Project description:The use of microbiological cultures for diagnosing bacterial infections in young febrile infants have substantial limitations, including false positive and false negative cultures, and non-ideal turn-around times. Analysis of host genomic expression patterns (“RNA biosignatures”) in response to the presence of specific pathogens, however, may provide an alternate and potentially improved diagnostic approach. This study was designed to define bacterial and non-bacterial RNA biosignatures to distinguish these infections in young febrile infants.
Project description:Bacteria release nano-sized membrane vesicles (MVs) into the extracellular milieu. Bacterial MVs contain molecular cargo originating from the parent bacterium and have important roles in bacterial survival and pathogenesis. Using 8-plex iTRAQ approaches, we profiled the MV proteome of two Escherichia coli strains - uropathogenic E. coli (UPEC) 536 and probiotic Nissle 1917. For these strains, we compared the difference in the MV proteome between a crude input MV prepared by ultracentrifugation alone with that from MVs that were further purified by either density gradient centrifugation (DGC) or size exclusion chromatography (SEC); and also compared MVs from bacterial cultures that were grown in iron-restricted (R) and ironsupplemented (RF) conditions. We found that overall, outer membrane components were highly enriched, and bacterial inner membrane components were significantly depleted in both UPEC and Nissle MVs, in keeping with an outer membrane origin. In addition, we found enrichment of ribosome-related Gene Ontology terms in UPEC MV, and proteins involved in glycolytic process and ligase activity in Nissle MV. We have identified that three proteins (RbsB of UPEC in R; YoeA of UPEC in RF; BamA of Nissle in R) were consistently enriched in the DGC- and SEC-purified MV samples in comparison to their crude input MV, whereas conversely the 60 kDa chaperonin GroEL was enriched in the crude input MVs for both UPEC and Nissle in R condition. Such proteins may have a potential utility as technical markers for assessing the purity from contaminating non-vesicular protein after MV purification. Several proteins were changed in their abundance depending on the iron availability in the media.
Project description:We investigated the effects of the crude extract of a South African medicinal plant, Cotyledon orbiculata, on cell survival of colon (HCT116) cancer cell lines. Using RNASeq, we discovered that the extract interfered with mRNA regulatory pathways. Here, we found that the extract of Cotyledon orbiculata, a South African medicinal plant, had an anti-proliferative effect in cancer cells, mediated by apoptosis induced by alternative splicing of hnRNPA2B1 and BCL2L1.
Project description:This dataset has been used to establish GroEL-proteotyping, a targeted proteotyping approach for assessing bacterial community compositions using the taxonomic marker protein GroEL. This dataset contains raw data of two experiments: 1. Pure cultures of T. aromatica (sample names: TA, TB, TC) and P. putida (sample names: PA, PB, PC) were analyzed individually after in-solution digestion 2. T. aromatica and P. putida were cultivated independently and crude extracts were mixed in defined ratios. GroEL was pre-separated by SDS-PAGE (sample names: LOD_..., in-gel digestion)
Project description:To assess the influences of a crude medicinal herb extract (MHE) on the immune composition and function in inguinal white adipose tissue (iWAT), we isolated iWAT stromal vascular fractions (SVFs) from control (PBS) and MHE-treated mice and performed RNA-seq analysis. iWAT SVF of mice treated with MHE was defined as the treatment group. Phosphate-buffered saline (PBS) treatment was used as the control group. Then RNA-Seq experiment was performed by OE Biotech Co., Ltd. (Shanghai, China) to analyze gene expression changes in iWAT SVF.
Project description:New and rapid antimicrobial susceptibility/resistance testing methods are required for bacteria from positive blood cultures. In the current study we developed and evaluated a targeted LC-MS/MS assay for the detection of beta-lactam, aminoglycoside and fluoroquinolone resistance mechanisms in blood cultures positive for E. coli or K. pneumoniae. Selected targets were the beta-lactamases SHV, TEM, OXA-1-like, CTX-M-1-like, CMY-2-like, chromosomal E. coli AmpC, OXA-48-like, NDM, VIM and KPC, the aminoglycoside modifying enzymes AAC(3)-Ia, AAC(3)-II, AAC(3)-IV, AAC(3)-VI, AAC(6’)-Ib, ANT(2”)-I and APH(3’)-VI, the 16S-RMTases ArmA, RmtB, RmtC and RmtF, the quinolone resistance mechanisms QnrA, QnrB, AAC(6’)-Ib-cr, the wildtype QRDR of GyrA, and for E. coli, the porins OmpC and OmpF. The developed assay was evaluated using 100 prospectively collected positive blood cultures, 100 negative blood cultures inoculated with isolates that were previously collected from blood cultures, and 48 isolates inoculated with isolates carrying genes of less prevalent resistance mechanisms.