge home....
Background: Activation of innate immunity via pathogen recognition receptors (PRR) modulates adaptive immune\r\nresponses. PRR ligands are being exploited as vaccine adjuvants and as therapeutics, but their utility in diagnostics has not\r\nbeen explored. Interferon-gamma (IFN-c) release assays (IGRAs) are functional T cell assays used to diagnose latent\r\ntuberculosis infection (LTBI); however, novel approaches are needed to improve their sensitivity.\r\nMethods: In vitro immunomodulation of a whole blood IGRA (QuantiFERONH-TB GOLD In-Tube) with Toll-like receptor\r\nagonists poly(I:C), LPS, and imiquimod was performed on blood from subjects with LTBI and negative controls.\r\nResults: In vitro immunomodulation significantly enhanced the response of T cells stimulated with M. tuberculosis antigens\r\nfrom subjects with LTBI but not from uninfected controls. Immunomodulation of IGRA revealed T cell responses in subjects\r\nwith LTBI whose T cells otherwise do not respond to in vitro stimulation with antigens alone. Similar to their in vivo\r\nfunctions, addition of poly(I:C) and LPS to whole blood induced secretion of inflammatory cytokines and IFN-a and\r\nenhanced the surface expression of antigen presenting and costimulatory molecules on antigen presenting cells.\r\nConclusions: In vitro immunomodulation of whole blood IGRA may be an effective strategy for enhancing the sensitivity of\r\nT cells for diagnosis of LTBI....
Gammaretroviruses related to murine leukemia virus (MLV) have variously been reported to be present or absent in blood\r\nfrom chronic fatigue syndrome/myalgic encephalomyelitis (CFS/ME) patients and healthy controls. Using subjects from New\r\nYork State, we have investigated by PCR methods whether MLV-related sequences can be identified in nucleic acids isolated\r\nfrom whole blood or from peripheral blood mononuclear cells (PBMCs) or following PBMC culture. We have also passaged\r\nthe prostate cancer cell line LNCaP following incubation with plasma from patients and controls and assayed nucleic acids\r\nfor viral sequences. We have used 15 sets of primers that can effectively amplify conserved regions of murine endogenous\r\nand exogenous retrovirus sequences. We demonstrate that our PCR assays for MLV-related gag sequences and for mouse\r\nDNA contamination are extremely sensitive. While we have identified MLV-like gag sequences following PCR on human DNA\r\npreparations, we are unable to conclude that these sequences originated in the blood samples....
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