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Abstract Astrocytes contribute to brain inflammation in neurological disorders but the molecular mechanisms controlling astrocyte reactivity and their relationship to neuroinflammatory endpoints are complex and poorly understood. In this study, we assessed the role of the calcium channel, Orai1, for astrocyte reactivity and inflammation-evoked depression behaviors in mice. Transcriptomics and metabolomics analysis indicated that deletion of Orai1 in astrocytes downregulates genes in inflammation and immunity, metabolism, and cell cycle pathways, and reduces cellular metabolites and ATP production. Systemic inflammation by peripheral lipopolysaccharide (LPS) increases hippocampal inflammatory markers in WT but not in astrocyte Orai1 knockout mice. Loss of Orai1 also blunts inflammation-induced astrocyte Ca2+ signaling and inhibitory neurotransmission in the hippocampus. In line with these cellular changes, Orai1 knockout mice showed amelioration of LPS-evoked depression-like behaviors including anhedonia and helplessness. These findings identify Orai1 as an important signaling hub controlling astrocyte reactivity and astrocyte-mediated brain inflammation that is commonly observed in many neurological disorders.

Laser–plasma interactions (LPIs) are an emerging source of a range of energetic radiation. LPI experiments drive ultra-short (< ps) and brilliant sources of X-rays from keV to MeV energies. Designing detectors to maximise the sensitivity and resolution achievable with these sources is paramount to optimising laser-driven accelerators. In this article, we explore the key parameters associated with laser-driven X-ray sources and the detector systems required to characterise them. We present a concise approach to modelling the sensitivity and resolution for indirect detector systems factoring in both the optical collection and the X-ray attenuation within the scintillator.

Patients with chronic lung disease, obesity, and other co-morbid conditions are at increased risk of severe illness and death when infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Hypercapnia, the elevation of CO2 in blood and tissue, commonly occurs in patients with severe acute and chronic lung disease, including those with pulmonary infections, and is also associated with high mortality risk. We previously reported that hypercapnia increases viral replication and mortality of influenza A virus infection in mice. We have also shown that culture in elevated CO2 upregulates expression of cholesterol synthesis genes in primary human bronchial epithelial cells. Interestingly, factors that increase the cholesterol content of lipid rafts and lipid droplets, platforms for viral entry and assembly, enhance SARS-CoV-2 infection. In the current study, we investigated the effects of hypercapnia on ACE2 expression and entry of SARS-CoV-2 pseudovirus (p-SARS-CoV-2) into airway epithelial cells. We found that hypercapnia increased ACE2 expression and p-SARS-CoV-2 uptake by airway epithelium in mice, and in cultured VERO and human bronchial epithelial cells. Hypercapnia also increased total cellular and lipid raft-associated cholesterol in epithelial cells. Moreover, reducing cholesterol synthesis with inhibitors of sterol regulatory element binding protein 2 (SREBP2) or statins, and depletion of cellular cholesterol, each blocked the hypercapnia-induced increases in ACE2 expression and p-SARS-CoV-2 entry into epithelial cells. Cigarette smoke extract (CSE) also increased ACE2 expression, p-SARS-CoV-2 entry and cholesterol accumulation in epithelial cells, an effect not additive to that of hypercapnia, but also inhibited by statins. These findings reveal a mechanism that may account, in part, for poor clinical outcomes of SARS-CoV-2 infection in patients with advanced lung disease and hypercapnia, and in those who smoke cigarettes. Further, our results suggest the possibility that cholesterol-lowering therapies may be of particular benefit in patients with hypercapnia when exposed to or infected with SARS-CoV-2.

OBJECTIVES:. To analyze the temporal trend in enrollment rates in a COVID-19 platform trial during the first three waves of the pandemic in the United States. DESIGN:. Secondary analysis of data from the I-SPY COVID randomized controlled trial (RCT). SETTING:. Thirty-one hospitals throughout the United States. PATIENTS:. Patients who were approached, either directly or via a legally authorized representative, for consent and enrollment into the I-SPY COVID RCT. INTERVENTIONS:. None. MEASUREMENTS AND MAIN RESULTS:. Among 1,338 patients approached for the I-SPY COVID trial from July 30, 2020, to February 17, 2022, the number of patients who enrolled (n = 1,063) versus declined participation (n = 275) was used to calculate monthly enrollment rates. Overall, demographic and baseline clinical characteristics were similar between those who enrolled versus declined. Enrollment rates fluctuated over the course of the COVID-19 pandemic, but there were no significant trends over time (Mann-Kendall test, p = 0.21). Enrollment rates were also comparable between vaccinated and unvaccinated patients. In multivariable logistic regression analysis, age, sex, region of residence, COVID-19 severity of illness, and vaccination status were not significantly associated with the decision to decline consent. CONCLUSIONS:. In this secondary analysis of the I-SPY COVID clinical trial, there was no significant association between the enrollment rate and time period or vaccination status among all eligible patients approached for clinical trial participation. Additional studies are needed to better understand whether the COVID-19 pandemic has altered clinical trial participation and to develop strategies for encouraging participation in future COVID-19 and critical care clinical trials.

The cell-surface targeting of neo-synthesized G protein-coupled receptors (GPCRs) involves the recruitment of receptors into COPII vesicles budding at endoplasmic reticulum exit sites (ERESs). This process is regulated for some GPCRs by escort proteins, which facilitate their export, or by gatekeepers that retain the receptors in the ER. PRAF2, an ER-resident four trans- membrane domain protein with cytoplasmic extremities, operates as a gatekeeper for the GB1 protomer of the heterodimeric GABAB receptor, interacting with a tandem di-leucine/RXR retention motif in the carboxyterminal tail of GB1. PRAF2 was also reported to interact in a two-hybrid screen with a peptide corresponding to the carboxyterminal tail of the chemokine receptor CCR5 despite the absence of RXR motifs in its sequence. Using a bioluminescence resonance energy transfer (BRET)-based subcellular localization system, we found that PRAF2 inhibits, in a concentration-dependent manner, the plasma membrane export of CCR5. BRET-based proximity assays and Co-IP experiments demonstrated that PRAF2/CCR5 interaction does not require the presence of a receptor carboxyterminal tail and involves instead the transmembrane domains of both proteins. The mutation of the potential di-leucine/RXR motif contained in the third intracellular loop of CCR5 does not affect PRAF2-mediated retention. It instead impairs the cell-surface export of CCR5 by inhibiting CCR5’s interaction with its private escort protein, CD4. PRAF2 and CD4 thus display opposite roles on the cell-surface export of CCR5, with PRAF2 inhibiting and CD4 promoting this process, likely operating at the level of CCR5 recruitment into COPII vesicles, which leave the ER.

Summary: Background: An urgent need exists to rapidly screen potential therapeutics for severe COVID-19 or other emerging pathogens associated with high morbidity and mortality. Methods: Using an adaptive platform design created to rapidly evaluate investigational agents, hospitalised patients with severe COVID-19 requiring ≥6 L/min oxygen were randomised to either a backbone regimen of dexamethasone and remdesivir alone (controls) or backbone plus one open-label investigational agent. Patients were enrolled to the arms described between July 30, 2020 and June 11, 2021 in 20 medical centres in the United States. The platform contained up to four potentially available investigational agents and controls available for randomisation during a single time-period. The two primary endpoints were time-to-recovery (

Abstract Aging in mammals leads to reduction in genes encoding the 45-subunit mitochondrial electron transport chain complex I. It has been hypothesized that normal aging and age-related diseases such as Parkinson’s disease are in part due to modest decrease in expression of mitochondrial complex I subunits. By contrast, diminishing expression of mitochondrial complex I genes in lower organisms increases lifespan. Furthermore, metformin, a putative complex I inhibitor, increases healthspan in mice and humans. In the present study, we investigated whether loss of one allele of Ndufs2, the catalytic subunit of mitochondrial complex I, impacts healthspan and lifespan in mice. Our results indicate that Ndufs2 hemizygous mice (Ndufs2 +/−) show no overt impairment in aging-related motor function, learning, tissue histology, organismal metabolism, or sensitivity to metformin in a C57BL6/J background. Despite a significant reduction of Ndufs2 mRNA, the mice do not demonstrate a significant decrease in complex I function. However, there are detectable transcriptomic changes in individual cell types and tissues due to loss of one allele of Ndufs2. Our data indicate that a 50% decline in mRNA of the core mitochondrial complex I subunit Ndufs2 is neither beneficial nor detrimental to healthspan.

Dental size variation in modern humans has been assessed from regional to worldwide scales, especially under microevolutionary and forensic contexts. Despite this, populations of mixed continental ancestry such as contemporary Latin Americans remain unexplored. In the present study we investigated a large Latin American sample from Colombia (N = 804) and obtained buccolingual and mesiodistal diameters and three indices for maxillary and mandibular teeth (except third molars). We evaluated the correlation between 28 dental measurements (and three indices) with age, sex and genomic ancestry (estimated using genome-wide SNP data). In addition, we explored correlation patterns between dental measurements and the biological affinities, based on these measurements, between two Latin American samples (Colombians and Mexicans) and three putative parental populations: Central and South Native Americans, western Europeans and western Africans through PCA and DFA. Our results indicate that Latin Americans have high dental size diversity, overlapping the variation exhibited by the parental populations. Several dental dimensions and indices have significant correlations with sex and age. Western Europeans presented closer biological affinities with Colombians, and the European genomic ancestry exhibited the highest correlations with tooth size. Correlations between tooth measurements reveal distinct dental modules, as well as a higher integration of postcanine dentition. The effects on dental size of age, sex and genomic ancestry is of relevance for forensic, biohistorical and microevolutionary studies in Latin Americans.

Sex assessment is a key part of forensic analysis to establish the identity of unknown deceased individuals. Previous studies have shown that canines are the most dimorphic teeth, but population-specific data are necessary for forensic methods. This study explores sex dimorphism in canine crown dimensions and morphology in a contemporary Croatian population. The material consisted of 302 dental casts (147 females, 155 males) of orthodontic patients and dental students (11–25 years). The distal accessory ridge (DAR) of the upper and lower canines was evaluated using the Arizona State University Dental Anthropology System. Mesiodistal (MD) and buccolingual (BL) crown dimensions were measured on 120 casts. Sex differences in MD and BL dimensions were significant (p < 0.05) for all the canines (upper and lower, left and right), while in DAR only for lower canines (p < 0.000001). When all variables were put into the model, backward stepwise discriminant function analysis isolated lower canine DAR and lower left canine MD as the two independent variables differentiating sex. Using these two variables, a discriminant function formula allowed for sex determination with an accuracy of 73.5%. This study shows that both canine crown morphology and dimensions are useful for sex determination, especially for lower canines. These methods can be applied to children, as lower canines erupt at about 9 years of age.

More than 12 years have passed since the seminal observation that meningococcus, a pathogen causing epidemic meningitis in humans, occasionally associated with infectious vasculitis and septic shock, can promote the translocation of β-arrestins to the cell surface beneath bacterial colonies. The cellular receptor used by the pathogen to induce signalling in host cells and allowing it to open endothelial cell junctions and reach meninges was unknown. The involvement of β-arrestins, which are scaffolding proteins regulating G protein coupled receptor signalling and function, incited us to specifically investigate this class of receptors. In this perspective article we will summarize the events leading to the discovery that the β2-adrenergic receptor is the receptor that initiates the signalling cascades induced by meningococcus in host cells. This receptor, however, cannot mediate cell infection on its own. It needs to be pre-associated with an “early” adhesion receptor, CD147, within a hetero-oligomeric complex, stabilized by the cytoskeletal protein α-actinin 4. It then required several years to understand how the pathogen actually activates the signalling receptor. Once bound to the N-terminal glycans of the β2-adrenergic receptor, meningococcus provides a mechanical stimulation that induces the biased activation of β-arrestin-mediated signalling pathways. This activating mechanical stimulus can be reproduced in the absence of any pathogen by applying equivalent forces on receptor glycans. Mechanical activation of the β2-adrenergic receptor might have a physiological role in signalling events promoted in the context of cell-to-cell interaction.

Basque population history has been examined through classic genetic markers, mtDNA, Y chromosome haplogroups, craniometrics, and recently dental morphology. Dental morphological data show Basques have a classic European dental pattern but fall as an outlier among European populations. Expanding on that work, Basque tooth size was examined to further evaluate the affinities of the Basque population. Mesiodistal and buccolingual maximum crown measurements were taken from medieval and post medieval skeletons from the Catedral de Santa María in Vitoria, Spain, along with living samples of modern Basques, Spanish, and Spanish Basques from dental students at the Universidad del País Vasco. A dental metric examination affirms the outlier status of Basques, as they exhibit smaller crown areas than neighboring populations. In biodistance analyses Basque populations group with linguistically and geographically distant populations. Even with gene flow from Spain, France, and North Africa, Basque individuals still demonstrate a unique pattern coincident with their ancient origins.

Objective of work: This paper provides an overview of crown and root morphology in Eskimo-Aleut populations of the American Arctic. For context, Eskimo-Aleut dental variation is compared to closely related American Indians and distantly related Europeans. Materials and methods: The characterization of dental trait frequency variation is based on observations made on approximately 10,000 dentitions scored by the late Christy G. Turner II and the author. Sixteen crown and five root traits were scored following the conventions outlined in the Arizona State University Dental Anthropology System. Results: Of the 21 dental traits considered, only three showed slight differences among Eskimo- Aleuts, American Indians, and Europeans (UM1 cusp 5, LM2 groove pattern, LM2 root number). For the remaining traits, there was typically a dramatic contrast between the two New World populations and Europeans. While generally similar, Eskimo-Aleuts and American Indians showed differences in UI1 winging, shoveling, and double shoveling, UM1 Carabelli trait, 2-cusped UM2, 3-rooted UM2, and especially 3-rooted LM1. Conclusion: The differences between the three groups are likely a product of genetic drift and founder effect although recent work on the EDAR V370A allele suggests some dental variables like shoveling and lower molar cusp number may indirectly reflect natural selection operating on other variables influenced by this allele.

Objective: Turner syndrome (TS) and Klinefelter syndrome (KS) represent the two most common X chromosome aneuploidies, each associated with systemic disruptions to growth and development. Effects of these conditions on tooth crown morphology are explored in a sample of Croatian individuals. Material and Methods: The sample included 57 TS, 37 KS and 88 control individuals. Dental crown morphology was scored on dental casts according to the Turner-Scott Dental Anthropology System. Results: Incisor shoveling and the hypocone were significantly different between TS individuals and both control and KS individuals. Individuals with TS exhibit lower grades of expression than either group. Furthermore, the number of lingual cusps on the mandibular premolars, the hypoconulid on the mandibular second molar, and cusp 7 on the mandibular first molar were significantly different, though pair-wise comparisons did not elucidate these differences. Tuberculum dentale, distal accessory ridge, and Carabelli’s trait were expressed similarly to the control. KS individuals were not significantly different from control individuals for any trait, though this may be related to sample size. Conclusions: Previous studies suggest the loss of an X chromosome has a reducing effect on dental crown morphology, which is confirmed in this research. TS individuals exhibit generally simpler dental morphology compared to the control sample, though some traits are expressed comparably to the control sample. The effects of KS are less clear. Though previous studies suggest that the presence of an extra X chromosome increases dental crown dimensions, there was no notable effect on crown morphology in this study.

The present study assesses the global distribution of marginal accessory tubercles of the maxillary premolars. This trait, despite constituting one of the variables standardized by Turner and colleagues (1991), has received little attention in morphological studies. Frequencies were calculated from data sheets collected by Christy G. Turner II for mesial, distal, and mesial + distal grades. Different geographic patterns were identified for both types of expression on the upper premolars. The patterned geographic distribution of these traits indicates their utility in biodistance investigations. In addition, the distinction between mesial and distal accessory tubercles specified by Scott and Irish (2017) is recommended, as these two traits exhibit different geographic patterns.

Objectives:. Determine the variation in outcomes and respiratory mechanics between the subjects who are intubated earlier versus later in their coronavirus disease 2019 course. Design:. Retrospective cohort study. Setting:. Northwestern Memorial Hospital ICUs. Patients:. All patients intubated for coronavirus disease 2019 between March 2020 and June 2020. Interventions:. Patients were stratified by time to intubation: 30 subjects were intubated 4–24 hours after presentation and 24 subjects were intubated 5–10 days after presentation. Baseline characteristics, hospitalization, ventilator mechanics, and outcomes were extracted and analyzed. Ten clinically available CT scans were manually reviewed to identify evidence of pulmonary vascular thrombosis and intussusceptive angiogenesis. Measurements and Main Results:. Median time from symptom onset to intubation was significantly different between the early and late intubation cohorts, with the latter being intubated later in the course of their illness (7.9 vs 11.8 d; p = 0.04). The early intubation cohort had a lower mortality rate than the late intubation cohort (6% vs 30%, p < 0.001) without significantly different respiratory mechanics at the time of intubation. The late intubation cohort was noted to have higher dead space ratio (0.40 vs 0.52; p = 0.03). On review of CT scans, the late intubation cohort also had more dilated peripheral segments on imaging (two segments vs five segments). Conclusions:. The question as to whether delaying intubation is beneficial or harmful for patients with coronavirus disease 2019-induced hypoxemic respiratory failure has yet to be answered. As our approaches to coronavirus disease 2019 continue to evolve, the decision of timing of intubation remains paramount. Although noninvasive ventilation may allow for delaying intubation, it is possible that there are downstream effects of delayed intubation that should be considered, including the potential for pulmonary vascular thrombosis and intussusceptive angiogenesis with delayed intubation.

The ectodysplasin receptor (EDAR) is a tumor necrosis factor receptor (TNF) superfamily member. A substitution in an exon of EDAR at position 370 (EDARV370A) creates a gain of function mutant present at high frequencies in Asian and Indigenous American populations but absent in others. Its frequency is intermediate in populations of Mexican ancestry. EDAR regulates the development of ectodermal tissues, including mammary ducts. Obesity and type 2 diabetes mellitus are prevalent in people with Indigenous and Latino ancestry. Latino patients also have altered prevalence and presentation of breast cancer. It is unknown whether EDARV370A might connect these phenomena. The goals of this study were to determine 1) whether EDARV370A is associated with metabolic phenotypes and 2) if there is altered breast anatomy in women carrying EDARV370A. Participants were from two Latino cohorts, the Arizona Insulin Resistance (AIR) registry and Sangre por Salud (SPS) biobank. The frequency of EDARV370A was 47% in the Latino cohorts. In the AIR registry, carriers of EDARV370A (GG homozygous) had significantly (p < 0.05) higher plasma triglycerides, VLDL, ALT, 2-hour post-challenge glucose, and a higher prevalence of prediabetes/diabetes. In a subset of the AIR registry, serum levels of ectodysplasin A2 (EDA-A2) also were associated with HbA1c and prediabetes (p < 0.05). For the SPS biobank, participants that were carriers of EDARV370A had lower breast density and higher HbA1c (both p < 0.05). The significant associations with measures of glycemia remained when the cohorts were combined. We conclude that EDARV370A is associated with characteristics of the metabolic syndrome and breast density in Latinos.

Abstract Idiopathic pulmonary fibrosis (IPF) is characterized by progressive scarring of the lung parenchyma, leading to respiratory failure and death. High resolution computed tomography of the chest is often diagnostic for IPF, but its cost and the risk of radiation exposure limit its use as a screening tool even in patients at high risk for the disease. In patients with lung cancer, investigators have detected transcriptional signatures of disease in airway and nasal epithelial cells distal to the site of disease that are clinically useful as screening tools. Here we assessed the feasibility of distinguishing patients with IPF from age-matched controls through transcriptomic profiling of nasal epithelial curettage samples, which can be safely and repeatedly sampled over the course of a patient’s illness. We recruited 10 patients with IPF and 23 age-matched healthy control subjects. Using 3′ messenger RNA sequencing (mRNA-seq), we identified 224 differentially expressed genes, most of which were upregulated in patients with IPF compared with controls. Pathway enrichment analysis revealed upregulation of pathways related to immune response and inflammatory signaling in IPF patients compared with controls. These findings support the concept that fibrosis is associated with upregulation of inflammatory pathways across the respiratory epithelium with possible implications for disease detection and pathobiology.

The aims of this study are to describe the frequency of occurrence and degree of expression of the parastyle in six different ethnic groups; to assess inter- and intra-observer errors when scoring the feature; and to compare the expression of the feature in a small number of twin pairs. Dental casts were examined for evidence of the parastyle from samples available in the Adelaide Dental School. A dental plaque developed by Katich & Turner was used to standardize scoring. The highest percentage frequency of parastyle occurrence was found in a sample of European twins with a value of 1.7%. The buccal aspect of the mesiobuccal cusp of the permanent maxillary right second molar was the most common site for the parastyle. Inter-observer reliability in scoring was lower than intra-observer reliability. In 10 pairs of twins (seven pairs of monozygotic (MZ) twins and three pairs of dizygotic (DZ) twins) only two pairs of MZ twins showed concordance for presence of the parastyle. The expression of parastyles likely results from a complex interaction of genetic, epigenetic, and environmental influences during dental crown development. There may be a relationship between parastyles and supernumerary teeth that are occasionally located buccally to the maxillary molar teeth.