ABSTRACT: Bligh and Dyer lipid extraction of Listeria monocytogenes cells previously stressed by the cultivation with nisin and CLPs in sublethal concentrations.
Project description:Bligh and Dyer lipid extraction of Listeria monocytogenes cells previously stressed by the cultivation with nisin and CLPs in sublethal concentrations.
Project description:Thermo .raw and centroided .mzXML files of a lipidomics LC-MS analyses ran on a Thermo Q-exactive instrument.
Pooled sampled were extracted using our regular lipid extraction protocol and with an adapted protocol for lipid extraction on the rest fraction after polar extraction, both performed six-fold.
Project description:Microbiome regulation of lipid metabolism Germfree male C57BL/6J mice were purchased at 8 weeks of age from Charles River Laboratories (L'Arbresle, France). Mice were treated with 10^8 CFU/mL E. coli M8 strain (isolated from the feces of an ob/ob mouse) in drinking water for 14 days. At the end of the treatment, mice were subjected to an oral lipid tolerance test (OLTT) (with 6ml/kg of corn oil) after overnight fasting (14h). All mice were sacrificed 6 hours after the lipid tolerance test and ileum samples were collected for further analysis.
Project description:A2780 ovarian cancer cells and a cisplatin resistant derivate of A2780 cells, obtained from ECACC, UK, No. 93112519 [A2780] and No. 93112517 [A2780 cis] were seeded out in T-75 flasks at a density of 2x106 for A2780sens and 3x106 for A2780cis cells in 15 ml of medium and preincubated overnight. Medium was removed and 15 ml fresh medium (37M-0C) with different concentrations of cisplatin, liposomal cisplatin or empty liposomes were added and incubated for 72 h at 37M-0C in a 5% CO2 incubator. In case of A2780sens cells, 1.72 M-5M cisplatin (IC50 concentration) and in case of A2780cis cells 8.94 M-5M cisplatin (IC50 concentration) were added. Liposomal formulations contained equal cisplatin concentrations. Empty liposomes were added in the same concentration as the liposomal cisplatin, to analyze the impact of liposomal lipids (A2780sens: 0.80M-5mol lipid, A2780cis: 4.15 M-5mol lipid). After incubation, medium was removed and cells were washed thrice with 10 ml PBS. 1 ml RLT-buffer was added and cells lysates were stored at -80M-0C until RNA extraction.
Project description:<p>Lipid lowering therapy using HMG-CoA reductase inhibitors (statins) is associated with an approximately 9-12% increase in the risk of new-onset type 2 diabetes (T2DM). The risk of diabetes could be increased by statins directly; however, genetic approaches have also implicated low LDL cholesterol (LDL-C) concentrations as a risk factor for T2DM. Mendelian randomization studies using functional variants in both HMGCR (<a href="https://www.ncbi.nlm.nih.gov/gene/?term=HMGCR" target="_blank">GeneID:3156</a>) and PCSK9 (<a href="https://www.ncbi.nlm.nih.gov/gene/?term=PCSK9" target="_blank">GeneID:255738</a>) genes found a higher risk of T2DM in individuals with variants associated with lower LDL-C concentrations. Since PCSK9 and HMGCR are involved in lipid metabolism through distinct molecular pathways, the altered glycemic effect associated with variants in both genes is likely to be the result of their common effect on LDL-C concentrations.</p> <p>Despite the findings from statin clinical trials and genetic studies, there is little direct evidence implicating low LDL-C concentrations with increased risk of T2DM. Individuals who have very low LDL-C concentrations not due to lipid lowering therapy can provide insights into the relationship between low LDL-C concentrations and T2DM. Here, we used de-identified electronic health records (EHRs) to test the hypothesis that low LDL-C concentrations are associated with T2DM. </p>
Project description:Transcriptional profiling of Asymptomatic Bacteriuria (ABU) Escherichia coli strain 83972 comparing the progenitor wild type strain ABU83972 with its re-isolates from human bladder colonization (PI-2, PII-4, PIII-4) and in vitro cultivation experiment (4.9).
Project description:The intermediate filament protein Nestin serves as a biomarker for stem cells and has been used to identify subsets of cancer stem-like cells. However, the mechanistic contributions of Nestin to cancer pathogenesis are not understood. Here we report that Nestin binds the hedgehog pathway transcription factor Gli3 to mediate the development of medulloblastomas of the hedgehog subtype. In a mouse model system, Nestin levels increased progressively during medulloblastoma formation resulting in enhanced tumor growth. Conversely, loss of Nestin dramatically inhibited proliferation and promoted differentiation. Mechanistic investigations revealed that the tumor-promoting effects of Nestin were mediated by binding to Gli3, a zinc finger transcription factor that negatively regulates hedgehog signaling. Nestin binding to Gli3 blocked Gli3 phosphorylation and its subsequent proteolytic processing, thereby abrogating its ability to negatively regulate the hedgehog pathway. Our findings show how Nestin drives hedgehog pathway-driven cancers and uncover in Gli3 a therapeutic target to treat these malignancies.
Project description:Transcriptional profiling of Asymptomatic Bacteriuria (ABU) Escherichia coli strain 83972 comparing the progenitor wild type strain ABU83972 with its re-isolates from human bladder colonization (PI-2, PII-4, PIII-4) and in vitro cultivation experiment (4.9). Wild type vs. re-isolate cells. Biological replicates: 3 wild type, 3 re-isolates, independently grown and harvested. One replicate per array.
Project description:C. elegans exhibits thermotaxis, where most of the animals that had been cultivated at a particular temperature ranging from 15°C to 25°C for a few hours with a food source and then placed on a thermal gradient for an hour migrate to the cultivation temperature. In addition, animals that were previously conditioned to migrate to a certain temperature are capable of migrating to a new cultivation temperature a few hours after the cultivation temperature was shifted to the new temperature To gain the detailed molecular insight into thermotactic behavior, the genome-wide microarray analysis during behavioral conditioning was performed. We compared the transcriptional profile of animals conditioned to migrate to the new temperature 17°C with that of animals conditioned to migrate to the previous temperature 23°C Synchronized adult wild-type animals cultivated at 23 degrees were selected for RNA extraction and hybridization on Affymetrix microarrays.
Project description:Scd1 is responsible for forming a carbon-carbon double bound at the 9-10th position from the C-terminus of saturated fatty acids such as palmitic acid and stearic acid (C16:0 and C18:0), to generate the products of palmitoleic acid (C16:1) and oleic acid (C18:1).Here, we found SCD1 is required for in vitro blastocyst embryo development, and one of the mechanisms is through regulating unsaturated fatty acid-mediated membrane fluidity and formation of apical domain. Overall, our study provides invaluable resources for lipid reprogramming in mammalian preimplantation embryo development and mechanistic insights on regulation of embryogenesis by lipid unsaturation.