CALX-CBD1 Ca2+-Binding Cooperativity Studied by NMR Spectroscopy and ITC with Bayesian Statistics.

The Na+/Ca2+ exchanger of Drosophila melanogaster, CALX, is the main Ca2+-extrusion mechanism in olfactory sensory neurons and photoreceptor cells. Na+/Ca2+ exchangers have two Ca2+ sensor domains, CBD1 and CBD2. In contrast to the mammalian homologs, CALX is inhibited by Ca2+ binding to CALX-CBD1, whereas CALX-CBD2 does not bind Ca2+ at physiological concentrations. CALX-CBD1 consists of a ß-sandwich and displays four Ca2+-binding sites at the tip of the domain. In this study, we used NMR spectroscopy and isothermal titration calorimetry (ITC) to investigate the cooperativity of Ca2+ binding to CALX-CBD1. We observed that this domain binds Ca2+ in the slow exchange regime at the NMR chemical shift timescale. Ca2+ binding restricts the dynamics in the Ca2+-binding region. Experiments of 15N chemical exchange saturation transfer and 15N R2 dispersion allowed the determination of Ca2+ dissociation rates (∼30 s-1). NMR titration curves of residues in the Ca2+-binding region were sigmoidal because of the contribution of chemical exchange to transverse magnetization relaxation rates, R2. Hence, a novel, to our knowledge, approach to analyze NMR titration curves was proposed. Ca2+-binding cooperativity was examined assuming two different stoichiometric binding models and using a Bayesian approach for data analysis. Fittings of NMR and ITC binding curves to the Hill model yielded nHill ∼2.9, near maximal cooperativity (nHill = 4). By assuming a stepwise model to interpret the ITC data, we found that the probability of binding from 2 up to 4 Ca2+ is approximately three orders of magnitude higher than that of binding a single Ca2+. Hence, four Ca2+ ions bind almost simultaneously to CALX-CBD1. Cooperative Ca2+ binding is key to enable this exchanger to efficiently respond to changes in the intracellular Ca2+ concentration in sensory neuronal cells.

Beyond species loss: How anthropogenic disturbances drive functional and phylogenetic homogenization of Neotropical dung beetles.

Anthropogenic activities drive tropical forest loss and biodiversity decay. However, few studies have addressed how the biodiversity response varies between disturbance-adapted species (i.e., winners) and those highly susceptible to disturbance (i.e., losers), or whether such responses differ between the taxonomic, functional, or phylogenetic dimensions of diversity. Understanding these dynamics can help prevent or buffer biotic homogenization processes. Using a meta-analytical approach with dung beetles as model organisms, we evaluated how anthropogenic habitat disturbances influence the multiple diversity dimensions of winner and loser species relative to conserved forest sites in the Neotropics. Habitats were organized according to a disturbance gradient ranging from second-growth forests, shaded agroforestry, lowly-shaded agroforestry, living fences, and pastures. Our database included 30 studies, from which we calculated nine metrics divided into three alfa diversity aspects: richness, evenness, and divergence. We also evaluated the beta-diversity response to disturbance and forest protection. All dimensions of dung beetle diversity decreased significantly with increasing disturbance levels, with phylogenetic diversity showing the highest losses, whereas evenness metrics increased in second-growth forests and agroforestry systems. Loser dung beetles showed high diversity loss as well as functional and phylogenetic clustering, reflecting a pervasive biotic homogenization in the most severely disturbed habitats, whereas winner species were insensitive to anthropogenic disturbances. Beta diversity increased significantly with disturbance and forest protection. Our study showed that heavy disturbances erode and homogenized all diversity dimensions of loser dung beetles. However, second-growth forests and agroforestry systems mitigated diversity loss and homogenization processes by favoring the coexistence between functional and phylogenetically distant species and maintaining assemblages compositionally similar to those in conserved forests, highlighting their importance for conservation. We encourage natural resource managers to consider protection of disturbed off-reserve forests in management schemes as these are essential for maintaining biodiversity in an increasingly anthropized world.

Acute toxicity of two insecticides on two species of Chagas disease vectors.

The control of triatomine vectors of Chagas disease is mainly based on the use of pyrethroid insecticides. Because chemical control is the primary method for managing these insects, it is crucial to diversify the range of products utilized to mitigate the risk of resistance development. This study evaluated the toxicity of two insecticides with different modes of action on Triatoma dimidiata Latreille and T. pallidipennis Stal first and third instar nymphs. Our study focused on the effects of two insecticides, buprofezin (a growth regulator) and flunocamid (an anti-feeder), on the mortality rate of triatomine bugs in a laboratory setting. Moreover, we investigated how direct and indirect (film method) exposure to these insecticides impacted the survival of the insects. Flonicamid emerged as a promising insecticide for triatomine control since it caused 100% mortality in first-instar nymphs 48 h after direct exposure. While, in third instar nymphs, the maximum mortality was 88% at 72 h after exposure. Our result can be used as a basis for future triatomine control plans.

Inter-rater reliability and prognostic value of baseline Radiographic Assessment of Lung Edema (RALE) scores in observational cohort studies of inpatients with COVID-19.

OBJECTIVES: To reliably quantify the radiographic severity of COVID-19 pneumonia with the Radiographic Assessment of Lung Edema (RALE) score on clinical chest X-rays among inpatients and examine the prognostic value of baseline RALE scores on COVID-19 clinical outcomes. SETTING: Hospitalised patients with COVID-19 in dedicated wards and intensive care units from two different hospital systems. PARTICIPANTS: 425 patients with COVID-19 in a discovery data set and 415 patients in a validation data set. PRIMARY AND SECONDARY OUTCOMES: We measured inter-rater reliability for RALE score annotations by different reviewers and examined for associations of consensus RALE scores with the level of respiratory support, demographics, physiologic variables, applied therapies, plasma host-response biomarkers, SARS-CoV-2 RNA load and clinical outcomes. RESULTS: Inter-rater agreement for RALE scores improved from fair to excellent following reviewer training and feedback (intraclass correlation coefficient of 0.85 vs 0.93, respectively). In the discovery cohort, the required level of respiratory support at the time of CXR acquisition (supplemental oxygen or non-invasive ventilation (n=178); invasive-mechanical ventilation (n=234), extracorporeal membrane oxygenation (n=13)) was significantly associated with RALE scores (median (IQR): 20.0 (14.1-26.7), 26.0 (20.5-34.0) and 44.5 (34.5-48.0), respectively, p<0.0001). Among invasively ventilated patients, RALE scores were significantly associated with worse respiratory mechanics (plateau and driving pressure) and gas exchange metrics (PaO2/FiO2 and ventilatory ratio), as well as higher plasma levels of IL-6, soluble receptor of advanced glycation end-products and soluble tumour necrosis factor receptor 1 (p<0.05). RALE scores were independently associated with 90-day survival in a multivariate Cox proportional hazards model (adjusted HR 1.04 (1.02-1.07), p=0.002). We replicated the significant associations of RALE scores with baseline disease severity and mortality in the independent validation data set. CONCLUSIONS: With a reproducible method to measure radiographic severity in COVID-19, we found significant associations with clinical and physiologic severity, host inflammation and clinical outcomes. The incorporation of radiographic severity assessments in clinical decision-making may provide important guidance for prognostication and treatment allocation in COVID-19.

Immersion-Induced Mitral Regurgitation: A Novel Risk Factor for Swimming-Induced Pulmonary Edema.

Immersion pulmonary edema, more commonly referred to as swimming-induced pulmonary edema (SIPE), is a well-documented condition believed to be a result of immersion physiologic condition that is characterized by a peripheral-to-central redistribution of blood volume. It disproportionally affects young, healthy athletes with no clinically overt cardiovascular or pulmonary conditions. We present four cases of healthy athletes with previously documented SIPE, who participated in Institutional Review Board-approved clinical studies that examined the pathophysiologic condition and prevention of SIPE. During standard recumbent echocardiography, trivial mitral regurgitation was observed in all four individuals. Acute exacerbation of their mitral regurgitation was observed during immersion with both immersed resting and immersed exercise echocardiography, contributing to the development of SIPE. These observations demonstrate that the occurrence of subclinical or trivial mitral valve regurgitation during dry rest is a novel risk factor for SIPE. We propose the use of immersion echocardiography as a useful investigative tool for otherwise healthy individuals with SIPE and no previously explainable cause.

Radiographic Assessment of Lung Edema (RALE) Scores are Highly Reproducible and Prognostic of Clinical Outcomes for Inpatients with COVID-19.

INTRODUCTION: Chest imaging is necessary for diagnosis of COVID-19 pneumonia, but current risk stratification tools do not consider radiographic severity. We quantified radiographic heterogeneity among inpatients with COVID-19 with the Radiographic Assessment of Lung Edema (RALE) score on Chest X-rays (CXRs). METHODS: We performed independent RALE scoring by ≥2 reviewers on baseline CXRs from 425 inpatients with COVID-19 (discovery dataset), we recorded clinical variables and outcomes, and measured plasma host-response biomarkers and SARS-CoV-2 RNA load from subjects with available biospecimens. RESULTS: We found excellent inter-rater agreement for RALE scores (intraclass correlation co-efficient=0.93). The required level of respiratory support at the time of baseline CXRs (supplemental oxygen or non-invasive ventilation [n=178]; invasive-mechanical ventilation [n=234], extracorporeal membrane oxygenation [n=13]) was significantly associated with RALE scores (median [interquartile range]: 20.0[14.1-26.7], 26.0[20.5-34.0] and 44.5[34.5-48.0], respectively, p<0.0001). Among invasively-ventilated patients, RALE scores were significantly associated with worse respiratory mechanics (plateau and driving pressure) and gas exchange metrics (PaO2/FiO2 and ventilatory ratio), as well as higher plasma levels of IL-6, sRAGE and TNFR1 levels (p<0.05). RALE scores were independently associated with 90-day survival in a multivariate Cox proportional hazards model (adjusted hazard ratio 1.04[1.02-1.07], p=0.002). We validated significant associations of RALE scores with baseline severity and mortality in an independent dataset of 415 COVID-19 inpatients. CONCLUSION: Reproducible assessment of radiographic severity revealed significant associations with clinical and physiologic severity, host-response biomarkers and clinical outcome in COVID-19 pneumonia. Incorporation of radiographic severity assessments may provide prognostic and treatment allocation guidance in patients hospitalized with COVID-19.

Mechanisms of diversity maintenance in dung beetle assemblages in a heterogeneous tropical landscape.

BACKGROUND: Anthropized landscapes play a crucial role in biodiversity conservation, as they encompass about 90% of the remaining tropical forest. Effective conservation strategies require a deep understanding of how anthropic disturbances determine diversity patterns across these landscapes. Here, we evaluated how attributes and assembly mechanisms of dung beetle communities vary across the Selva El Ocote Biosphere Reserve (REBISO) landscape. METHODS: Community attributes (species diversity, abundance, and biomass) were assessed at the landscape scale, using spatial windows and vegetation classes. Windows were categorized as intact, variegated, or fragmented based on their percent cover of tropical forest. The vegetation classes analyzed were tropical forest, second-growth forest, and pastures. RESULTS: We collected 15,457 individuals and 55 species. Variegated windows, tropical forests, and second-growth forests showed the highest diversity values, while the lowest values were found in intact windows and pastures. Landscape fragmentation was positively and strongly related to dung beetle diversity and negatively related to their abundance; biomass was positively associated with forest cover. Beta diversity was the primary driver of the high dung beetle diversity in the landscape analyzed. DISCUSSION: The landscape heterogeneity and its biodiversity-friendly matrix facilitate the complementarity of dung beetle assemblages in the Selva El Ocote Biosphere Reserve. Random processes govern beta diversity patterns in intact and variegated windows. Therefore, vegetation cover in the region is sufficient to maintain a continuous flow of dung beetles between forested landscape segments. However, intense anthropic disturbances acted as deterministic environmental filters in fragmented windows and pastures sites, leading to biotic homogenization processes. Our results suggest that increasing habitat variegation in highly fragmented sites is an effective strategy to prevent or buffer homogenization processes in the REBISO landscape.