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Abstract Bacterial spores owe their incredible resistance capacities to molecular structures that protect the cell content from external aggressions. Among the determinants of resistance are the quaternary structure of the chromosome and an extracellular shell made of proteinaceous layers (the coat), the assembly of which remains poorly understood. Here, in situ cryo-electron tomography on lamellae generated by cryo-focused ion beam micromachining provides insights into the ultrastructural organization of Bacillus subtilis sporangia. The reconstructed tomograms reveal that early during sporulation, the chromosome in the forespore adopts a toroidal structure harboring 5.5-nm thick fibers. At the same stage, coat proteins at the surface of the forespore form a stack of amorphous or structured layers with distinct electron density, dimensions and organization. By analyzing mutant strains using cryo-electron tomography and transmission electron microscopy on resin sections, we distinguish seven nascent coat regions with different molecular properties, and propose a model for the contribution of coat morphogenetic proteins.

Heat waves (HWs) and cold waves (CWs) can have considerable impact on people. Mapping risks of extreme temperature at local scale, accounting for the interactions between hazard, exposure, and vulnerability, remains a challenging task. In this study, we quantify risks from HWs and CWs for the Trentino-Alto Adige region of Italy from 1980 to 2018 at high spatial resolution. We use the Heat Wave Magnitude Index daily (HWMId) and the Cold Wave Magnitude Index daily (CWMId) as the hazard indicators. To obtain HWs and CW risk maps we combined the following: (i) occurrence probability maps of the hazard obtained using the zero-inflated Tweedie distribution (accounting directly for the absence of events for certain years), (ii) normalized population density maps, and (iii) normalized vulnerability maps based on eight socioeconomic indicators. The methodology allowed us to disentangle the contributions of each component of the risk relative to total change in risk. We find a statistically significant increase in HW hazard and exposure, while CW hazard remained stagnant in the analyzed area over the study period. A decrease in vulnerability to extreme temperature spells is observed through the region except in the larger cities where vulnerability increased. HW risk increased in 40 % of the region, with the increase being greatest in highly populated areas. Stagnant CW hazard and declining vulnerability result in reduced CW risk levels overall, except for the four main cities where increased vulnerability and exposure increased risk levels. These findings can help to steer investments in local risk mitigation, and this method can potentially be applied to other regions where there are sufficient detailed data.

BACKGROUND AND OBJECTIVES: To expand the clinical knowledge of GPAA1-related glycosylphosphatidylinositol (GPI) deficiency. METHODS: An international case series of 7 patients with biallelic GPAA1 variants were identified. Clinical, biochemical, and neuroimaging data were collected for comparison. Where possible, GPI-anchored proteins were assessed using flow cytometry. RESULTS: Ten novel variants were identified in 7 patients. Flow cytometry samples of 3 available patients confirmed deficiency of several GPI-anchored proteins on leukocytes. Extensive phenotypic information was available for each patient. The majority experienced developmental delay, seizures, and hypotonia. Neuroimaging revealed cerebellar anomalies in the majority of the patients. Alkaline phosphatase was within the normal range in 5 individuals and low in 1 individual, as has been noted in other transamidase defects. We notably describe individuals either less affected or older than the ones published previously. DISCUSSION: Clinical features of the cases reported broaden the spectrum of the known phenotype of GPAA1-related GPI deficiency, while outlining the importance of using functional studies such as flow cytometry to aid in variant classification.

ABSTRACTA serum-free, highly purified rabies vaccine produced in Vero cells is under development. The initial formulation, PVRV-NG, was evaluated in five Phase II studies and subsequently reformulated (PVRV-NG2). This multicenter, observer-blinded Phase II study investigated the safety and immune response of three different doses (antigen content) of PVRV-NG2 versus a licensed human diploid cell rabies vaccine (HDCV; Imovax rabies®). Healthy adults (N = 320) were randomized to receive PVRV-NG2 (low, medium, or high dose), PVRV-NG, or HDCV (2:2:2:1:1 ratio), according to a five-dose Essen simulated post-exposure regimen (Days [D] 0, 3, 7, 14, and 28). All participants received human rabies immunoglobulin intramuscularly on D0. Immunogenicity was assessed at D0, 14, 28, 42, and 6 months after the final injection using the rapid fluorescent focus inhibition test. Seroconversion rates were calculated as the percentage of participants achieving rabies virus neutralizing antibody titers ≥0.5 IU/mL. All analyses were descriptive. At each timepoint, geometric mean titers (GMTs) increased with antigen content (measured using an enzyme-linked immunosorbent assay). High-dose PVRV-NG2 GMTs were the highest at all timepoints, medium-dose PVRV-NG2 GMTs were similar to those with HDCV, and low-dose PVRV-NG2 GMTs were similar to PVRV-NG. The safety profile of PVRV-NG2 was comparable to PVRV-NG; however, fewer injection site reactions were reported with PVRV-NG2 or PVRV-NG (range 36.7–47.5%) than with HDCV (61.5%). This study demonstrated a dose–effect of antigen content at all timepoints. As post-exposure prophylaxis, the safety and immunogenicity profiles of the high-dose PVRV-NG2 group compared favorably with HDCV. Clinicaltrials.gov number: NCT03145766.

La investigación se realizó para evaluar el desempeño de parametrizaciones físicas y las condiciones de frontera estática del modelo de predicción e investigación meteorológica, WRF, para el estudio de los topoclimas en la región oriental de Cuba. En tres etapas, se realizaron simulaciones del clima local a corto plazo con combinaciones de parametrizaciones cúmulo, microfísica y de capa límite planetaria. En la tercera etapa se evaluó, además, el aporte al desempeño del modelo de la integración de bases de datos espaciales nacionales de alta resolución como condiciones de frontera estática. Como conjunto de datos de condiciones iniciales y de contorno se utilizó ERA5, con tres dominios anidados a resoluciones espaciales de 30, 10 y 3.33 km. El estudio se realizó para la temperatura y precipitación de los periodos lluvioso y poco lluvioso del año 2018. Los resultados obtenidos mostraron que, de los elementos climáticos en estudio, la precipitación mostró la mayor incertidumbre en todos los experimentos y casos de estudio. Análogamente, las bases de datos espaciales de alta resolución que se integraron al modelo como condiciones de frontera estática, incidió en el desempeño de WRF a escala local. El mejor desempeño del WRF para el estudio de los topoclimas en la región oriental de Cuba se alcanzó con las parametrizaciones de Kain-Fritcsh como cúmulo, la microfísica de Thompson y Yonsei University como opción de capa límite planetaria, activo viento topográfico por el método de corrección por geoformas.

In recent decades, the area affected and the severity of forest fires have increased in many regions of the world. Without anticipatory adjustments, climate change is likely to increase their frequency and intensity. This study aims to assess the possible impact of climate change on the initiation of forest fires in Las Tunas, The purpose of this study was to assess the possible impact of the climate change in forest fires initiation in Las Tunas, which is a most deforested region in Cuba thus a vulnerable region to aridity, drought and prone to forest fires. Taking into account the characteristics of the meteorological variables, the aridity conditions and the behavior of forest fires, a long-term risk index was used. It allows determining by months the areas with favorable conditions for the start and development of fires. Based on the possible changes in the meteorological variables in the RCP2.6 and RCP8.5 scenarios, this index was evaluated, which allowed obtaining the areas with the highest fire risk for the periods 2020-2049, 2050-2079 and 2080-2099. In general, in the territory of Las Tunas, there is a tendency to warmer conditions with reductions in rainfall, especially in the north coast. Besides, there is a decrease in relative humidity, which will lead to more areas affected by aridity and drought. Also, there will be larger areas with the presence of fire danger conditions for vegetation.

The paper presents a high-resolution (-3km) gridded dataset for daily precipitation across Cuba for 1961-2008, called CubaPrec1. The dataset was built using the information from the data series of 630 stations from the network operated by the National Institute of Water Resources. The original station data series were quality controlled using a spatial coherence process of the data, and the missing values were estimated on each day and location independently. Using the filled data series, a grid of 3 × 3 km spatial resolution was constructed by estimating daily precipitation and their corresponding uncertainties at each grid box. This new product represents a precise spatiotemporal distribution of precipitation in Cuba and provides a useful baseline for future studies in hydrology, climatology, and meteorology. The data collection described is available on zenodo: https://doi.org/10.5281/zenodo.7847844

Abstract Background Genetic tumor risk syndromes are responsible for at least five to ten percent of the 4 million cases of cancer diagnosed in Europe every year. Currently, the care of oncological patients suffers from a lack of specialists in medical genetics and also a lack of access to genetic care in rural areas and structured care pathways between oncologists and medical geneticists. As a result, genetic tumor risk syndromes are underdiagnosed with potentially fatal consequences for patients and their families. Methods The OnkoRiskNET study is supported by a grant from the Federal Joint Committee of the Federal Republic of Germany. The study will include 2,000 oncological index patients from oncology practices in Lower Saxony and Saxony after the start of the study in July 2021. Randomization is carried out by means of a stepped wedge design at the level of the practices. Patients either go through routine care or the new form of care with structured cooperation between medical geneticists and oncologists, case management and the use of telemedical genetic counseling. Using a mixed-methods approach, the following parameters will be evaluated in the control and intervention group: (1) Conducted genetic counseling sessions by patients with suspected tumor risk syndrome and their first degree relatives; (2) Patient satisfaction and psychological distress after genetic counseling and testing; (3) Factors influencing the acceptance and experience of telemedical genetic counseling; (4) Satisfaction of oncologists and medical genetics with the structured pathway; (5) Cost efficiency of the new form of care. Discussion OnkoRiskNET aims to close the gap in care through the formation of a cooperation network between practicing oncologists and specialists in medical genetics and the use of telemedical genetic counseling, thereby, increasing the diagnostic rate in genetic tumor risk syndromes and serving as a model for future genetic care in Germany. Trial registration Trial was registered on 01.12.2021 in the German Clinical Trial Register ( https://trialsearch.who.int/ ) with the DRKS-ID: DRKS00026679 . Title: Cooperation network for the provision of local care for patients and families with a genetic tumour risk syndrome. Trial acronym: OnkoRiskNET. Protocol version 1.1.

Abstract The World Health Organization protocol for rabies post-exposure prophylaxis (PEP) recommends extensive wound washing, immediate vaccination, and administration of rabies immunoglobulin (RIG) in severe category III exposures. Some studies have shown that RIG can interfere with rabies vaccine immunogenicity to some extent. We investigated the interference of RIG on a next generation highly purified Vero cell rabies vaccine candidate (PVRV-NG) versus standard-of-care vaccines in a previously described hamster model. The interference of either human (h) or equine (e) RIG on the immune response elicited by PVRV-NG, Verorab® (purified Vero cell rabies vaccine, PVRV), and Imovax® Rabies (human diploid cell rabies vaccine; HDCV) was evaluated using the 4-dose Essen PEP regimen. The anti-rabies seroneutralizing titers and specific serum IgM titers were measured by fluorescent antibody virus neutralization test and enzyme-linked immunosorbent assay, respectively, for the vaccines administered with or without RIG. The RIG interference on PVRV-NG, observed transiently at Day 7, was similar to that on PVRV and tended to be lower than that on HDCV using both read-outs. In summary, the results generated in the hamster model showed that RIG induced similar or less interference on PVRV-NG than the standard-of-care vaccines.

Ante la evidencia que confirma el calentamiento del sistema climático y la preocupación por las insuficientes medidas de respuesta, el Manejo de la Radiación Solar (SRM) ha sido considerado como una acción adicional para limitar el aumento de la temperatura global. A pesar de que el SRM es muy controvertido, una evaluación de los efectos potenciales en los extremos climáticos podría proporcionar elementos adicionales sobre sus implicaciones. En el presente trabajo se realiza un análisis de los posibles efectos que tendría el SRM comparando las proyecciones sobre el clima futuro de Cuba bajo escenarios SRM, con respecto a un escenario de emisiones de gases de efecto invernadero. Se utilizaron los datos diarios de precipitación y temperaturas máximas y mínimas correspondientes a las salidas del modelo HadGEM2-ES para el escenario RCP4.5 y los dos esquemas SRM G3 y G4. Con esta información se calcularon y contrastaron 10 indicadores de extremos climáticos en el futuro 2020-2070 con respecto al período de referencia 1970-2000. Los resultados muestran que la posible implementación de SRM con aerosoles estratosféricos podría mejorar levemente el escenario futuro proyectado bajo RCP4.5 en relación a las temperaturas, al atenuar el aumento o disminución de extremos relacionados con el régimen térmico. Sin embargo, en el caso de precipitaciones extremas, los cambios estimados para los escenarios G3 y G4 no son significativamente diferentes de los proyectados bajo RCP4.5.

Se presenta una valoración de las tendencias hacia el aumento de la temperatura y la salinidad en las aguas aledañas a Cuba. Se utilizan proyecciones de los modelos climáticos globales HadGEM2-AO, HadGEM2-ES, HadGEM2-CC y el CNRC-CM5 en el escenario de forzamiento de CPR 4.5. La validez de los cambios identificados en las salidas de los modelos globales utilizados, se analiza en comparación con observaciones de expediciones en aguas cubanas, realizadas en 1966-2000. Los resultados muestran que los cambios proyectados para el siglo XXI pueden tener implicaciones adversas para el régimen de inundaciones costeras y los ecosistemas marinos en aguas cubanas.

Summary: Fluorescence microscopy is a method of choice for studying peptidoglycan assembly, but it presents two major challenges: the peptidoglycan must be labeled with a probe that will not perturb the physiological process, and the spatial resolution must reach the nanometer scale to reveal fine details of the synthesis process. This protocol meets both challenges by combining biorthogonal metabolic labeling of peptidoglycan in Streptococcus pneumoniae with super-resolution fluorescence microscopy (dSTORM), also providing cues to adapt it to other bacteria.For complete details on the use and execution of this protocol, please refer to Trouve et al. (2021).

How organisms develop into specific shapes is a central question in biology. The maintenance of bacterial shape is connected to the assembly and remodelling of the cell envelope. In endospore-forming bacteria, the pre-spore compartment (the forespore) undergoes morphological changes that result in a spore of defined shape, with a complex, multi-layered cell envelope. However, the mechanisms that govern spore shape remain poorly understood. Here, using a combination of fluorescence microscopy, quantitative image analysis, molecular genetics and transmission electron microscopy, we show that SsdC (formerly YdcC), a poorly-characterized new member of the MucB / RseB family of proteins that bind lipopolysaccharide in diderm bacteria, influences spore shape in the monoderm Bacillus subtilis. Sporulating cells lacking SsdC fail to adopt the typical oblong shape of wild-type forespores and are instead rounder. 2D and 3D-fluorescence microscopy suggest that SsdC forms a discontinuous, dynamic ring-like structure in the peripheral membrane of the mother cell, near the mother cell proximal pole of the forespore. A synthetic sporulation screen identified genetic relationships between ssdC and genes involved in the assembly of the spore coat. Phenotypic characterization of these mutants revealed that spore shape, and SsdC localization, depend on the coat basement layer proteins SpoVM and SpoIVA, the encasement protein SpoVID and the inner coat protein SafA. Importantly, we found that the ΔssdC mutant produces spores with an abnormal-looking cortex, and abolishing cortex synthesis in the mutant largely suppresses its shape defects. Thus, SsdC appears to play a role in the proper assembly of the spore cortex, through connections to the spore coat. Collectively, our data suggest functional diversification of the MucB / RseB protein domain between diderm and monoderm bacteria and identify SsdC as an important factor in spore shape development.