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Hepatitis E virus (HEV) is the causative agent of hepatitis E in humans and the leading cause for acute viral hepatitis worldwide. The virus is classified as a member of the genus Orthohepevirus A within the Hepeviridae family. Due to the absence of a robust cell culture model for HEV infection, the analysis of the viral life cycle, the development of effective antivirals and a vaccine is severely limited. In this study, we established a protocol based on the HEV genotype 3 p6 (Kernow C-1) and the human hepatoma cell lines HepG2 and HepG2/C3A with different media conditions to produce intracellular HEV cell culturederived particles (HEVcc) with viral titers between 10⁵ and 10⁶ FFU/mL. Viral titers could be further enhanced by an HEV variant harboring a mutation in the RNA-dependent RNA polymerase. These HEVcc particles were characterized in density gradients and allowed the trans-complementation of subgenomic reporter HEV replicons. In addition, in vitro produced intracellular-derived particles were infectious in liver-humanized mice with high RNA copy numbers detectable in serum and feces. Efficient infection of primary human and swine hepatocytes using the developed protocol could be observed and was inhibited by ribavirin. Finally, RNA sequencing studies of HEV-infected primary human hepatocytes demonstrated a temporally structured transcriptional defense response. In conclusion, this robust cell culture model of HEV infection provides a powerful tool for studying viral–host interactions that should facilitate the discovery of antiviral drugs for this important zoonotic pathogen

Achieving an energy-neutral building stock by 2050 requires sustainable energy-efficient renovation of facades. Conventional renovation with thick insulation reduces heat losses, but demands large material quantities, particularly when meeting high thermal performance and low U-value standards. Sustainable solutions should combine energy efficiency with resource conservation, low-carbon material cycles, and reduced gray energy. Dynamic exterior wall temperature control offers such a solution by active heat flow management. The EnergySkin project is developing a facade module for energy-efficient renovation. This module will integrate an electrically conductive coating for heating and photovoltaic in an insulating glass unit, which will be mounted on an existing opaque facade as part of a modular system for serial installation. The configuration enables active external wall temperature control whilst generating the necessary electrical power itself, thereby minimizing the heating demand and primary energy consumption of the building without additional CO 2 emissions. The system is being extensively investigated with respect to its thermal and electrical functionality, operating modes, and the integration of an electrical storage unit. Preliminary laboratory and component-scale tests have confirmed the feasibility of coupling transparent conductive coatings with insulating glass to achieve both passive insulation and active heating. Simulations demonstrate that integrated photovoltaic can cover the overall operational energy demand, enabling energy self-sufficiency. These findings highlight the potential of dynamic facade systems to surpass conventional insulation approaches by incorporating adaptability, renewable heat generation and circular design principles. Consequently, the project contributes to sustainable refurbishment strategies and supports climate-neutral building targets.

In order to minimize re-discovery of already known anti-infective compounds, we focused our screening approach on understudied, almost untapped marine environments including marine invertebrates and their associated bacteria. Therefore, two sea cucumber species, Holothuria leucospilota and Stichopus vastus, were collected from Lampung (Indonesia), and 127 bacterial strains were identified by partial 16S rRNA-gene sequencing analysis and compared with the NCBI database. In addition, the overall bacterial diversity from tissue samples of the sea cucumbers H. leucospilota and S. vastus was analyzed using the cultivation-independent Illumina MiSEQ analysis. Selected bacterial isolates were grown to high densities and the extracted biomass was tested against a selection of bacteria and fungi as well as the hepatitis C virus (HCV). Identification of putative bioactive bacterial-derived compounds were performed by analyzing the accurate mass of the precursor/parent ions (MS1) as well as product/daughter ions (MS2) using high resolution mass spectrometry (HRMS) analysis of all active fractions. With this attempt we were able to identify 23 putatively known and two previously unidentified precursor ions. Moreover, through 16S rRNA-gene sequencing we were able to identify putatively novel bacterial species from the phyla Actinobacteria, Proteobacteria and also Firmicutes. Our findings suggest that sea cucumbers like H. leucospilota and S. vastus are promising sources for the isolation of novel bacterial species that produce compounds with potentially high biotechnological potential.

Equine hepacivirus (EqHV) is the closest known genetic homologue of hepatitis C virus. An effective prophylactic vaccine is currently not available for either of these hepaciviruses. The equine as potential surrogate model for hepacivirus vaccine studies was investigated, while equine host responses following vaccination with EqHV E2 recombinant protein and subsequent EqHV inoculation were elucidated. Four ponies received prime and booster vaccinations (recombinant protein, adjuvant) four weeks apart (day −55 and −27). Two control ponies received adjuvant only. Ponies were inoculated with EqHV RNA-positive plasma on day 0. Blood samples and liver biopsies were collected over 26 weeks (day −70 to +112). Serum analyses included detection of EqHV RNA, isotypes of E2-specific immunoglobulin G (IgG), nonstructural protein 3-specific IgG, haematology, serum biochemistry, and metabolomics. Liver tissue analyses included EqHV RNA detection, RNA sequencing, histopathology, immunohistochemistry, and fluorescent in situ hybridization. Al-though vaccination did not result in complete protective immunity against experimental EqHV inoculation, the majority of vaccinated ponies cleared the serum EqHV RNA earlier than the control ponies. The majority of vaccinated ponies appeared to recover from the EqHV-associated liver insult earlier than the control ponies. The equine model shows promise as a surrogate model for future hepacivirus vaccine research.

Hepatitis E virus (HEV) is a member of the genus Orthohepevirus in the family Hepeviridae and the causative agent of hepatitis E in humans. HEV is a major health problem in developing countries, causing mortality rates up to 25% in pregnant women. However, these cases are mainly reported for HEV genotype (gt)1, while gt3 infections are usually associated with subclinical courses of disease. The pathogenic mechanisms of adverse maternal and fetal outcome during pregnancy in HEV‐infected pregnant women remain elusive. In this study, we observed that HEV is capable of completing the full viral life cycle in placental‐derived cells (JEG‐3). Following transfection of JEG‐3 cells, HEV replication of both HEV gts could be observed. Furthermore, determination of extracellular and intracellular viral capsid levels, infectivity, and biophysical properties revealed production of HEV infectious particles with similar characteristics as in liver‐derived cells. Viral entry was analyzed by infection of target cells and detection of either viral RNA or staining for viral capsid protein by immunofluorescence. HEV gt1 and gt3 were efficiently inhibited by ribavirin in placental as well as in human hepatoma cells. In contrast, interferon‐α sensitivity was lower in the placental cells compared to liver cells for gt1 but not gt3 HEV. Simultaneous determination of interferon‐stimulated gene expression levels demonstrated an efficient HEV‐dependent restriction in JEG‐3. Conclusion: We showed differential tissue‐specific host responses to HEV genotypes, adding to our understanding of the mechanisms contributing to fatal outcomes of HEV infections during pregnancy. Using this cell‐culture system, new therapeutic options for HEV during pregnancy can be identified and evaluated. (Hepatology Communications 2018;2:173–187) ‐ HEV genotype 1 and 3 strains are replicating in placental‐derived cell lines ‐ Placental‐derived JEG‐3 cells support full‐length HEV replication, viral assembly and release and are permissive for infection with HEVcc ‐ A differential tissue‐specific host response to HEV genotypes could be observed as the genotype 1 interferon‐α sensitivity was lower in the placental cells as compared to liver cells, which was not the case for HEV genotype 3 ‐ The interferon‐stimulated gene expression is downregulated by HEV in placental‐derived JEG‐3 cells

The anaerobic biodegradability of municipal primary and secondary sludge with increasing levels of partially dewatered fat, oil, and grease (FOG) was assessed using a mixed methanogenic culture at 35 °C. Under batch conditions with an acclimated and enriched microbial population, the sludge loading was 3 kg volatile solids/m 3 and the highest FOG loading tested was 1.5 kg volatile solids/m 3 , resulting in a methane yield of 245 mL methane/g sludge volatile solids added at 35 °C and 1010 mL methane/g FOG volatile solids added at 35 °C. Under semicontinuous feeding conditions, the sludge and sludge plus FOG loading tested were 3 and 3.75 kg volatile solids/m 3 -d, respectively. Within 23 days of operation, the volatile fatty acid concentrations were reduced below 200 mg chemical oxygen demand/L (187 mg/L as acetic acid). Enhancement of sludge digestion was observed in those reactors where codigestion of sludge and FOG took place, which was attributed to a higher level of microbial activity maintained in these reactors as a result of FOG degradation. The results of this study demonstrate that beneficial use of FOG through codigestion with municipal sludge is feasible.

The World Health Organization (WHO) published 2 alcohol-based formulations to be used in healthcare settings and for outbreak-associated infections, but inactivation efficacies of these products have not been determined against (re-)emerging viruses. In this study, we evaluated the virucidal activity of these WHO products in a comparative analysis. Zika virus (ZIKV), Ebola virus (EBOV), severe acute respiratory syndrome coronavirus (SARS-CoV), and Middle East respiratory syndrome coronavirus (MERS-CoV) as (re-)emerging viral pathogens and other enveloped viruses could be efficiently inactivated by both WHO formulations, implicating their use in healthcare systems and viral outbreak situations.

Hepatitis C virus (HCV) and human immunodeficiency virus (HIV-1) transmissions among people who inject drugs (PWID) continue to pose a challenging global health problem. Here, we aimed to analyse a universally applicable inactivation procedure, namely microwave irradiation, as a safe and effective method to reduce the risk of viral transmission. The exposure of HCV from different genotypes to microwave irradiation resulted in a significant reduction of viral infectivity. Furthermore, microwave irradiation reduced viral infectivity of HIV-1 and of HCV/HIV-1 suspensions indicating that this inactivation may be effective at preventing co-infections. To translate microwave irradiation as prevention method to used drug preparation equipment, we could further show that HCV as well as HIV-1 infectivity could be abrogated in syringes and filters. This study demonstrates the power of microwave irradiation for the reduction of viral transmission and establishment of this safety strategy could help reduce the transmission of blood-borne viruses.

The anaerobic biodegradability of municipal primary sludge, thickened waste activated sludge (TWAS), and fat, oil, and grease (FOG) was assessed using semi-continuous-feed, laboratory-scale anaerobic digesters and compared with the ultimate degradability obtained from 120-day batch digestion at 35°C. In run 1, combined primary sludge and TWAS (40/60%, volatile solids [VS] basis) were fed to digesters operated at mesophilic (35°C) and thermophilic (52°C) temperatures at loading rates of 0.99 and 1.46 g-VS/L.d for primary sludge and TWAS, respectively, and a hydraulic retention time (HRT) of 12 days. The volatile solids destruction values were 25.3 and 30.7% (69 and 83% biodegradable volatile solids destruction) at 35°C and 52°C, respectively. The methane (CH₄) yields were 159 and 197 mL at the standard temperature and pressure (STP) conditions of 0°C and 1 atm/g-VS added or 632 and 642 mL @ STP/g-VS destroyed at 35°C and 52°C, respectively. In run 2, a mix of primary sludge, TWAS, and FOG (21/31/48%, volatile solids basis) was fed to an acid digester operated at a 1-day HRT, at 35°C, and a loading rate of 52.5 g-VS/Ld. The acidreactor effluent was fed to two parallel methane-phase reactors operated at an HRT of 12 days and maintained at 35°C and 52°C, respectively. After an initial period of 20 days with near-zero gas production in the acid reactor, biogas production increased and stabilized to approximately 2 mL CH₄ @ STP/g-VS added, corresponding to a volatile solids destruction of 0.4%. The acid-phase reactor achieved a 43% decrease in nonsaturated fat and a 16, 26, and 20% increase of soluble COD, volatile fatty acids, and ammonia, respectively. The methane-phase volatile solids destruction values in run 2 were 45 and 51% (85 and 97% biodegradable volatile solids destruction) at 35°C and 52°C, respectively. The methane yields for the methane-phase reactors were 473 and 551 mL @ STP/g-VS added, which is approximately 3 times larger compared with run 1, or 1040 and 1083 mL @ STP/g-VS destroyed, at 35°C and 52°C, respectively. The results indicate that, when co-digesting municipal sludge and FOG, a large FOG organic load fraction could have a profound effect on the methane gas yield.

The anaerobic biodegradability of municipal primary and secondary sludge with increasing levels of partially dewatered fat, oil and grease (FOG) using a mixed methanogenic culture at 35 degree C was assessed. Measurement of pH was performed using the potentiometric method with a digital pH/milivolt meter and a gel-filled combination pH electrode. Volatile fatty acids were not detected in any of the six reactors at the end of the incubation. The overall volatile solids and chemical oxygen demand (COD) destruction increased as the FOG loading increased. The highest methane production rate was observed between 6 and 9 days of incubation. It was concluded that codigestion of municipal sludge and FOG was an efficient way of significantly increasing methane gas production and allowed for the beneficial use of FOG.