Tension-voltage relationship in membrane fusion and its implication in exocytosis

AbstractIn this study, new methods are used to control cellular membrane tension to evaluate the role it plays in electrofusion. The data show that membrane tension present during the application of an electric field facilitates electro-induced membrane fusion. No enhancement was detected if the strain was applied after the pulse. Analysis of the electromechanical process of fusion revealed a synergy between the two kinds of constraints in the membrane fusion. Both mechanical and electrical constraints apparently play a key role in membrane fusion between the granule membrane and the plasma membrane, i.e. the exocytosis process

Spontaneous lipid vesicle fusion with electropermeabilized cells

AbstractFusion is obtained between electropermeabilized mammalian cells and intact large unilamellar lipid vesicles. This is monitored by a fluorescence assay. Prepulse contact is obtained by Ca2+ when negatively charged lipids are present in the liposomes. The mixing of the liposome content in the cell cytoplasm is observed under conditions preserving cell viability. Electric conditions are such that free liposomes are not affected by the external field. Therefore destabilization of only one of the two membranes of the partners is sufficient for fusion. The comparison between the efficiency of dye delivery for different liposome preparations (multilamellar vesicles, large unilamellar vesicles, small unilamellar vesicles) is indicative that more metastable liposomes are more fusable with electropulsated cells. This observation is discussed within the framework of the recent hypothesis that occurrence of a contact induced electrostatic destabilization of the plasma membrane is a key step in the exocytosis process

Climate Change and IMF Surveillance. The Need for Ambition

El Fondo Monetario Internacional (FMI) necesita diseñar rápidamente una estrategia de cambio climático que ayude a sus miembros a cumplir sus objetivos colectivos de cambio climático y desarrollo. En este breve informe se describen los aspectos macrocríticos del cambio climático que deberían incorporarse a la actividad de supervisión del FMI y se examina en qué medida los riesgos climáticos han formado parte de la supervisión del FMI en los últimos años. Nuestra investigación muestra que el FMI ha prestado una atención mínima y desigual a los riesgos climáticos en los informes del Artículo IV, y aún menos en sus Programas de Evaluación del Sector Financiero (PESF), aunque en ambos casos el FMI ha experimentado con análisis que pueden servir de base para trabajar. Como resultado de las revisiones del Artículo IV y los PESF en curso, es imperativo que el FMI reconozca que tanto los riesgos físicos como los de transición dentro de los países y entre ellos son amenazas macrocríticas para los sistemas financieros y fiscales; que garantice que los análisis de los riesgos físicos y de transición dentro de los países y entre ellos sea obligatorio, sistemático y universal para todos los informes del Artículo IV y PESF de aquí en adelante; y que el FMI trabaje con el personal, los Estados miembros, las partes interesadas, la Red para la Ecologización del Sistema Financiero (NGFS) y los/as expertos/as para aprovechar su experiencia inicial y alinear la política de supervisión del FMI con los ambiciosos objetivos del cambio climático.The International Monetary Fund (IMF) needs to rapidly devise a climate change strategy that helps its members meet their collective climate change and development goals. This policy brief outlines the macro-critical aspects of climate change that will need to be incorporated into IMF surveillance activity and examines the extent to which climate risks have been a part of IMF surveillance in recent years. Our research shows that the IMF has paid minimal and uneven attention to climate risks in Article IV reports, and even less so in its Financial Sector Assessment Programs (FSAPs)—though in each case the IMF has experimented with analyses that can be built upon. As a result of the reviews of Article IV surveillance and FSAPs currently underway, it is imperative that the IMF: recognize that both physical and transition risks within and across countries are macro-critical threats to financial and fiscal systems; ensure that physical and transitions risks analysis within and across countries is compulsory, systematic, and universal for all Article IV and FSAPs moving forward; and that the IMF work with staff, member states, stakeholders, the NGFS, and experts to build on its early experience and align IMF surveillance policy with ambitious climate change goals.Facultad de Ciencias Jurídicas y Sociale

Functional diversity and redundancy across fish gut, sediment and water bacterial communities.

This article explores the functional diversity and redundancy in a bacterial metacommunity constituted of three habitats (sediment, water column and fish gut) in a coastal lagoon under anthropogenic pressure. Comprehensive functional gene arrays covering a wide range of ecological processes and stress resistance genes to estimate the functional potential of bacterial communities were used. Then, diversity partitioning was used to characterize functional diversity and redundancy within (α), between (β) and across (γ) habitats. It was showed that all local communities exhibit a highly diversified potential for the realization of key ecological processes and resistance to various environmental conditions, supporting the growing evidence that macro-organisms microbiomes harbour a high functional potential and are integral components of functional gene dynamics in aquatic bacterial metacommunities. Several levels of functional redundancy at different scales of the bacterial metacommunity were observed (within local communities, within habitats and at the metacommunity level). The results suggested a high potential for the realization of spatial ecological insurance within this ecosystem, that is, the functional compensation among microorganisms for the realization and maintenance of key ecological processes, within and across habitats. Finally, the role of macro-organisms as dispersal vectors of microbes and their potential influence on marine metacommunity dynamics were discussed

The efficacy of whole human genome capture on ancient dental calculus and dentin

Objectives: Dental calculus is among the richest known sources of ancient DNA in the archaeological record. Although most DNA within calculus is microbial, it has been shown to contain sufficient human DNA for the targeted retrieval of whole mitochondrial genomes. Here, we explore whether calculus is also a viable substrate for whole human genome recovery using targeted enrichment techniques. Materials and methods: Total DNA extracted from 24 paired archaeological human dentin and calculus samples was subjected to whole human genome enrichment using in-solution hybridization capture and high-throughput sequencing. Results: Total DNA from calculus exceeded that of dentin in all cases, and although the proportion of human DNA was generally lower in calculus, the absolute human DNA content of calculus and dentin was not significantly different. Whole genome enrichment resulted in up to fourfold enrichment of the human endogenous DNA content for both dentin and dental calculus libraries, albeit with some loss in complexity. Recovering more on-target reads for the same sequencing effort generally improved the quality of downstream analyses, such as sex and ancestry estimation. For nonhuman DNA, comparison of phylum-level microbial community structure revealed few differences between precapture and postcapture libraries, indicating that off-target sequences in human genome-enriched calculus libraries may still be useful for oral microbiome reconstruction. Discussion: While ancient human dental calculus does contain endogenous human DNA sequences, their relative proportion is low when compared with other skeletal tissues. Whole genome enrichment can help increase the proportion of recovered human reads, but in this instance enrichment efficiency was relatively low when compared with other forms of capture. We conclude that further optimization is necessary before the method can be routinely applied to archaeological samples

Enabling countries to manage outbreaks: statistical, operational, and contextual analysis of the early warning and response system (EWARS-csd) for dengue outbreaks

IntroductionDengue is currently the fastest-spreading mosquito-borne viral illness in the world, with over half of the world's population living in areas at risk of dengue. As dengue continues to spread and become more of a health burden, it is essential to have tools that can predict when and where outbreaks might occur to better prepare vector control operations and communities' responses. One such predictive tool, the Early Warning and Response System for climate-sensitive diseases (EWARS-csd), primarily uses climatic data to alert health systems of outbreaks weeks before they occur. EWARS-csd uses the robust Distribution Lag Non-linear Model in combination with the INLA Bayesian regression framework to predict outbreaks, utilizing historical data. This study seeks to validate the tool's performance in two states of Colombia, evaluating how well the tool performed in 11 municipalities of varying dengue endemicity levels.MethodsThe validation study used retrospective data with alarm indicators (mean temperature and rain sum) and an outbreak indicator (weekly hospitalizations) from 11 municipalities spanning two states in Colombia from 2015 to 2020. Calibrations of different variables were performed to find the optimal sensitivity and positive predictive value for each municipality.ResultsThe study demonstrated that the tool produced overall reliable early outbreak alarms. The median of the most optimal calibration for each municipality was very high: sensitivity (97%), specificity (94%), positive predictive value (75%), and negative predictive value (99%; 95% CI).DiscussionThe tool worked well across all population sizes and all endemicity levels but had slightly poorer results in the highly endemic municipality at predicting non-outbreak weeks. Migration and/or socioeconomic status are factors that might impact predictive performance and should be further evaluated. Overall EWARS-csd performed very well, providing evidence that it should continue to be implemented in Colombia and other countries for outbreak prediction

Exploring the capacity for anaerobic biodegradation of polycyclic aromatic hydrocarbons and naphthenic acids by microbes from oil-sands-process-affected waters

Both polycyclic aromatic hydrocarbons (PAHs) and naphthenic acids (NAs) are natural components of fossil fuels, but they are also widespread toxic and environmentally persistent pollutants. They are the major cause of environmental toxicity in oil-sands-process waters (OSPW). This study aimed to investigate the anaerobic biodegradation of the PAHs pyrene and 2-methylnaphthalene, and the NAs adamantane-1-carboxylic acid and a "natural" NA mixture (i.e., acid-extractable NAs from OSPW) under sulfate-reducing and methanogenic conditions by a microbial community derived from an oil sands tailings pond. Using gas-chromatography mass spectrometry (GC-MS), the rate of biodegradation was measured in relation to changes in bacterial community composition. Only 2-methylnaphthalene was significantly degraded after 260 days, with significantly more degradation under sulfate-reducing (40%) than methanogenic conditions (25%). During 2-methylnaphthalene biodegradation, a major metabolite was produced and tentatively identified as 2-naphthoic acid. Denaturing gradient gel electrophoresis (DGGE) demonstrated an increase in intensity of bands during the anaerobic biodegradation of 2-methylnaphalene, which derived from species of the genera Fusibacter, Alkaliphilus, Desulfobacterium, Variovorax, Thaurea, and Hydrogenophaga. Despite the biodegradation of 2-methylnaphthalene, this study demonstrates that, under anaerobic conditions, NAs and high-molecular-weight PAHs are the predominant molecules likely to persist in OSPW. Therefore alternative remediation strategies are required

Payers' views of the changes arising through the possible adoption of adaptive pathways

Payers are a major stakeholder in any considerations and initiatives concerning adaptive licensing of new medicinal products, also referred to as Medicines Adaptive Pathways to patients (MAPPs). Firstly, the scope and necessity of MAPPs need further scrutiny, especially with regard to the definition of unmet need. Conditional approval pathways already exist for new medicines for seriously debilitating or life-threatening diseases and only a limited number of new medicines are innovative. Secondly, MAPPs will result in new medicines on the market with limited evidence about their effectiveness and safety. Additional data are to be collected after approval. Consequently, adaptive pathways may increase the risk of exposing patients to ineffective or unsafe medicines. We have already seen medicines approved conventionally that subsequently proved ineffective or unsafe amongst a wider, more co-morbid population as well as medicines that could have been considered for approval under MAPPs but subsequently proved ineffective or unsafe in Phase III trials and were never licensed. Thirdly, MAPPs also put high demands on payers. Routine collection of patient level data is difficult with high transaction costs. It is not clear who will fund these. Other challenges for payers include shifts in the risk governance framework, implications for evaluation and HTA, increased complexity of setting prices, difficulty with ensuring equity in the allocation of resources, definition of responsibility and liability and implementation of stratified use. Exit strategies also need to be agreed in advance, including price reductions, rebates, or reimbursement withdrawals when price premiums are not justified. These issues and concerns will be discussed in detail including potential ways forward

Orally Available Selective Melanocortin-4 Receptor Antagonists Stimulate Food Intake and Reduce Cancer-Induced Cachexia in Mice

BACKGROUND: Cachexia is among the most debilitating and life-threatening aspects of cancer. It represents a metabolic syndrome affecting essential functional circuits involved in the regulation of homeostasis, and includes anorexia, fat and muscle tissue wasting. The anorexigenic peptide alpha-MSH is believed to be crucially involved in the normal and pathologic regulation of food intake. It was speculated that blockade of its central physiological target, the melanocortin (MC)-4 receptor, might provide a promising anti-cachexia treatment strategy. This idea is supported by the fact that in animal studies, agouti-related protein (AgRP), the endogenous inverse agonist at the MC-4 receptor, was found to affect two hallmark features of cachexia, i.e. to increase food intake and to reduce energy expenditure. METHODOLOGY/PRINCIPAL FINDINGS: SNT207707 and SNT209858 are two recently discovered, non peptidic, chemically unrelated, orally active MC-4 receptor antagonists penetrating the blood brain barrier. Both compounds were found to distinctly increase food intake in healthy mice. Moreover, in mice subcutaneously implanted with C26 adenocarcinoma cells, repeated oral administration (starting the day after tumor implantation) of each of the two compounds almost completely prevented tumor induced weight loss, and diminished loss of lean body mass and fat mass. CONCLUSIONS/SIGNIFICANCE: In contrast to the previously reported peptidic and small molecule MC-4 antagonists, the compounds described here work by the oral administration route. Orally active compounds might offer a considerable advantage for the treatment of cachexia patients