De novo assembly and annotation of the Patagonian toothfish (Dissostichus eleginoides) genome.

BACKGROUND: Patagonian toothfish (Dissostichus eleginoides) is an economically and ecologically important fish species in the family Nototheniidae. Juveniles occupy progressively deeper waters as they mature and grow, and adults have been caught as deep as 2500 m, living on or in just above the southern shelves and slopes around the sub-Antarctic islands of the Southern Ocean. As apex predators, they are a key part of the food web, feeding on a variety of prey, including krill, squid, and other fish. Despite its importance, genomic sequence data, which could be used for more accurate dating of the divergence between Patagonian and Antarctic toothfish, or establish whether it shares adaptations to temperature with fish living in more polar or equatorial climes, has so far been limited. RESULTS: A high-quality D. eleginoides genome was generated using a combination of Illumina, PacBio and Omni-C sequencing technologies. To aid the genome annotation, the transcriptome derived from a variety of toothfish tissues was also generated using both short and long read sequencing methods. The final genome assembly was 797.8 Mb with a N50 scaffold length of 3.5 Mb. Approximately 31.7% of the genome consisted of repetitive elements. A total of 35,543 putative protein-coding regions were identified, of which 50% have been functionally annotated. Transcriptomics analysis showed that approximately 64% of the predicted genes (22,617 genes) were found to be expressed in the tissues sampled. Comparative genomics analysis revealed that the anti-freeze glycoprotein (AFGP) locus of D. eleginoides does not contain any AFGP proteins compared to the same locus in the Antarctic toothfish (Dissostichus mawsoni). This is in agreement with previously published results looking at hybridization signals and confirms that Patagonian toothfish do not possess AFGP coding sequences in their genome. CONCLUSIONS: We have assembled and annotated the Patagonian toothfish genome, which will provide a valuable genetic resource for ecological and evolutionary studies on this and other closely related species.

Socrates: A Novel N-Ethyl-N-nitrosourea-Induced Mouse Mutant with Audiogenic Epilepsy.

Epilepsy is one of the common neurological diseases that affects not only adults but also infants and children. Because epilepsy has been studied for a long time, there are several pharmacologically effective anticonvulsants, which, however, are not suitable as therapy for all patients. The genesis of epilepsy has been extensively investigated in terms of its occurrence after injury and as a concomitant disease with various brain diseases, such as tumors, ischemic events, etc. However, in the last decades, there are multiple reports that both genetic and epigenetic factors play an important role in epileptogenesis. Therefore, there is a need for further identification of genes and loci that can be associated with higher susceptibility to epileptic seizures. Use of mouse knockout models of epileptogenesis is very informative, but it has its limitations. One of them is due to the fact that complete deletion of a gene is not, in many cases, similar to human epilepsy-associated syndromes. Another approach to generating mouse models of epilepsy is N-Ethyl-N-nitrosourea (ENU)-directed mutagenesis. Recently, using this approach, we generated a novel mouse strain, soc (socrates, formerly s8-3), with epileptiform activity. Using molecular biology methods, calcium neuroimaging, and immunocytochemistry, we were able to characterize the strain. Neurons isolated from soc mutant brains retain the ability to differentiate in vitro and form a network. However, soc mutant neurons are characterized by increased spontaneous excitation activity. They also demonstrate a high degree of Ca2+ activity compared to WT neurons. Additionally, they show increased expression of NMDA receptors, decreased expression of the Ca2+-conducting GluA2 subunit of AMPA receptors, suppressed expression of phosphoinositol 3-kinase, and BK channels of the cytoplasmic membrane involved in protection against epileptogenesis. During embryonic and postnatal development, the expression of several genes encoding ion channels is downregulated in vivo, as well. Our data indicate that soc mutation causes a disruption of the excitation-inhibition balance in the brain, and it can serve as a mouse model of epilepsy.

Srsf1 and Elavl1 act antagonistically on neuronal fate choice in the developing neocortex by controlling TrkC receptor isoform expression.

The seat of higher-order cognitive abilities in mammals, the neocortex, is a complex structure, organized in several layers. The different subtypes of principal neurons are distributed in precise ratios and at specific positions in these layers and are generated by the same neural progenitor cells (NPCs), steered by a spatially and temporally specified combination of molecular cues that are incompletely understood. Recently, we discovered that an alternatively spliced isoform of the TrkC receptor lacking the kinase domain, TrkC-T1, is a determinant of the corticofugal projection neuron (CFuPN) fate. Here, we show that the finely tuned balance between TrkC-T1 and the better known, kinase domain-containing isoform, TrkC-TK+, is cell type-specific in the developing cortex and established through the antagonistic actions of two RNA-binding proteins, Srsf1 and Elavl1. Moreover, our data show that Srsf1 promotes the CFuPN fate and Elavl1 promotes the callosal projection neuron (CPN) fate in vivo via regulating the distinct ratios of TrkC-T1 to TrkC-TK+. Taken together, we connect spatio-temporal expression of Srsf1 and Elavl1 in the developing neocortex with the regulation of TrkC alternative splicing and transcript stability and neuronal fate choice, thus adding to the mechanistic and functional understanding of alternative splicing in vivo.

Exploring the association of Brownfield remediation status with socioeconomic conditions in Wayne County, MI.

Urban neighborhoods with locations of environmental contamination, known as brownfields, impact entire neighborhoods, but corrective environmental remedial action on brownfields is often tracked on an individual property basis, neglecting the larger neighborhood-level impact. This study addresses this impact by examining spatial differences between brownfields with unmitigated environmental concerns (open site) and sites that are considered fully mitigated or closed in urban neighborhoods (closed site) on the US census tract scale in Wayne County, MI. Michigan's Department of Environment, Great Lakes, and Energy's leaking underground storage tank (LUST) database provided brownfield information for Wayne County. Local indicators of spatial association (LISA) produced maps of spatial clustering and outliers. A McNemar's test demonstrated significant discordances in LISA categories between LUST open and closed sites (p < 0.001). Geographically weighted regressions (GWR) evaluated the association between open and closed site spatial density (open-closed) with socioeconomic variables (population density, proportion of White or Black residents, proportion of college educated populations, the percentage of owner-occupied units, vacant units, rented units, and median household value). Final multivariate GWR showed that population density, being Black, college education, vacant units, and renter occupied units were significantly associated (p < 0.05) with open-closed, and that those associations varied across Wayne County. Increases in Black population was associated with increased open-closed. Increases in vacant units, renter-occupied units, and college education were associated with decreased open-closed. These results provide input for environmental justice research to identify inequalities and discover the distribution of environmental hazards among urban neighborhoods.

A population of ultraviolet-dim protoclusters detected in absorption.

Galaxy protoclusters, which will eventually grow into the massive clusters we see in the local Universe, are usually traced by locating overdensities of galaxies1. Large spectroscopic surveys of distant galaxies now exist, but their sensitivity depends mainly on a galaxy's star-formation activity and dust content rather than its mass. Tracers of massive protoclusters that do not rely on their galaxy constituents are therefore needed. Here we report observations of Lyman-α absorption in the spectra of a dense grid of background galaxies2,3, which we use to locate a substantial number of candidate protoclusters at redshifts 2.2 to 2.8 through their intergalactic gas. We find that the structures producing the most absorption, most of which were previously unknown, contain surprisingly few galaxies compared with the dark-matter content of their analogues in cosmological simulations4,5. Nearly all of the structures are expected to be protoclusters, and we infer that half of their expected galaxy members are missing from our survey because they are unusually dim at rest-frame ultraviolet wavelengths. We attribute this to an unexpectedly strong and early influence of the protocluster environment6,7 on the evolution of these galaxies that reduced their star formation or increased their dust content.

Acute association between heatwaves and stillbirth in six US states.

BACKGROUND: Heatwaves are becoming more frequent and may acutely increase the risk of stillbirth, a rare and severe pregnancy outcome. OBJECTIVES: Examine the association between multiple heatwave metrics and stillbirth in six U.S. states. METHODS: Data were collected from fetal death and birth records in California (1996-2017), Florida (1991-2017), Georgia (1994-2017), Kansas (1991-2017), New Jersey (1991-2015), and Oregon (1991-2017). Cases were matched to controls 1:4 based on maternal race/ethnicity, maternal education, and county, and exposure windows were aligned (gestational week prior to stillbirth). County-level temperature data were obtained from Daymet and linked to cases and controls by residential county and the exposure window. Five heatwave metrics (1 categorical, 3 dichotomous, 1 continuous) were created using different combinations of the duration and intensity of hot days (mean daily temperature exceeding the county-specific 97.5th percentile) during the exposure window, as well as a continuous measure of mean temperature during the exposure window modeled using natural splines to allow for nonlinear associations. State-specific odds ratios (ORs) and 95% confidence intervals (CI) were estimated using conditional logistic regression models. State-specific results were pooled using a fixed-effects meta-analysis. RESULTS: In our data set of 140,428 stillbirths (553,928 live birth controls), three of the five heatwave metrics examined were not associated with stillbirth. However, four consecutive hot days during the previous week was associated with a 3% increase in stillbirth risk (CI: 1.01, 1.06), and a 1 °C average increase over the threshold was associated with a 10% increase in stillbirth risk (CI: 1.04, 1.17). In continuous temperature analyses, there was a slight increased risk of stillbirth associated with extremely hot temperatures (≥ 35 °C). DISCUSSION: Most heat wave definitions examined were not associated with acute changes in stillbirth risk; however, the most extreme heatwave durations and temperatures were associated with a modest increase in stillbirth risk.

How Did the COVID-19 Pandemic Affect Trends in Facial Trauma?

Study Design: Retrospective cohort analysis. Objective: To examine the impact the COVID-19 pandemic and its accompanying societal measures had on the incidence, characteristics, and management of maxillofacial traumatic injuries. Methods: This cohort analysis compared facial trauma injuries presenting to the highest-volume Level I Trauma Center in New Jersey, USA from January 1 to July 31 in 2020 and 2019. Differences in demographics, mechanisms, and interventions were compared between the pandemic period (March 16-July 31, 2020) and the equivalent pre-pandemic date period in 2019 using X 2, Fishers Exact, and Mann-Whitney U testing. Results: In total, 616 subjects were included. The daily incidence of facial trauma consults during the 2020 pandemic (1.81 ± 1.1) decreased compared to 2019 (2.15 ± 1.3) (p = 0.042). During the outbreak, there was an increase in the proportion of subjects with positive urine drug screens (21.5% vs. 12.2%; p = 0.011) and injuries related to domestic violence (10.2% vs. 4.5%; p = 0.023). Patients were 30% less likely to be transferred from local hospitals (RR, 0.70 [0.53-0.93]; p = 0.014). Although subjects had a 25% increased risk of presenting with injuries deemed procedural (RR, 1.25 [95% CI, 1.05-1.56]; p = 0.048), a greater proportion were discharged with operative procedures scheduled as outpatients (16.0% vs. 4.9%; p = 0.005). Conclusions: The COVID-19 pandemic has impacted both the epidemiology and management of maxillofacial traumatic injuries, perhaps secondary to modifications in personal and community behaviors or the effects on healthcare systems in our region.

Pathogenic Mechanisms of Cytosolic and Membrane-Enriched α-Synuclein Converge on Fatty Acid Homeostasis.

α-Synuclein (αS) plays a key role in Parkinson's disease. Although Parkinson's disease is typically "sporadic," inherited αS missense mutations provide crucial insights into molecular mechanisms. Here, we examine two clinical mutants, E46K and G51D, which are both in the conserved N-terminus that mediates transient αS-membrane interactions. However, E46K increases and G51D decreases αS-membrane interactions. Previously, we amplified E46K via the 11-residue repeat motifs, creating "3K" (E35K+E46K+E61K). Here, we engineered these motifs to amplify G51D (V40D+G51D+V66D = "3D") and systematically compared E46K/3K versus G51D/3D. We found that G51D increased cytosolic αS in neural cells and 3D aggravates this. G51D, and 3D even more, reduced αS multimer-to-monomer (αS60:αS14) ratio. Both amplified variants caused cellular stress in rat primary neurons and reduced growth in human neuroblastoma cells. Importantly, both 3K- and 3D-induced stress was ameliorated by pharmacologically inhibiting stearoyl-CoA desaturase or by conditioning the cells in palmitic (16:0) or myristic (14:0) acid. SCD inhibition lowered lipid-droplet accumulation in both 3D- and 3K-expressing cells and benefitted G51D by normalizing multimer:monomer ratio, as reported previously for E46K. Our findings suggest that, despite divergent cytosol/membrane partitioning, both G51D and E46K neurotoxicity can be prevented by decreasing fatty-acid unsaturation as a common therapeutic approach.SIGNIFICANCE STATEMENT α-Synuclein (αS) dyshomeostasis is linked to Parkinson's disease. Here we focus on two contrasting familial-Parkinson's disease αS mutants, E46K and G51D, that alter αS membrane association in opposite directions (E46K increases, G51D decreases it). Taking advantage of αS repeat structure, here we designed αS "3D," an amplified G51D variant (V40D+G51D+V66D). αS 3D further enhanced G51D's cytosolic enrichment. Systematic comparison of G51D/3D with membrane-enriched E46K/its amplified variant 3K revealed that both can elicit stress in human neural cells and primary rodent neurons. This toxicity can be ameliorated by inhibiting stearoyl-CoA desaturase or by saturated fatty acid conditioning. Thus, despite divergent membrane binding, both G51D and E46K αS dyshomeostasis are mitigated by altering fatty acid saturation as a shared target.

Estimating Heat-Related Exposures and Urban Heat Island Impacts: A Case Study for the 2012 Chicago Heatwave.

Accelerated urbanization increases both the frequency and intensity of heatwaves (HW) and urban heat islands (UHIs). An extreme HW event occurred in 2012 summer that caused temperatures of more than 40°C in Chicago, Illinois, USA, which is a highly urbanized city impacted by UHIs. In this study, multiple numerical models, including the High Resolution Land Data Assimilation System (HRLDAS) and Weather Research and Forecasting (WRF) model, were used to simulate the HW and UHI, and their performance was evaluated. In addition, sensitivity testing of three different WRF configurations was done to determine the impact of increasing model complexity in simulating urban meteorology. Model performances were evaluated based on the statistical performance metrics, the application of a multi-layer urban canopy model (MLUCM) helps WRF to provide the best performance in this study. HW caused rural temperatures to increase by ∼4°C, whereas urban Chicago had lower magnitude increases from the HW (∼2-3°C increases). Nighttime UHI intensity (UHII) ranged from 1.44 to 2.83°C during the study period. Spatiotemporal temperature fields were used to estimate the potential heat-related exposure and to quantify the Excessive Heat Factor (EHF). The EHF during the HW episode provides a risk map indicating that while urban Chicago had higher heat-related stress during this event, the rural area also had high risk, especially during nighttime in central Illinois. This study provides a reliable method to estimate spatiotemporal exposures for future studies of heat-related health impacts.

Targeting Cbx3/HP1γ Induces LEF-1 and IL-21R to Promote Tumor-Infiltrating CD8 T-Cell Persistence.

Immune checkpoint blockade (ICB) relieves CD8+ T-cell exhaustion in most mutated tumors, and TCF-1 is implicated in converting progenitor exhausted cells to functional effector cells. However, identifying mechanisms that can prevent functional senescence and potentiate CD8+ T-cell persistence for ICB non-responsive and resistant tumors remains elusive. We demonstrate that targeting Cbx3/HP1γ in CD8+ T cells augments transcription initiation and chromatin remodeling leading to increased transcriptional activity at Lef1 and Il21r. LEF-1 and IL-21R are necessary for Cbx3/HP1γ-deficient CD8+ effector T cells to persist and control ovarian cancer, melanoma, and neuroblastoma in preclinical models. The enhanced persistence of Cbx3/HP1γ-deficient CD8+ T cells facilitates remodeling of the tumor chemokine/receptor landscape ensuring their optimal invasion at the expense of CD4+ Tregs. Thus, CD8+ T cells heightened effector function consequent to Cbx3/HP1γ deficiency may be distinct from functional reactivation by ICB, implicating Cbx3/HP1γ as a viable cancer T-cell-based therapy target for ICB resistant, non-responsive solid tumors.

Angiolymphoid Hyperplasia with Eosinophilia: Many Syllables, Many Unanswered Questions.

Angiolymphoid hyperplasia with eosinophilia (ALHE) is an uncommon, benign inflammatory vasoproliferation. The literature is divided regarding whether it embodies a vascular neoplasm or a reactive process secondary to various stimuli. ALHE presents as solitary or clustered papules or nodules primarily on the head and neck, especially on or around the auricle. Histologically, ALHE is characterized by a proliferation of blood vessels lined by plump epithelioid endothelial cells and a prominent perivascular infiltrate rich in lymphocytes and eosinophils. ALHE follows a benign clinical course, yet treatment is challenging because of its high recurrence rate. We present the case of a 37-year-old Filipino man with lesions located on the central face. Kimura disease was considered due to his age, sex, and ethnicity; however, his clinical features-specifically, the presence of discrete papules and lack of lymphadenopathy-and his histological findings were consistent with ALHE. He reported trauma prior to the onset of the lesions, suggesting a reactive etiology.

Excess membrane binding of monomeric alpha-, beta- and gamma-synuclein is invariably associated with inclusion formation and toxicity.

α-Synuclein (αS) has been well-documented to play a role in human synucleinopathies such as Parkinson's disease (PD) and dementia with Lewy bodies (DLB). First, the lesions found in PD/DLB brains-Lewy bodies and Lewy neurites-are rich in aggregated αS. Second, genetic evidence links missense mutations and increased αS expression to familial forms of PD/DLB. Third, toxicity and cellular stress can be caused by αS under certain experimental conditions. In contrast, the homologs ß-synuclein (ßS) and γ-synuclein (γS) are not typically found in Lewy bodies/neurites, have not been clearly linked to brain diseases and have been largely non-toxic in experimental settings. In αS, the so-called non-amyloid-ß component of plaques (NAC) domain, constituting amino acids 61-95, has been identified to be critical for aggregation in vitro. This domain is partially absent in ßS and only incompletely conserved in γS, which could explain why both homologs do not cause disease. However, αS in vitro aggregation and cellular toxicity have not been firmly linked experimentally, and it has been proposed that excess αS membrane binding is sufficient to induce neurotoxicity. Indeed, recent characterizations of Lewy bodies have highlighted the accumulation of lipids and membranous organelles, raising the possibility that ßS and γS could also become neurotoxic if they were more prone to membrane/lipid binding. Here, we increased ßS and γS membrane affinity by strategic point mutations and demonstrate that these proteins behave like membrane-associated monomers, are cytotoxic and form round cytoplasmic inclusions that can be prevented by inhibiting stearoyl-CoA desaturase.

Time-series analysis of daily ambient temperature and emergency department visits in five US cities with a comparison of exposure metrics derived from 1-km meteorology products.

BACKGROUND: Ambient temperature observations from single monitoring stations (usually located at the major international airport serving a city) are routinely used to estimate heat exposures in epidemiologic studies. This method of exposure assessment does not account for potential spatial variability in ambient temperature. In environmental health research, there is increasing interest in utilizing spatially-resolved exposure estimates to minimize exposure measurement error. METHODS: We conducted time-series analyses to investigate short-term associations between daily temperature metrics and emergency department (ED) visits for well-established heat-related morbidities in five US cities that represent different climatic regions: Atlanta, Los Angeles, Phoenix, Salt Lake City, and San Francisco. In addition to airport monitoring stations, we derived several exposure estimates for each city using a national meteorology data product (Daymet) available at 1 km spatial resolution. RESULTS: Across cities, we found positive associations between same-day temperature (maximum or minimum) and ED visits for heat-sensitive outcomes, including acute renal injury and fluid and electrolyte imbalance. We also found that exposure assessment methods accounting for spatial variability in temperature and at-risk population size often resulted in stronger relative risk estimates compared to the use of observations at airports. This pattern was most apparent when examining daily minimum temperature and in cities where the major airport is located further away from the urban center. CONCLUSION: Epidemiologic studies based on single monitoring stations may underestimate the effect of temperature on morbidity when the station is less representative of the exposure of the at-risk population.

Using logic regression to characterize extreme heat exposures and their health associations: a time-series study of emergency department visits in Atlanta.

BACKGROUND: Short-term associations between extreme heat events and adverse health outcomes are well-established in epidemiologic studies. However, the use of different exposure definitions across studies has limited our understanding of extreme heat characteristics that are most important for specific health outcomes or subpopulations. METHODS: Logic regression is a statistical learning method for constructing decision trees based on Boolean combinations of binary predictors. We describe how logic regression can be utilized as a data-driven approach to identify extreme heat exposure definitions using health outcome data. We evaluated the performance of the proposed algorithm in a simulation study, as well as in a 20-year time-series analysis of extreme heat and emergency department visits for 12 outcomes in the Atlanta metropolitan area. RESULTS: For the Atlanta case study, our novel application of logic regression identified extreme heat exposure definitions that were associated with several heat-sensitive disease outcomes (e.g., fluid and electrolyte imbalance, renal diseases, ischemic stroke, and hypertension). Exposures were often characterized by extreme apparent minimum temperature or maximum temperature over multiple days. The simulation study also demonstrated that logic regression can successfully identify exposures of different lags and duration structures when statistical power is sufficient. CONCLUSION: Logic regression is a useful tool for identifying important characteristics of extreme heat exposures for adverse health outcomes, which may help improve future heat warning systems and response plans.

Acute associations between heatwaves and preterm and early-term birth in 50 US metropolitan areas: a matched case-control study.

BACKGROUND: The effect of heatwaves on adverse birth outcomes is not well understood and may vary by how heatwaves are defined. The study aims to examine acute associations between various heatwave definitions and preterm and early-term birth. METHODS: Using national vital records from 50 metropolitan statistical areas (MSAs) between 1982 and 1988, singleton preterm (< 37 weeks) and early-term births (37-38 weeks) were matched (1:1) to controls who completed at least 37 weeks or 39 weeks of gestation, respectively. Matching variables were MSA, maternal race, and maternal education. Sixty heatwave definitions including binary indicators for exposure to sustained heat, number of high heat days, and measures of heat intensity (the average degrees over the threshold in the past 7 days) based on the 97.5th percentile of MSA-specific temperature metrics, or the 85th percentile of positive excessive heat factor (EHF) were created. Odds ratios (OR) for heatwave exposures in the week preceding birth (or corresponding gestational week for controls) were estimated using conditional logistic regression adjusting for maternal age, marital status, and seasonality. Effect modification by maternal education, age, race/ethnicity, child sex, and region was assessed. RESULTS: There were 615,329 preterm and 1,005,576 early-term case-control pairs in the analyses. For most definitions, exposure to heatwaves in the week before delivery was consistently associated with increased odds of early-term birth. Exposure to more high heat days and more degrees above the threshold yielded higher magnitude ORs. For exposure to 3 or more days over the 97.5th percentile of mean temperature in the past week compared to zero days, the OR was 1.027 for early-term birth (95%CI: 1.014, 1.039). Although we generally found null associations when assessing various heatwave definitions and preterm birth, ORs for both preterm and early-term birth were greater in magnitude among Hispanic and non-Hispanic black mothers. CONCLUSION: Although associations varied across metrics and heatwave definitions, heatwaves were more consistently associated with early-term birth than with preterm birth. This study's findings may have implications for prevention programs targeting vulnerable subgroups as climate change progresses.

psiCLIP reveals dynamic RNA binding by DEAH-box helicases before and after exon ligation.

RNA helicases remodel the spliceosome to enable pre-mRNA splicing, but their binding and mechanism of action remain poorly understood. To define helicase-RNA contacts in specific spliceosomal states, we develop purified spliceosome iCLIP (psiCLIP), which reveals dynamic helicase-RNA contacts during splicing catalysis. The helicase Prp16 binds along the entire available single-stranded RNA region between the branchpoint and 3'-splice site, while Prp22 binds diffusely downstream of the branchpoint before exon ligation, but then switches to more narrow binding in the downstream exon after exon ligation, arguing against a mechanism of processive translocation. Depletion of the exon-ligation factor Prp18 destabilizes Prp22 binding to the pre-mRNA, suggesting that proofreading by Prp22 may sense the stability of the spliceosome during exon ligation. Thus, psiCLIP complements structural studies by providing key insights into the binding and proofreading activity of spliceosomal RNA helicases.

Crowded organelles, lipid accumulation, and abnormal membrane tubulation in cellular models of enhanced α-synuclein membrane interaction.

Previous work from our group showed that certain engineered missense mutations to the α-synuclein (αS) KTKEGV repeat motifs abrogate the protein's ability to form native multimers. The resultant excess monomers accumulate in lipid-membrane-rich inclusions associated with neurotoxicity exceeding that of natural familial Parkinson's disease mutants such as E46K. We presented an initial characterization of the lipid-rich inclusions and found similarities to the αS- and vesicle-rich inclusions that form in baker's yeast when αS is expressed. We also discussed, with some caution, a possible role of membrane-rich inclusions as precursors to filamentous Lewy bodies, the widely accepted hallmark pathology of Parkinson's disease and other synucleinopathies. In the meantime, advances in the microscopic characterization of Lewy bodies have highlighted the presence of crowded organelles and lipid membranes in addition to αS accumulation. This prompted us to revisit the αS inclusions caused by our repeat motif variants in neuroblastoma cells. In addition to our previous characterization, we found that these inclusions can often be seen by brightfield microscopy, overlap with endogenous vesicle markers in immunofluorescence experiments, stain positive for lipid dyes, and can be found to be closely associated with mitochondria. We also observed abnormal tubulation of membranes, which was subtle in inducible lines and pronounced in cells that transiently expressed high amounts of the highly disruptive KTKEGV motif mutant "KLKEGV". Membrane tubulation had been reported before as an αS activity in reductionist systems. Our in-cellulo demonstration now suggests that this mechanism could possibly be a relevant aspect of aberrant αS behavior in cells.

Infective Endocarditis Observed With Cryoglobulinemic Vasculitis.

Cryoglobulinemic vasculitis is most commonly observed in the setting of hepatitis C infection. However, many other etiologies have been identified as well. We herein report a case of cryoglobulinemic vasculitis in a 56-year-old male secondary to infective endocarditis. The patient presented with a five-week history of a painful, purpuric rash and was found to have positive blood cultures, elevated cryoglobulins, and vasculitis on histology. He recovered after treatment with intravenous antibiotics and steroids. Although rarely identified as a cause of cryoglobulinemic vasculitis, infective endocarditis must be considered on the differential diagnosis as a delay in treatment can worsen the prognosis for the patient.