The use of human sewage screening for community surveillance of hepatitis E virus in the UK

Hepatitis E virus sequences were detected by RT‐PCR in 14/15 (93%) of untreated sewage samples from Edinburgh, Scotland, UK. Phylogenetic analysis of amplicons at limiting dilution revealed the co‐circulation of multiple variants of HEV‐3, with a pattern of diversity matching that observed in a local cohort of HEV‐infected hepatitis patients. J. Med. Virol. 88:915–918, 2016. © 2015 The Authors. Journal of Medical Virology Published by Wiley Periodicals, Inc

Mechanochemically Synthetized PAN-Based Co-N-Doped Carbon Materials as Electrocatalyst for Oxygen Evolution Reaction

We report a new class of polyacrylonitrile (PAN)-based Co-N-doped carbon materials that can act as suitable catalyst for oxygen evolution reactions (OER). Different Co loadings were mechanochemically added into post-consumed PAN fibers. Subsequently, the samples were treated at 300 °C under air (PAN-A) or nitrogen (PAN-N) atmosphere to promote simultaneously the Co3O4 species and PAN cyclization. The resulting electrocatalysts were fully characterized and analyzed by X-ray diffraction (XRD) and photoelectron spectroscopy (XPS), transmission (TEM) and scanning electron (SEM) microscopies, as well as nitrogen porosimetry. The catalytic performance of the Co-N-doped carbon nanomaterials were tested for OER in alkaline environments. Cobalt-doped PAN-A samples showed worse OER electrocatalytic performance than their homologous PAN-N ones. The PAN-N/3% Co catalyst exhibited the lowest OER overpotential (460 mV) among all the Co-N-doped carbon nanocomposites, reaching 10 mA/cm2. This work provides in-depth insights on the electrocatalytic performance of metal-doped carbon nanomaterials for OER

Akt phosphorylation of HCV NS5B regulates polymerase activity and HCV infection

Hepatitis C virus (HCV) is a single-stranded RNA virus of positive polarity [ssRNA(+)] that replicates its genome through the activity of one of its proteins, called NS5B. This viral protein is responsible for copying the positive-polarity RNA genome into a negative-polarity RNA strand, which will be the template for new positive-polarity RNA genomes. The NS5B protein is phosphorylated by cellular kinases, including Akt. In this work, we have identified several amino acids of NS5B that are phosphorylated by Akt, with positions S27, T53, T267, and S282 giving the most robust results. Site-directed mutagenesis of these residues to mimic (Glu mutants) or prevent (Ala mutants) their phosphorylation resulted in a reduced NS5B in vitro RNA polymerase activity, except for the T267E mutant, the only non-conserved position of all those that are phosphorylated. In addition, in vitro transcribed RNAs derived from HCV complete infectious clones carrying mutations T53E/A and S282E/A were transfected in Huh-7.5 permissive cells, and supernatant viral titers were measured at 6 and 15 days post-transfection. No virus was rescued from the mutants except for T53A at 15 days post-transfection whose viral titer was statistically lower as compared to the wild type. Therefore, phosphorylation of NS5B by cellular kinases is a mechanism of viral polymerase inactivation. Whether this inactivation is a consequence of interaction with cellular kinases or a way to generate inactive NS5B that may have other functions are questions that need further experimental workMinisterio de Ciencia, Innovación y Universidades (MCIU), PI18/00210 from Instituto de Salud Carlos III, and S2018/BAA-4370 (PLATESA2 from Comunidad de Madrid/FEDER). CP was supported by the Miguel Servet program of the Instituto de Salud Carlos III (CPII19/00001), cofinanced by the European Regional Development Fund (ERDF). CG-C was supported by the predoctoral contract PRE2018-083422 from MCIU. CIBERehd (Centro de Investigación en Red de Enfermedades Hepáticas y Digestivas) was funded by the Instituto de Salud Carlos III. Institutional grants from the Fundación Ramón Areces and Banco Santander to the CBMS

In vivo targeting of adoptively transferred T-cells with antibody- and cytokine-conjugated liposomes

In adoptive cell therapy (ACT), autologous tumor-specific T-cells isolated from cancer patients are activated and expanded ex vivo, then infused back into the individual to eliminate metastatic tumors. A major limitation of this promising approach is the rapid loss of ACT T-cell effector function in vivo due to the highly immunosuppressive environment in tumors. Protection of T-cells from immunosuppressive signals can be achieved by systemic administration of supporting adjuvant drugs such as interleukins, chemotherapy, and other immunomodulators, but these adjuvant treatments are often accompanied by serious toxicities and may still fail to optimally stimulate lymphocytes in all tumor and lymphoid compartments. Here we propose a novel strategy to repeatedly stimulate or track ACT T-cells, using cytokines or ACT-cell-specific antibodies as ligands to target PEGylated liposomes to transferred T-cells in vivo. Using F(ab′)[subscript 2] fragments against a unique cell surface antigen on ACT cells (Thy1.1) or an engineered interleukin-2 (IL-2) molecule on an Fc framework as targeting ligands, we demonstrate that > 95% of ACT cells can be conjugated with liposomes following a single injection in vivo. Further, we show that IL-2-conjugated liposomes both target ACT cells and are capable of inducing repeated waves of ACT T-cell proliferation in tumor-bearing mice. These results demonstrate the feasibility of repeated functional targeting of T-cells in vivo, which will enable delivery of imaging contrast agents, immunomodulators, or chemotherapy agents in adoptive cell therapy regimens.National Institutes of Health (U.S.) (CA140476)National Institutes of Health (U.S.) (CA172164)United States. Dept. of Defense (Contract W81XWH-10-1-0290)National Cancer Institute (U.S.) (Koch Institute Support (core) Grant P30-CA14051

Hepatitis E Virus Epidemiology in Industrialized Countries

To determine the prevalence of Hepatitis E virus (HEV) in industrialized nations, we analyzed the excretion of HEV strains by the populations of Spain, France, Greece, Sweden, and the United States. Twenty of 46 (43.5%) urban sewage samples collected in Barcelona from 1994 to 2002 tested positive for HEV. We identified 15 HEV strains, which were similar to two HEV isolates previously described in Barcelona in clinical samples and to strains from diverse geographic HEV-nonendemic areas. We also identified two HEV strains in sewage samples from Washington, D.C., and Nancy, France; these samples were also positive for Hepatitis A virus. In addition, we studied the role of pigs as a reservoir for HEV and identified one new swine HEV strain. Our results suggest that HEV may be more prevalent than previously considered in industrialized countries and that variants of the virus circulate simultaneously in one region

Environmental Surveillance. An Additional/Alternative Approach for Virological Surveillance in Greece?

The detection of viruses in the sewage of an urban city by nucleic acid amplification techniques allows the identification of the viral strains that are circulating in the community. The aim of the study was the application of such detection which gives useful data on the distribution, spread, and frequency of these viruses, supporting epidemiological studies of the related viral infections. A two year (2007–2009) survey was conducted in order to evaluate the presence of human adenoviruses (hAdV), hepatitis A viruses (HAV), hepatitis E viruses (HEV), noroviruses (NoV), and human polyomaviruses (hPyV) in sewage samples collected from the inlet of a municipal biological wastewater treatment plant located in southwestern Greece. PCR methods were used for this survey. In total, viruses have been detected in 87.5% (42/48) of the analyzed sewage samples. Analytically, DNA viruses, hAdVs and hPyVs have been detected in 45.8% (22/48) and 68.8% (33/48) of the samples, respectively. As it concerns RNA viruses, HAV was detected in 8.3% (4/48), NoVs in 6.3% (3/48), while HEV has not been detected at all. After sequencing, AdVs were typed as Ad8, Ad40 and Ad41, while both JC and BK hPyVs have been recognized. All NoVs have been identified as GII4, while HAV was typed as genotype IA. Similar long-term studies could be undertaken in countries such as Greece in order to offer a valuable and complementary tool to current problematic epidemiological surveillance systems. This study demonstrates the advantages of environmental surveillance as a tool to determine the epidemiology of viruses circulating in a given community. To our knowledge this was the first of its kind study performed in Greece in order to establish this new way of surveillance

Sources of Hepatitis E Virus Genotype 3 in the Netherlands

Four subtypes have been detected, and pathogenicity, zoonotic potential, or stability may vary between subtypes

Non-travel related Hepatitis E virus genotype 3 infections in the Netherlands; A case series 2004 – 2006

<p>Abstract</p> <p>Background</p> <p>Human hepatitis E virus (HEV) infections are considered an emerging disease in industrialized countries. In the Netherlands, Hepatitis E virus (HEV) infections have been associated with travel to high-endemic countries. Non-travel related HEV of genotype 3 has been diagnosed occasionally since 2000. A high homology of HEV from humans and pigs suggests zoonotic transmission but direct molecular and epidemiological links have yet to be established. We conducted a descriptive case series to generate hypotheses about possible risk factors for non-travel related HEV infections and to map the genetic diversity of HEV.</p> <p>Methods</p> <p>A case was defined as a person with HEV infection laboratory confirmed (positive HEV RT-PCR and/or HEV IgM) after 1 January 2004, without travel to a high-endemic country three months prior to onset of illness. For virus identification 148 bp of ORF2 was sequenced and compared with HEV from humans and pigs. We interviewed cases face to face using a structured questionnaire and collected information on clinical and medical history, food preferences, animal and water contact.</p> <p>Results</p> <p>We interviewed 19 cases; 17 were male, median age 50 years (25–84 y), 12 lived in the North-East of the Netherlands and 11 had preexisting disease. Most common symptoms were dark urine (n = 16) and icterus (n = 15). Sixteen ate pork ≥ once/week and six owned dogs. Two cases had received blood transfusions in the incubation period. Seventeen cases were viremic (genotype 3 HEV), two had identical HEV sequences but no identified relation. For one case, HEV with identical sequence was identified from serum and surface water nearby his home.</p> <p>Conclusion</p> <p>The results show that the modes of transmission of genotype-3 HEV infections in the Netherlands remains to be resolved and that host susceptibility may play an important role in development of disease.</p