Welfare Benefits of Intradermal Vaccination of Piglets

Vaccination is reported as a stressful and painful event for animals. This study investigated whether needle-free intradermal vaccination improves the welfare of weaned pigs through the reduction of stress and pain biomarkers and improvement of behavioural parameters compared to traditional intramuscular injection with a needle. A total of 339 weaned piglets were allocated to 3 treatment groups: Intradermal Application of Liquids (IDAL) pigs, vaccinated against Porcine Circovirus type 2 (PCV2) by means of intradermal vaccination using a needle-free device Porcilis® PCV ID; Intramuscular (IM) pigs vaccinated against PCV2 with Porcilis® PCV intramuscularly with a needle; CONTROL pigs were managed identically but did not receive any vaccine injection. At the time of the injection, the reaction of IDAL piglets was similar to control piglets, whereas a greater percentage of piglets vaccinated intramuscularly displayed high-pitch vocalizations (7% CONTROL, 7% IDAL, 32% IM) and retreat attempts (3% CONTROL, 7% IDAL, 39% IM). The day after vaccination, IDAL piglets did not differ from the control piglets for any of the behavioural variables studied through scan samplings. IM piglets showed a lower frequency of social negative interactions (p = 0.001) and rope manipulation (p = 0.04) compared to the CONTROL group. Resting postures did not differ between treatments. At 28 h post-vaccination, IDAL piglets presented lower blood C-reactive protein levels (CONTROL = 20 μg/mL; IDAL = 39 μg/mL; IM = 83 μg/mL, p < 0.0001) and blood Haptoglobin (CONTROL = 1.8 mg/mL; IDAL = 1.9 mg/mL vs. IM = 3.1 mg/mL, p < 0.0001) compared to IM piglets. Salivary chromogranin A and alpha-amylase did not differ between treatment groups when measured 25 min post-vaccination. The method of vaccination did not affect the growth of the piglets or their rectal temperature. These results support that needle-free intradermal vaccination reduces vaccination-related pain in growing pigs.info:eu-repo/semantics/publishedVersio

Welfare benefits of intradermal vaccination of piglets

Vaccination is reported as a stressful and painful event for animals. This study investigated whether needle-free intradermal vaccination improves the welfare of weaned pigs through the reduction of stress and pain biomarkers and improvement of behavioural parameters compared to traditional intramuscular injection with a needle. A total of 339 weaned piglets were allocated to 3 treatment groups: Intradermal Application of Liquids (IDAL) pigs, vaccinated against Porcine Circovirus type 2 (PCV2) by means of intradermal vaccination using a needle-free device Porcilis® PCV ID; Intramuscular (IM) pigs vaccinated against PCV2 with Porcilis® PCV intramuscularly with a needle; CONTROL pigs were managed identically but did not receive any vaccine injection. At the time of the injection, the reaction of IDAL piglets was similar to control piglets, whereas a greater percentage of piglets vaccinated intramuscularly displayed high-pitch vocalizations (7% CONTROL, 7% IDAL, 32% IM) and retreat attempts (3% CONTROL, 7% IDAL, 39% IM). The day after vaccination, IDAL piglets did not differ from the control piglets for any of the behavioural variables studied through scan samplings. IM piglets showed a lower frequency of social negative interactions (p = 0.001) and rope manipulation (p = 0.04) compared to the CONTROL group. Resting postures did not differ between treatments. At 28 h post-vaccination, IDAL piglets presented lower blood C-reactive protein levels (CONTROL = 20 µg/mL; IDAL = 39 µg/mL; IM = 83 µg/mL, p < 0.0001) and blood Haptoglobin (CONTROL = 1.8 mg/mL; IDAL = 1.9 mg/mL vs. IM = 3.1 mg/mL, p < 0.0001) compared to IM piglets. Salivary chromogranin A and alpha-amylase did not differ between treatment groups when measured 25 min post-vaccination. The method of vaccination did not affect the growth of the piglets or their rectal temperature. These results support that needle-free intradermal vaccination reduces vaccination-related pain in growing pigs

Identification of genes under dynamic post-transcriptional regulation from time-series epigenomic data

Aim: Prediction of genes under dynamic post-transcriptional regulation from epigenomic data. Materials & methods: We used time-series profiles of chromatin immunoprecipitation-seq data of histone modifications from differentiation of mesenchymal progenitor cells toward adipocytes and osteoblasts to predict gene expression levels at five time points in both lineages and estimated the deviation of those predictions from the RNA-seq measured expression levels using linear regression. Results & conclusion: The genes with biggest changes in their estimated stability across the time series are enriched for noncoding RNAs and lineage-specific biological processes. Clustering mRNAs according to their stability dynamics allows identification of post-transcriptionally coregulated mRNAs and their shared regulators through sequence enrichment analysis. We identify miR-204 as an early induced adipogenic microRNA targeting Akr1c14 and Il1rl1

Temporal enhancer profiling of parallel lineages identifies AHR and GLIS1 as regulators of mesenchymal multipotency

Temporal data on gene expression and context-specific open chromatin states can improve identification of key transcription factors (TFs) and the gene regulatory networks (GRNs) controlling cellular differentiation. However, their integration remains challenging. Here, we delineate a general approach for data-driven and unbiased identification of key TFs and dynamic GRNs, called EPIC-DREM. We generated time-series transcriptomic and epigenomic profiles during differentiation of mouse multipotent bone marrow stromal cell line (ST2) toward adipocytes and osteoblasts. Using our novel approach we constructed time-resolved GRNs for both lineages and identifed the shared TFs involved in both differentiation processes. To take an alternative approach to prioritize the identified shared regulators, we mapped dynamic super-enhancers in both lineages and associated them to target genes with correlated expression profiles. The combination of the two approaches identified aryl hydrocarbon receptor (AHR) and Glis family zinc finger 1 (GLIS1) as mesenchymal key TFs controlled by dynamic cell type-specific super-enhancers that become repressed in both lineages. AHR and GLIS1 control differentiation-induced genes and their overexpression can inhibit the lineage commitment of the multipotent bone marrow-derived ST2 cells

ATP1A1/BCL2L1 predicts the response of myelomonocytic and monocytic acute myeloid leukemia to cardiac glycosides.

peer reviewedMyelomonocytic and monocytic acute myeloid leukemia (AML) subtypes are intrinsically resistant to venetoclax-based regimens. Identifying targetable vulnerabilities would limit resistance and relapse. We previously documented the synergism of venetoclax and cardiac glycoside (CG) combination in AML. Despite preclinical evidence, the repurposing of cardiac glycosides (CGs) in cancer therapy remained unsuccessful due to a lack of predictive biomarkers. We report that the ex vivo response of AML patient blasts and the in vitro sensitivity of established cell lines to the hemi-synthetic CG UNBS1450 correlates with the ATPase Na+/K+ transporting subunit alpha 1 (ATP1A1)/BCL2 like 1 (BCL2L1) expression ratio. Publicly available AML datasets identify myelomonocytic/monocytic differentiation as the most robust prognostic feature, along with core-binding factor subunit beta (CBFB), lysine methyltransferase 2A (KMT2A) rearrangements, and missense Fms-related receptor tyrosine kinase 3 (FLT3) mutations. Mechanistically, BCL2L1 protects from cell death commitment induced by the CG-mediated stepwise triggering of ionic perturbation, protein synthesis inhibition, and MCL1 downregulation. In vivo, CGs showed an overall tolerable profile while impacting tumor growth with an effect ranging from tumor growth inhibition to regression. These findings suggest a predictive marker for CG repurposing in specific AML subtypes

Pyruvate dehydrogenase fuels a critical citrate pool that is essential for Th17 cell effector functions

Pyruvate dehydrogenase (PDH) is the central enzyme connecting glycolysis and the tricarboxylic acid (TCA) cycle. The importance of PDH function in T helper 17 (Th17) cells still remains to be studied. Here, we show that PDH is essential for the generation of a glucose-derived citrate pool needed for Th17 cell proliferation, survival, and effector function. In vivo, mice harboring a T cell-specific deletion of PDH are less susceptible to developing experimental autoimmune encephalomyelitis. Mechanistically, the absence of PDH in Th17 cells increases glutaminolysis, glycolysis, and lipid uptake in a mammalian target of rapamycin (mTOR)-dependent manner. However, cellular citrate remains critically low in mutant Th17 cells, which interferes with oxidative phosphorylation (OXPHOS), lipid synthesis, and histone acetylation, crucial for transcription of Th17 signature genes. Increasing cellular citrate in PDH-deficient Th17 cells restores their metabolism and function, identifying a metabolic feedback loop within the central carbon metabolism that may offer possibilities for therapeutically targeting Th17 cell-driven autoimmunity

APPROACHES FOR IDENTIFICATION OF TRANSCRIPTIONAL AND POST-TRANSCRIPTIONAL REGULATORS OF MESENCHYMAL STEM CELL DIFFERENTIATION USING TIME-SERIES EPIGENOMIC DATA

Gene regulatory networks (GRNs) control cellular differentiation and development and recapitulate the physical interactions between transcription factors (TFs) and their influence on their target genes that ultimately results into a defined cell phenotype. In addition, cellular differentiation represents the path a cell undergoes through multiple stages before reaching a terminally differentiated state and is by nature dynamic. Moreover, epigenetic regulation as well as post-transcriptional control of gene expression are critical for faithful cellular phenotype. Cellular differentiation of progenitor cells into their daughter cells provide a dynamic controllable system to study the epigenetic mechanisms as well as the transcriptional output that take place towards cellular specifications, and the TFs and non-coding RNAs that dictate their differentiation. Here, we have generated time-series transcriptomic and epigenomic data during the differentiation of bone marrow stromal cells towards adipocytes and osteoblasts and characterized a novel approach called EPIC-DREM to construct dynamic GRNs of adipocytes and osteoblasts. In order to focus on shared transcriptional regulators of early commitment of bone marrow stromal cells towards adipocytes and osteoblasts, we have concentrated our analysis on dynamic super-enhancers to prioritize the identified TFs and discovered aryl hydrocarbon receptor (AHR) as a transcriptional regulator of the multipotent state. In addition, the generated of time-series epigenomic data were used as input for linear regression analysis that allowed to predict genes that are dynamically controlled by post-transcriptional regulators such as microRNAs (miRs). Indeed, genes that differ from their predicted expression level as assessed by the residuals of the linear regression model can be informative about their mRNA stability. In order to decipher genes that are under dynamic post-transcriptional control, the standard deviation of gene’s residuals was taken as a dynamic measure of changes in mRNA stability and clustering analysis coupled to microRNA motifs enrichment analysis allowed to identify post-transcriptionally co-regulated mRNAs. Based on the linear regressions analysis, miR-204 was identified as a potential regulator of adipogenesis. Integration of these types of data can contribute to the understanding of transcriptional and post-transcriptional control of cell differentiation and the here established approaches for key regulators identification can be widely applied to study other cell states transitions

L'immunopathologie de la maladie de Crohn et les traitements envisageables

PARIS-BIUP (751062107) / SudocSudocFranceF

Assessment of Mitochondrial Cell Metabolism by Respiratory Chain Electron Flow Assays

Cellular energy metabolism is regulated by complex metabolic pathways. Although anaerobic glycolysis was reported as a primary source of energy in cancer leading to a high rate of lactate production, current evidence shows that the main energy source supporting cancer cell metabolism relies on mitochondrial metabolism. Mitochondria are the key organelle maintaining optimal cellular energy levels. MitoPlate™ S-1 provides a highly reproducible bioenergetics tool to analyze the electron flow rate in live cells. Measuring the rates of electron flow into and through the electron transport chain using different NADH and FADH2-producing metabolic substrates enables the assessment of mitochondrial functionality. MitoPlate™ S-1 are 96-well microplates pre-coated with different substrates used as probes to examine the activity of mitochondrial metabolic pathways based on a colorimetric assay. A comparative metabolic analysis between cell lines or primary cells allows to establish a specific metabolic profile and to detect possible alterations of the mitochondrial function of a tumor cell. Moreover, the direct measurements of electron flux triggered by metabolic pathway activation could highlight targets for potential drug candidates

Welfare benefits of intradermal vaccination of piglets

Vaccination is reported as a stressful and painful event for animals. This study investigated whether needle-free intradermal vaccination improves the welfare of weaned pigs through the reduction of stress and pain biomarkers and improvement of behavioural parameters compared to traditional intramuscular injection with a needle. A total of 339 weaned piglets were allocated to 3 treatment groups: Intradermal Application of Liquids (IDAL) pigs, vaccinated against Porcine Circovirus type 2 (PCV2) by means of intradermal vaccination using a needle-free device Porcilis® PCV ID; Intramuscular (IM) pigs vaccinated against PCV2 with Porcilis® PCV intramuscularly with a needle; CONTROL pigs were managed identically but did not receive any vaccine injection. At the time of the injection, the reaction of IDAL piglets was similar to control piglets, whereas a greater percentage of piglets vaccinated intramuscularly displayed high-pitch vocalizations (7% CONTROL, 7% IDAL, 32% IM) and retreat attempts (3% CONTROL, 7% IDAL, 39% IM). The day after vaccination, IDAL piglets did not differ from the control piglets for any of the behavioural variables studied through scan samplings. IM piglets showed a lower frequency of social negative interactions (p = 0.001) and rope manipulation (p = 0.04) compared to the CONTROL group. Resting postures did not differ between treatments. At 28 h post-vaccination, IDAL piglets presented lower blood C-reactive protein levels (CONTROL = 20 µg/mL; IDAL = 39 µg/mL; IM = 83 µg/mL, p < 0.0001) and blood Haptoglobin (CONTROL = 1.8 mg/mL; IDAL = 1.9 mg/mL vs. IM = 3.1 mg/mL, p < 0.0001) compared to IM piglets. Salivary chromogranin A and alpha-amylase did not differ between treatment groups when measured 25 min post-vaccination. The method of vaccination did not affect the growth of the piglets or their rectal temperature. These results support that needle-free intradermal vaccination reduces vaccination-related pain in growing pigs