Altered vacuole membrane protein 1 (VMP1) expression is associated with increased NLRP3 inflammasome activation and mitochondrial dysfunction.

BACKGROUND: Altered expression of vacuole membrane protein 1 (VMP1) has recently been observed in the context of multiple sclerosis and Parkinson's disease (PD). However, how changes in VMP1 expression may impact pathogenesis has not been explored. OBJECTIVE: This study aimed to characterize how altered VMP1 expression affects NLRP3 inflammasome activation and mitochondrial function. METHODS: VMP1 expression was depleted in a monocytic cell line using CRISPR-Cas9. The effect of VMP1 on NLRP3 inflammasome activation was examined by stimulating cells with LPS and ATP or α-synuclein fibrils. Inflammasome activation was determined by caspase-1 activation using both a FLICA assay and a biosensor as well as by the release of proinflammatory molecules measured by ELISA. RNA-sequencing was utilized to define global gene expression changes resulting from VMP1 deletion. SERCA activity and mitochondrial function were investigated using various fluorescence microscopy-based approaches including a novel method that assesses the function of individual mitochondria in a cell. RESULTS: Here, we report that genetic deletion of VMP1 from a monocytic cell line resulted in increased NLRP3 inflammasome activation and release of proinflammatory molecules. Examination of the VMP1-dependent changes in these cells revealed that VMP1 deficiency led to decreased SERCA activity and increased intracellular [Ca2+]. We also observed calcium overload in mitochondria in VMP1 depleted cells, which was associated with mitochondrial dysfunction and release of mitochondrial DNA into the cytoplasm and the extracellular environment. CONCLUSIONS: Collectively, these studies reveal VMP1 as a negative regulator of inflammatory responses, and we postulate that decreased expression of VMP1 can aggravate the inflammatory sequelae associated with neurodegenerative diseases like PD.

Vacuole Membrane Protein 1 (VMP1) Restricts NLRP3 Inflammasome Activation by Modulating SERCA Activity and Autophagy.

Altered expression of vacuole membrane protein 1 (VMP1) has recently been observed in the context of multiple sclerosis and Parkinson's disease (PD). However, how changes in VMP1 expression may impact pathogenesis has not been explored. Here, we report that genetic deletion of VMP1 from a monocytic cell line resulted in increased NLRP3 inflammasome activation and release of proinflammatory molecules. Examination of the VMP1 dependent changes in these cells revealed that VMP1 deficiency led to decreased SERCA activity and increased intracellular [Ca2+]. We also observed calcium overload in mitochondria in VMP1 depleted cells, which was associated with mitochondrial dysfunction and release of mitochondrial DNA into the cytoplasm and the extracellular environment. Autophagic defects were also observed in VMP1 depleted macrophages. Collectively, these studies reveal VMP1 as a negative regulator of inflammatory responses, and we postulate that decreased expression of VMP1 can aggravate the inflammatory sequelae associated with neurodegenerative diseases like PD.

GISS-E2.1: Configurations and Climatology.

This paper describes the GISS-E2.1 contribution to the Coupled Model Intercomparison Project, Phase 6 (CMIP6). This model version differs from the predecessor model (GISS-E2) chiefly due to parameterization improvements to the atmospheric and ocean model components, while keeping atmospheric resolution the same. Model skill when compared to modern era climatologies is significantly higher than in previous versions. Additionally, updates in forcings have a material impact on the results. In particular, there have been specific improvements in representations of modes of variability (such as the Madden-Julian Oscillation and other modes in the Pacific) and significant improvements in the simulation of the climate of the Southern Oceans, including sea ice. The effective climate sensitivity to 2 × CO2 is slightly higher than previously at 2.7-3.1°C (depending on version) and is a result of lower CO2 radiative forcing and stronger positive feedbacks.

Cargo and cell-specific differences in extracellular vesicle populations identified by multiplexed immunofluorescent analysis.

Extracellular vesicles (EVs) have been implicated in a wide variety of biological activities, have been implicated in the pathogenesis of numerous diseases, and have been proposed to serve as potential biomarkers of disease in human patients and animal models. However, characterization of EV populations is often performed using methods that do not account for the heterogeneity of EV populations and require comparatively large sample sizes to facilitate analysis. Here, we describe an imaging-based method that allows for the multiplexed characterization of EV populations at the single EV level following centrifugation of EV populations directly onto cover slips, allowing comprehensive analysis of EV populations with relatively small samples. We observe that canonical EV markers are present on subsets of EVs which differ substantially in a producer cell and cargo specific fashion, including differences in EVs containing different HIV-1 proteins previously reported to be incorporated into pathogenic EVs. We also describe a lectin binding assay to interrogate EVs based on their glycan content, which we observe to change in response to pharmacological modulation of secretory autophagy pathways. These studies collectively reveal that a multiplexed analysis of EV populations using fluorescent microscopy can reveal differences in specific EV populations that may be used to understand the biogenesis of specific EV populations and/or to interrogate small subsets of EVs of interest within larger EV populations in biological samples.

Photoemission sources and beam blankers for ultrafast electron microscopy.

Observing atomic motions as they occur is the dream goal of ultrafast electron microscopy (UEM). Great progress has been made so far thanks to the efforts of many scientists in developing the photoemission sources and beam blankers needed to create short pulses of electrons for the UEM experiments. While details on these setups have typically been reported, a systematic overview of methods used to obtain a pulsed beam and a comparison of relevant source parameters have not yet been conducted. In this report, we outline the basic requirements and parameters that are important for UEM. Different types of imaging modes in UEM are analyzed and summarized. After reviewing and analyzing the different kinds of photoemission sources and beam blankers that have been reported in the literature, we estimate the reduced brightness for all the photoemission sources reviewed and compare this to the brightness in the continuous and blanked beams. As for the problem of pulse broadening caused by the repulsive forces between electrons, four main methods available to mitigate the dispersion are summarized. We anticipate that the analysis and conclusions provided in this manuscript will be instructive for designing an UEM setup and could thus push the further development of UEM.

Disparities in mental health service use of racial and ethnic minority elderly adults.

OBJECTIVES: To apply the Institute of Medicine definition of healthcare disparities, to measure disparities in different aspects of episodes of mental health care and to identify disparities in types of mental health services used. DESIGN: Four 2-year longitudinal datasets from Panels 9 to 13 (2004-2009) of the Medical Expenditure Panel Surveys were combined. SETTING: Large-scale surveys of families and individuals and their medical providers across the United States. PARTICIPANTS: One thousand six hundred fifty-eight participants (981 white, 303 black, and 374 Latino) aged 60 and older with probable mental healthcare needs. MEASUREMENTS: Mental healthcare need was defined as a Kessler-6 Scale score >12 and a Patient Health Questionnaire-2 score >2. Five aspects of mental healthcare episodes were analyzed: treatment initiation, adequacy of care, duration of care, number of visits, and expenditures. Whether episodes of care included only prescription drug fills, only outpatient visits, or both was assessed. RESULTS: Treatment initiation and adequacy were lower for blacks and Latinos than whites. Latinos experienced episodes of longer duration, more visits, and higher expenditures. Blacks and Latinos had significantly lower rates of episodes that consisted of only medication refills. Blacks had significantly greater rates of episodes with only outpatient care visits. Latinos had significantly higher rates of medication plus outpatient visits. CONCLUSION: Low mental health treatment initiation and poor adequacy suggest the need for culturally appropriate interventions to engage older blacks and Latinos in mental health care. The surprising findings in blacks (higher rates of outpatient care visits) and Latinos (higher rates of medication plus outpatient visits) highlight the complexities of the older adult population and suggest new avenues for disparities research.

Measuring performance in skin cancer practice: the SCARD initiative.

BACKGROUND: The Skin Cancer College of Australia and New Zealand (SCCANZ) has developed a unique project named SCARD - the Skin Cancer Audit and Research Database. Designed initially as a self-audit tool for primary care skin cancer practitioners, SCARD acts as a tracking tool to enhance practice safety, and it also creates practice performance reports. Pooling of de-identified data enables participating practitioners to confidentially compare their own practice to that of their peers. Additionally, this creates a large database with significant research potential, as SCARD records for every lesion de-identified practitioner and patient data, and extensive details of location, provisional and histological diagnosis, and the procedure(s) performed in its treatment. METHODS: Preliminary data collected in the database have been presented in this study. RESULTS: An initial pool of data from 177 practitioners contains 77,553 specimens from 41,006 individual patients. CONCLUSIONS: The data presented are being analyzed for further studies, and additional data continues to be collected from this ongoing project. SCARD is a useful tool at practice level, and substantial uptake by Australian primary care skin cancer practitioners has provided a unique opportunity for research into skin cancer and its management. SCCANZ, a professional college of predominantly primary care medical practitioners, with a commitment to the management of skin cancer in Australia and New Zealand, has formed a partnership with the School of Medicine at the University of Queensland to ensure that these data are managed and analyzed appropriately.

Social determinants of mental health treatment among Haitian, African American, and White youth in community health centers.

We examine adequate mental health treatment, emergency room (ER) use, and early treatment dropout for Haitian, African American and White youth with a psychiatric diagnosis treated in community health centers in the United States. We present associations with possible socioeconomic determinants of care. Adequate treatment was less likely among Haitian youth from areas with greater poverty and among all youth from areas with more female-headed households. Medicaid-insured youth had more ER visits, especially African Americans. The relative impact of poverty on adequate care was higher for Haitians than Whites, and the relative impact of Medicaid coverage on ER use was higher for African Americans than for Whites. Early dropout was more likely among youth who were uninsured or from areas with more female-headed households. Socioeconomic factors and insurance status were significant determinants of care. Haitians living in poverty in the U.S. may face barriers to mental health services relative to other racial/ethnic groups.

Improving the energy spread and brightness of thermal-field (Schottky) emitters with PHAST--PHoto Assisted Schottky Tip.

Using a relatively simple model of photoemission we derive an expression for the reduced on axis brightness of a thermal-photofield emitter. We then show that it is theoretically possible to reduce the energy spread of a Schottky (thermal field) emitter whilst increasing the reduced brightness. This can be achieved by the illumination of the tip with a high intensity laser light. We call the source PHAST-PHoto Assisted Schottky Tip. We find that due to the strong E-fields applied PHAST may operate at photon energies below the (Schottky reduced) work function. Thus removing the need for UV lasers, we will show that it is in fact preferable to work in the red, or in the green. The necessary laser intensities probably limit the application to pulsed operation.