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Keywords: coagulation; red blood cells; viscoelastic assays EN coagulation red blood cells viscoelastic assays 329 331 3 01/27/23 20230201 NES 230201 Blood is a non-Newtonian fluid, a colloidal solution with the partial behavior of a suspension. In this issue of I Critical Care Medicine i , Roh et al (11) have published their investigation into the effects of hemoglobin concentration ([Hb]) on viscoelastic tests performed on the ROTEM (https://www.werfen.com/uk/en/haemostasis-diagnostics/rotem-sigma) and TEG-5000 (https://teg.haemonetics.com/en-gb) platforms. However, the effect of RBC concentration abnormalities (anemia and polycythemia) on viscoelastic tests is less clear, though laboratory investigations to date have suggested an anomalous paradoxical effect on the assays, compared with the clinical state. [Extracted from the article]

Following acute HCV infection, the virus establishes a chronic disease in the majority of patients whilst few individuals clear the infection spontaneously. The precise mechanisms that determine chronic HCV infection or spontaneous clearance are not completely understood but are proposed to be driven by host and viral genetic factors as well as HCV encoded immunomodulatory proteins. Using the HIV-1 LTR as a tool to measure NF-κB activity, we identified that the HCV E1E2 glycoproteins and more so the E2 protein down-modulates HIV-1 LTR activation in 293T, TZM-bl and the more physiologically relevant Huh7 liver derived cell line. We demonstrate this effect is specifically mediated through inhibiting NF-κB binding to the LTR and show that this effect was conserved for all HCV genotypes tested. Transcriptomic analysis of 293T cells expressing the HCV glycoproteins identified E1E2 mediated stimulation of the endoplasmic reticulum (ER) stress response pathway and upregulation of stress response genes such as ATF3. Through shRNA mediated inhibition of ATF3, one of the components, we observed that E1E2 mediated inhibitory effects on HIV-1 LTR activity was alleviated. Our in vitro studies demonstrate that HCV Env glycoprotein activates host ER Stress Pathways known to inhibit NF-κB activity. This has potential implications for understanding HCV induced immune activation as well as oncogenesis. [ABSTRACT FROM AUTHOR]

Development of a prophylactic hepatitis C virus (HCV) vaccine will require accurate and reproducible measurement of neutralizing breadth of vaccine-induced antibodies. Currently available HCV panels may not adequately represent the genetic and antigenic diversity of circulating HCV strains, and the lack of standardization of these panels makes it difficult to compare neutralization results obtained in different studies. Here, we describe the selection and validation of a genetically and antigenically diverse reference panel of 15 HCV pseudoparticles (HCVpps) for neutralization assays. We chose 75 envelope (E1E2) clones to maximize representation of natural polymorphisms observed in circulating HCV isolates, and 65 of these clones generated functional HCVpps. Neutralization sensitivity of these HCVpps varied widely. HCVpps clustered into 15 distinct groups based on patterns of relative sensitivity to 7 broadly neutralizing monoclonal antibodies. We used these data to select a final panel of 15 antigenically representative HCVpps. Both the 65 and 15 HCVpp panels span 4 tiers of neutralization sensitivity, and neutralizing breadth measurements for 7 broadly neutralizing monoclonal antibodies were nearly equivalent using either panel. Differences in neutralization sensitivity between HCVpps were independent of genetic distances between E1E2 clones. Neutralizing breadth of HCV antibodies should be defined using viruses spanning multiple tiers of neutralization sensitivity rather than panels selected solely for genetic diversity. We propose that this multitier reference panel could be adopted as a standard for the measurement of neutralizing antibody potency and breadth, facilitating meaningful comparisons of neutralization results from vaccine studies in different laboratories. [Display omitted] A panel of hepatitis C virus pseudoparticles with 4 tiers of antibody resistance was developed. These pseudoparticles can be used to measure neutralizing breadth of antibodies induced by candidate vaccines. [ABSTRACT FROM AUTHOR]

Screw piles potentially offer quieter installation and enhanced axial tensile capacity over straight-shafted driven piles. As such, they have been suggested as a possible foundation solution for offshore jacket-supported wind turbines in deeper water. To investigate the feasibility of their use in this setting, centrifuge testing of six model screw piles of different designs was conducted to measure the installation requirements and ultimate axial capacity of the piles in very dense and medium-dense sand. The screw piles were designed to sustain loads generated by an extreme design scenario using published axial capacity and torque prediction formulae. Single- and double-helix designs, including an optimised design, intended to minimise installation requirements, with reduced geometry were installed and tested in-flight. Piles in the medium-dense sand, for example, had significant installation requirements of up to 18·4 MNm (torque) and 28·8 MN (vertical force), which were accurately predicted using correlations with cone resistance data (cone penetration test). Existing axial capacity design methods did not perform well for these large-scale screw piles, overestimating compressive and tensile capacities. Revised analytical methods for installation and axial capacity estimates are proposed here based on the centrifuge test results. [ABSTRACT FROM AUTHOR]

Nosocomial severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infections have severely affected bed capacity and patient flow. We utilized whole-genome sequencing (WGS) to identify outbreaks and focus infection control resources and intervention during the United Kingdom's second pandemic wave in late 2020. Phylogenetic analysis of WGS and epidemiological data pinpointed an initial transmission event to an admission ward, with immediate prior community infection linkage documented. High incidence of asymptomatic staff infection with genetically identical viral sequences was also observed, which may have contributed to the propagation of the outbreak. WGS allowed timely nosocomial transmission intervention measures, including admissions ward point-of-care testing and introduction of portable HEPA14 filters. Conversely, WGS excluded nosocomial transmission in 2 instances with temporospatial linkage, conserving time and resources. In summary, WGS significantly enhanced understanding of SARS-CoV-2 clusters in a hospital setting, both identifying high-risk areas and conversely validating existing control measures in other units, maintaining clinical service overall. [ABSTRACT FROM AUTHOR]

Persistent hepatitis C virus (HCV) infection is a major cause of chronic liver disease, worldwide. With the development of direct-acting antivirals, treatment of chronically infected patients has become highly effective, although a subset of patients responds less well to therapy. Sofosbuvir is a common component of current de novo or salvage combination therapies, that targets the HCV NS5B polymerase. We use pre-treatment whole-genome sequences of HCV from 507 patients infected with HCV subtype 3a and treated with sofosbuvir containing regimens to detect viral polymorphisms associated with response to treatment. We find three common polymorphisms in non-targeted HCV NS2 and NS3 proteins are associated with reduced treatment response. These polymorphisms are enriched in post-treatment HCV sequences of patients unresponsive to treatment. They are also associated with lower reductions in viral load in the first week of therapy. Using in vitro short-term dose-response assays, these polymorphisms do not cause any reduction in sofosbuvir potency, suggesting an indirect mechanism of action in decreasing sofosbuvir efficacy. The identification of polymorphisms in NS2 and NS3 proteins associated with poor treatment outcomes emphasises the value of systematic genome-wide analyses of viruses in uncovering clinically relevant polymorphisms that impact treatment. Sofosbuvir is a common therapy in hepatitis C virus infection, which targets the NS5B polymerase. Here, Smith et al. analyze the association between whole genome HCV polymorphisms and sofosbuvir treatment failure and identify three common polymorphisms present in non-targeted NS2 and NS3 proteins associated with reduced treatment response. [ABSTRACT FROM AUTHOR]