Project description:Aim: To explore the potentially important role of circulating miRNAs in systemic inflammation during sepsis. Method: 8 to 10-week old C57BL/6 mice underwent cecal ligation and puncture (CLP) or laparotomy (Sham). Plasma RNA was isolated using Trizol LS reagent 24h post-surgery. NGS cDNA libraries were prepared using Norgen Biotek Small RNA Library Prep Kit. Library quality was validated prior to sequencing on an Illumina NextSeq 500 platform.
Project description:In this study, we performed deep sequencing and bioinformatics analyses of tea plant leaves to identify and characterize known and novel miRNAs. A total of 26,876,261 raw reads were produced from 2 libraries. We detected 422 known miRNAs belonging to 125 families, and 68 putative novel miRNAs.
Project description:AimsAim was to elucidate the specific role of pattern recognition receptors in vascular dysfunction during polymicrobial sepsis (colon ascendens stent peritonitis, CASP).Methods and resultsVascular contractility of C57BL/6 (wildtype) mice and mice deficient for Toll-like receptor 2/4/9 (TLR2-D, TLR4-D, TLR9-D) or CD14 (CD14-D) was measured 18 h following CASP. mRNA expression of pro- (Tumor Necrosis Factor-α (TNFα), Interleukin (IL)-1β, IL-6) and anti-inflammatory cytokines (IL-10) and of vascular inducible NO-Synthase (iNOS) was determined using RT-qPCR. Wildtype mice exhibited a significant loss of vascular contractility after CASP. This was aggravated in TLR2-D mice, blunted in TLR4-D animals and abolished in TLR9-D and CD14-D animals. TNF-α expression was significantly up-regulated after CASP in wildtype and TLR2-D animals, but not in mice deficient for TLR4, -9 or CD14. iNOS was significantly up-regulated in TLR2-D animals only. TLR2-D animals showed significantly higher levels of TLR4, -9 and CD14. Application of H154-ODN, a TLR9 antagonist, attenuated CASP-induced cytokine release and vascular dysfunction in wildtype mice.ConclusionsWithin our model, CD14 and TLR9 play a decisive role for the development of vascular dysfunction and thus can be effectively antagonized using H154-ODN. TLR2-D animals are more prone to polymicrobial sepsis, presumably due to up-regulation of TLR4, 9 and CD14.
Project description:<p>There are currently few datasets describing baseline expression levels for total cell-free circulating RNA from healthy control subjects. Total extracellular RNA was isolated from plasma, urine, and saliva samples from healthy controls. We sequenced small RNAs from 183 plasma samples, 204 urine samples and 46 saliva samples from 55 college athletes ages 18-25 years. Many of the participants provided more than one sample, weeks or months apart, allowing us to assess variability in an individual's exRNA expression levels over time. Several individuals provided all three biofluid types at one time, producing data on individual expression levels across several biofluid types. Here we provide a systematic analysis of small exRNAs present in each biofluid, as well as an analysis of exogenous RNAs. We find that a large number of RNA fragments in plasma (63%) and urine (54%) have sequences that are assigned to YRNA and tRNA fragments respectively. Surprisingly, while many miRNAs can be detected, there are few miRNAs that are consistently detected in all samples from a single biofluid.</p>
Project description:Description of the global expression of microRNAs (miRNAs) and proteins in healthy human term placentas may increase our knowledge of molecular biological pathways that are important for normal fetal growth and development in term pregnancy. The aim of this study was to explore the global expression of miRNAs and proteins, and to point out functions of importance in healthy term placentas. Placental samples (n = 19) were identified in a local biobank. All samples were from uncomplicated term pregnancies with vaginal births and healthy, normal weight newborns. Next-generation sequencing and nano-scale liquid chromatographic tandem mass spectrometry were used to analyse miRNA and protein expression, respectively. A total of 895 mature miRNAs and 6,523 proteins were detected in the placentas, of which 123 miRNAs and 346 proteins were highly abundant. The miRNAs were in high degree mapped to chromosomes 19, 14 and X. Analysis of the highly abundant miRNAs and proteins showed several significantly predicted functions in common, including immune and inflammatory response, lipid metabolism and development of the nervous system. The predicted function inflammatory response may reflect normal vaginal delivery, while lipid metabolism and neurodevelopment may be important processes for the term fetus. The data presented in this study, including supplementary information with complete miRNA and protein findings, will enhance the knowledge base for future research in the field of placental function and pathology.
Project description:We used next-generation sequencing (NGS) to sequence and differentially quantitate miRNAs in 10 pools of plasma derived from individuals with sepsis and SIRS.Plasma pools were preferred to individual samples because they decrease the impact of individual outliers on the analysis. Total RNA was then extracted from equal volumes of plasma and technical duplicates of cDNA libraries for Illumina NGS created. Results from 10 pools representative of 89 individuals (including no-SIRS controls) are uploaded to the repository.
Project description:To describe the transcriptional changes associated with polymicrobial-sepsis induced myocardial depression in wild type and iNOS deficient mice. Keywords: myocardium, contractility, differential gene expression, nitric oxide synthase, infection We compared the transcriptional profile of C57/BL6 WT mice and congenic B6 129P2-Nos2tm1Lau/J mice after 48 hrs of polymicrobial sepsis induced by caecal ligation and perforation. 48 hours after surgery, mice were anaesthetised (intraperitoneal 100 mg/kg ketamine and 10 mg/kg xylazine). The right common carotid artery was cannulated (Millar Mikro-Tip pressure transducing catheter: 1.4F sensor, 2F catheter; Houston TX). Pressure tracings from the aorta and left ventricle were recorded (SonoLAB software; Sonometrics Corp., London Ontario Canada) and analysed using Cardiosoft and Origin 6.0 (Sonometrics Corp., and Microcal Software, Northampton MA). The heart was removed, emptied of blood, and snap frozen.