The hepatitis B virus PRE contains a splicing regulatory element

The posttranscriptional regulatory element (PRE) is considered to enhance hepatitis B virus (HBV) gene expression by facilitating the nuclear export of intronless viral subgenomic RNAs. Its role in the RNA metabolism of the viral pregenomic RNA (pgRNA) is currently unknown. We identified a positively cis-acting splicing regulatory element (SRE-1) and present two lines of evidence for its functionality. Firstly, in a heterologous context SRE-1 functionally substitutes for a retroviral bidirectional exonic splicing enhancer (ESE). As expected, SRE-1 is a splicing enhancer also in its natural viral sequence context, since deletion of SRE-1 reduces splicing of pgRNA in cell culture experiments. Secondly, we show that stimulation of HBV RNA splicing by the splicing factor PSF was repressed by the PRE. Analysis of a variety of PSF mutants indicated that RNA-binding and protein–protein interaction were required to enhance splicing. In addition, we show that the PRE contributed to pgRNA stability, but has little influence on its nuclear export. Herein, we report for the first time that the PRE harbors splicing stimulating and inhibiting regulatory elements controlling processing of the viral pregenome. We discuss a model in which the regulation of pgRNA splicing depends on cellular factors interacting with the PRE

Neue Methoden zur Bekämpfung des Echten Mehltaus im Ökologischen Landbau

Blattoberflächen-modifizierende Salzapplikationen sollten als Bekämpfungsmittel gegen Echten Mehltau an Tomaten und Gurken getestet werden hinsichtlich folgender Eigenschaften: - Wirkung verschiedener Salze - Blattverträglichkeit - Untersuchung der unterschiedlichen Wirkungsmechanismen mittels REM - Übertragbarkeit in praxisnahe Versuche. Die verschiedenen Salzlösungen und Kombinationen aus diesen Salzen unterschieden sich deutlich in ihrer Wirkung auf Gurkenmehltau. Die beste Wirkung ergab sich mit allenfalls geringer Schädigung des Blattes mit 0,1% MnCl2, 1 % Patentkali (K2SO4/MgSO4) und 1% Knöterichextrakt. MnCl2 liegt auf Grund seines niedrigen Deliqueszenzpunktes ständig gelöst auf der Blattoberfläche vor, wogegen sich Knöterichextrakt und Patentkali auf der Blattoberfläche in einer Art „Salzkriechen“ verteilen durch häufigen Wechsel zwischen Trocknen und Wiederverflüssigung. Salzschäden ließen sich verringern durch Zusatz von Netzmitteln zur Sprühlösung. Rasterelektronische Visualisierung zeigte für alle untersuchten Lösungen einen direkt toxischen Effekt auf die Mehltausporen und ihre Hyphen in jeder Entwicklungsphase des Pilzes. Eine Verbesserung des Nährstoffstatus der Pflanzen scheidet als Wirkungsmechanismus aus, da alle Pflanzen optimal ernährt waren und die Behandlungen zu keinen signifikanten Veränderungen der Nährstoffgehalte in den Blättern führten. Besonders chloridische Salze führten z.T. zu erheblicher Nekrotisierung der Blätter. Da diese Salzlösungen jedoch auch in geringer Konzentration in der Lage waren, die Ausbreitung des Echten Mehltau effektiv zu verhindern, sollten Möglichkeiten zur Mikrodosierung dieser Salze geprüft werden. Extreme Witterungsbedingungen im Sommer 2003 führten dazu, dass Praxistests nicht ausgeführt oder normal beendet werden konnten. Es gilt jedoch als sicher, dass einige der untersuchten Salzmischungen in geringen Konzentrationen (zw. 0,1 und 1%) den Befall mit Echten Mehltaupilzen effektiv reduzieren können, ohne Blattoberflächen zu schädigen

In situ investigation of the bainitic transformation from deformed austenite during continuous cooling in a low carbon Mn-Si-Cr-Mo steel

The effects of hot deformation on the bainitic transformation of a low carbon steel during continuous cooling were comprehensively studied through in situ high-energy synchrotron X-ray diffraction, dilatometry, and ex situ microstructural characterizations. The obtained results indicated that the prior deformation of austenite at 950 C accelerates the bainite formation at the early stages. During the ongoing of the transformation, both the overall kinetics of bainite and carbon enrichment of austenite are lower in deformed austenite. The bainitic microstructure developed from deformed austenite is more refined and presents the same retained austenite content at room temperature with slightly lower carbon content when compared with the undeformed sample. Besides, a significant higher dilatation strain was measured during the bainitic transformation in the deformed sample, which can be explained by the crystallographic texture in hot deformed austenite. The evolution of the peak broadening of the {220}c and {211}a reflections during bainitic transformation are discussed in detail

rac-4-{(E)-[1-Cyano-1-cyclo­hexyl-2-(1H-indol-3-yl)eth­yl]imino­meth­yl}benzonitrile

A phosphine-catalysed addition of gramine to an alkyl­idene­amino­nitrile gives the title compound, C25H24N4, in good yield. In the crystal, pairs of mol­ecules are connected via N—H⋯N hydrogen bonds into inversion dimers. The mol­ecules are characterized by a planar indole moiety [maximum deviation = 0.012 (1) Å], a chair conformation of the cyclo­hexane ring and an anti­periplanar conformation of the H atom on the cyclo­hexane and the adjacent cyano group

Hepatitis E virus genome detection in commercial pork livers and pork meat products in Germany

The hepatitis E virus (HEV) is one of the most common causes of hepatitis worldwide. HEV is also widespread in many developed countries, where the number of infections is steadily increasing. In those countries, the virus is transmitted mainly through consumption of undercooked or raw food or through contact with animals. Especially, pigs serve as a main reservoir of HEV. Here, we investigated the prevalence of HEV RNA in pork livers and pork meat products to assess the actual risk of HEV infection through food consumption in Germany. A total of 131 pork products were collected from grocery stores and butcher shops between October 2019 and February 2020 and screened for HEV RNA using nested PCR and subsequent sequencing. Overall, 10% of the samples were positive for HEV, including pork livers (5%), spreadable liver sausages (13%) and liver pâté samples (15%). Sequence analyses indicated that the large majority of HEV strains belonged to subtype HEV-3c, representing the most frequent subtype in Germany. One sample belonged to subtype HEV-3f. Further sequence analysis revealed large sequence variation between the samples; however, most of the mutations identified were synonymous. Although infectivity of the virus was not tested, the results suggest a considerable risk of HEV infection through food consumption. Therefore, preventive measures should be taken according to a One Health approach.Peer Reviewe

Predictors of response to intra-arterial vasodilatory therapy of non-occlusive mesenteric ischemia in patients with severe shock: results from a prospective observational study

Background: Non-occlusive mesenteric ischemia (NOMI) is a life-threatening condition occurring in patients with shock and is characterized by vasoconstriction of the mesenteric arteries leading to intestinal ischemia and multi-organ failure. Although minimal invasive local intra-arterial infusion of vasodilators into the mesenteric circulation has been suggested as a therapeutic option in NOMI, current knowledge is based on retrospective case series and it remains unclear which patients might benefit. Here, we prospectively analyzed predictors of response to intra-arterial therapy in patients with NOMI. Methods: This is a prospective single-center observational study to analyze improvement of ischemia (indicated by reduction of blood lactate > 2 mmol/l from baseline after 24 h, primary endpoint) and 28-day mortality (key secondary endpoint) in patients with NOMI undergoing intra-arterial vasodilatory therapy. Predictors of response to therapy concerning primary and key secondary endpoint were identified using a) clinical parameters as well as b) data from 2D-perfusion angiography and c) experimental biomarkers of intestinal injury. Results: A total of 42 patients were included into this study. At inclusion patients had severe shock, indicated by high doses of norepinephrine (NE) (median (interquartile range (IQR)) 0.37 (0.21-0.60) μg/kg/min), elevated lactate concentrations (9.2 (5.2-13) mmol/l) and multi-organ failure. Patients showed a continuous reduction of lactate following intra-arterial prostaglandin infusion (baseline: (9.2 (5.2-13) mmol/l vs. 24 h: 4.4 (2.5-9.1) mmol/l, p 2 mmol/l at 24 h following intervention. Initial higher lactate concentrations and lower NE doses at baseline were independent predictors of an improvement of ischemia. 28-day mortality was 59% in patients with a reduction of lactate > 2 mmol/l 24 h after inclusion, while it was 85% in all other patients (hazard ratio 0.409; 95% CI, 0.14-0.631, p = 0.005). Conclusions: A reduction of lactate concentrations was observed following implementation of intra-arterial therapy, and lactate reduction was associated with better survival. Our findings concerning outcome predictors in NOMI patients undergoing intra-arterial prostaglandin therapy might help designing a randomized controlled trial to further investigate this therapeutic approach. Trial registration Retrospectively registered on January 22, 2020, at clinicaltrials.gov (REPERFUSE, NCT04235634), https://clinicaltrials.gov/ct2/show/NCT04235634?cond=NOMI&draw=2&rank=1 . Keywords: Intestinal failure; Non-occlusive mesenteric ischemia; Sepsis; Shoc

Absence of chronic hepatitis E in a German cohort of common variable immunodeficiency patients

Cases of chronic or prolonged hepatitis E virus (HEV) infections have been described in solid organ transplant recipients, HIV infected patients and in patients with malignancies or idiopathic CD4+ T lymphopenia. It is unknown if HEV infection also takes chronic courses in patients with common variable immunodeficiency (CVID). We studied a cohort of 73 CVID patients recruited in a low endemic Central European country. None of the subjects tested positive for HEV RNA or anti-HEV IgG. Immunoglobulin transfusions (n=10) tested negative for HEV RNA but all were anti-HEV positive. To verify that such pooled blood products contain anti-HEV protective antibodies we measured the anti-HEV IgG optical density (OD) values in patients before and after transfusion. Anti-HEV OD values increased after infusion but did not reach the cut-off considered as positive. Thus, chronic HEV infections seem to be rare events in CVID patients in Germany. Commercially available immuno globulin infusions contain anti HEV antibodies and may contribute to protection from HEV infectio

PROTEINCHALLENGE: Crowd sourcing in proteomics analysis and software development

AbstractIn large-scale proteomics studies there is a temptation, after months of experimental work, to plug resulting data into a convenient—if poorly implemented—set of tools, which may neither do the data justice nor help answer the scientific question. In this paper we have captured key concerns, including arguments for community-wide open source software development and “big data” compatible solutions for the future. For the meantime, we have laid out ten top tips for data processing. With these at hand, a first large-scale proteomics analysis hopefully becomes less daunting to navigate.However there is clearly a real need for robust tools, standard operating procedures and general acceptance of best practises. Thus we submit to the proteomics community a call for a community-wide open set of proteomics analysis challenges—PROTEINCHALLENGE—that directly target and compare data analysis workflows, with the aim of setting a community-driven gold standard for data handling, reporting and sharing. This article is part of a Special Issue entitled: New Horizons and Applications for Proteomics [EuPA 2012]

Antiretroviral penetration into the CNS and incidence of AIDS-defining neurologic conditions

Objective: The link between CNS penetration of antiretrovirals and AIDS-defining neurologic disorders remains largely unknown. Methods: HIV-infected, antiretroviral therapy-naive individuals in the HIV-CAUSAL Collaboration who started an antiretroviral regimen were classified according to the CNS Penetration Effectiveness (CPE) score of their initial regimen into low (,8), medium (8-9), or high (.9) CPE score. We estimated "intention-to-treat" hazard ratios of 4 neuroAIDS conditions for baseline regimens with high and medium CPE scores compared with regimens with a low score. We used inverse probability weighting to adjust for potential bias due to infrequent follow-up. Results: A total of 61,938 individuals were followed for a median (interquartile range) of 37 (18, 70) months. During follow-up, there were 235 cases of HIV dementia, 169 cases of toxoplasmosis, 128 cases of cryptococcal meningitis, and 141 cases of progressive multifocal leukoencephalopathy. The hazard ratio (95% confidence interval) for initiating a combined antiretroviral therapy regimen with a high vs low CPE score was 1.74 (1.15, 2.65) for HIV dementia, 0.90 (0.50, 1.62) for toxoplasmosis, 1.13 (0.61, 2.11) for cryptococcal meningitis, and 1.32 (0.71, 2.47) for progressive multifocal leukoencephalopathy. The respective hazard ratios (95% confidence intervals) for a medium vs low CPE score were 1.01 (0.73, 1.39), 0.80 (0.56, 1.15), 1.08 (0.73, 1.62), and 1.08 (0.73, 1.58). Conclusions: We estimated that initiation of a combined antiretroviral therapy regimen with a high CPE score increases the risk of HIV dementia, but not of other neuroAIDS conditions