Structural and Biophysical Dynamics of Fungal Plasma Membrane Proteins and Implications for Echinocandin Action in Candida glabrata.

Fungal plasma membrane proteins represent key therapeutic targets for antifungal agents, yet their structure and spatial distribution in the native context remain poorly characterized. Herein, we employ an integrative multimodal approach to elucidate the structural and functional organization of plasma membrane protein complexes in Candida glabrata , focusing on prominent and essential membrane proteins, the polysaccharide synthase ß-(1,3)-glucan synthase (GS) and the proton pump Pma1. Cryo-electron tomography (cryo-ET) and live cell imaging reveal that GS and Pma1 are heterogeneously distributed into distinct plasma membrane microdomains. Treatment with caspofungin, an echinocandin antifungal that targets GS, alters the plasma membrane and disrupts the native distribution of GS and Pma1. Based on these findings, we propose a model for echinocandin action that considers how drug interactions with the plasma membrane environment lead to inhibition of GS. Our work underscores the importance of interrogating the structural and dynamic characteristics of fungal plasma membrane proteins in situ to understand function and facilitate precisely targeted development of novel antifungal therapies.

Beyond MPOWER: a systematic review of population-level factors that affect European tobacco smoking rates.

BACKGROUND: Population-level factors within and beyond the scope of the World Health Organization's (WHO) MPOWER policy package have significant impacts on smoking rates. However, no synthesis of the existing evidence exists. This systematic review identifies population-level factors that influence cigarette smoking rates in European countries. METHODS: We searched the ProQuest database collection for original, peer-reviewed quantitative evaluations that investigated the effects of population-level exposures on smoking rates in European countries. Of the 3122 studies screened, 62 were ultimately included in the review. A standardized data extraction form was used to identify key characteristics of each study including publication year, years evaluated, countries studied, population characteristics, study design, data sources, analytic methods, exposure studied, relevant covariates and effects on tobacco smoking outcomes. RESULTS: One hundred and fifty-five population-level exposures were extracted from the 62 studies included in the review, 99 of which were related to WHO MPOWER measures. An additional 56 exposures fell into eight policy realms: economic crises, education policy, macro-economic factors, non-MPOWER tobacco regulations, population welfare, public policy, sales to minors and unemployment rates. About one-half of the MPOWER exposures affected smoking rates (55/99) and did so in an overwhelmingly positive way (55/55). Over three-quarters of the non-MPOWER exposures were associated with statistically significant changes in smoking outcomes (43/56), with about two-thirds of these exposures leading to a decrease in smoking (29/43). CONCLUSIONS: Population-level factors that fall outside of the WHO's MPOWER measures are an understudied research area. The impacts of these factors on tobacco control should be considered by policymakers.

Quasi-experimental evaluation of national border closures on COVID-19 transmission.

With over 200 pandemic threats emerging every year, the efficacy of closing national borders to control the transmission of disease in the first months of a pandemic remains a critically important question. Previous studies offer conflicting evidence for the potential effects of these closures on COVID-19 transmission and no study has yet empirically evaluated the global impact of border closures using quasi-experimental methods and real-world data. We triangulate results from interrupted time-series analysis, meta-regression, coarsened exact matching, and an extensive series of robustness checks to evaluate the effect of 166 countries' national border closures on the global transmission of COVID-19. Total border closures banning non-essential travel from all countries and (to a lesser extent) targeted border closures banning travel from specific countries had some effect on temporarily slowing COVID-19 transmission in those countries that implemented them. In contrast to these country-level impacts, the global sum of targeted border closures implemented by February 5, 2020 was not sufficient to slow global COVID-19 transmission, but the sum of total border closures implemented by March 19, 2020 did achieve this effect. Country-level results were highly heterogeneous, with early implementation and border closures so broadly targeted that they resemble total border closures improving the likelihood of slowing the pandemic's spread. Governments that can make productive use of extra preparation time and cannot feasibly implement less restrictive alternatives might consider enacting border closures. However, given their moderate and uncertain impacts and their significant harms, border closures are unlikely to be the best policy response for most countries and should only be deployed in rare circumstances and with great caution. All countries would benefit from global mechanisms to coordinate national decisions on border closures during pandemics.

Classifying European cigarette consumption trajectories from 1970 to 2015.

OBJECTIVES: To systematically code and classify longitudinal cigarette consumption trajectories in European countries since 1970. DESIGN: Blinded duplicate qualitative coding of periods of year-over-year relative increase, plateau, and decrease of national per capita cigarette consumption and categorisation of historical cigarette consumption trajectories based on longitudinal patterns emerging from the data. SETTING: 41 countries or former countries in the European region for which data are available between 1970 and 2015. RESULTS: Regional trends in longitudinal consumption patterns identify stable or decreasing consumption throughout Northern, Western and Southern European countries, while Eastern and Southeastern European countries experienced much greater instability. The 11 emergent classes of historical cigarette consumption trajectories were also regionally clustered, including a distinctive inverted U or sine wave pattern repeatedly emerging from former Soviet and Southeastern European countries. CONCLUSIONS: The open-access data produced by this study can be used to conduct comparative international evaluations of tobacco control policies by separating impacts likely attributable to gradual long-term trends from those more likely attributable to acute short-term events. The complex, regionally clustered historical trajectories of cigarette consumption in Europe suggest that the enduring normative frame of a gently sloping downward curve in cigarette consumption can offer a false sense of security among policymakers and can distract from plausible causal mechanisms among researchers. These multilevel and multisectoral causal mechanisms point to the need for a greater understanding of the political economy of regional and global determinants of cigarette consumption.

Long-Term Prognosis and Treatment of Crop Impaction in Chickens via Ingluviotomy with Local Infiltration Anesthetic: Case Report.

Two hens from two backyard chicken flocks were presented to a private veterinary practice specializing in poultry, both with a chief complaint of crop distention. Crop impaction was diagnosed based on physical examinations and radiographic findings. The hens were admitted for ingluviotomy surgery to remove the impacted fibrous material under local anesthesia. Even though one of the hens was diagnosed with pendulous crop, both hens recovered uneventfully after ingluviotomy and postoperative care. Follow-up examinations and interviews with the owners confirmed excellent long-term recovery.

Reporte de caso- Pronóstico a largo plazo y tratamiento de la impactación del buche en pollos mediante ingluviotomía con anestesia local por infiltración: Reporte de un caso. Dos gallinas de dos parvadas de pollos de traspatio fueron presentadas a una clínica veterinaria privada especializada en aves, ambas aves mostraban como problema principal la distensión del buche. La impactación del buche se diagnosticó con base en exámenes físicos y hallazgos radiográficos. Las gallinas fueron ingresadas para cirugía de inluviotomía para remover el material fibroso impactado bajo anestesia local. A pesar de que a una de las gallinas se le diagnosticó buche pendular, ambas gallinas se recuperaron sin incidentes después de la ingluviotomía y el cuidado postoperatorio. Los exámenes de seguimiento y las entrevistas con los propietarios confirmaron una excelente recuperación a largo plazo.

International treaties have mostly failed to produce their intended effects.

There are over 250,000 international treaties that aim to foster global cooperation. But are treaties actually helpful for addressing global challenges? This systematic field-wide evidence synthesis of 224 primary studies and meta-analysis of the higher-quality 82 studies finds treaties have mostly failed to produce their intended effects. The only exceptions are treaties governing international trade and finance, which consistently produced intended effects. We also found evidence that impactful treaties achieve their effects through socialization and normative processes rather than longer-term legal processes and that enforcement mechanisms are the only modifiable treaty design choice with the potential to improve the effectiveness of treaties governing environmental, human rights, humanitarian, maritime, and security policy domains. This evidence synthesis raises doubts about the value of international treaties that neither regulate trade or finance nor contain enforcement mechanisms.

Principles and methods of global legal epidemiology.

Although the theory and methods of legal epidemiology-the scientific study and deployment of law as a factor in the cause, distribution, and prevention of disease and injury in a population-have been well developed in the context of domestic law, the challenges posed by shifting the frame of analysis to the global legal space have not yet been fully explored. While legal epidemiology rests on the foundational principles that law acts as an intervention, that law can be an object of scientific study and that law has impacts that should be evaluated, its application to the global level requires the recognition that international laws, policies and norms can cause effects independently from their legal implementation within countries. The global legal space blurs distinctions between 'hard' and 'soft' law, often operating through pathways of global agenda setting, legal language, political pressures, social mobilisation and trade pressures to have direct impacts on people, places and products. Despite these complexities, international law has been overwhelmingly studied as operating solely through national policy change, with only one global quasi-experimental evaluation of an international law's impact on health published to date. To promote greater adoption of global legal epidemiology, we expand on an existing typology of public health law studies with examples of policymaking, mapping, implementation, intervention and mechanism studies. Global legal epidemiology holds great promise as a way to produce rigorous and impactful research on the international laws, policies and norms that shape our collective health, equity and well-being.

Paromomycin Sulfate Treatment in Histomoniasis Outbreaks in Three Commercial Turkey Flocks in the Fraser Valley of British Columbia, Canada.

Over the last couple of years, the number of histomoniasis cases in commercial turkeys has increased substantially in British Columbia, particularly in the Fraser Valley. Due to a lack of approved efficacious preventive or curative drugs in Canada, histomoniasis outbreaks have had significant economic and animal welfare impacts on the commercial turkey industry. In July 2020, Health Canada conditionally approved the treatment use of paromomycin sulfate on a case-by-case basis via an emergency drug release authorization. Three flocks infected with Histomonas meleagridis were treated with labeled-dose paromomycin sulfate in the feed shortly after presumptive diagnosis. Despite the treatment, two out of three flocks suffered significant losses. One flock suffered over 67% mortality by the eighth day of treatment. Due to significant production loss and animal welfare concerns, the flock was shipped early for mercy cull; thus sustained 100% production loss. Another flock experienced over 38% mortality by the end of the fourth week of treatment and was slaughtered early to minimize production loss. The treatment response in two out of three cases suggests that any curative effect of paromomycin is limited. Thus, future field evaluation should carefully consider the prophylactic use of paromomycin sulfate, especially on farms with recurrent outbreaks.

Interfacial dilatational deformation accelerates particle formation in monoclonal antibody solutions.

Protein molecules are amphiphilic moieties that spontaneously adsorb at the air/solution (A/S) interface to lower the surface energy. Previous studies have shown that hydrodynamic disruptions to these A/S interfaces can result in the formation of protein aggregates that are of concern to the pharmaceutical industry. Interfacial hydrodynamic stresses encountered by protein therapeutic solutions under typical manufacturing, filling, and shipping conditions will impact protein stability, prompting a need to characterize the contribution of basic fluid kinematics to monoclonal antibody (mAb) destabilization. We demonstrate that dilatational surface deformations are more important to antibody stability when compared to constant-area shear of the A/S interface. We have constructed a dilatational interfacial rheometer that utilizes simultaneous pressure and bubble shape measurements to study the mechanical stability of mAbs under interfacial aging. It has a distinct advantage over methods utilizing the Young-Laplace equation, which incorrectly describes viscoelastic interfaces. We provide visual evidence of particle ejection from dilatated A/S interfaces and spectroscopic data of ejected mAb particles. These rheological studies frame a molecular understanding of the protein-protein interactions at the complex-fluid interface.

Multiplexed Fluid Flow Device to Study Cellular Response to Tunable Shear Stress Gradients.

Endothelial cells (ECs) line the interior of blood and lymphatic vessels and experience spatially varying wall shear stress (WSS) as an intrinsic part of their physiological function. How ECs, and mammalian cells generally, sense spatially varying WSS remains poorly understood, due in part to a lack of convenient tools for exposing cells to spatially varying flow patterns. We built a multiplexed device, termed a 6-well impinging flow chamber, that imparts controlled WSS gradients to a six-well tissue culture plate. Using this device, we investigated the migratory response of lymphatic microvascular ECs, umbilical vein ECs, primary fibroblasts, and epithelial cells to WSS gradients on hours to days timescales. We observed that lymphatic microvascular ECs migrate upstream, against the direction of flow, a response that was unique among all the cells types investigated here. Time-lapse, live cell imaging revealed that the microtubule organizing center relocated to the upstream side of the nucleus in response to the applied WSS gradient. To further demonstrate the utility of our device, we screened for the involvement of canonical signaling pathways in mediating this upstream migratory response. These data highlight the importance of WSS magnitude and WSS spatial gradients in dictating the cellular response to fluid flow.