Project description:Mice were challneged with Mycobacterium abscessus cell wall particles intra- tracheally or saline. Lung was harvested after 3 weeks. RNA extracted from lungs and RNASeq was performed to identify differentially expressed genes.
Project description:We report the first use of ex vivo lung perfusion (EVLP) in the genetic and physiologic modification of lungs from deceased pulmonary arterial hypertension (PAH) patients and propose this as a translational platform to both (1) derive clinically relevant mechanistic insights into pulmonary pathophysiology and (2) to test treatments on human lungs. The EVLP consist in the perfusion of the lungs out of the body during 6 hours. It is a well established protocol in where basically lungs are on a table connected to a close circuit containing a special perfusion solution that is circulated through the pulmonary vein and artery using a pump. The circuit contains also a deoxygenator. The perfusion temperature and flow are adjusted gradually and after 20 mins of perfusion the ventilation is initiated. Every hour lungs are recruited in order to assess pulmonary function and collect perfusate samples. In addition to perfusate, tissue samples from the lower lobe of the left lung and bronchial alveolar lavage (BAL) are collected at times T0, 3 and 6 hr.
Project description:In-vivo Gene Signatures of Mycobacterium tuberculosis In C3HeB/FeJ Mice In this experiment we have compared Mtb transcriptomics in-vivo, using samples derived from chronically infected C3HeB/FeJ mice, which produce human like caseating lesions, unlike other murine species, to Mtb cultured in-vitro. Similarly, the genome-wide expression of MtbdeldosR, MtbdeldosS and MtbdeldosT mutants is also compared between lungs from C3HeB/FeJ mice and in-vitro culturing.
Project description:Neutrophils are accumulated in mouse lungs following Mycobacterium tuberculosis infections. Increased influx of neutrophils is associated with increased susceptibility to disease, but the underlying mechanisms of neutrophil mediated Tuberculosis disease pathogenesis is poorly understood. To understand the transcriptomic differences between resistant (C57 BL/6) and susceptible (C3HeB/FeJ) mouse lung neutrophils, we sorted them at Day 29 post infection and analyzed them by bulk mRNA sequencing.
Project description:Detection of species-specific proteotypic peptides for accurate and easy characterization of infectious non-tuberculous mycobacteria such as Mycobacterium avium subsp. paratuberculosis, Mycobacterium marinum and Mycobacterium vaccae is essential. Therefore, we carried out reanalysis of publicly available M. avium subsp. paratuberculosis, M. marinum and M. vaccae proteomic dataset PXD027444, PXD003766 and PASS00954 by proteome database search and followed by spectral library generation. The raw DDA data were searched against their respective reference proteome databases using Proteome Discoverer and FragPipe. The resulting peptide spectrum matches were converted into a spectral library using BiblioSpec.
Project description:A growing body of evidence suggests that epithelial cells have special roles during pneumonia. The purpose of this study was to elucidate epithelial-specific responses during lung infection. Mice received intratracheal instillations of Streptococcus pneumoniae (10^6 CFU) into the left lung. 15 hours after the challenge, left lungs were collected. Single cell supensions were generated and sorted into 2 separate populations, epithelial cells (CD45-EpCAM+) and others (all non-epithelial cells). Epithelial cells from uninfected mice were also collected.
Project description:The “ship of the desert”, the one-humped Arabian camel (Camelus dromedarius), has a remarkable capacity to survive in conditions of extreme heat without needing to drink water. One of the ways that this is achieved is through the actions of the antidiuretic hormone vasopressin (AVP) and the natriuretic hormone oxytocin (OXT), both of which are made in a specialised part of the brain called the hypothalamo-neurohypophyseal system (HNS), but exert their effects at the level of the kidney to, respectively, provoke water conservation and salt excretion. Interestingly, our electron microscopy studies have shown that the ultrastructure of the camel HNS changes according to season, suggesting that in the arid conditions of summer the dromedary’s HNS is in a state of permanent activation, in preparation for the likely prospect of water deprivation. Based on our camel genome sequence, we have carried out an RNAseq analysis of the camel HNS in summer and winter.
