RyR1-targeted drug discovery pipeline integrating FRET-based high-throughput screening and human myofiber dynamic Ca2+ assays.

Elevated cytoplasmic [Ca2+] is characteristic in severe skeletal and cardiac myopathies, diabetes, and neurodegeneration, and partly results from increased Ca2+ leak from sarcoplasmic reticulum stores via dysregulated ryanodine receptor (RyR) channels. Consequently, RyR is recognized as a high-value target for drug discovery to treat such pathologies. Using a FRET-based high-throughput screening assay that we previously reported, we identified small-molecule compounds that modulate the skeletal muscle channel isoform (RyR1) interaction with calmodulin and FK506 binding protein 12.6. Two such compounds, chloroxine and myricetin, increase FRET and inhibit [3H]ryanodine binding to RyR1 at nanomolar Ca2+. Both compounds also decrease RyR1 Ca2+ leak in human skinned skeletal muscle fibers. Furthermore, we identified compound concentrations that reduced leak by > 50% but only slightly affected Ca2+ release in excitation-contraction coupling, which is essential for normal muscle contraction. This report demonstrates a pipeline that effectively filters small-molecule RyR1 modulators towards clinical relevance

Climate change, water and agriculture in the Greater Mekong subregion

Climate change / Adaptation / Indicators / Water resource management / River basins / Water availability / Water quality / Groundwater / Fisheries / Ecosystems / Water power / Population growth / Land use / Biofuels / Sea level / South East Asia / Cambodia / Laos / Myanmar / Thailand / Vietnam / China / Greater Mekong Subregion / Yunnan Province

Excess Body Weight and Gait Influence Energy Cost of Walking in Older Adults

Purpose: To study how excess body weight influences the energy cost of walking (Cw) and determine if overweight and obese older adults self-select stride frequency to minimize Cw. Methods: Using body mass index (BMI) men and women between the ages of 65–80 yr were separated into normal weight (NW, BMI ≤ 24.9 kg m−2, n = 13) and overweight-obese groups (OWOB, BMI ≥25.0 kg m−2, n = 13). Subjects walked at 0.83 m s−1 on an instrumented treadmill that recorded gait parameters, and completed three, six-minute walking trials; at preferred stride frequency (PSF), at +10% PSF, and at −10% PSF. Cw was determined by indirect calorimetry. Repeated measures analysis of variance was used to compare groups, and associations were tested with Pearson correlations, α = 0.05. Results: OWOB had 62% greater absolute Cw (301 ± 108 vs. 186 ± 104 J m−1, P \u3c 0.001) and 20% greater relative Cwkg (3.48 ± 0.95 vs. 2.91 ± 0.94 J kg−1 m−1, P = 0.046) than NW. Although PSF was not different between OWOB and NW (P = 0.626), Cw was 8% greater in OWOB at +10% PSF (P \u3c 0.001). At PSF OWOB spent less time in single-limb support (33.1 ± 1.5 vs. 34.9 ± 1.6 %GC, P = 0.021) and more time in double-limb support (17.5 ± 1.6 vs. 15.4 ± 1.4 %GC, P = 0.026) than NW. In OWOB, at PSF, Cw was correlated to impulse (r = −0.57, P = 0.027) and stride frequency (r = 0.51, P = 0.046). Conclusions: Excess body weight is associated with greater Cw in older adults, possibly contributing to reduced mobility in overweight and obese older persons

Scoping study on natural resources and climate change in Southeast Asia with a focus on agriculture. Final report

Climate change / Natural resources / Environmental effects / Agroecology / Agricultural production / Crops / Cropping systems / Farming systems / Livestock / Fisheries / Food security / Water management / Economic aspects / Rural poverty / Policy / Nutrient management / South East Asia / Cambodia / Laos / Thailand / Vietnam / Myanmar / China / Greater Mekong Subregion / Tonle Sap / Yunnan

Knocking on closed doors: Host interferons dynamically regulate blood-brain barrier function during viral infections of the central nervous system

The central nervous system (CNS) is among the most important organ systems, integrating information inputs and coordinating the activity of all other body systems. Like many organ systems, the CNS is susceptible to infection by pathogenic microorganisms, including many arboviruses that are considered neurotropic because they are able to achieve robust replicatio

CCR5 limits cortical viral loads during West Nile virus infection of the central nervous system

BACKGROUND: Cell-mediated immunity is critical for clearance of central nervous system (CNS) infection with the encephalitic flavivirus, West Nile virus (WNV). Prior studies from our laboratory have shown that WNV-infected neurons express chemoattractants that mediate recruitment of antiviral leukocytes into the CNS. Although the chemokine receptor, CCR5, has been shown to play an important role in CNS host defense during WNV infection, regional effects of its activity within the infected brain have not been defined. METHODS: We used CCR5-deficient mice and an established murine model of WNV encephalitis to determine whether CCR5 activity impacts on WNV levels within the CNS in a region-specific fashion. Statistical comparisons between groups were made with one- or two-way analysis of variance; Bonferroni’s post hoc test was subsequently used to compare individual means. Survival was analyzed by the log-rank test. Analyses were conducted using Prism software (GraphPad Prism). All data were expressed as means ± SEM. Differences were considered significant if P ≤ 0.05. RESULTS: As previously shown, lack of CCR5 activity led to increased symptomatic disease and mortality in mice after subcutaneous infection with WNV. Evaluation of viral burden in the footpad, draining lymph nodes, spleen, olfactory bulb, and cerebellum derived from WNV-infected wild-type, and CCR5(−/−) mice showed no differences between the genotypes. In contrast, WNV-infected, CCR5(−/−) mice exhibited significantly increased viral burden in cortical tissues, including the hippocampus, at day 8 post-infection. CNS regional studies of chemokine expression via luminex analysis revealed significantly increased expression of CCR5 ligands, CCL4 and CCL5, within the cortices of WNV-infected, CCR5(−/−) mice compared with those of similarly infected WT animals. Cortical elevations in viral loads and CCR5 ligands in WNV-infected, CCR5(−/−) mice, however, were associated with decreased numbers of infiltrating mononuclear cells and increased permeability of the blood-brain barrier. CONCLUSIONS: These data indicate that regional differences in chemokine expression occur in response to WNV infection of the CNS, and that cortical neurons require CCR5 activity to limit viral burden in this brain region

Regional astrocyte IFN signaling restricts pathogenesis during neurotropic viral infection

Type I IFNs promote cellular responses to viruses, and IFN receptor (IFNAR) signaling regulates the responses of endothelial cells of the blood-brain barrier (BBB) during neurotropic viral infection. However, the role of astrocytes in innate immune responses of the BBB during viral infection of the CNS remains to be fully elucidated. Here, we have demonstrated that type I IFNAR signaling in astrocytes regulates BBB permeability and protects the cerebellum from infection and immunopathology. Mice with astrocyte-specific loss of IFNAR signaling showed decreased survival after West Nile virus infection. Accelerated mortality was not due to expanded viral tropism or increased replication. Rather, viral entry increased specifically in the hindbrain of IFNAR-deficient mice, suggesting that IFNAR signaling critically regulates BBB permeability in this brain region. Pattern recognition receptors and IFN-stimulated genes had higher basal and IFN-induced expression in human and mouse cerebellar astrocytes than did cerebral cortical astrocytes, suggesting that IFNAR signaling has brain region–specific roles in CNS immune responses. Taken together, our data identify cerebellar astrocytes as key responders to viral infection and highlight the existence of distinct innate immune programs in astrocytes from evolutionarily disparate regions of the CNS

Reliability of Standardized Assessment for Adults who are Deafblind

This study assessed the reliability of the interRAI Community Health Assessment (interRAI CHA) and Deafblind Supplement (DbS). The interRAI CHA and DbS represents a multidimensional, standardized assessment instrument for use with adults (18 and older) who are deafblind. The interrater reliability of the instrument was tested through the completion of dual assessments with 44 individuals who were deafblind in the province of Ontario, Canada. Overall, nearly 50% of items had a kappa value of at least 0.60, indicating fair to substantial agreement for these items. Several items related to psychosocial well-being, mood, and sense of involvement had kappa scores of less than 0.40. However, among these items with low kappa values, most (78%) showed at least 70% agreement between the two assessors. The internal consistency of several health subscales, embedded within the assessment, was also very good and ranged from 0.63 to 0.93. The interRAI was also very good and ranged from 0.63 to 0.93. The interRAI CHA and DbS represents a reliable instrument for assessing adults with deafblindness to better understand their needs, abilities, and preferences

Risk factors for human brucellosis in northern Tanzania

Little is known about the epidemiology of human brucellosis in sub-Saharan Africa. This hampers prevention and control efforts at the individual and population levels. To evaluate risk factors for brucellosis in northern Tanzania, we conducted a study of patients presenting with fever to two hospitals in Moshi, Tanzania. Serum taken at enrollment and at 4–6 week follow-up was tested by Brucella microagglutination test. Among participants with a clinically compatible illness, confirmed brucellosis cases were defined as having a ≥ 4-fold rise in agglutination titer between paired sera or a blood culture positive for Brucella spp., and probable brucellosis cases were defined as having a single reciprocal titer ≥ 160. Controls had reciprocal titers < 20 in paired sera. We collected demographic and clinical information and administered a risk factor questionnaire. Of 562 participants in the analysis, 50 (8.9%) had confirmed or probable brucellosis. Multivariable analysis showed that risk factors for brucellosis included assisting goat or sheep births (Odds ratio [OR] 5.9, 95% confidence interval [CI] 1.4, 24.6) and having contact with cattle (OR 1.2, 95% CI 1.0, 1.4). Consuming boiled or pasteurized dairy products was protective against brucellosis (OR 0.12, 95% CI 0.02, 0.93). No participants received a clinical diagnosis of brucellosis from their healthcare providers. The under-recognition of brucellosis by healthcare workers could be addressed with clinician education and better access to brucellosis diagnostic tests. Interventions focused on protecting livestock keepers, especially those who assist goat or sheep births, are needed