Inactivation of yellow fever virus by WHO-recommended hand rub formulations and surface disinfectants.

Despite continued outbreaks of yellow fever virus (YFV) in endemic regions, data on its environmental stability or guidelines for its effective inactivation is limited. Here, we evaluated the susceptibility of the YFV 17D vaccine strain to inactivation by ethanol, 2-propanol, World Health Organization (WHO)-recommended hand rub formulations I and II, as well as surface disinfectants. In addition, two pathogenic strains were tested to compare inactivation kinetics by WHO-recommended hand rub formulations I and II. Furthermore, environmental stability of the vaccine strain was assessed. YFV 17D particles displayed infectivity half-life decay profiles of ~13 days at room temperature. Despite this extended environmental stability, YFV was efficiently inactivated by alcohols, WHO-recommended hand formulations, and four out of five tested surface disinfectants. These results are useful in defining disinfection protocols to prevent non-vector borne YFV transmission.

TMPRSS2-mediated SARS-CoV-2 uptake boosts innate immune activation, enhances cytopathology, and drives convergent virus evolution.

The accessory protease transmembrane protease serine 2 (TMPRSS2) enhances severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) uptake into ACE2-expressing cells, although how increased entry impacts downstream viral and host processes remains unclear. To investigate this in more detail, we performed infection assays in engineered cells promoting ACE2-mediated entry with and without TMPRSS2 coexpression. Electron microscopy and inhibitor experiments indicated TMPRSS2-mediated cell entry was associated with increased virion internalization into endosomes, and partially dependent upon clathrin-mediated endocytosis. TMPRSS2 increased panvariant uptake efficiency and enhanced early rates of virus replication, transcription, and secretion, with variant-specific profiles observed. On the host side, transcriptional profiling confirmed the magnitude of infection-induced antiviral and proinflammatory responses were linked to uptake efficiency, with TMPRSS2-assisted entry boosting early antiviral responses. In addition, TMPRSS2-enhanced infections increased rates of cytopathology, apoptosis, and necrosis and modulated virus secretion kinetics in a variant-specific manner. On the virus side, convergent signatures of cell-uptake-dependent innate immune induction were recorded in viral genomes, manifesting as switches in dominant coupled Nsp3 residues whose frequencies were correlated to the magnitude of the cellular response to infection. Experimentally, we demonstrated that selected Nsp3 mutations conferred enhanced interferon antagonism. More broadly, we show that TMPRSS2 orthologues from evolutionarily diverse mammals facilitate panvariant enhancement of cell uptake. In summary, our study uncovers previously unreported associations, linking cell entry efficiency to innate immune activation kinetics, cell death rates, virus secretion dynamics, and convergent selection of viral mutations. These data expand our understanding of TMPRSS2's role in the SARS-CoV-2 life cycle and confirm its broader significance in zoonotic reservoirs and animal models.

Targeting cellular cathepsins inhibits hepatitis E virus entry.

BACKGROUND AND AIMS: HEV is estimated to be responsible for 70,000 deaths annually, yet therapy options remain limited. In the pursuit of effective antiviral therapies, targeting viral entry holds promise and has proven effective for other viruses. However, the precise mechanisms and host factors required during HEV entry remain unclear. Cellular proteases have emerged as host factors required for viral surface protein activation and productive cell entry by many viruses. Hence, we investigated the functional requirement and therapeutic potential of cellular protease during HEV infection. APPROACH AND RESULTS: Using our established HEV cell culture model and subgenomic HEV replicons, we found that blocking lysosomal cathepsins (CTS) with small molecule inhibitors impedes HEV infection without affecting replication. Most importantly, the pan-cathepsin inhibitor K11777 suppressed HEV infections with an EC 50 of ~0.02 nM. Inhibition by K11777, devoid of notable toxicity in hepatoma cells, was also observed in HepaRG and primary human hepatocytes. Furthermore, through time-of-addition and RNAscope experiments, we confirmed that HEV entry is blocked by inhibition of cathepsins. Cathepsin L (CTSL) knockout cells were less permissive to HEV, suggesting that CTSL is critical for HEV infection. Finally, we observed cleavage of the glycosylated ORF2 protein and virus particles by recombinant CTSL. CONCLUSIONS: In summary, our study highlights the pivotal role of lysosomal cathepsins, especially CTSL, in the HEV entry process. The profound anti-HEV efficacy of the pan-cathepsin inhibitor K11777, especially with its notable safety profile in primary cells, further underscores its potential as a therapeutic candidate.

Unraveling the dynamics of hepatitis C virus adaptive mutations and their impact on antiviral responses in primary human hepatocytes.

Hepatitis C virus (HCV) infection progresses to chronicity in the majority of infected individuals. Its high intra-host genetic variability enables HCV to evade the continuous selection pressure exerted by the host, contributing to persistent infection. Utilizing a cell culture-adapted HCV population (p100pop) which exhibits increased replicative capacity in various liver cell lines, this study investigated virus and host determinants that underlie enhanced viral fitness. Characterization of a panel of molecular p100 clones revealed that cell culture adaptive mutations optimize a range of virus-host interactions, resulting in expanded cell tropism, altered dependence on the cellular co-factor micro-RNA 122 and increased rates of virus spread. On the host side, comparative transcriptional profiling of hepatoma cells infected either with p100pop or its progenitor virus revealed that enhanced replicative fitness correlated with activation of endoplasmic reticulum stress signaling and the unfolded protein response. In contrast, infection of primary human hepatocytes with p100pop led to a mild attenuation of virion production which correlated with a greater induction of cell-intrinsic antiviral defense responses. In summary, long-term passage experiments in cells where selective pressure from innate immunity is lacking improves multiple virus-host interactions, enhancing HCV replicative fitness. However, this study further indicates that HCV has evolved to replicate at low levels in primary human hepatocytes to minimize innate immune activation, highlighting that an optimal balance between replicative fitness and innate immune induction is key to establish persistence. IMPORTANCE: Hepatitis C virus (HCV) infection remains a global health burden with 58 million people currently chronically infected. However, the detailed molecular mechanisms that underly persistence are incompletely defined. We utilized a long-term cell culture-adapted HCV, exhibiting enhanced replicative fitness in different human liver cell lines, in order to identify molecular principles by which HCV optimizes its replication fitness. Our experimental data revealed that cell culture adaptive mutations confer changes in the host response and usage of various host factors. The latter allows functional flexibility at different stages of the viral replication cycle. However, increased replicative fitness resulted in an increased activation of the innate immune system, which likely poses boundary for functional variation in authentic hepatocytes, explaining the observed attenuation of the adapted virus population in primary hepatocytes.

Repurposing screen identifies novel candidates for broad-spectrum coronavirus antivirals and druggable host targets.

Libraries composed of licensed drugs represent a vast repertoire of molecules modulating physiological processes in humans, providing unique opportunities for the discovery of host-targeting antivirals. We screened the Repurposing, Focused Rescue, and Accelerated Medchem (ReFRAME) repurposing library with approximately 12,000 molecules for broad-spectrum coronavirus antivirals and discovered 134 compounds inhibiting an alphacoronavirus and mapping to 58 molecular target categories. Dominant targets included the 5-hydroxytryptamine receptor, the dopamine receptor, and cyclin-dependent kinases. Gene knock-out of the drugs' host targets including cathepsin B and L (CTSB/L; VBY-825), the aryl hydrocarbon receptor (AHR; Phortress), the farnesyl-diphosphate farnesyltransferase 1 (FDFT1; P-3622), and the kelch-like ECH-associated protein 1 (KEAP1; Omaveloxolone), significantly modulated HCoV-229E infection, providing evidence that these compounds inhibited the virus through acting on their respective host targets. Counter-screening of all 134 primary compound candidates with SARS-CoV-2 and validation in primary cells identified Phortress, an AHR activating ligand, P-3622-targeting FDFT1, and Omaveloxolone, which activates the NFE2-like bZIP transcription factor 2 (NFE2L2) by liberating it from its endogenous inhibitor KEAP1, as antiviral candidates for both an Alpha- and a Betacoronavirus. This study provides an overview of HCoV-229E repurposing candidates and reveals novel potentially druggable viral host dependency factors hijacked by diverse coronaviruses.

Traceable determination of metal composition of tyres using tandem ICP-MS and benchmarking of emissions inventories.

Non-exhaust emissions are becoming of increasing significance with respect to total particulate matter (PM) concentrations in ambient air. Of particular interest is the metal content of this PM since metallic compounds are well known to have toxic effects on human health and the environment. In this study, 'bottom-up' annual tyre wear emission rates were estimated and compared to top-down' emissions declared by the UK; it was calculated that between 14 and 25 tonnes of Zn entered the atmosphere in PM10 in 2020. The emission rates were estimated using a cost-effective, simple but robust validated method for analysis of the metals in tyres using tandem inductively coupled plasma mass spectrometry (ICP-MS/MS) for the first time, involving minimal offline sample preparation. This method was applied to five different tyre makes and brands, all available for sale in the UK, and the uncertainty of each measurement was determined. Traceability was ensured in all methods and novel validation techniques were applied due to lack of available reference materials. Zn was found to be the largest metal component in all tyres with a mass fraction of approximately 10 mg g-1. The mean mass fractions of metals in the tyres decreased in the order of Zn > Al > Fe > Mg > Ti > Pb > Cu > Ba > Ni. Significant differences in composition were found between the five tyres. The relative expanded uncertainties of the metals measurements ranged from 4 to 21%, with elements of higher mass fraction resulting in lower uncertainties. These findings will contribute to assessing current and future air quality challenges and will help to inform regulation surrounding non-exhaust emissions.

Spatiotemporal analysis of SARS-CoV-2 infection reveals an expansive wave of monocyte-derived macrophages associated with vascular damage and virus clearance in hamster lungs.

IMPORTANCE: We present the first study of the 3D kinetics of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection and the early host response in a large lung volume using a combination of tissue imaging and transcriptomics. This approach allowed us to make a number of important findings: Spatially restricted antiviral response is shown, including the formation of monocytic macrophage clusters and upregulation of the major histocompatibility complex II in infected epithelial cells. The monocyte-derived macrophages are linked to SARS-CoV-2 clearance, and the appearance of these cells is associated with post-infection endothelial damage; thus, we shed light on the role of these cells in infected tissue. An early onset of tissue repair occurring simultaneously with inflammatory and necrotizing processes provides the basis for longer-term alterations in the lungs.

Hepatitis C virus cell culture adaptive mutations enhance cell culture propagation by multiple mechanisms but boost antiviral responses in primary human hepatocytes.

Hepatitis C virus (HCV) infection progresses to chronicity in the majority of infected individuals. Its high intra-host genetic variability enables HCV to evade the continuous selection pressure exerted by the host, contributing to persistent infection. Utilizing a cell culture adapted HCV population (p100pop) which exhibits increased replicative capacity in various liver cell lines, this study investigated virus and host determinants which underlie enhanced viral fitness. Characterization of a panel of molecular p100 clones revealed that cell culture adaptive mutations optimize a range of virus-host interactions, resulting in expanded cell tropism, altered dependence on the cellular co-factor micro-RNA 122 and increased rates of virus spread. On the host side, comparative transcriptional profiling of hepatoma cells infected either with p100pop or its progenitor virus revealed that enhanced replicative fitness correlated with activation of endoplasmic reticulum stress signaling and the unfolded protein response. In contrast, infection of primary human hepatocytes with p100pop led to a mild attenuation of virion production which correlated with a greater induction of cell-intrinsic antiviral defense responses. In summary, long-term passage experiments in cells where selective pressure from innate immunity is lacking improves multiple virus-host interactions, enhancing HCV replicative fitness. However, this study further indicates that HCV has evolved to replicate at low levels in primary human hepatocytes to minimize innate immune activation, highlighting that an optimal balance between replicative fitness and innate immune induction is key to establishing persistence.

A robust regression analysis method to determine the significance of trends in concentrations of heavy metals in UK ambient air and improve network design and emission inventories.

A novel application of the Theil-Sen robust regression method for determining the temporal trends in the concentration of heavy metals in UK ambient air over the period 2005-2020 is presented and compared to other regression methods. We have demonstrated improvements over non-robust methods of regression, proving the ability to tease out trends that are small with respect to the variability of the concentration measurement. The method is used to identify, in general, large and significant trends in the concentrations of Ni, As, Pb and V over the period 2005-2020, either across the UK as a whole or at groupings of site classifications in the UK. These trends have been compared to trends in emission data determined in the same manner. Although the results for most metals provide confidence that the UK metal network of monitoring sites is successful in appropriately capturing changes in emissions, a key finding of this work is the disagreement between trends in measured concentrations and emissions for Cu, Mn and Ni, for which we suggest improvements in future network design. The results also indicate that UK emission data for V should be reviewed, as we propose that the rate of reduction of V emissions is likely to have been overestimated.

Hazardous particles during diesel engine cold-start and warm-up: Characterisation of particulate mass and number under the impact of biofuel and lubricating oil.

The increasing share of using biofuels in vehicles (mandated by current regulations) leads to a reduction in particle size, resulting in increased particle toxicity. However, existing regulations disregarded small particles (sub-23 nm) that are more toxic. This impact is more significant during vehicle cold-start operation, which is an inevitable frequent daily driving norm where after-treatment systems prove ineffective. This study investigates the impact of biofuel and lubricating oil (as a source of nanoparticles) on the concentration, size distribution, median diameter of PN and PM, and their proportion at size ranges within accumulation and nucleation modes during four phases of cold-start and warm-up engine operation (diesel-trucks/busses application). The fuels used were 10% and 15% biofuel and with the addition of 5% lubricating oil to the fuel. Results show that as the engine warms up, PN for all the fuels increases and the size of particles decreases. PN concentration with a fully warmed-up engine was up to 132% higher than the cold-start. Sub-23 nm particles accounted for a significant proportion of PN (9%) but a smaller proportion of PM (0.1%). The fuel blend with 5% lubricating oil showed a significant increase in PN concentration and a decrease in particle size during cold-start.

Mouse Liver-Expressed Shiftless Is an Evolutionarily Conserved Antiviral Effector Restricting Human and Murine Hepaciviruses.

Mice are refractory to infection with human-tropic hepatitis C virus (HCV), although distantly related rodent hepaciviruses (RHV) circulate in wild rodents. To investigate whether liver intrinsic host factors can exhibit broad restriction against these distantly related hepaciviruses, we focused on Shiftless (Shfl), an interferon (IFN)-regulated gene (IRG) which restricts HCV in humans. Unusually, and in contrast to selected classical IRGs, human and mouse SHFL orthologues (hSHFL and mSHFL, respectively) were highly expressed in hepatocytes in the absence of viral infection, weakly induced by IFN, and highly conserved at the amino acid level (>95%). Replication of both HCV and RHV subgenomic replicons was suppressed by ectopic expression of mSHFL in human or rodent hepatoma cell lines. Gene editing of endogenous mShfl in mouse liver tumor cells increased HCV replication and virion production. Colocalization of mSHFL protein with viral double-stranded RNA (dsRNA) intermediates was confirmed and could be ablated by mutational disruption of the SHFL zinc finger domain, concomitant with a loss of antiviral activity. In summary, these data point to an evolutionarily conserved function for this gene in humans and rodents: SHFL is an ancient antiviral effector which targets distantly related hepaciviruses via restriction of viral RNA replication. IMPORTANCE Viruses have evolved ways to evade or blunt innate cellular antiviral mechanisms within their cognate host species. However, these adaptations may fail when viruses infect new species and can therefore limit cross-species transmission. This may also prevent development of animal models for human-pathogenic viruses. HCV shows a narrow species tropism likely due to distinct human host factor usage and innate antiviral defenses limiting infection of nonhuman liver cells. Interferon (IFN)-regulated genes (IRGs) partially inhibit HCV infection of human cells by diverse mechanisms. Here, we show that mouse Shiftless (mSHFL), a protein that interferes with HCV replication factories, inhibits HCV replication and infection in human and mouse liver cells. We further report that the zinc finger domain of SHFL is important for viral restriction. These findings implicate mSHFL as a host factor that impairs HCV infection of mice and provide guidance for development of HCV animal models needed for vaccine development.

Correspondence on "An International Study Evaluating Elemental Analysis".

This Correspondence provides a brief commentary on a recent ACS Central Science article that evaluated the performance of different laboratories in elemental analysis and suggests that a broader conclusion should be drawn instead, recognizing the benefits of metrology and the international quality infrastructure.

Falling nickel concentrations in ambient air in South Wales - 50 years of progress.

Measurement of the composition of ambient air has become increasingly widespread over the last 50 years as the detrimental health effects of some air pollutants have become clearer and requirements for these measurements has been embedded in national and international legislation. The aim of this has been not only to assess exposure of the general population to air pollutants but also to assess the effectiveness of abatement strategies to reduce emissions of these pollutants at source. With a rich industrial heritage, the Swansea Valley (South Wales, UK) has long been associated with the refining and production of metal products, especially nickel. Despite a decline in output during the latter part of the twentieth century there is still sufficient activity to prompt a requirement for targeted air monitoring in the area. This is most important for nickel where there is a local history of measured concentrations exceeding legislative target values. This work demonstrates the effectiveness of nickel emissions abatement strategies over the last 50 years by tracking the falling air concentration of nickel over this period. It also demonstrates how the monitoring network in the Swansea Valley has expanded over this time and become significantly more sensitive to nickel emissions. The data presented represents a significant public health achievement - it is likely that the exposure to nickel in air of the population in the Swansea Valley has decreased more than 100-fold over the last 50 years - and reflects the progress in regulation, industrial efficiency, emissions abatement technology and air quality monitoring science achieved during this period.

Progress in pre-treatment and extraction of organic and inorganic pollutants by layered double hydroxide for trace-level analysis.

Continuous release of pollutants into the environment poses serious threats to environmental sustainability and human health. For trace-level analysis of pollutants, layered double hydroxide (LDH) is an attractive option to impart enhanced sorption capability and sensitivity toward pollutants because of its unique layered structure, tunable interior architecture, high anion-exchange capacities, and high porosity (e.g., Zn/Cr LDH/DABCO-IL, Ni/Al LDH, CS-Ni/Fe LDH, SDS-Fe3O4@SiO2@Mg-Al LDH, Boeh/Mg/Al LDH/pC, and Fe@NiAl LDH). In concert with the well-defined analytical methodologies (e.g., HPLC and GC), the LDH materials can be employed to detect trace-level targets (e.g., as low as âˆ¼ 20 fg/L for phenols) in aqueous environments. This review highlights LDH as a promising material for pre-treatment of a variety of organic and inorganic target pollutants in complex real matrices. Challenges and future requirements for research into LDH-based analytical methods are also discussed.

Vaccine-associated enhanced respiratory pathology in COVID-19 hamsters after TH2-biased immunization.

Vaccine-associated enhanced respiratory disease (VAERD) is a severe complication for some respiratory infections. To investigate the potential for VAERD induction in coronavirus disease 2019 (COVID-19), we evaluate two vaccine leads utilizing a severe hamster infection model: a T helper type 1 (TH1)-biased measles vaccine-derived candidate and a TH2-biased alum-adjuvanted, non-stabilized spike protein. The measles virus (MeV)-derived vaccine protects the animals, but the protein lead induces VAERD, which can be alleviated by dexamethasone treatment. Bulk transcriptomic analysis reveals that our protein vaccine prepares enhanced host gene dysregulation in the lung, exclusively up-regulating mRNAs encoding the eosinophil attractant CCL-11, TH2-driving interleukin (IL)-19, or TH2 cytokines IL-4, IL-5, and IL-13. Single-cell RNA sequencing (scRNA-seq) identifies lung macrophages or lymphoid cells as sources, respectively. Our findings imply that VAERD is caused by the concerted action of hyperstimulated macrophages and TH2 cytokine-secreting lymphoid cells and potentially links VAERD to antibody-dependent enhancement (ADE). In summary, we identify the cytokine drivers and cellular contributors that mediate VAERD after TH2-biased vaccination.

Mesoporous silica imprinted carbon dots for the selective fluorescent detection of triclosan.

A molecularly imprinted fluorescence sensor built as a mesoporous structured silica imprinted layer on the surface of carbon dots (CDs@m-MIP) was employed for the selective detection of triclosan (TRI). The fluorescence of this CDs@m-MIP was affected sensitively and selectively by TRI via an electron transfer-induced fluorescence quenching mechanism with a detection limit of TRI at 1.08 nM (range 1.72-138 nM) under the optimum setup (e.g., pH, response time, and CDs@m-MIP dose). This approach was used successfully to detect TRI in real water samples (e.g., sewage, river, and tap water). The recoveries of TRI were satisfactory in spiked river and tap water (in 94.7-99.5 %). The outcome of this research is thus expected to help develop highly efficient fluorescent sensing systems towards diverse hazardous compounds including TRI.

Construction of novel luminescent thermometers based on dual-emission centers of rare-earth and bismuth ions.

Optical thermometry based on fluorescence intensity ratio (FIR) technology has several advantages for industrial and medical applications such as remote signaling, non-invasiveness, and excellent spatial resolution. Here, an approach to the construction of luminescent thermometers is proposed based on high-temperature solid-state reactions through doping of rare earth (RE) elements (e.g., samarium (Sm3+) or europium (Eu3+)) into Ca2Y0.97Bi0.03SbO6 (CYBS) phosphors. The tuning of the CYBS:Eu3+ and CYBS:Sm3+ ratios in the phosphors provided a wide range of color changes from purplish blue to red and from purplish blue to pink, respectively. The superiority of optical thermometer is validated by higher values of absolute sensitivity (Sa) and relative sensitivity (Sr). As such, both phosphors exhibit excellent temperature sensing performance with Sa/Sr values (at 483 K) of 4.945 × 10-2/0.968 × 10-2 K-1 (CYBS:0.05Eu3+) and 2.964 × 10-2/0.864 × 10-2 K-1 (CYBS:0.05 Sm3+). Thus, RE-doped CYBS materials with color tuning properties and superior temperature sensing performance are recommended for the construction of novel luminescent optical thermometers.