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.
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.
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).
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.
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.
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.
The novel coronavirus SARS‐Cov‐2 or COVID‐19 became a global pandemic and currently few medically approved curative treatments exist. SARS‐Cov‐2 acts similarly to SARS‐CoV‐1 from where it may have ...evolved. The COVID‐19 virus can survive ~3 hours in air and < 72 hours on distinct surfaces. COVID‐19 mutates by introducing sequence errors in the host’s RNA genome or by modifying proteins and enzymes. Vaccination, including booster shots, social distancing and isolation are the most generally practiced guidelines in the global management of COVID‐19. Globally the most frequently utilized pharmacologic treatments include ivermectin, hydroxychloroquine, glucocorticoids, the anti‐viral agent remdesivir, monoclonal antibodies, and convalescent plasma in combination with an antimicrobial agent such as azithromycin to minimize secondary microbial infection and nutritional supplementation (vitamins C, D3, and zinc) to enhance cellular immune responses and are included in the routine protocols of some emergency rooms. COVID‐19 viral transmission occurs via respiratory microdroplets, by inhaling COVID‐19 laden airborne particles and contact with contaminated surfaces on which these droplets have been deposited. SARS‐CoV‐2 targets ACE2 receptors in the upper and lower respiratory tracts in addition to the heart, brain, and gastrointestinal tract, and may cause thromboses in the liver, heart, and kidney.
Significant risk factors for the severity of COVID‐19 infection include advanced age and pre‐existing comorbid conditions obesity, hypertension, cardiovascular disease, diabetes, and compromised immune status, which all correlate with greater mortality. Outcome‐effecting factors include viral load, viral mutations, and pre‐existing conditions. Since its origination, genomic studies of the virus have identified numerous variants which have become regionally prevalent in different countries. Infective factors, comorbidities and viral load strongly affect outcomes; patients infected with the greatest viral load showed a higher mortality. It is opined that the number of deaths attributed to COVID‐19 may be inaccurate due to errors in diagnosing and reporting, since other similar illnesses may exhibit similar symptoms.
Future research should focus on prevention practices, comorbidities, genetic prevalence, reliable systematic and consistency in country‐by‐country testing and reporting procedures, further scrutiny regarding the efficacy of current vaccines and protocols, and the pursuit for innovative therapies for Coronaviruses and variants including biophotonics and exploration of emerging bioenergetic, nutritional, pharmacological, immunotherapeutic and vaccination‐preventive applications for eradication of COVID‐19.
Refs: 1. Cheng, RZ. (2020a). Med Drug Disc, 5, 100028; 2. Shankar, AH, & Prasad, AS. (1998). ACJN, 68(2), 447S‐463S; 3. Petrilli, CM, Jones, SA, et al. (2020); medRxiv; 4.van Doremalen, N, Bushmaker, T et al. (2020). NEJM, 382(16), 1564‐1567
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.
Covid‐19 vaccination and social barrier efforts worldwide have deescalated the death and morbidity toll due to Sars CoV‐2 infection in most countries. However, intramuscular vaccination leads to a ...variable antibody response insufficient to provide airway mucosal immunity that prevents contagion and reinfection in asymptomatic carriers. Nasal vaccines could provide this missing piece and are currently being tested with encouraging results, but large‐scale availability and efficacy in humans are unknown. Moreover, previous exposure to the virus would render nasal spray vaccines less effective. Past literature on upper airway infections has shown that water steam inhalation therapy (WSIT) may modulate airway mucosa immunity responses. Airway mucosal immunity in COVID‐19 infection is a critical aspect to consider with WSIT. Based on a small but slowly growing body of evidence, we propose that WSIT may be a relevant home remedy worth revisiting to help halt mass contagion. Here, we review current and past studies on WSIT use in upper respiratory infections (URI). Previous studies reported mixed results on the efficacy of steam therapy on viral respiratory diseases. Safety concerns with steam therapy include possible nosocomial infections associated and burns. There is a paucity of studies with high methodological quality exploring the outcomes of this ancient therapy. Upper airway blood flow increases substantially after 15‐min of steam inhalation with temperatures of 42 ℃. Still, methods to measure bronchial blood flow are lacking, and this may be an essential variable for the analysis of how steam may impact mucosal immune responses. Studies with better methodologies demonstrate that nebulized steam at 80°C for 20 min reduces neutrophils compared to controls in COPD patients indicating an anti‐inflammatory effect. The best evidence for using WSIT for upper airway viral disease shows that 60% of asymptomatic carriers increased viral shedding immediately post steam inhalation, and Rt‐PCR testing confirmed viral negativity at 10‐days post‐treatment. Subjects used steam for a total of 20 minutes per hour with steam temperatures ranging from 55 to 65℃. Efficacy rates decline with research methodologies that use WSIT for less than 20 minutes. In summary, more recent studies using WSIT for URI show encouraging results and may help to disentangle former controversies. Lessons emerging from previous studies indicate that exposure of WSIT less than 20 minutes may be ineffective. Adopting WSIT as an inexpensive public health measure may lead to secondary prevention of airway viral infections, especially in new viral strands that have the potential to become pandemic.
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Introduction
Recent research suggests that endothelial activation plays a role in COVID‐19 pathogenesis by promoting a pro‐coagulative and pro‐inflammatory state. However, the mechanism ...by which the endothelium is activated in COVID‐19 is unclear.
Objective
To investigate the mechanism by which COVID‐19 activates the pulmonary endothelium.
Hypothesis
The pulmonary endothelium generates reactive oxygen species (ROS) upon exposure to the “inflammatory load” of the systemic circulation.
Methods
COVID‐19 was recreated
in vitro
and
ex vivo
, by exposing human lung endothelial cells (EC) or donor human lung slices (human precision‐cut lung slices or huPCLS) to medium supplemented with serum from COVID‐19 affected subjects. Sera were acquired from patients with COVID‐19 infection admitted to the Intensive Care Unit of the Hospital at the University of Pennsylvania. ROS (fluorescent dye, CellROX) and intercellular adhesion molecule (ICAM‐1) levels were assessed by fluorescence labeling and imaging.
Results
Both EC activation (as monitored by ROS production) and pro‐inflammatory phenotype (as assessed by ICAM‐1), were significantly higher with COVID‐19 as compared to normal subjects.
Conclusions
The endothelium is activated with COVID‐19 via ROS production; thus, the ROS produced drive a pro‐inflammatory phenotype by inducing the expression of ICAM‐1, a pivotal marker of endothelium inflammation. As ROS mediates EC activation and inflammation during COVID‐19, ROS blockade could be a therapeutic target in maintaining vascular health.
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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.
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.
The emergence of novel, zoonotic betacoronavirus, SARS‐CoV‐2, demands quantification of infectious virus rather than viral RNA to provide more accurate assessment of transmission risk for COVID‐19. ...To maximize investigator safety, similar in morphology and inactivation profile, human betacoronavirus OC43 (OC43) was used as a surrogate to refine infectious virus recovery methods for SARS‐CoV‐2, a biosafety level (BSL)‐3 pathogen. Using OC43 and SARS‐CoV‐2, we examined the ability of InnovaPrep Mano Surface Samplers (MANOs), made of hydrophobic, open‐cell polymeric foam, to recover virus from large stainless‐steel surfaces hypothesizing that they would outperform hydrophilic cellulose sponges (sponges), used for environmental sampling.
In triplicate, 1.0 x 105 TCID50 of OC43 and SARS‐CoV‐2 were applied to the inner surface of a biological safety cabinet (BSC) within the literature or vendor specified optimal surface area, 1267.36 cm2 (MANO) and 100 cm2 (sponges). The areas were sampled with eluant presoaked MANOs and sponges, and samples aliquoted and stored at ‐80ºC until batch titration by indirect immunofluorescence TCID50 assay, using human ileocecal colorectal adenocarcinoma cells (HRT‐18 CCL‐244, ATCC) for OC43 samples and crystal violet detection‐based TCID50 using African green monkey kidney epithelial cells (VeroE6 CRL‐1586, ATCC) for SARS‐CoV‐2. Tested eluants were a beef extract buffer, pH 7 (BEB7), BEB7 with 0.05% Tween‐20 (BEB7/T) and (MANOs only) phosphate buffered saline with 0.05% Tween‐20 (PBS/T), the vendor default. Additionally, an expanded surface area (0.77m2 for MANOs, 300cm2 for sponges) was tested with OC43 as described above, except with 1.0x104 TCID50 inoculum.
For OC43, recovery for MANOs with BEB7 was 2.66x105 TCID50 compared to PBS/T’s 4.40x104 TCID50. BEB7 MANOs had a higher average percent recovery (102%) than BEB7 sponges (84%). By one‐way ANOVA (α=0.05), MANO BEB7’s and BEB7/T’s average recovery percentages were significantly greater than PBS/T’s. Samples taken from the larger surface areas for both samplers were below the assay’s limits of detection for BEB7; however, OC43 eluted with BEB7/T from sponges was detectable (3.47x102 TCID50, 32% recovery). For SARS‐CoV‐2, MANO BEB7’s average recovery percentage (27%) was again shown to be significantly greater than PBS/T’s (0.38%). However, decreased recovery of infectious virus was seen across both tools and all eluants, perhaps indicating a difference between SARS‐CoV‐2 and OC43 for BEB7.
Use of BEB7 improved OC43 and SARS‐CoV‐2 recovery from optimal surface areas with MANOs. Surprisingly, when the surface area was enlarged, all samples except those from BEB7/T sponges were below the limit of detection. This might be explained by additional eluant loss on the larger surface during mechanical recovery or virus inactivation due to desiccation from BSC airflow or other environmental factors. Similar factors might explain the SARS‐CoV‐2 results. Quantification by RT‐qPCR or flow virometry, and planned experiments in BSL‐3 agricultural space, where the room is primary containment, may provide further insights.
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