Kartierungsstudien und funktionelle Kandidatengenanalyse für Tot- und Schwergeburt in der deutschen Fleckviehpopulation

It is very important to work against the increase of stillbirth and calving difficulties in the German Simmental population from breeding, economics and animal protections point of view. The igf2r, esr1 and sod2 were chosen from earlier analysis as candidate genes for stillbirth. igf2r was excluded as a candidate gene during analysis because the inactivation of the paternal Allel was proven with the microsatellite LMU0901 in comparison between genomic DNA and cDNA. The candidate genes esr1 and sod2 were tested for variations in their sequences and in their expression. Three mutations in the esr1 Exons I, IV and VIII were detected, but none of them leads to an amino acid change. A little change in the expression rate of esr1 could be observed by Real Time PCR analyses which suggest its involvement in the signal cascade of the causal gene for the stillbirth trait. But it is not directly involved. Three mutations could be detected in the mRNA sequence of sod2. One of them causes an amino acid change. However, it could not be associated with the trait of stillbirth. In both RNA samples, heart and liver, no change in expression rate of sod2 through Real Time analyses was detected. Four QTLs were found in the genome wide association and mapping studies. They are localised on BTA01, BTA05, BTA06 and BTA21

Diagnostic applications of next generation sequencing: working towards quality standards

Over the past 6 years, next generation sequencing (NGS) has been established as a valuable high-throughput method for research in molecular genetics and has successfully been employed in the identification of rare and common genetic variations. All major NGS technology companies providing commercially available instruments (Roche 454, Illumina, Life Technologies) have recently marketed bench top sequencing instruments with lower throughput and shorter run times, thereby broadening the applications of NGS and opening the technology to the potential use for clinical diagnostics. Although the high expectations regarding the discovery of new diagnostic targets and an overall reduction of cost have been achieved, technological challenges in instrument handling, robustness of the chemistry and data analysis need to be overcome. To facilitate the implementation of NGS as a routine method in molecular diagnostics, consistent quality standards need to be developed. Here the authors give an overview of the current standards in protocols and workflows and discuss possible approaches to define quality criteria for NGS in molecular genetic diagnostics

Extracellular MRP8/14 is a regulator of β2 integrin-dependent neutrophil slow rolling and adhesion

Myeloid-related proteins (MRPs) 8 and 14 are cytosolic proteins secreted from myeloid cells as proinflammatory mediators. Currently, the functional role of circulating extracellular MRP8/14 is unclear. Our present study identifies extracellular MRP8/14 as an autocrine player in the leukocyte adhesion cascade. We show that E-selectin-PSGL-1 interaction during neutrophil rolling triggers Mrp8/14 secretion. Released MRP8/14 in turn activates a TLR4-mediated, Rap1-GTPase-dependent pathway of rapid beta 2 integrin activation in neutrophils. This extracellular activation loop reduces leukocyte rolling velocity and stimulates adhesion. Thus, we identify Mrp8/14 and TLR4 as important modulators of the leukocyte recruitment cascade during inflammation in vivo

Extracellular MRP8/14 is a regulator of β2 integrin-dependent neutrophil slow rolling and adhesion

Myeloid-related proteins (MRPs) 8 and 14 are cytosolic proteins secreted from myeloid cells as proinflammatory mediators. Currently, the functional role of circulating extracellular MRP8/14 is unclear. Our present study identifies extracellular MRP8/14 as an autocrine player in the leukocyte adhesion cascade. We show that E-selectin-PSGL-1 interaction during neutrophil rolling triggers Mrp8/14 secretion. Released MRP8/14 in turn activates a TLR4-mediated, Rap1-GTPase-dependent pathway of rapid beta 2 integrin activation in neutrophils. This extracellular activation loop reduces leukocyte rolling velocity and stimulates adhesion. Thus, we identify Mrp8/14 and TLR4 as important modulators of the leukocyte recruitment cascade during inflammation in vivo

Potential of high dimensional radiomic features to assess blood components in intraaortic vessels in non-contrast CT scans

Background: To assess the potential of radiomic features to quantify components of blood in intraaortic vessels to non-invasively predict moderate-to-severe anemia in non-contrast enhanced CT scans. Methods: One hundred patients (median age, 69 years; range, 19–94 years) who received CT scans of the thoracolumbar spine and blood-testing for hemoglobin and hematocrit levels ± 24 h between 08/2018 and 11/2019 were retrospectively included. Intraaortic blood was segmented using a spherical volume of interest of 1 cm diameter with consecutive radiomic analysis applying PyRadiomics software. Feature selection was performed applying analysis of correlation and collinearity. The final feature set was obtained to differentiate moderate-to-severe anemia. Random forest machine learning was applied and predictive performance was assessed. A decision-tree was obtained to propose a cut-off value of CT Hounsfield units (HU). Results: High correlation with hemoglobin and hematocrit levels was shown for first-order radiomic features (p < 0.001 to p = 0.032). The top 3 features showed high correlation to hemoglobin values (p) and minimal collinearity (r) to the top ranked feature Median (p < 0.001), Energy (p = 0.002, r = 0.387), Minimum (p = 0.032, r = 0.437). Median (p < 0.001) and Minimum (p = 0.003) differed in moderate-to-severe anemia compared to non-anemic state. Median yielded superiority to the combination of Median and Minimum (p(AUC) = 0.015, p(precision) = 0.017, p(accuracy) = 0.612) in the predictive performance employing random forest analysis. A Median HU value ≤ 36.5 indicated moderate-to-severe anemia (accuracy = 0.90, precision = 0.80). Conclusions: First-order radiomic features correlate with hemoglobin levels and may be feasible for the prediction of moderate-to-severe anemia. High dimensional radiomic features did not aid augmenting the data in our exemplary use case of intraluminal blood component assessment

Impact of rescanning and repositioning on radiomic features employing a multi-object phantom in magnetic resonance imaging

Our purpose was to analyze the robustness and reproducibility of magnetic resonance imaging (MRI) radiomic features. We constructed a multi-object fruit phantom to perform MRI acquisition as scan-rescan using a 3 Tesla MRI scanner. We applied T2-weighted (T2w) half-Fourier acquisition single-shot turbo spin-echo (HASTE), T2w turbo spin-echo (TSE), T2w fluid-attenuated inversion recovery (FLAIR), T2 map and T1-weighted (T1w) TSE. Images were resampled to isotropic voxels. Fruits were segmented. The workflow was repeated by a second reader and the first reader after a pause of one month. We applied PyRadiomics to extract 107 radiomic features per fruit and sequence from seven feature classes. We calculated concordance correlation coefficients (CCC) and dynamic range (DR) to obtain measurements of feature robustness. Intraclass correlation coefficient (ICC) was calculated to assess intra- and inter-observer reproducibility. We calculated Gini scores to test the pairwise discriminative power specific for the features and MRI sequences. We depict Bland Altmann plots of features with top discriminative power (Mann-Whitney U test). Shape features were the most robust feature class. T2 map was the most robust imaging technique (robust features (rf), n=84). HASTE sequence led to the least amount of rf (n=20). Intra-observer ICC was excellent (>= 0.75) for nearly all features (max-min; 99.1-97.2%). Deterioration of ICC values was seen in the inter-observer analyses (max-min; 88.7-81.1%). Complete robustness across all sequences was found for 8 features. Shape features and T2 map yielded the highest pairwise discriminative performance. Radiomics validity depends on the MRI sequence and feature class. T2 map seems to be the most promising imaging technique with the highest feature robustness, high intra-/inter-observer reproducibility and most promising discriminative power

Analysis of and function predictions for previously conserved hypothetical or putative proteins in -0

<p><b>Copyright information:</b></p><p>Taken from "Analysis of and function predictions for previously conserved hypothetical or putative proteins in "</p><p>BMC Microbiology 2006;6():1-1.</p><p>Published online 9 Jan 2006</p><p>PMCID:PMC1360075.</p><p>Copyright © 2006 Gaudermann et al; licensee BioMed Central Ltd.</p>nclude: a MFS family transporter (Bfl240, IolF-like, top right; exact substrate specificity not known), ADP-heptose synthase (Bfl063, IolC-like), an acetolactate synthase II, large subunit (Bfl593; IolD-like) and its small subunit, neighbouring protein Bfl 592 (both proteins should physically interact), fructose 1,6-bisphosphate aldolase (Bfl255, IolJ-like; this enzyme is also involved in glycolysis) and predicted myo-inositol-1(or 4)-monophosphatase (Bfl535; related to the Archaeal fructose-1,6-bisphosphatase and related enzymes of inositol monophosphatase family). Bfl601 (triosephosphate isomerase) finishes the pathway converting created dihydroxyacetone phosphate into glyceraldehyde 3-phosphate