Project description:Studies of expression of mechanims of defense of the Acinetobacter sp.5-2Ac.02 from airborne hospital environment under stress conditions, such as SOS response (ROS response, heavy metals resistant mechanisms, peptides), as well as Quorum network (acetoin cluster and aromatics biodegradation cluster). Characterization functional of AcoN-like as negative regulator protein from acetoin cluster in Acinetobacter spp. Strains
Project description:Members of the genus Acinetobacter drag attention due to their importance in microbial pathology and biotechnology. OmpA is a porin with multifaceted functions in different species of Acinetobacter. In this study we identified this protein in Acinetobacter sp. SA01, an efficient phenol degrader strain, in different cellular and sub-cellular compartments (such as OM, OMV, biofilm and extracellular environment). Differential expression of proteins, including OmpA, under two conditions of phenol and ethanol supplementation was assessed using shotgun proteomics.
Project description:MBL-1 is an RNA-binding protein important for the differentiation and development of many animal tissues. An evolutionarily conserved alternative splicing mechanism generates MBL-1 isoforms that are either expressed in the nucleus or show a more cytoplasmic distribution. We used small RNA-seq to investigate the individual contribution of evolutionarily conserved mbl-1 isoform expression of C. elegans miRNA expression. The analysis was done on L4 stage animals
Project description:Cerebrospinal fluid (CSF) plays an important role in brain tumours, including medulloblastoma (MBL). CSF proteomics studies have mainly focused on extracellular vesicle (EV) proteins rather than the total CSF proteome. Recent advancements in mass spectrometry systems and �쁎mics�� data analysis methods enable unbiased, high proteome depth research. We conducted proteomic profiling of the total CSF in MBL patients with the purpose of finding a potential diagnostic biomarker for MBL. Methods CSF samples from patients with MBL (n=31) and hydrocephalus (HC, control, n=19) were analysed using liquid chromatography-tandem mass spectrometry (LC�묺S/MS). Enrichment analysis of canonical pathways and differentially expressed proteins (DEPs) was performed. Feature selection of the proteins was conducted using the analysis of variance F value and fast correlation-based filter methods. The selected proteins were validated through ELISA and EV characterization using ExoView systems. Results We quantified 1,112 proteins per CSF sample. We further investigated 273 DEPs extracted from a comparison of MBL and HC. Hierarchical clustering of DEPs between MBL and HC clearly showed classification into two groups. Feature selection identified four elevated soluble proteins (SPTBN1, HSP90AA1, TKT, and NME1-NME2) in MBL CSF. Validation with ELISA confirmed that TKT was significantly elevated in MBL compared to HC, as well as in Group 4 MBL and leptomeningeal seeding (LMS). Additionally, TKT-positive EVs were significantly enriched in MBL CSF compared to HC CSF and correlated with the burden of LMS. Conclusions Our results provide insights into the proteomics data of the total CSF of MBL patients. Furthermore, we identified the significance of TKT within the total CSF and its presence within circulating EVs in the CSF. We suggest that TKT may serve as a biomarker for MBL, particularly for the diagnosis of LMS.
Project description:MBL-1 is an RNA-binding protein important for the differentiation and development of many animal tissues. Our aim was to identify the binding targets of the MBL-1 protein in C. elegans. RNA from a CRISPR/CAS9-generated strain expressing a 3xFLAG-tagged MBL-1 protein was immunopreciprecipitated with an anti-FLAG antibody. RNA-seq was then performed to identify binding targets of the RNA-binding protein MBL-1. Experiments were performed at the L4 stage.
Project description:Homozygous masterblind (mbl-/-) zebrafish exhibit reduced or absent eyes and telecephalon, and the expansion of the diencephalic fates to the front of the brain. A missense mutation in the GSK3-binding domain of zebrafish axin1, a scaffolding protein in the Wnt signaling pathway, results in the mbl phenotype. In an effort to identify and study the genes affected by Wnt signaling, we used a 14,000-oligonucleotide-gene microarray to determine differentially expressed genes in mbl/axin1 (-/-) and wild type control zebrafish embryos and larvae. Keywords: zebrafish, Danio rerio, wild-type mbl, axin1, development
Project description:Ozone is a common pollutant and a potent oxidant in industrialized nations. The mechanisms of ozone-induced lung injury and differential susceptibility are not fully understood. Ozone-induced lung inflammation is mediated, in part, by the innate immune system. We hypothesized that mannose binding lectin (MBL), which has a central role in the activation of the complement pathway of innate immunity, is a necessary component of the pro-inflammatory events caused by ozone-mediated activation of the innate immune system. Wild-type (Mbl+/+) and MBL deficient (Mbl-/-) mice were exposed to ozone (0.3 ppm) for 24, 48, and 72 hours, and bronchoalveolar lavage fluid (BALF) was examined for inflammatory markers. Compared to Mbl+/+ mice, significantly greater mean BALF eosinophils, neutrophils and neutrophil attractants CXCL2 (MIP-2) and CXCL5 (LIX) were found in Mbl-/- mice exposed to ozone. Using genome-wide mRNA microarray analyses, we identified significant differences in expression response profiles and networks at baseline (e.g. NRF2 mediated oxidative stress response) and after exposure (e.g. humoral immune response) between Mbl+/+ and Mbl-/- mice. The microarray data were further analyzed using a pattern recognition analysis for Extracting Patterns and Identifying co-expressed Genes (EPIG), and discovered several informative differential response patterns and subsequent gene sets, including antimicrobial response and inflammatory response. These novel findings demonstrate that targeted deletion of Mbl caused differential expression of inflammation-related gene sets basally and after exposure to ozone, and significantly reduced pulmonary inflammation thus indicating an important innate immunomodulatory role of the gene in this model.