Project description:As the importance of transcriptional variation and regulation for Plasmodium becomes more apparent, advances for non-falciparum species are hindered by our reliance on natural infections to study parasite biology. Untargeted transcriptomic research is also complicated by low parasite densities and high proportions of human genetic material, highlighting the need for optimized sample processing protocols. In this study, we used a P. knowlesi culture diluted in whole blood as a mock P. vivax natural infection to compare white blood cell, rRNA-, and globin depletion methods and RNA-seq library preparation kits to create an optimized protocol for low-volume sample processing.
Project description:Untargeted Temporal Analysis of Serum using FPSE after 8 Gy radiation
Time points of room temperature serum analyzed: 1, 3, 24, 72, and 120 hours.
Project description:<p>Blood microsampling (BµS) devices collect less than 100 µL of blood, offering a less invasive and more cost-effective alternative to venipuncture. However, its metabolomic comparability to conventional samples remains unclear, and standardized BµS metabolomic workflows are lacking. This study compared BµS extraction methods and assessed the metabolite coverage of three BµS devices (Mitra®, Capitainer®, and Whatman™ 903) as an alternative to conventional samples (plasma and whole blood) for human biomonitoring. Venous blood from 10 adults (5 males, 5 females) was sampled onto the three devices. First, three extraction techniques (ultrasound, shaker, and homogenizer) were evaluated at three blood concentrations (1.5%, 5.5%, and 11%). The optimized method was then used to compare the metabolite profiles between BµS devices, whole blood, and plasma. Reverse-phase and hydrophilic-interaction chromatography, in positive and negative ionization modes, were combined for Liquid Chromatography–Mass Spectrometry (LC-MS) analysis. All extraction techniques and concentrations proved suitable for BµS untargeted metabolomics. Combining different analytical modes and fragmentation ranges proved useful for maximizing metabolite coverage. BµS-derived metabolite profiles aligned more closely with whole blood than plasma. Some metabolites were more characteristic of a sample type, whereas others were common across sample types. All sample types enabled sex-based differentiation, with metabolites such as testosterone sulfate and hydroxyisovaleric acid driving the separation. These findings enhance our understanding of BµS metabolite coverage and highlight its potential in human biomonitoring. The choice of device depends on the application and the metabolites of interest, offering flexibility for clinical use and research. </p>