Guanylate binding proteins (GBPs) are an interferon (IFN)-inducible subfamily of guanosine triphosphatases (GTPases) with well-established activity against intracellular bacteria and parasites. Here ...we show that GBP5 potently restricts HIV-1 and other retroviruses. GBP5 is expressed in the primary target cells of HIV-1, where it impairs viral infectivity by interfering with the processing and virion incorporation of the viral envelope glycoprotein (Env). GBP5 levels in macrophages determine and inversely correlate with infectious HIV-1 yield over several orders of magnitude, which may explain the high donor variability in macrophage susceptibility to HIV. Antiviral activity requires Golgi localization of GBP5, but not its GTPase activity. Start codon mutations in the accessory vpu gene from macrophage-tropic HIV-1 strains conferred partial resistance to GBP5 inhibition by increasing Env expression. Our results identify GBP5 as an antiviral effector of the IFN response and may explain the increased frequency of defective vpu genes in primary HIV-1 strains.
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•GBP5 reduces HIV-1 infectivity by interfering with Env processing and incorporation•GBP5 expression levels determine infectious HIV-1 production in macrophages•Antiretroviral activity of GBP5 requires Golgi localization but not GTPase activity•Mutations in vpu reduce HIV-1 susceptibility to GBP5 by increasing Env expression
GBPs are IFN-inducible guanosine triphosphatases important for cell-intrinsic immunity against bacteria and parasites. Krapp et al. demonstrate that GBP5, expressed in HIV-1 target cells, interferes with HIV-1 Env glycoprotein to potently suppress infectious virus yield. Vpu mutations in macrophage-tropic HIV-1 strains increase Env expression and confer partial resistance to GBP5.
Autophagy is a cellular homeostatic pathway with functions ranging from cytoplasmic protein turnover to immune defense. Therapeutic modulation of autophagy has been demonstrated to positively impact ...the outcome of autophagy-dysregulated diseases such as cancer or microbial infections. However, currently available agents lack specificity, and new candidates for drug development or potential cellular targets need to be identified. Here, we present an improved method to robustly detect changes in autophagy in a high-throughput manner on a single cell level, allowing effective screening. This method quantifies eGFP-LC3B positive vesicles to accurately monitor autophagy. We have significantly streamlined the protocol and optimized it for rapid quantification of large numbers of cells in little time, while retaining accuracy and sensitivity. Z scores up to 0.91 without a loss of sensitivity demonstrate the robustness and aptness of this approach. Three exemplary applications outline the value of our protocols and cell lines: (I) Examining autophagy modulating compounds on four different cell types. (II) Monitoring of autophagy upon infection with e.g. measles or influenza A virus. (III) CRISPR/Cas9 screening for autophagy modulating factors in T cells. In summary, we offer ready-to-use protocols to generate sensitive autophagy reporter cells and quantify autophagy in high-throughput assays.
Tetherin (or BST-2) is an antiviral host restriction factor that suppresses the release of HIV-1 and other enveloped viruses by tethering them to the cell surface. Recently, it has been demonstrated ...that tetherin also acts as an innate sensor of HIV-1 assembly that induces NF-κB-dependent proinflammatory responses. Furthermore, it has been reported that polymorphisms in the promoter and 3' untranslated region of the bst2 gene may affect the clinical outcome of HIV-1 infection. However, non-synonymous polymorphisms in the bst2 open reading frame have not yet been described or functionally characterized.
Mining of the Exome Variant Server database identified seven very rare naturally occurring missense variants of tetherin (Y8H, R19H, N49S, D103N, E117A, D129E and V146L) in human populations. Functional analyses showed that none of these sequence variants significantly affects the ability of tetherin to inhibit HIV-1 virion release or its sensitivity to antagonism by HIV-1 Vpu or SIVtan Env, although Y8H alters a potential YxY endocytic motif proposed to play a role in virion uptake. Thus, these variants do most likely not represent an evolutionary advantage in directly controlling HIV-1 replication or spread. Interestingly, however, the R19H variant selectively abrogated the signaling activity of tetherin.
Restriction of HIV-1 virion release and immune sensing are two separable functions of human tetherin and the latter activity is severely impaired by a single amino acid variant (R19H) in the cytoplasmic part of tetherin.
Background
Internet‐based participatory surveillance systems, such as the German GrippeWeb, monitor the frequency of acute respiratory illnesses on population level. In order to interpret syndromic ...information better, we devised a microbiological feasibility study (GrippeWeb‐Plus) to test whether self‐collection of anterior nasal swabs is operationally possible, acceptable for participants and can yield valid data.
Methods
We recruited 103 GrippeWeb participants (73 adults and 30 children) and provided them with a kit, instructions and a questionnaire for each sample. In the first half of 2016, participants took an anterior nasal swab and sent it to the Robert Koch Institute whenever an acute respiratory illness occurred. Reporting of illnesses through the GrippeWeb platform continued as usual. We analysed swabs for the presence of human c‐myc‐DNA and 22 viral and bacterial pathogens. After the study, we sent participants an evaluation questionnaire. We analysed timeliness, completeness, acceptability and validity.
Results
One hundred and two participants submitted 225 analysable swabs. Ninety per cent of swabs were taken within 3 days of symptom onset. Eighty‐nine per cent of swabs had a corresponding reported illness in the GrippeWeb system. Ninety‐nine per cent of adults and 96% of children would be willing to participate in a self‐swabbing scheme for a longer period. All swabs contained c‐myc‐DNA. In 119 swabs, we identified any of 14 viruses but no bacteria. The positivity rate of influenza was similar to that in the German physician sentinel.
Conclusion
Self‐collection of anterior nasal swabs proofed to be feasible, was well accepted by participants, gave valid results and was an informative adjunct to syndromic data.
Microplastics (MP) enter the aquatic environment via several pathways. Many research groups have focused on municipal discharge, while research on industrial sources is rare. This study provides one ...of the first insights into MP occurrence and distribution in the wastewater systems of industrial parks (IPs) and their wastewater treatment plants (IPWWTPs). The effluents from production plants as well as influent, effluent, and internal samples from the IPWWTPs were assessed. Sampling methods for parallel MP mass and number analyses were developed for varying conditions. The total item emissions of MP (≥10 μm) into the environment were analyzed using μ-Raman spectroscopy and ranged from 3 · 102 to 8 · 104 MP m−3, with a median of 6 · 103 MP m−3 per IPWWTP. Masses analyzed using differential scanning calorimetry showed an MP mass discharge into the environment of 0.2 to 11 mg m−3 with a median of 3.7 mg m−3 per IPWWTP. MP item concentrations within an IPWWTP varied by two to three log levels over several days. Fibers were rare in all samples. Polymer types varied depending on the types of industrial sites and the production plants discharging into the IPWWTP. Within an IP, MP could be allocated to its dischargers, which could be useful for future regulatory requirements. Further research is needed to include different types of IPs producing various polymers and additional processing plants to expand this data set.
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•German industrial parks as MP emitters to the environment were researched in detail.•Detection of mass- and number concentrations with a simultaneous sampling technique•MP deriving from plastic producers is detectable throughout the wastewater system.•Total MP numbers per day within IPWWTPs fluctuate greatly over time.•In- and effluent MP concentrations of industrial and municipal WWTPs are comparable.
Designästhetik Oliver Ruf, Stefan Neuhaus / Oliver Ruf, Stefan Neuhaus
2020
eBook
Design ist allgegenwärtig - es durchtränkt gleichsam das Leben, oftmals unbewusst, doch immer durchscheinend und folgenhaft. Es ist Bestandteil des Habitus und unverzichtbarer Teil einer jeden ...Identität. Selbst im bewussten Verzicht auf Design kommt eine spezifische Designästhetik zum Ausdruck, die anders sein möchte. Doch welche Normierungen greifen hier und dienen als Orientierung für Absetzbewegungen? Dies zeigt sich vor allem in der Prägung durch massenmediale Diskurse.Die Beiträge des Bandes entwickeln zu diesem zentralen Dispositiv einen theoretischen wie praktischen Rahmen und reflektieren Indikatoren für entsprechende Leistungen.
House dust mite (HDM) allergens play an important role in the pathogenesis of perennial allergy and bronchial asthma. There is broad consensus that assessment of exposure to HDM allergens is mainly ...based on measurements of allergen concentrations in dust and that the "threshold" level for sensitization in atopic children is approximately 2000 ng of group I allergen (sum of Der p 1 and Der f 1) per gram of dust. Many data are available about exposure to HDM allergens for persons in private homes and public buildings (eg, schools and day care centers) but hardly any for military personnel at their places of duty. In this article we present the results of a pilot study of HDM allergens in German submarines.
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