Tracking the molecular patterns of a common response across tissues in murine sepsis to refine treatment strategies
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ABSTRACT: Transcriptional effects in liver, lung, spleen and blood samples from mice challenged to Sham and Sepsis by Peritoneal Contamination and Infection (PCI) were monitored after 6 and 24 hours 4 Tissues x 2 Time Resolved Treatment Groups x 4 Replicates, 4 Tissues x Sham Control x 3 Replicates
Project description:Transcriptional effects in liver, lung and blood samples from mice after intratracheal challenge with either Streptococcus pneumoniae serotype 19 (lobar-pneumonia) or serotype 2 (sepsis) were monitored after 6 and 24 hours and compared to sham (vehicle control). We gratefully acknowledge the BMBF grant within the “Promoting global research excellence in severe sepsis” (PROGRESS) study (01KI07111). Three tissues x two serotypes x two time resolved treatment groups x four replicates, three tissues x Sham Control x three replicates.
Project description:Cyclophosphamide (CPAm) is a widely used chemotherapeutic agent that exhibits potent anti-cancer properties but is often associated with debilitating side effects. Despite its efficacy, the management of CPAm-induced toxicities remains a significant clinical challenge. There has been growing interest in exploring complementary and alternative therapies to mitigate these adverse effects in recent years, and this may be a chance for the earthworm-derived preparation, Venetin 1. Its rich composition of bioactive compounds has demonstrated promising pharmacological properties, including anti-inflammatory, antioxidant, and immunomodulatory effects. These properties suggest its potential to counteract the various systemic toxicities induced by CP. To investigate the effect of Venetin 1 on cyclophosphamide-induced toxicity, we conducted a comprehensive study. Mice were administered CPAm for four days, followed by the earthworm preparation in two doses (50 mg/kg and 100 mg/kg b.w). Importantly, the preparation did not cause any side effects in all mice, ensuring the safety of the intervention. We then determined global changes in the proteome using proteomics and quantitative SWATH-MS analysis, a robust and reliable method. This allowed us to identify up- and downregulated proteins in each studied group, providing valuable insights into the mechanism of action of Venetin 1. According to the obtained results, Venetin 1 significantly affected the proteome of mouse lung tissue. It was possible to determine quantitative changes for 400 proteins, and the analysis after administration of Venetin 1 showed a change in the global proteomic profile from upregulated to down-regulated. The stimulating properties of the preparation concerning the complement system have also been confirmed in a separate validation experiment. It has been shown that Venetin 1 has the potential to eliminate the toxic effect of cyclophosphamide in lung tissue. It also initiates regenerative processes, inhibits inflammatory processes, supports autophagy and stimulates the immune system. Further research is required to understand and to fully describe the effects of Venetin 1 .
Project description:Microarray analysis was performed 24 hr after status epilepticus in mice that had received previously either siezure preconditioning (tolerance) or sham-preconditioning (injury). Transcriptional changes in the CA3 subfield of the hippocampus were analyzed using the Affymetric Mouse 430_2 Genechip.
Project description:Total RNA was extracted from monocyte and macrophages isolated from the peritoneal cavity of un-inflamed mice or 4, 24, 48 an 72h after mice had been injected with 0.1 or 10mg zymosan
Project description:We wanted to investigate the impact of macronutrient composition on blood gene expression in obese men and relate it to low-grade systemic inflammation and onset of lifestyle diseases. Additionally, we wanted to compare blood cell gene expression pattern with subcutaneous adipose tissue cell gene expression pattern. The current study was designed as a pilot to a more comprehensive main study (manuscript in preparation). We wanted to gain experience to optimize sample size, diet intervention length, tissue for gene expression analysis, and the type of food to be used for diet intervention in the main study. Three final gene expression matrices are provided for this study. The first is a 3 persons x 6 time points blood cell RNA hybridizations (time profiling of blood cell gene expression). The second is blood cell RNA hybridizations normalized and the third is adipose tissue cell RNA hybridizations normalized.
Project description:The impact of PDF supplementation with alanyl-glutamine (AlaGln) on peritoneal immune-competence in 6 patients in an open-label, randomized, crossover pilot trial (EudraCT 2012-004004-36) was tested by relating functional test results to transcriptome changes (RNAseq and miRNA analysis) in PD effluent cells.
Project description:Explore DNA methylation in traumatic brain injury model of epilepsy and its relationship to gene expression. Examination of expression changes in stimulated rats compared to sham operated animals in traumatic brain injury model of epilepsy.
Project description:To analyze gene expression we isolated total mRNA from the core, periinfarct and contralateral cortex of 60 sprague-dawley rats after 24 hours or 3 days of permanent middle cerebral artery occlusion (pMCAo). Also ipsilateral and contralateral cortex was obtained from sham-operated and healthy animals. A total of 44 microarrays were performed with RNA pooled from 3 independent animals. For each experimental condition the total n number is of 12. Microarray analysis of different areas of the rat brain (core, periinfarct and contralateral cortex) after 24h and 3days of permanent ischemia
Project description:Acute kidney injury (AKI) is associated with an abrupt loss of kidney function that results in significant morbidity and mortality. Considerable effort has focused around the identification of diagnostic biomarkers and the analysis of molecular events. Most studies have adopted organ-wide approaches that do not fully capture the interplay among different cell types in the pathophysiology of AKI. To extend our understanding of molecular and cellular events in AKI, we developed a mouse line that enables the identification of translational profiles in specific cell types by CRE recombinase-dependent activation of an eGFP-tagged L10a ribosomal protein subunit, and consequently, translating ribosome affinity purification (TRAP) of mRNA populations. By utilizing cell-type specific CRE-driver lines, in this study we identify distinct cellular responses in an ischemia reperfusion injury (IRI) model of AKI. Cell-specific translational expression profiles were uncovered 24 hours after IRI from four populations enriched for distinct anatomical and cellular subgroups: nephron, interstitial cell populations, vascular endothelium, and macrophages/monocytes by Affymetrix microarray. A construct containing the CAGGS promoter driving eGFP-L10a, with a loxP-site flanked triple SV40 polyA cassette between promoter and eGFP-L10a cassette was targeted into the ubiquitously active Rosa26 locus. The upstream polyA cassette is designed to block activity of the downstream eGFP-L10a cassette. CRE-dependent removal of this transcriptional block activates eGFP::L10a production within the CRE-producing cell, and all of its descendants. Mice carrying the conditional eGFP-L10a allele, referred to as L10a, were maintained in a homozygous state. L10a mice were crossed to four CRE strains to activate eGFP::L10a expression in four predominantly non-overlapping cellular compartments in the kidney. A Six2-Tet-GFP::CRE allele is active exclusively within nephron progenitors; consequently, historical labeling results in eGFP::L10a expression throughout the main body of the nephron. A Foxd1-GFP::CRE allele is active in the progenitors of many of the interstitial cell lineages including those generating mesangial and non-glomerular pericytes. In addition, Foxd1 is normally expressed in podocytes. Cdh5-CRE is reported to be active throughout the vascular endothelium, and finally, Lyz2-CRE specifically labels cells of the myeloid lineage, notably macrophages, monocytes and dendritic cells. Mice carrying any CRE allele and the L10a allele are designated generically CRE-L10a. six2-L10a, foxd1-L10a, cdh5-L10a and lyz2-L10a denote specifically mice that are compound heterozygotes for the indicated CRE driver and L10a. CRE-L10a, L10a heterozygous littermates without CRE allele, C57BL/6 wild type mice were subjected to renal bilateral warm ischemia 28 minutes followed by 24-hour reperfusion when the kidney TRAP RNA and total RNA were isolated and subjected to Affymetrix microarray. Biological triplicates for each CRE-L10a line underwent no Surgery; sham Surgery and IRI treatment.
Project description:We assayed leukocyte global gene expression for a prospective validation cohort of 221 adult patients admitted to UK intensive care units with sepsis due to community acquired pneumonia or faecal peritonitis. 10 samples from patients scheduled for elective cardiac surgery were also assayed as non-septic controls. We assigned all samples to sepsis response signature groups after performing unsupervised analysis of the transcriptomic data.