Requirement of Activation for Hepatitis B Virus Infection

Although _in vitro_ models of human hepatitis B virus replication are established, so far none could approximate infection efficiency as expected from _in vivo_ observations. Susceptibility for HBV infections has only been reported for primary hepatocytes of human, chimpanzee or Tupaia belangeri and the cell line HepaRG. Here we show that the insusceptible human hepatoma cell line HepG2 can be infected, when the virus was beforehand activated by passage over whole duck liver cell cultures. That suggests an activation step to be performed by specialized liver cells

Requirement of Activation for Hepatitis B Virus Infection

Although _in vitro_ models of human hepatitis B virus replication are established, so far none could approximate infection efficiency as expected from _in vivo_ observations. Susceptibility for HBV infections has only been reported for primary hepatocytes of human, chimpanzee or Tupaia belangeri and the cell line HepaRG. Here we show that the insusceptible human hepatoma cell line HepG2 can be infected, when the virus was beforehand activated by passage over whole duck liver cell cultures. That suggests an activation step to be performed by specialized liver cells

A comprehensive flow-cytometric analysis of graft infiltrating lymphocytes, draining lymph nodes and serum during the rejection phase in a fully allogeneic rat cornea transplant model

Purpose: To establish a cornea transplant model in a pigmented rat strain and to define the immunologic reaction toward corneal allografts, by studying the cellular and humoral immune response after keratoplasty. Methods:Full thickness penetrating keratoplasty was performed on Brown Norway (RT1n) recipients using fully major histocompatibility complex (MHC)-mismatched Piebald-Viral-Glaxo (PVG; RT1c) donors. Using multicolor flow cytometry (FACS) we quantified and compared the cellular composition of draining versus non-draining lymph nodes (LN). Furthermore, we developed an isolation method to release viable graft infiltrating lymphocytes (GIL) and subjected them to phenotypic analysis and screened serum from transplanted animals for allo-antibodies. Results:Assessing ipsi-lateral submandibular LN we find ample evidence for post surgical inflammation such as elevated absolute numbers of cluster of differentiation (CD)4+, CD8+, B-cells, and differential expression of CD134. However, we could not unequivocally identify an allo-antigen-specific immune response. FACS analysis of lymphocytes isolated from collagenase digested rejected corneas revealed the following six distinct subpopulations: MHC-2+ cells, CD4+ T-cells, CD8+ T-cells, CD161dull large granular lymphocytes, CD3+ CD8+ CD161dull natural killer (NK)-T-cells and CD161high CD3- NK cells. At post-operation day (POD)-07 only CD161dull MHC-2neg large granular lymphocytes (LGLs) were detected in syngeneic and allo-grafts. In concordance with an increase in B-cell numbers we often detected copious amounts of allo-antibodies in serum of rejecting animals, in particular immunoglobulin (Ig) M (IgM), immunoglobulin (Ig) G1 (IgG1), and IgG2a. Conclusions:Our results demonstrate that despite its immune privileged status and low-responder characteristics of the strain combination, allogeneic corneal grafts mount a full fledged T helper1 (Th1) and Th2 response. The presence of NK-T-cells and NK-cells in rejecting corneas shows the synergy between innate and adaptive immunity during allograft destruction

Template assisted surface micro microstructuring of flowable dental composites and its effect on the microbial adhesion properties

Despite their various advantages, such as good esthetic properties, absence of mercury and adhesive bonding to teeth, modern dental composites still have some drawbacks, e.g., a relatively high rate of secondary caries on teeth filled with composite materials. Recent research suggests that microstructured biomaterials surfaces may reduce microbial adhesion to materials due to unfavorable physical material–microbe interactions. The objectives of this study were, therefore, to test the hypotheses that (i) different surface microstructures can be created on composites by a novel straightforward approach potentially suitable for clinical application and (ii) that these surface structures have a statistically significant effect on microbial adhesion properties.Peer ReviewedPostprint (author's final draft

Spritzgießtechnische Herstellung duroplastgebundener Dauermagnete

Kunststoffgebundene Dauermagnete kommen bereits seit einem dreiviertel Jahrhundert in diversen Anwendungen zum Einsatz. Dabei werden die gängigen Kunststoffverarbeitungsverfahren wie beispielsweise Spritzgießen, Extrudieren, Pressen und Kalandrieren angewandt. Ähnlich zu der Aufteilung in der Verarbeitung von herkömmlichen Kunststoffen, werden auch für die kunststoffgebundenen Dauermagnete nur selten duroplastische Matrixmaterialien im Spritzgießverfahren eingesetzt. Dabei bringt gerade diese Werkstoffklasse hervorragende Eigenschaften mit sich, die eine Verbesserung der Eigenschaften von kunststoffgebundenen Dauermagneten zur Folge haben können. Neben der Medien- und Temperaturbeständigkeit dieser Werkstoffklasse spielt dabei auch deren Viskosität eine bedeutende Rolle für diese Anwendung. Im Rahmen der vorliegenden Arbeit soll geprüft werden, ob der Einsatz duroplastischer Matrixmaterialien zu einer Verbesserung der Eigenschaften kunststoffgebundener Dauermagnete führen kann. Hierfür werden anisotrope Magnetpartikel in den Kunststoff eincompoundiert und während der Verarbeitung im Spritzgießverfahren orientiert. Dies geschieht durch ein integriertes Magnetfeld im Spritzgießwerkzeug. Insbesondere sollen der Einfluss des Matrixmaterials, der Partikelgeometrie, der Fließrichtung sowie des Richtfeldes auf die Partikelorientierung untersucht werden. Zusammen mit grundlegenden analytischen Untersuchungen der Ausgangsmaterialien sollen abschließend Richtlinien für eine Bauteilauslegung definiert und erste Ansätze für eine Simulation der magnetischen Bauteileigenschaften erarbeitet werden.Polymer bonded magnets are used for different applications for over 70 years yet. These materials are produced within known polymer processing procedures as injection molding, extruding, compression molding and calendaring. However, as for standard plastics thermoset materials are seldom used in injection molding even though they show great potential regarding temperature and media resistance as well as in terms of a low viscosity. These properties could be of use in case of polymer bonded magnets. Within this work the use of thermoset matrix materials for polymer bonded magnets should be evaluated and thus their properties shall be improved. Therefore anistotropic magnetic particles will be incorporated in the thermoset matrix and they will be oriented during the injection molding process which requires a magnetic field in the cavity during processing. Especially the influences of the matrix material, the particle geometry, the flow direction and the aligning field on the particle orientation should be investigated and together with fundamental analytical procedures guidelines for engineering of parts should be generated as well as first approaches for a simulation of the part properties shall be given

Characteristics and utilization of canola seed fractions in ruminant feeds a processing and value added research

Non-Peer Reviewe

Characterization of the human DYRK1A promoter and its regulation by the transcription factor E2F1

<p>Abstract</p> <p>Background</p> <p>Overexpression of the human <it>DYRK1A </it>gene due to the presence of a third gene copy in trisomy 21 is thought to play a role in the pathogenesis of Down syndrome. The observation of gene dosage effects in transgenic mouse models implies that subtle changes in expression levels can affect the correct function of the <it>DYRK1A </it>gene product. We have therefore characterized the promoter of the human <it>DYRK1A </it>gene in order to study its transcriptional regulation.</p> <p>Results</p> <p>Transcription start sites of the human <it>DYRK1A </it>gene are distributed over 800 bp within a region previously identified as an unmethylated CpG island. We have identified a new alternative noncoding 5'-exon of the <it>DYRK1A </it>gene which is located 772 bp upstream of the previously described transcription start site. Transcription of the two splicing variants is controlled by non-overlapping promoter regions that can independently drive reporter gene expression. We found no evidence of cell- or tissue-specific promoter usage, but the two promoter regions differed in their activity and their regulation. The sequence upstream of exon 1A (promoter region A) induced about 10-fold higher reporter gene activity than the sequence upstream of exon 1B (promoter region B). Overexpression of the transcription factor E2F1 increased <it>DYRK1A </it>mRNA levels in Saos2 and Phoenix cells and enhanced the activity of promoter region B three- to fourfold.</p> <p>Conclusion</p> <p>The identification of two alternatively spliced transcripts whose transcription is initiated from differentially regulated promoters regions indicates that the expression of the <it>DYRK1A </it>gene is subject to complex control mechanisms. The regulatory effect of E2F1 suggests that DYRK1A may play a role in cell cycle regulation or apoptosis.</p

Effect of feeding broilers diets differing in susceptible phytate content

Measurements of total phytate phosphorus content of diets may be deceptive as they do not indicate substrate availability for phytase; it may be that measurements of phytate susceptible to phytase effects are a more accurate measure of phosphorus (P) availability to the bird. To verify this hypothesis, an experiment was conducted to compare diets formulated to contain either high or low susceptible phytate, supplemented with either 0 or 500 FTU/kg phytase. Susceptible phytate was determined by exposing the feed samples to conditions that mimicked the average pH of the proximal gastrointestinal tract (pH 4.5) and the optimum temperature for phytase activity (37 °C) and then measuring phytate dissolved. Ross 308 birds (n = 240) were fed one of 4 dietary treatments in a 2 × 2 factorial design; 2 diets with high (8.54 g/kg, 57.90% of total phytate) or low (5.77 g/kg, 46.33% of total phytate) susceptible phytate, containing 0 or 500 FTU/kg phytase. Diets were fed to broilers (12 replicate pens of 5 birds per pen) from d 0 to 28 post hatch. Birds fed diets high in susceptible phytate had greater phytate hydrolysis in the gizzard (P < 0.001), jejunum (P < 0.001) and ileum (P < 0.001) and resulting greater body weight gain (BWG) (P = 0.015) and lower FCR (P = 0.003) than birds fed the low susceptible phytate diets, irrespective of phytase presence. Birds fed the high susceptible diets also had greater P solubility in the gizzard and Ca and P solubility in the jejunum and ileum (P < 0.05) and resulting greater tibia and femur Ca and P (P < 0.05) content than those fed the low susceptible diets. All the susceptible phytate was fully degraded in the tract in the absence of added phytase, suggesting the assay used in this study was able to successfully estimate the amount of total dietary phytate that was susceptible to the effects of phytase when used at standard levels. No interactions were observed between susceptible phytate and phytase on phytate hydrolysis. Hydrolysis of phytate was greater (P < 0.05) in the gizzard of birds fed the diets supplemented with phytase, regardless of the concentration of susceptible phytate in the diet. Phytase supplementation resulted in improved BWG (P < 0.001) and FCR (P = 0.001), increased P solubility (P < 0.001) in the gizzard, Ca and P solubility (P < 0.001) in the jejunum and ileum and Ca and P concentration (P < 0.001) and strength (P < 0.001) in the tibia and femur. Pepsin activity was higher in birds fed the diets supplemented with phytase (P < 0.001) and was greater (P = 0.031) in birds fed the high susceptible phytate diets compared with the low susceptible phytate diets. Findings from this study suggest that there may be a measure more meaningful to animal nutritionists than measurements of total phytate

A multicenter phase 4 geriatric assessment directed trial to evaluate gemcitabine +/− nab-paclitaxel in elderly pancreatic cancer patients (GrantPax)

Background: In the group of elderly patients (≥70 years) with metastatic pancreatic ductal adenocarcinoma (mPDAC), it is not known who benefits from intensive 1st line nab-paclitaxel/gemcitabine (nab-p/gem) combination chemotherapy or who would rather suffer from increased toxicity. We aim to determine whether treatment individualization by comprehensive geriatric assessments (CGAs) improves functional outcome of the patients. Methods/Design: GrantPax is a multicenter, open label phase 4 interventional trial. We use a CGA to stratify elderly patients into three parallel treatment groups (n = 45 per arm): 1) GOGO (nab-p/gem), 2) SLOWGO (gem mono) or 3) FRAIL (best supportive care). After the 1st cycle of chemotherapy (or 4 weeks in FRAIL group) another CGA and safety assessment is performed. CGA-stratified patients may not decline in their CGA performance in response to the first cycle of chemotherapy (primary objective), measured as a loss of 5 points or less in Barthels activities of daily living. Based on the second CGA, patients are re-assigned to their definite treatment arm and undergo further CGAs to monitor the course of treatment. Secondary endpoints include CGA scores during the course of therapy (CGA1–4), response rates, safety and survival rates. Discussion: GrantPax is the first trial implementing a CGA-driven treatment to personalize therapy for elderly patients with pancreatic cancer. This may lead to standardization of therapy decisions for elderly patients and may optimize standard of care for this increasing group of patients. Trial registration: NCT02812992 , registered 24.06.2016