IMPORTANCE: Attention-deficit/hyperactivity disorder (ADHD) is a heritable neurodevelopmental disorder. It has been linked to reductions in total brain volume and subcortical abnormalities. However, ...owing to heterogeneity within and between studies and limited sample sizes, findings on the neuroanatomical substrates of ADHD have shown considerable variability. Moreover, it remains unclear whether neuroanatomical alterations linked to ADHD are also present in the unaffected siblings of those with ADHD. OBJECTIVE: To examine whether ADHD is linked to alterations in whole-brain and subcortical volumes and to study familial underpinnings of brain volumetric alterations in ADHD. DESIGN, SETTING, AND PARTICIPANTS: In this cross-sectional study, we included participants from the large and carefully phenotyped Dutch NeuroIMAGE sample (collected from September 2009-December 2012) consisting of 307 participants with ADHD, 169 of their unaffected siblings, and 196 typically developing control individuals (mean age, 17.21 years; age range, 8-30 years). MAIN OUTCOMES AND MEASURES: Whole-brain volumes (total brain and gray and white matter volumes) and volumes of subcortical regions (nucleus accumbens, amygdala, caudate nucleus, globus pallidus, hippocampus, putamen, thalamus, and brainstem) were derived from structural magnetic resonance imaging scans using automated tissue segmentation. RESULTS: Regression analyses revealed that relative to control individuals, participants with ADHD had a 2.5% smaller total brain (β = −31.92; 95% CI, −52.69 to −11.16; P = .0027) and a 3% smaller total gray matter volume (β = −22.51; 95% CI, −35.07 to −9.96; P = .0005), while total white matter volume was unaltered (β = −10.10; 95% CI, −20.73 to 0.53; P = .06). Unaffected siblings had total brain and total gray matter volumes intermediate to participants with ADHD and control individuals. Significant age-by-diagnosis interactions showed that older age was linked to smaller caudate (P < .001) and putamen (P = .01) volumes (both corrected for total brain volume) in control individuals, whereas age was unrelated to these volumes in participants with ADHD and their unaffected siblings. Attention-deficit/hyperactivity disorder was not significantly related to the other subcortical volumes. CONCLUSIONS AND RELEVANCE: Global differences in gray matter volume may be due to alterations in the general mechanisms underlying normal brain development in ADHD. The age-by-diagnosis interaction in the caudate and putamen supports the relevance of different brain developmental trajectories in participants with ADHD vs control individuals and supports the role of subcortical basal ganglia alterations in the pathophysiology of ADHD. Alterations in total gray matter and caudate and putamen volumes in unaffected siblings suggest that these volumes are linked to familial risk for ADHD.
Toll-like receptors (TLRs) are discovered as crucial pattern recognition receptors (PRRs) involved in the recognition of pathogen-associated molecular patterns (PAMPs). Later studies showed their ...involvement in the recognition of various damage/danger-associated molecular patterns (DAMPs) generated by host itself. Thus, TLRs are capable of recognizing wide-array of patterns/molecules derived from pathogens and host as well and initiating a proinflammatory immune response through the activation of NF-κB and other transcription factors causing synthesis of proinflammatory molecules. The process of neuroinflammation is seen under both sterile and infectious inflammatory diseases of the central nervous system (CNS) and may lead to the development of neurodegeneration. The present article is designed to highlight the importance of TLRs in the pathogenesis of neuroinflammation under diverse conditions. TLRs are expressed by various immune cells present in CNS along with neurons. However out of thirteen TLRs described in mammals, some are present and active in these cells, while some are absent and are described in detail in main text. The role of various immune cells present in the brain and their role in the pathogenesis of neuroinflammation depending on the type of TLR expressed is described. Thereafter the role of TLRs in bacterial meningitis, viral encephalitis, stroke, Alzheimer's disease (AD), Parkinson's disease (PD), and autoimmune disease including multiple sclerosis (MS) is described. The article is designed for both neuroscientists needing information regarding TLRs in neuroinflammation and TLR biologists or immunologists interested in neuroinflammation.
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•TLRs are crucial PRRs to recognize both intracellular and extracellular PAMPs or DAMPs.•TLRs are also expressed by brain microglia, astrocytes, oligodendrocytes, and neurons.•Activation of TLRs plays a crucial role in generating neuroinflammatory immune response.•Both MyD88-dependent and MyD88-independent TLR signaling pathways generate neuroinflammation.•Neuroinflammation is involved in both sterile (stroke, AD, PD, and MS) and infectious diseases of the brain.
SARS‐CoV‐2 is responsible for the ongoing COVID‐19 pandemic, which causes respiratory failure and damage to multiple organ systems. Emergence of new variants of concern (VOCs), including Omicron can ...potentially render the current vaccines ineffective. However, our understanding of COVID‐19 pathophysiology and molecular basis of SARS‐CoV‐2 infection is very limited. The role of the Hippo signaling pathway, an evolutionarily conserved organogenesis circuitry, in tissue inflammation and innate immune response is beginning to be understood. Given the complexity of COVID‐19 associated cell injury and immunopathogenesis processes, we investigated this Hippo pathway dynamic in SARS‐CoV‐2 infection by utilizing COVID‐19 lung samples, transcriptome and human cell models based on pluripotent stem cell‐derived cardiomyocytes (PSC‐CMs) and human primary lung air‐liquid interface (ALI) culture. The SARS‐CoV‐2 infection resulted in stoppage of cardiomyocyte beating and extensive apoptotic cell death. Especially the infection caused activation of Hippo signaling pathway in cardiomyocytes, as shown by increased level of phosphorylated form of YAP, a downstream transcriptional co‐factor involved in tissue growth, mitochondrial biogenesis and innate immunity. Similar activation was noted in SARS‐CoV‐2 infected lung ALI epithelial cells and COVID‐19 lung autopsy samples. The shRNA‐mediated partial knockdown and pharmacological inhibitor of YAP/TAZ resulted in significantly reduced SARS‐CoV‐2 replication, whereas inhibition of Hippo pathway upstream LATS1 and MST1 kinases led to enhanced virus replication. These results indicate a direct role of Hippo signaling in SARS‐CoV‐2 mediated disease pathogenesis and this pathway can be pharmacologically targeted to treat COVID‐19.
Scientists continue to study SARS‐CoV‐2, the virus currently responsible for a worldwide pandemic, to determine the best methods to inactivate the virus on surfaces and reduce the risk of contact ...transmission. Its classification as a Biosafety Level (BSL) ‐3 pathogen complicates research since few facilities have the capacity for this level of containment. Human betacoronavirus OC43 (OC43) is a good surrogate for SARS‐CoV‐2 based on its genetic similarity and OC43’s BSL‐2 classification. Two industry specific surfaces were inoculated with OC43 and sampled at timepoints that reflect typical industry timelines for sanitation. The objective was to determine the stability of OC43 on typical meat facility surfaces.
OC43 was propagated on human ileocecal colorectal adenocarcinoma cells (HRT‐18, ATCC CCL‐244). Infectious virus was quantified by indirect immunofluorescence TCID50 assays on HRT‐18 cells. Stainless steel grade 316 and polyethylene (poly top) coupons were inoculated with 100 ul of 2.51 x 106 TCID50/ml virus by pipette in a grid pattern at room temperature (20ºC) in an active biosafety cabinet in triplicate for each timepoint. Coupons were rinsed with 5 ml of sterile filtered beef extract buffer (BEB7), composed of Tris base, glycine, magnesium chloride, and beef extract pH adjusted to 7.0, at 0, 1, 4, 8, and 16 hours post inoculation (hpi). The collected rinsate was aliquoted and stored at ‐80ºC until the TCID50 assays were conducted in batch. The 16 hpi stainless steel samples were created at the time of the poly top experiment. The TCID50 assays were run in duplicate for each sample and TCID50/ml was calculated and averaged for each replicate. Percent recovery of infectious virus was determined for each timepoint.
Virus was observed to have partially dried at 1 hpi and was completely dry at all following timepoints on both surfaces. Few infectious virions were lost between inoculation and 0 hpi collection from both surfaces. Mean percent recovery (MPR) of infectious OC43 from stainless steel and poly top at 0 hpi was 153% and 90%, respectively. At 1 hpi, the MPR from stainless steel was 65% while poly top was 123%. The variability of the assay explains these findings. Stainless steel and poly top had average recoveries of 60% and 25% at 4 hours, respectively. The lowest MPR for stainless steel was 17% at 8 hpi while the lowest MPR for poly top was 7% at 16 hpi. A higher MPR of 43% from stainless steel at 16 hpi reflects this timepoint being conducted on a different day.
Infectious OC43 is still detectable on common meat facility surfaces several hours after inoculation, but our current knowledge of enveloped viruses asserts that they are inviable after drying. A maximum decrease of only 1 log TCID50 between inoculation and the 16 hpi samples means that a person could potentially be infected between cleanings if it is possible to transfer dried infectious virus from a surface to a mucous membrane. However, this is a conjecture that requires further study since the infectious dose of SARS‐CoV‐2 is unknown and no experiment to determine if this contact transfer is possible has been conducted. Currently, similar experiments with SARS are underway.
Severe viral pneumonia caused by the severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) is characterized by a hyperinflammatory state typified by elevated circulating pro‐inflammatory ...cytokines, frequently leading to potentially lethal vascular complications including thromboembolism, disseminated intracellular coagulopathy and vasculitis. Though endothelial infection and subsequent endothelial damage have been described in patients with fatal COVID‐19, the mechanism by which this occurs remains elusive, particularly given that, under naïve conditions, pulmonary endothelial cells demonstrate minimal cell surface expression of the SARS‐CoV‐2 binding receptor ACE2. Herein we describe SARS‐CoV‐2 infection of the pulmonary endothelium in postmortem lung samples from individuals who died of COVID‐19, demonstrating both heterogeneous ACE2 expression and endothelial damage (Figure). In primary endothelial cell cultures, we show that SARS‐CoV‐2 infection is dependent on the induction of ACE2 protein expression and that this process is facilitated by type 1 interferon‐alpha (IFNα) or ‐beta(β) ‐ two of the main anti‐viral cytokines induced in severe SARS‐CoV‐2 infection ‐ but not significantly by other cytokines (including interleukin 6 and interferon g /λ). Our findings suggest that the stereotypical anti‐viral interferon response may paradoxically facilitate the propagation of COVID‐19 from the respiratory epithelium to the vasculature, raising concerns regarding the use of exogenous IFNα/β in the treatment of patients with COVID‐19.
Introduction
After its emergence in China, the COVID‐19 pandemic has spread rapidly around the world, affecting all of our lives. Over the past few months, different vaccines have been issued to the ...public following FDA approval. Since their release, the number of COVID‐19 cases in the United States have gone down significantly. However, there is a long way to go until the eradication of the virus. There has also been a strong emergence of delta variants across the world, which (for the most part) can still be partially blocked by vaccines. This is why it is so important everyone gets a vaccine for COVID‐19.
Purpose
The purpose of this project was to determine the public’s opinion of COVID‐19 and the COVID‐19 vaccines, while also determining correlation between different demographics and these opinions.
Hypothesis
There are many different factors within various demographics that cause a correlation to a certain belief regarding COVID‐19 and COVID‐19 vaccines.
Materials and Methods
This study was conducted in collaboration with Loyola University Chicago. Survey questions about COVID‐19 and COVID‐19 vaccinations have been sent electronically to the respondents. Initial results based on a pilot study where the answers from 250 respondents were compiled. Basic descriptive statistics were used to characterize the sample and study variables. Univariate analysis of associations between categorical variables and vaccination intent and acceptance were assessed by chi‐square statistics. Multivariate analysis performed by regression analysis to determine significant independent predictors. A p‐value of less than 0.05 was considered to indicate statistical significance; all tests were 2‐tailed.
Results
In our study population vaccine willingness have been found as 94,4%, the rate of receiving at least one dose of vaccination have been found as 90,8%. In univariate analysis, gender, believing that vaccinations protect others and having enough information about safety and efficacy of vaccines have been found significant factors for vaccine willingness. For receiving at least one dose of vaccination, gender, previous COVID‐19 infection, believing that vaccinations protect others, having enough information about safety and efficacy of vaccines, the degree of COVID‐19 related effects on person’s life, knowledge that COVID‐19 can cause blood clots and the degree of concerns about delta variant have been found significant factors for vaccine willingness. In multivariate analysis only believing that vaccinations protect others have been found as an independent factor for vaccine willingness. For receiving at least one dose of vaccination, believing that vaccinations protect others, the degree of COVID‐19 related effects on person’s life and having enough information about safety and efficacy of vaccines have been found independent factors in multivariate analysis.
Conclusion
There are several vaccines currently available in the United States. All of these vaccines provide immunity against COVID‐19 and its variants which have been identified in 2020 and 2021. The level of immunity and the reported adverse events vary with each of the vaccines. Despite minor issues the vaccination outweighs the benefits and provides a clear approach to control this pandemic.
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Neuropathological complications are frequently observed in SARS‐CoV‐2 infection and brain autopsies from human subjects who died from COVID‐19 have revealed significant pathology, ...including wide‐spread neuroinflammation, hypoxic‐ischemic injury, and microhemorrhages. To begin to understand the neuropathogenesis of SARS‐CoV‐2 infection, we investigated brain from infected non‐human primates (NHP)s for pathological changes consistent with that seen among humans. Eight aged NHPs were inoculated with the 2019‐nCoV/USA‐WA1/2020 strain of SARS‐CoV‐2 via a multi‐route mucosal or aerosol challenge. Hematoxylin and eosin (H&E) and immunohistochemistry (IHC) staining was done on seven brain regions to elucidate general pathology, microhemorrhages, platelet derived thrombi, neuronal apoptosis, microglia and astrocyte morphology, hypoxia, and virus present. Similar to humans, pathology was variable but included wide‐spread neuroinflammation, nodular lesions, neuronal degeneration, and microhemorrhages. Neuronal degeneration was most often seen in the cerebellum and brainstem of infected animals. Neuronal death was confirmed through FluorJade C and cleaved (active) caspase 3 IHC, which showed foci of positivity, particularly among Purkinje cells of the cerebellum. Importantly, this was seen among infected animals that did not develop severe respiratory disease. Hypoxia inducible factor‐1α (HIF‐1α) was observed at a higher intensity around the vasculature within deep brain regions of the infected animals. Microhemorrhages were prevalent among all animals but were less frequently associated with platelet derived thrombi in the infected animals, as compared to mock‐infected controls. Sparse virus was detected in brain endothelial cells but did not associate with the severity of CNS injury. Increased HIF‐1α suggests that brain hypoxia may promote neuronal degeneration within infected brain. Wide‐spread neuroinflammation may also contribute to neuronal injury/death and neurological manifestations seen in the context of infection.
Observational studies have overwhelmingly demonstrated that vitamin D deficiency is a risk factor for coronavirus disease‐19 (COVID‐19) infection and severity. However, serum 25(OH)D may act as a ...negative acute phase reactant and therefore an unreliable marker for vitamin D status post‐inflammatory insult. This study evaluated the serum levels of 25(OH)D, 34 cytokines and chemokines in 220 participants (82 control and 138 SARS‐CoV‐2 patients). Serum 25(OH)D levels were significantly lower in the SARS‐CoV‐2 group than controls. Serum IP‐10, MCP‐1, CRP, IFNγ, IL‐10, IL‐13, IL‐17α, IL‐23, and IL‐6 were significantly higher in COVID‐19 patients compared to healthy control. Results showed that serum levels of VEGF, IFNγ, IL‐13, and IL‐5 were significantly higher in male patients compared to females. 25(OH)D was significantly correlated with EFG (R=0.39, p<0.05) and IL‐15 (R=0.39, p<0.05) in male patients, while inversely correlated with CRP (R=‐0.51, p<0.05) in female patients. In conclusion, we recommend 25(OH)D supplementation among high‐risk individuals and SARS‐CoV‐2‐infected individuals. Additionally, the upregulated cytokines might serve as therapeutic targets to modulate the heightened inflammation and disease severity, moreover, they could be helpful in the early screening of critical illness, diagnosis and treatment of SARS‐CoV‐2.
Background
With the spread of highly infectious strains such as the Delta variant of SARS‐CoV‐2, rapid antigen testing and variant tracking are gaining more attention from infectious disease ...specialists and the general public as essentials for disease control and prevention amidst the COVID‐19 pandemic. The lateral flow based antigen tests provide fast turnaround of results within minutes, but it’s open ‐faced assay format means that the test operators are more prone to accidental exposure to pathogen‐containing samples. The viral inactivation transport media (ITMs) provide much needed protection against exposure to highly transmissive live viruses during sample collection, transport, and storage. However, the current ITMs were designed for PCR tests and share a harsh chemical formulation that denatures protein analytes, making them incompatible with antigen tests.
We developed an innovative viral transport media that is designed for a variety of tests, such as antigen testing and PCR. The new formulation can be used to safely inactivate SARS‐CoV‐2 virus while maintaining compatibility with many different formats beyond rt‐PCR.
Design
To evaluate the viral inactivation performance of the novel inactivation transport media formulation, a cytopathic effect assay was conducted using VERO E6 cells in presence of SARS‐CoV‐2 samples incubated in our novel formulation and log reduction were reported.
To assess its compatibility with different SARS‐CoV‐2 related assays, the novel formulation was used as inactivation transport media in antigen, PCR, and NGS based Swab‐Seq tests with known controls.
Results
The results for the inactivation study confirmed > 3.0 log reduction in viral titer after 30 min exposure in novel transport media formulation and demonstrated 99.5% effectiveness in SARS‐CoV‐2 inactivation. The results for the PCR and antigen test compatibility studies showed comparable and/or superior performance in detection limits for the novel formulation when compared to other sample media. The Swab‐Seq result offers preliminary support on the novel formulation’s compatibility with NGS based assays.
Conclusion
Using a novel viral transport medium, we were able to completely inactivate the SARS‐CoV‐2 virus. The new formulation was compatible with both nucleic acid and protein assays while protecting cells from infection. This viral transport media could allow more test types to be done from a single specimen, resulting in safer and reliable outcomes. Additionally, it would reduce the need for multiple samples collected from patients while supporting SARS‐CoV‐2 variant tracking via NGS.
Severe Acute Respiratory Syndrome Coronavirus 2 (SARS‐CoV‐2), more commonly known as COVID‐19, is a virus that has spread at a rapid rate beginning in 2019 causing a pandemic. Currently, we remain in ...this pandemic, and as this virus progresses it is creating mutations of itself, called variants. With these variants, it has become increasingly difficult to control COVID‐19 and keep track of new variants that continue to infect people everywhere. In Orange County, California, COVID‐19 and its variants have enabled county wide shut downs and thousands of infections and deaths. To investigate its devastating effect, five positive COVID‐19 nasal samples were collected between February 2021 to July 2021, and the RNA was extracted. After extraction, a Reverse Transcription Polymerase Chain Reaction (RT‐PCR) was done to amplify 1,727 base pairs of the S‐ Gene within the COVID‐19 genome and was sent to sequencing. These sequences were then aligned using the Basic Local Alignment Search Tool (BLAST) and compared to the original COVID‐19 S‐Gene genome to analyze possible variations. Within the five samples, 30 different mutations were detected and a phylogenetic tree was generated. Awareness of the genetic variation of SARS‐CoV‐2 provides potential future development for more effective targeted research.
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