High density Huh7.5 cell hollow fiber bioreactor culture for high-yield production of hepatitis C virus and studies of antivirals

Abstract Chronic hepatitis C virus (HCV) infection poses a serious global public health burden. Despite the recent development of effective treatments there is a large unmet need for a prophylactic vaccine. Further, antiviral resistance might compromise treatment efficiency in the future. HCV cell culture systems are typically based on Huh7 and derived hepatoma cell lines cultured in monolayers. However, efficient high cell density culture systems for high-yield HCV production and studies of antivirals are lacking. We established a system based on Huh7.5 cells cultured in a hollow fiber bioreactor in the presence or absence of bovine serum. Using an adapted chimeric genotype 5a virus, we achieved peak HCV infectivity and RNA titers of 7.6 log10 FFU/mL and 10.4 log10 IU/mL, respectively. Bioreactor derived HCV showed high genetic stability, as well as buoyant density, sensitivity to neutralizing antibodies AR3A and AR4A, and dependency on HCV co-receptors CD81 and SR-BI comparable to that of HCV produced in monolayer cell cultures. Using the bioreactor platform, treatment with the NS5A inhibitor daclatasvir resulted in HCV escape mediated by the NS5A resistance substitution Y93H. In conclusion, we established an efficient high cell density HCV culture system with implications for studies of antivirals and vaccine development

HbA1c levels in non-diabetic older adults - No J-shaped associations with primary cardiovascular events, cardiovascular and all-cause mortality after adjustment for confounders in a meta-analysis of individual participant data from six cohort studies

Background:To determine the shape of the associations of HbA1c with mortality and cardiovascular outcomes in non-diabetic individuals and explore potential explanations. Methods: The associations of HbA1c with all-cause mortality, cardiovascular mortality and primary cardiovascular events (myocardial infarction or stroke) were assessed in non-diabetic subjects ≥50 years from six population-based cohort studies from Europe and the USA and meta-analyzed. Very low, low, intermediate and increased HbA1c were defined as <5.0, 5.0 to <5.5, 5.5 to <6.0 and 6.0 to <6.5 % (equals <31, 31 to <37, 37 to <42 and 42 to <48 mmol/mol), respectively, and low HbA1c was used as reference in Cox proportional hazards models. Results:Overall, 6,769 of 28,681 study participants died during a mean follow-up of 10.7 years, of whom 2,648 died of cardiovascular disease. Furthermore, 2,493 experienced a primary cardiovascular event. A linear association with primary cardiovascular events was observed. Adjustment for cardiovascular risk factors explained about 50 % of the excess risk and attenuated hazard ratios (95 % confidence interval) for increased HbA1c to 1.14 (1.03–1.27), 1.17 (1.00–1.37) and 1.19 (1.04–1.37) for all-cause mortality, cardiovascular mortality and cardiovascular events, respectively. The six cohorts yielded inconsistent results for the association of very low HbA1c levels with the mortality outcomes and the pooled effect estimates were not statistically significant. In one cohort with a pronounced J-shaped association of HbA1c levels with all-cause and cardiovascular mortality (NHANES), the following confounders of the association of very low HbA1c levels with mortality outcomes were identified: race/ethnicity; alcohol consumption; BMI; as well as biomarkers of iron deficiency anemia and liver function. Associations for very low HbA1c levels lost statistical significance in this cohort after adjusting for these confounders.Conclusions: A linear association of HbA1c levels with primary cardiovascular events was observed. For cardiovascular and all-cause mortality, the observed small effect sizes at both the lower and upper end of HbA1c distribution do not support the notion of a J-shaped association of HbA1c levels because a certain degree of residual confounding needs to be considered in the interpretation of the results. Keywords: Glycated hemoglobin, Cardiovascular disease, Myocardial infarction, Stroke, Mortality, Cohort study, Meta-analysi

The more the better: on the formation of single-phase high entropy alloy nanoparticles as catalysts for the oxygen reduction reaction.

High entropy alloys (HEAs) are an important new material class with significant application potential in catalysis and electrocatalysis. The entropy-driven formation of HEA materials requires high temperatures and controlled cooling rates. However, catalysts in general also require highly dispersed materials, i.e., nanoparticles. Only then a favorable utilization of the expensive raw materials can be achieved. Several recently reported HEA nanoparticle synthesis strategies, therefore, avoid the high-temperature regime to prevent particle growth. In our work, we investigate a system of five noble metal single-source precursors with superior catalytic activity for the oxygen reduction reaction. Combining in situ X-ray powder diffraction with multi-edge X-ray absorption spectroscopy, we address the fundamental question of how single-phase HEA nanoparticles can form at low temperatures. It is demonstrated that the formation of HEA nanoparticles is governed by stochastic principles and the inhibition of precursor mobility during the formation process favors the formation of a single phase. The proposed formation principle is supported by simulations of the nanoparticle formation in a randomized process, rationalizing the experimentally found differences between two-element and multi-element metal precursor mixtures

On the History and Future of 100% Renewable Energy Systems Research

Research on 100% renewable energy systems is a relatively recent phenomenon. It was initiated in the mid-1970s, catalyzed by skyrocketing oil prices. Since the mid-2000s, it has quickly evolved into a prominent research field encompassing an expansive and growing number of research groups and organizations across the world. The main conclusion of most of these studies is that 100% renewables is feasible worldwide at low cost. Advanced concepts and methods now enable the field to chart realistic as well as cost- or resource-optimized and efficient transition pathways to a future without the use of fossil fuels. Such proposed pathways in turn, have helped spur 100% renewable energy policy targets and actions, leading to more research. In most transition pathways, solar energy and wind power increasingly emerge as the central pillars of a sustainable energy system combined with energy efficiency measures. Cost-optimization modeling and greater resource availability tend to lead to higher solar photovoltaic shares, while emphasis on energy supply diversification tends to point to higher wind power contributions. Recent research has focused on the challenges and opportunities regarding grid congestion, energy storage, sector coupling, electrification of transport and industry implying power-to-X and hydrogen-to-X, and the inclusion of natural and technical carbon dioxide removal (CDR) approaches. The result is a holistic vision of the transition towards a net-negative greenhouse gas emissions economy that can limit global warming to 1.5°C with a clearly defined carbon budget in a sustainable and cost-effective manner based on 100% renewable energy-industry-CDR systems. Initially, the field encountered very strong skepticism. Therefore, this paper also includes a response to major critiques against 100% renewable energy systems, and also discusses the institutional inertia that hampers adoption by the International Energy Agency and the Intergovernmental Panel on Climate Change, as well as possible negative connections to community acceptance and energy justice. We conclude by discussing how this emergent research field can further progress to the benefit of society

On the history and future of 100% renewable energy systems research

Research on 100% renewable energy systems is a relatively recent phenomenon. It was initiated in the mid-1970s, catalyzed by skyrocketing oil prices. Since the mid-2000s, it has quickly evolved into a prominent research field encompassing an expansive and growing number of research groups and organizations across the world. The main conclusion of most of these studies is that 100% renewables is feasible worldwide at low cost. Advanced concepts and methods now enable the field to chart realistic as well as cost- or resource-optimized and efficient transition pathways to a future without the use of fossil fuels. Such proposed pathways in turn, have helped spur 100% renewable energy policy targets and actions, leading to more research. In most transition pathways, solar energy and wind power increasingly emerge as the central pillars of a sustainable energy system combined with energy efficiency measures. Cost-optimization modeling and greater resource availability tend to lead to higher solar photovoltaic shares, while emphasis on energy supply diversification tends to point to higher wind power contributions. Recent research has focused on the challenges and opportunities regarding grid congestion, energy storage, sector coupling, electrification of transport and industry implying power-to-X and hydrogen-to-X, and the inclusion of natural and technical carbon dioxide removal (CDR) approaches. The result is a holistic vision of the transition towards a net-negative greenhouse gas emissions economy that can limit global warming to 1.5˚C with a clearly defined carbon budget in a sustainable and cost-effective manner based on 100% renewable energy-industry-CDR systems. Initially, the field encountered very strong skepticism. Therefore, this paper also includes a response to major critiques against 100% renewable energy systems, and also discusses the institutional inertia that hampers adoption by the International Energy Agency and the Intergovernmental Panel on Climate Change, as well as possible negative connections to community acceptance and energy justice. We conclude by discussing how this emergent research field can further progress to the benefit of society

Platform trials

Platform trials focus on the perpetual testing of many interventions in a disease or a setting. These trials have lasting organizational, administrative, data, analytic, and operational frameworks making them highly efficient. The use of adaptation often increases the probabilities of allocating participants to better interventions and obtaining conclusive results. The COVID-19 pandemic showed the potential of platform trials as a fast and valid way to improved treatments. This review gives an overview of key concepts and elements using the Intensive Care Platform Trial (INCEPT) as an example.</p

Risk Factors, Subsequent Disease Onset, and Prognostic Impact of Myocardial Infarction and Atrial Fibrillation

Background Although myocardial infarction (MI) and atrial fibrillation (AF) are frequent comorbidities and share common cardiovascular risk factors, the direction and strength of the association of the risk factors with disease onset, subsequent disease incidence, and mortality are not completely understood.Methods and Results In pooled multivariable Cox regression analyses, we examined temporal relations of disease onset and identified predictors of MI, AF, and all-cause mortality in 108 363 individuals (median age, 46.0 years; 48.2% men) free of MI and AF at baseline from 6 European population-based cohorts. During a maximum follow-up of 10.0 years, 3558 (3.3%) individuals were diagnosed exclusively with MI, 1922 (1.8%) with AF but no MI, and 491 (0.5%) individuals developed both MI and AF. Association of sex, systolic blood pressure, antihypertensive treatment, and diabetes appeared to be stronger with incident MI than with AF, whereas increasing age and body mass index showed a higher risk for incident AF. Total cholesterol and daily smoking were significantly related to incident MI but not AF. Combined population attributable fraction of cardiovascular risk factors was >70% for incident MI, whereas it was only 27% for AF. Subsequent MI after AF (hazard ratio [HR], 1.68; 95% CI, 1.03-2.74) and subsequent AF after MI (HR, 1.75; 95% CI, 1.31-2.34) both significantly increased overall mortality risk.Conclusions We observed different associations of cardiovascular risk factors with both diseases indicating distinct pathophysiological pathways. Subsequent diagnoses of MI and AF significantly increased mortality risk.</p

Genome-Wide Association Study and Functional Characterization Identifies Candidate Genes for Insulin-Stimulated Glucose Uptake

Distinct tissue-specific mechanisms mediate insulin action in fasting and postprandial states. Previous genetic studies have largely focused on insulin resistance in the fasting state, where hepatic insulin action dominates. Here we studied genetic variants influencing insulin levels measured 2 h after a glucose challenge in \u3e55,000 participants from three ancestry groups. We identified ten new loci (P \u3c 5 × 10-8) not previously associated with postchallenge insulin resistance, eight of which were shown to share their genetic architecture with type 2 diabetes in colocalization analyses. We investigated candidate genes at a subset of associated loci in cultured cells and identified nine candidate genes newly implicated in the expression or trafficking of GLUT4, the key glucose transporter in postprandial glucose uptake in muscle and fat. By focusing on postprandial insulin resistance, we highlighted the mechanisms of action at type 2 diabetes loci that are not adequately captured by studies of fasting glycemic traits