Seroprevalence and Incidence of hepatitis E in Blood Donors in Upper Austria

<div><p>Background</p><p>In recent years various studies showed, that hepatitis E virus (HEV) is a growing public health problem in many developed countries. Therefore, HEV infections might bear a transmission risk by blood transfusions. The clinical relevance still requires further investigations. The aim of this study was to provide an overview of acute HEV infections in Upper Austrian blood donors as well as a risk estimation of this transfusion-related infection.</p><p>Methods and Findings</p><p>A total of 58,915 blood donors were tested for HEV RNA using a commercial HEV RT-PCR Kit. 7 of these donors (0.01%) were PCR-positive with normal laboratory parameters in absence of clinical signs of hepatitis. Viral load determined by quantitative real-time PCR showed a HEV nucleic acid concentration of 2,217 293,635 IU/ml. At follow-up testing (2–11 weeks after donation) all blood donors had negative HEV RNA results. Additionally, genotyping was performed by amplification and sequencing of the ORF1 or ORF2 region of the HEV genome. All HEV RNA positive donor samples revealed a genotype 3 isolate. For the antibody screening, anti-HEV IgM and IgG were detected by ELISA. Follow up serological testing revealed that no donor was seropositive for HEV IgM or IgG antibodies at time of donation. Moreover, we verified the prevalence of anti-HEV IgG in 1,203 of the HEV RNA negative tested blood donors. Overall 13.55% showed positive results for anti-HEV IgG.</p><p>Conclusions</p><p>In the presented study, we investigated HEV infections in blood donations of Upper Austria over 1 year. We concluded that 1 out of 8,416 blood donations is HEV RNA positive. Seroprevalence of anti HEV IgG results in an age-related increase of 13.55%. Therefore, based on this data, we recommend HEV-PCR screening to prevent transmission of hepatitis E virus by transfusion.</p></div

Comparison of HEV seroprevalence in all districts of Upper Austria.

<p>Comparison of HEV seroprevalence in all districts of Upper Austria.</p

Prevalence of anti-HEV IgG.

<p>Estimated prevalence of anti-HEV IgG in 1,203 of Upper Austrian blood donors by age group.</p

Phylogenetic relationship between genotype 3 HEV strains derived from Austrian blood donors (#1–6) and HEV reference strains (GenBank sequences are cited by their respective accession numbers).

<p>The tree was constructed by the Clustal W method on the basis of a 304-nt sequence fragment of ORF2 region.</p

Sequence alignment.

<p>Comparison of a partial genome sequence in ORF2 from HEV positive samples. Dots indicate identity to the consensus sequence.</p

Geographical map of Upper Austria.

<p>A detailed analysis of seroprevalences [%] marked in green, yellow, orange and red of all districts is shown. Areas in grey are those with no available data. Copyright: The map of Austria is adapted from Statistik Austria and modified by Niklas N., <a href="http://www.statistik.at/web_de/services/interaktive_karten" target="_blank">http://www.statistik.at/web_de/services/interaktive_karten</a>; The map of Upper Austria is adapted from wikipedia, author AleXXw and modified by Hofmann M., <a href="http://de.wikipedia.org/wiki/Datei:Karte_A_Ooe_ohne.svg" target="_blank">http://de.wikipedia.org/wiki/Datei:Karte_A_Ooe_ohne.svg</a></p

Multiphoton-Polymerized 3D Protein Assay

Multiphoton polymerization (MPP) enables 3D fabrication of micro- and nanoscale devices with complex geometries. Using MPP, we create a 3D platform for protein assays. Elevating the protein-binding sites above the substrate surface allows an optically sectioned readout, minimizing the inevitable background signal from nonspecific protein adsorption at the substrate surface. Two fluorescence-linked immunosorbent assays are demonstrated, the first one relying on streptavidin–biotin recognition and the second one on antibody recognition of apolipoprotein A1, a major constituent of high-density lipoprotein particles. Signal-to-noise ratios exceeding 1000 were achieved. The platform has high potential for 3D multiplexed recognition assays with an increased binding surface for on-chip flow cells

Apoptotic MNC-secretomes in experimental stroke

<p>Mixed Model Analysis (SAS output)</p> <p>The data were analyzed using linear mixed models for the neuroscore on treatment group and time-point with the factor animal included as a random effect. The MIXED procedure in SAS 9.3 was used to perform the calculations. The raw output contains information on the model specifications, the estimated error variance and random effects variance, the estimated regression coefficients, the covariance structure of the model coefficients and type III F-tests for the hypotheses of no effect of either fixed effect or their interactions. An interaction plot was drawn using the GLM procedure. This plot shows the individual observations and their sample mean values in each group and for each time-point. The group labels 0,1,2 and 3 in the raw output refer to the treatment group in setting 1, the control group in setting 1, the treatment group in setting 2 and the control group in setting 2, respectively.</p> <p>Original Western blots to Figure 5 </p> <p>Expression of proteins involved in cytoprotective pathways in human Astrocytes and Schwann Cells </p> <p>Astrocytes (page 1) or Schwann Cells (page 2) were stimulated with hMNCapo sec, control medium (served as control to treatment) or positive control (control to the measured protein). Original blots for all measured proteins are given in this raw data set (pages 1 and 2). For each blot, lanes (1), (2), and (3) correspond to the groups medium control [(1)=control to treatment], human apoptotic MNC-secretomes [(2)=treatment] and positive control [(3)=recombinant protein].<br>Bands in each blot are shown for phosphorylated CREB, total-CREB, phosphorylated Erk1/2, total-Erk 1/2, phosphorylated HSP27, total-HSP27, phosphorylated cJun, total-cJun, phosphorylated Akt, and total-Akt. The molecular weight (kDa) for each protein can be seen under each blot.<br>Ponceau staining was used as loading control for each group (1), (2), and (3) and suggest equal loading.</p> <p>Original Western blots to Figure 6 </p> <p>Expression of Phosphorylated CREB in Astrocytes and Neurons after stimulation with the active compound hMNCapo sec or control medium: </p> <p>Cultured human Astrocytes and Neurons were incubated with hMNCapo sec or control (cell culture-) medium at indicated concentrations. Original blots can be seen here.<br>Ponceau staining shows equal loading.</p> <p> </p

Immune Repertoire Profiling Reveals that Clonally Expanded B and T Cells Infiltrating Diseased Human Kidneys Can Also Be Tracked in Blood

<div><p>Recent advances in high-throughput sequencing allow for the competitive analysis of the human B and T cell immune repertoire. In this study we compared Immunoglobulin and T cell receptor repertoires of lymphocytes found in kidney and blood samples of 10 patients with various renal diseases based on next-generation sequencing data. We used Biomed-2 primer panels and ImmunExplorer software to sequence, analyze and compare complementarity determining regions and V-(D)-J elements. While generally an individual’s renal receptor repertoire is different from the repertoire present in blood, 94% (30/32) of the lymphocytes with clonal expansion in kidney can also be traced in blood however, not all of these clonotypes are equally abundant. Summarizing the data of all analyzed patients, 68% of highly expanded T cell clonotypes and 30% of the highly expanded B cell clonotypes that have infiltrated the kidney can be found amongst the five most abundant clonotypes in blood. In addition, complementarity determining region 3 sequences of the immunoglobulin heavy chains are on average more diverse than T cell receptor beta chains. Immune repertoire analysis of tissue infiltrating B and T cells adds new approaches to the assessment of adaptive immune response in kidney diseases. Our data suggest that expanded clonotypes in the tissues might be traceable in blood samples in the course of treatment or the natural history of the disease.</p></div

Additional file 2: of cFinder: definition and quantification of multiple haplotypes in a mixed sample

Archive containing test data. Include annotations and exemplary alignment files from AVA and CLC for sample 5, as well as simulated test reads for overlapping amplicons