Context-dependent role of ATG4B as target for autophagy inhibition in prostate cancer therapy

ATG4B belongs to the autophagin family of cysteine proteases required for autophagy, an emerging target of cancer therapy. Developing pharmacological ATG4B inhibitors is a very active area of research. However, detailed studies on the role of ATG4B during anticancer therapy are lacking. By analyzing PC-3 and C4-2 prostate cancer cells overexpressing dominant negative ATG4B^C74Ain vitro and in vivo, we show that the effects of ATG4B^C74A are cell type, treatment, and context-dependent. ATG4B^C74A expression can either amplify the effects of cytotoxic therapies or contribute to treatment resistance. Thus, the successful clinical application of ATG4B inhibitors will depend on finding predictive markers of response.     

Taking into account water use impacts in the LCA of biofuels: an Argentinean case study

Purpose  

The assessment of biofuels has until now mainly focused on energy demand and greenhouse gas emissions. Only little attention has been given to other impacts, although the general importance of water use for the life cycle assessment (LCA) of agricultural products has been recognized in recent publications. The aim of this work is to assess in detail the water consumption along a biofuel production chain taking into account irrigation efficiencies, levels of water scarcity, and type of feedstock, and to integrate those results in a full LCA. Furthermore, we compare the results for biofuels from various feedstocks and regions with conventional petrol.     

Differential efficacies of Cas nucleases on microsatellites involved in human disorders and associated off-target mutations

Microsatellite expansions are the cause of >20 neurological or developmental human disorders. Shortening expanded repeats using specific DNA endonucleases may be envisioned as a gene editing approach. Here, we measured the efficacy of several CRISPR–Cas nucleases to induce recombination within disease-related microsatellites, in Saccharomyces cerevisiae. Broad variations in nuclease performances were detected on all repeat tracts. Wild-type Streptococcus pyogenes Cas9 (SpCas9) was more efficient than Staphylococcus aureus Cas9 on all repeats tested, except (CAG)₃₃. Cas12a (Cpf1) was the most efficient on GAA trinucleotide repeats, whereas GC-rich repeats were more efficiently cut by SpCas9. The main genetic factor underlying Cas efficacy was the propensity of the recognition part of the sgRNA to form a stable secondary structure, independently of its structural part. This suggests that such structures form in vivo and interfere with sgRNA metabolism. The yeast genome contains 221 natural CAG/CTG and GAA/CTT trinucleotide repeats. Deep sequencing after nuclease induction identified three of them as carrying statistically significant low frequency mutations, corresponding to SpCas9 off-target double-strand breaks.     

LCA of energetic biomass utilization: actual projects and new developments—April 23, 2012, Berne, Switzerland

Introduction

In the last years, the use of biomass for energy purposes has been seen as a promising option to reduce the use of nonrenewable energy sources and the emissions of fossil carbon. However, LCA studies have shown that the energetic use of biomass also causes impacts on climate change and, furthermore, that different environmental issues arise, such as land use and agricultural emissions. While biomass is renewable, it is not an unlimited resource. Its use, to whatever purpose, must therefore be well studied to promote the most efficient option with the least environmental impacts. The 47th LCA Discussion Forum gathered several national and international speakers who provided a broad and qualified view on the topic.

Summary of the topics presented in DF 47

Several aspects of energetic biomass use from a range of projects financed by the Swiss Federal Office of Energy (SFOE) were presented in this Discussion Forum. The first session focused on important aspects of the agricultural biogas production like the use of high energy crops or catch crops as well as the influence of plant size on the environmental performance of biogas. In the second session, other possibilities of biomass treatment like direct combustion, composting, and incineration with municipal waste were presented. Topic of the first afternoon session was the update and harmonization of biomass inventories and the resulting new assessment of biofuels. The short presentations investigated some further aspects of the LCA of bioenergy like the assessment of spatial variation of greenhouse gas (GHG) emissions from bioenergy production in a country, the importance of indirect land use change emissions on the overall results, the assessment of alternative technologies to direct spreading of digestate or the updates of the car operation datasets in ecoinvent.

Conclusions

One main outcome of this Discussion Forum is that bioenergy is not environmentally friendly per se. In many cases, energetic use of biomass allows a reduction of GHG and fossil energy use. However, there is often a tradeoff with other environmental impacts linked to agricultural production like eutrophication or ecotoxicity. Methodological challenges still exist, like the assessment of direct and indirect land use change emissions and their attribution to the bioenergy production, or the influence of heavy metal flows on the bioenergy assessment. Another challenge is the implementation of a life cycle approach in certification or legislation schemes, as shown by the example of the Renewable Energy Directive of the European Union.     

Anti-prothrombin autoantibodies enriched after infection with SARS-CoV-2 and influenced by strength of antibody response against SARS-CoV-2 proteins

Antiphospholipid antibodies (aPL), assumed to cause antiphospholipid syndrome (APS), are notorious for their heterogeneity in targeting phospholipids and phospholipid-binding proteins. The persistent presence of Lupus anticoagulant and/or aPL against cardiolipin and/or β2-glycoprotein I have been shown to be independent risk factors for vascular thrombosis and pregnancy morbidity in APS. aPL production is thought to be triggered by–among other factors–viral infections, though infection-associated aPL have mostly been considered non-pathogenic. Recently, the potential pathogenicity of infection-associated aPL has gained momentum since an increasing number of patients infected with Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) has been described with coagulation abnormalities and hyperinflammation, together with the presence of aPL. Here, we present data from a multicentric, mixed-severity study including three cohorts of individuals who contracted SARS-CoV-2 as well as non-infected blood donors. We simultaneously measured 10 different criteria and non-criteria aPL (IgM and IgG) by using a line immunoassay. Further, IgG antibody response against three SARS-CoV-2 proteins was investigated using tripartite automated blood immunoassay technology. Our analyses revealed that selected non-criteria aPL were enriched concomitant to or after an infection with SARS-CoV-2. Linear mixed-effects models suggest an association of aPL with prothrombin (PT). The strength of the antibody response against SARS-CoV-2 was further influenced by SARS-CoV-2 disease severity and sex of the individuals. In conclusion, our study is the first to report an association between disease severity, anti-SARS-CoV-2 immunoreactivity, and aPL against PT in patients with SARS-CoV-2.     

Life Cycle Inventories for the Nuclear and Natural Gas Energy Systems,and Examples of Uncertainty Analysis (14 pp)

Goal, Scope and Background The energy systems included in the ecoinvent database v1.1 describe the situation around year 2000 of Swiss and Western European power plants and boilers with the associated energy chains. The addressed nuclear systems concern Light Water Reactors (LWR) with mix of open and closed fuel cycles. The system model ‘Natural Gas’ describes production, distribution, and combustion of natural gas. Methods Comprehensive life cycle inventories of the energy systems were established and cumulative results calculated within the ecoinvent framework. Swiss conditions for the nuclear cycle were extrapolated to major nuclear countries. Long-term radon emissions from uranium mill tailings have been estimated with a simplified model. Average natural gas power plants were analysed for different countries considering specific import/export of the gas, with seven production regions separately assessed. Uncertainties have been estimated quantitatively. Results and Discussion Different radioactive emission species and wastes are produced from different steps of the nuclear cycle. Emissions of greenhouse gases from the nuclear cycle are mostly from the upstream chain, and the total is small and decreasing with increasing share of centrifuge enrichment. The results for natural gas show the importance of transport and low pressure distribution network for the methane emissions, whereas energy is mostly invested for production and long-distance pipeline transportation. Because of significant differences in power plant efficiencies and gas supply, country specific averages differ greatly. Conclusion The inventory describes average worldwide supply of nuclear fuel and average nuclear reactors in Western Europe. Although the model for nuclear waste management was extrapolated from Swiss conditions, the ranges obtained for cumulative results can represent the average in Europe. Emissions per kWh electricity are distributed very differently over the natural gas chain for different species. Modern combined cycle plants show better performance for several burdens like cumulative greenhouse gas emissions compared to average plants. Recommendation and Perspective Comparison of country-specific LWRs or LWR types on the basis of these results is not recommended. Specific issues on different strategies for the nuclear fuel cycle or location-specific characteristics would require extension of analysis. Results of the gas chain should not be directly applied to areas other than those modelled because emission factors and energy requirements may differ significantly. A future update of inventory data should reconsider production and transport from Russia, as it is a major producer and exporter to Europe. The calculated ranges of uncertainty factors in ecoinvent provide useful information but they are more indications of uncertainties rather than strict 95% intervals, and should therefore be applied carefully.