Current Issue : April - June Volume : 2019 Issue Number : 2 Articles : 5 Articles
Measuring oxidative stress has become increasingly valuable in ecological studies, especially\nwhen different markers are measured on the same individual. However, many of the\ncurrent methods lack sensitivity for analysis of low blood volume samples, which represent\na challenge for longitudinal field studies of small organisms. Small blood volumes can usually\nonly be analysed by using a single assay, therefore providing limited information on individualâ??s\noxidative profile. In this study, we used blood collected from a population of wild\neastern chipmunks (Tamias striatus) and modified methods presented in the literature to\nimprove analytical selectivity and sensitivity required for small blood volumes. Specifically,\nwe proposed a modified malondialdehyde (MDA) analysis protocol by HPLC and also optimized\nboth the uric acid independent ferric reducing antioxidant power (FRAP) and hypochlorous\nacid shock capacity (HASC) assays. Development of the three modified methods\nwas achieved with a sensitivity and repeatability that meets standards of field ecology while\nallowing measurement of all three assays in duplicate using less than..............
In metabolic diagnostics, there is an emerging need for a comprehensive test to acquire\na complete view of metabolite status. Here, we describe a non-quantitative direct-infusion\nhigh-resolution mass spectrometry (DI-HRMS) based metabolomics method and evaluate the method\nfor both dried blood spots (DBS) and plasma. 110 DBS of 42 patients harboring 23 different inborn\nerrors of metabolism (IEM) and 86 plasma samples of 38 patients harboring 21 different IEM were\nanalyzed using DI-HRMS. A peak calling pipeline developed in R programming language provided\nZ-scores for -1875 mass peaks corresponding to-3835 metabolite annotations (including isomers)\nper sample. Based on metabolite Z-scores, patients were assigned a â??most probable diagnosisâ?? by\nan investigator blinded for the known diagnoses of the patients. Based on DBS sample analysis,\n37/42 of the patients, corresponding to 22/23 IEM, could be correctly assigned a â??most probable\ndiagnosisâ??. Plasma sample analysis, resulted in a correct â??most probable diagnosisâ?? in 32/38 of the\npatients, corresponding to 19/21 IEM. The added clinical value of the method was illustrated by\na case wherein DI-HRMS metabolomics aided interpretation of a variant of unknown significance\n(VUS) identified by whole-exome sequencing. In summary, non-quantitative DI-HRMS metabolomics\nin DBS and plasma is a very consistent, high-throughput and nonselective method for investigating\nthe metabolome in genetic disease....
The cell-free genomic DNA (gDNA) concentration in serum ranges from 1500 to\n7500 copies/mL within 2 h after phlebotomy (6â??24 times the concentration observed in plasma).\nHere, we aimed to evaluate the gDNA size distribution in serum with time after coagulation and\nto test if crude serum can be directly used as a source of gDNA for qPCR. Next, we investigated if\nsingle nucleotide polymorphisms (SNPs) could be genotyped directly from the crude serum isolated\nfrom capillary blood using a hand-powered paper centrifuge. All tested PCR targets (65, 100, 202 and\n688 base pairs) could be successfully amplified from DNA extracted from serum, irrespective of\ntheir amplicon size. The observed qPCR quantitation cycles suggested that the genomic DNA yield\nincreased in serum with incubation at room temperature. Additionally, only 65 and 101 base pair\nqPCR targets could be amplified from crude serum soon after the coagulation. Incubation for 4 days at\nroom temperature was necessary for the amplification of PCR targets of 202 base pairs. The 688 base\npair qPCR target could not be amplified from serum directly. Lastly, serum was successfully separated\nfrom capillary blood using the proposed paper centrifuge and the genotypes were assigned by testing\nthe crude serum using allele-specific qPCR, producing small amplicon sizes in complete agreement\nwith the genotypes assigned by testing the DNA extracted from whole blood. The serum can be used\ndirectly as the template in qPCR for SNP genotyping, especially if small amplicon sizes are applied.\nThis shortcut in the SNP genotyping process could further molecular point-of-care diagnostics due to\nelimination of the DNA extraction step....
Guinep is traditionally used in the management of cardiovascular ailments. This study\naims to evaluate its medicinal constituents and effects in the management of myocardial injury in\nan experimental isoproterenol (ISO) rat model. Sprague-Dawley rats were randomly assigned to four\ngroups: Group 1 was the control group; Group 2 received M. bijugatus extract (100 mg/Kg; MB) for\nsix weeks; Group 3 was given ISO (85 mg/Kg) i.p. twice during a 24-hour period; and Group 4 was\ngiven ISO (85 mg/Kg) i.p. and MB extract (100 mg/Kg) for six weeks. The MB was administered\norally by gavage, daily. The blood pressure of conscious animals was measured, while ECG was\nperformed under anesthesia. Blood and serum were collected for biochemical and hematological\nanalysis. The ISO group treated with MB showed a significant decrease............
In sperm proteomic experiments round cells and leukocyte proteins are profiled along with\nsperm proteome. The influence of round cell and leukocyte proteins on the sperm proteome has\nnot been investigated. The objective of this study was to identify if the proteins from round cells,\nincluding leukocytes, interfere with the proteomic analysis of spermatozoa in frozen semen samples.\nProteomic profiling of sperm was performed using liquid chromatography-tandem mass spectrometry\nin four groups: Group 1 contained neat semen with round cells and leukocytes.................
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