Stroke disrupts neuronal functions in both local and remotely connected regions, leading to network-wide deficits that can hinder recovery. The thalamus is particularly affected, with progressive ...development of neurodegeneration accompanied by inflammatory responses. However, the complexity of the involved inflammatory responses is poorly understood. Herein we investigated the spatiotemporal changes in the secondary degenerative thalamus after cortical stroke, using targeted transcriptome approach in conjunction with histology and flow cytometry.
Cortical ischemic stroke was generated by permanent occlusion of the left middle cerebral artery in male C57BL6J mice. Neurodegeneration, neuroinflammatory responses, and microglial activation were examined in naive and stroke mice at from poststroke days (PD) 1 to 84, in both ipsilesional somatosensory cortex and ipsilesional thalamus. NanoString neuropathology panel (780 genes) was used to examine transcriptome changes at PD7 and PD28. Fluorescence activated cell sorting was used to collect CD11c
microglia from ipsilesional thalamus, and gene expressions were validated by quantitative real-time polymerase chain reaction.
Neurodegeneration in the thalamus was detected at PD7 and progressively worsened by PD28. This was accompanied by rapid microglial activation detected as early as PD1, which preceded the neurodegenerative changes. Transcriptome analysis showed higher number of differentially expressed genes in ipsilesional thalamus at PD28. Notably, neuroinflammation was the top activated pathway, and microglia was the most enriched cell type.
(CD11c) was the most significantly increased gene, and its expression was highly detected in microglia. Flow-sorted CD11c
microglia from degenerative thalamus indicated molecular signatures similar to neurodegenerative disease-associated microglia; these included downregulated Tmem119 and CX3CR1 and upregulated ApoE, Axl, LpL, CSF1, and Cst7.
Our findings demonstrate the dynamic changes of microglia after stroke and highlight the importance of investigating stroke network-wide deficits. Importantly, we report the existence of a unique subtype of microglia (CD11c
) with neurodegenerative disease-associated microglia features in the degenerative thalamus after stroke.
Stroke is one of the major causes of chronic disability worldwide and increasing efforts have focused on studying brain repair and recovery after stroke. Following stroke, the primary injury site can ...disrupt functional connections in nearby and remotely connected brain regions, resulting in the development of secondary injuries that may impede long-term functional recovery. In particular, secondary degenerative injury occurs in the connected ipsilesional thalamus following a cortical stroke. Although secondary thalamic injury was first described decades ago, the underlying mechanisms still remain unclear. We performed a systematic literature review using the NCBI PubMed database for studies that focused on the secondary thalamic degeneration after cortical ischemic stroke. In this review, we discussed emerging studies that characterized the pathological changes in the secondary degenerative thalamus after stroke; these included excitotoxicity, apoptosis, amyloid beta protein accumulation, blood-brain-barrier breakdown, and inflammatory responses. In particular, we highlighted key findings of the dynamic inflammatory responses in the secondary thalamic injury and discussed the involvement of several cell types in this process. We also discussed studies that investigated the effects of blocking secondary thalamic injury on inflammatory responses and stroke outcome. Targeting secondary injuries after stroke may alleviate network-wide deficits, and ultimately promote stroke recovery.
To evaluate associations between childhood obesity and post-adenotonsillectomy complications, informing guidelines for postoperative management.
The retrospective review assessed outpatient pediatric ...tonsillectomy/adenoidectomy cases performed at 2 ambulatory surgery centers in 2020. Complications in the recovery unit and within 2 weeks of surgical discharge were reviewed along with clinical and demographic variables. Obesity was defined as sex-specific body mass index-for-age, or weight-for-age if height data were unavailable, at/above the 95th percentile. The 99th percentile served as the threshold for severe obesity. Analyses used Chi-square/Fisher's exact tests and independent-samples t-tests with relative risk or effect sizes.
The review included 707 cases (180 patients with obesity). Overall incidence of complications in the recovery unit was 9.1%. Patients with obesity were significantly more likely to require supplemental blow-by oxygen (P = .02); relative risk was 1.65 (95% CI: 1.16–2.35) times greater in the cohort with obesity. Obesity had a small effect on postoperative oxygen saturation nadirs, which were significantly lower among patients with obesity (d = −0.34; P < .001). No differences emerged between cohorts with and without obesity in the incidence of any other complications before or after surgical discharge. Overall incidence of post-discharge returns was 7.9%. Incidence of complications did not vary by obesity severity.
From this cohort, childhood obesity without other significant comorbidities may not warrant routine inpatient care following adenotonsillectomy. Patients with obesity should receive additional monitoring for oxygen desaturation events during the first hours of recovery. Further prospective studies should continue to address this important topic.
•Childhood obesity alone may not warrant routine inpatient care after tonsillectomy.•Outpatient surgery can be safe for children with obesity if other risks are absent.•Oxygen desaturation in the first hours of recovery was more likely given obesity.•Obesity was not associated with incidence of any other postoperative complications.•Incidence of postoperative complications did not vary by obesity severity.
Abstract only Objective: Post-stroke brain stimulations have been shown to successfully enhance functional recovery. Previously we have demonstrated that optogenetic pan-neuronal stimulations in the ...ipsilesional primary motor cortex (iM1) promote functional recovery after stroke, however, the specific neuronal cell types involved (excitatory or inhibitory) remain unclear. Here we use optogenetic techniques to specifically stimulate excitatory neurons in layer V ipsilesional motor cortex and investigate the effects on functional recovery. Methods: C57BL/6 wild type mice were used (7-9 weeks). Mice underwent stereotaxic surgery to inject AAV1-CAMKIIa-ChR2-eYFP and implant a fiber cannula in the iM1. Six weeks later, all mice were subjected to an intraluminal middle cerebral artery suture occlusion (30 minutes). Optogenetic stimulation began at post-stroke (PD) day 5 and continued until PD14. Sensorimotor tests were used to assess behavioral recovery at PD4, 7 and 14. Mice were sacrificed at PD14 and their brains were processed for immunohistochemistry with CD68 and MAP2 for infarct analysis. Results: High expression of ChR2-YFP was detected in excitatory neurons of ipsilesional layer V motor cortex. Optogenetic excitatory stimulations in iM1 resulted in significant forelimb movements. Horizontal rotating beam test demonstrated that iM1-stimulated mice recovered significantly faster than non-stimulated mice in both speed and distance at PD14 after stroke. In particular, iM1-stimulated mice exhibited robust recovery in distance traveled at PD14 (p<0.01), with most mice performed similar to pre-stroke baseline. Furthermore, iM1-stimulated mice also exhibited faster regain of body weight loss after stroke. Infarct analysis using immunohistochemistry showed that there was no difference in lesion size between groups. Conclusions: These data highlight excitatory neurons as key cell type for brain stimulation-induced functional recovery after stroke. Current studies examine the molecular and circuit mechanisms underlying this recovery, including the role of neurotrophins in both the ipsilesional and contralesional sensory and motor cortices.
The purpose of this paper was to summarize the findings of a literature review on the use of peroxide-based tooth-whitening agents in children and adolescents. Safety considerations, including ...localized adverse effects and toxicological concerns, are described. Oral findings include:
(1) 1 in every 2 to 3 patients may experience tooth sensitivity and/or gingival irritation after bleaching treatment, which may be more traumatic an experience for children than adults; (2) depending on dose, duration, frequency, and route, studies indicate excessive exposure to peroxide can
be potentially harmful; (3) degree of potential toxicity and harmful outcomes increases in those who overuse whiteners-a concern in teenagers; (4) careful case selection using stringent criteria is suggested for primary teeth whitening; (5) whitening in healthy adolescents is a case-by-case
determination that must include the weighing of risks (oral health and age) vs benefits (improved esthetic perception). It is hoped that the present review will lead to a better understanding of the health implications of tooth whitening in children and adolescents, and offer guidance for
treatment that provides satisfactory outcomes externally (enamel and gingiva) and internally (endodontic tissues and systemic health).
Climate change and decadal variability are impacting marine fish and invertebrate species worldwide and these impacts will continue for the foreseeable future. Quantitative approaches have been ...developed to examine climate impacts on productivity, abundance, and distribution of various marine fish and invertebrate species. However, it is difficult to apply these approaches to large numbers of species owing to the lack of mechanistic understanding sufficient for quantitative analyses, as well as the lack of scientific infrastructure to support these more detailed studies. Vulnerability assessments provide a framework for evaluating climate impacts over a broad range of species with existing information. These methods combine the exposure of a species to a stressor (climate change and decadal variability) and the sensitivity of species to the stressor. These two components are then combined to estimate an overall vulnerability. Quantitative data are used when available, but qualitative information and expert opinion are used when quantitative data is lacking. Here we conduct a climate vulnerability assessment on 82 fish and invertebrate species in the Northeast U.S. Shelf including exploited, forage, and protected species. We define climate vulnerability as the extent to which abundance or productivity of a species in the region could be impacted by climate change and decadal variability. We find that the overall climate vulnerability is high to very high for approximately half the species assessed; diadromous and benthic invertebrate species exhibit the greatest vulnerability. In addition, the majority of species included in the assessment have a high potential for a change in distribution in response to projected changes in climate. Negative effects of climate change are expected for approximately half of the species assessed, but some species are expected to be positively affected (e.g., increase in productivity or move into the region). These results will inform research and management activities related to understanding and adapting marine fisheries management and conservation to climate change and decadal variability.
Severe Acute Respiratory Syndrome coronavirus 2 (SARS-CoV-2) infects cells by binding to the host cell receptor ACE2 and undergoing virus-host membrane fusion. Fusion is triggered by the protease ...TMPRSS2, which processes the viral Spike (S) protein to reveal the fusion peptide. SARS-CoV-2 has evolved a multibasic site at the S1-S2 boundary, which is thought to be cleaved by furin in order to prime S protein for TMPRSS2 processing. Here we show that CRISPR-Cas9 knockout of furin reduces, but does not prevent, the production of infectious SARS-CoV-2 virus. Comparing S processing in furin knockout cells to multibasic site mutants reveals that while loss of furin substantially reduces S1-S2 cleavage it does not prevent it. SARS-CoV-2 S protein also mediates cell-cell fusion, potentially allowing virus to spread virion-independently. We show that loss of furin in either donor or acceptor cells reduces, but does not prevent, TMPRSS2-dependent cell-cell fusion, unlike mutation of the multibasic site that completely prevents syncytia formation. Our results show that while furin promotes both SARS-CoV-2 infectivity and cell-cell spread it is not essential, suggesting furin inhibitors may reduce but not abolish viral spread.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused millions of human infections, and an effective vaccine is critical to mitigate coronavirus-induced disease 2019 (COVID-19). ...Previously, we developed a replication-competent vesicular stomatitis virus (VSV) expressing a modified form of the SARS-CoV-2 spike gene in place of the native glycoprotein gene (VSV-eGFP-SARS-CoV-2). Here, we show that vaccination with VSV-eGFP-SARS-CoV-2 generates neutralizing immune responses and protects mice from SARS-CoV-2. Immunization of mice with VSV-eGFP-SARS-CoV-2 elicits high antibody titers that neutralize SARS-CoV-2 and target the receptor binding domain that engages human angiotensin-converting enzyme-2 (ACE2). Upon challenge with a human isolate of SARS-CoV-2, mice that expressed human ACE2 and were immunized with VSV-eGFP-SARS-CoV-2 show profoundly reduced viral infection and inflammation in the lung, indicating protection against pneumonia. Passive transfer of sera from VSV-eGFP-SARS-CoV-2-immunized animals also protects naive mice from SARS-CoV-2 challenge. These data support development of VSV-SARS-CoV-2 as an attenuated, replication-competent vaccine against SARS-CoV-2.
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•A replicating VSV-SARS-CoV-2 vaccine induces high-titer neutralizing antibodies•Infectious SARS-CoV-2 is undetectable in the lung of vaccinated mice post-challenge•SARS-CoV-2-induced lung inflammation and pathology is decreased in vaccinated mice•Transfer of vaccine-derived immune sera to naive mice protects against SARS-CoV-2
Case, Rothlauf et al. report the efficacy of a replicating VSV-based SARS-CoV-2 vaccine. Immunized hACE2-expressing mice challenged with SARS-CoV-2 are protected against lung infection, inflammation, and pneumonia. Neutralizing antibodies are a correlate of this protection, as passive transfer of vaccine-derived immune sera protects naive mice from subsequent SARS-CoV-2 challenge.
1.8 million Veterans are estimated to need legal services, such as for housing eviction prevention, discharge upgrades, and state and federal Veterans benefits. While having one's legal needs met is ...known to improve one's health and its social determinants, many Veterans' legal needs remain unmet. Public Law 116-315 enacted in 2021 authorizes VA to fund legal services for Veterans (LSV) by awarding grants to legal service providers including nonprofit organizations and law schools' legal assistance programs. This congressionally mandated LSV initiative will award grants to about 75 competitively selected entities providing legal services. This paper describes the protocol for evaluating the initiative. The evaluation will fulfill congressional reporting requirements, and inform continued implementation and sustainment of LSV over time.
Our protocol calls for a prospective, mixed-methods observational study with a repeated measures design, aligning to the Reach Effectiveness Adoption Implementation Maintenance (RE-AIM) and Integrated Promoting Action on Research Implementation in Health Services (i-PARIHS) frameworks. In 2023, competitively selected legal services-providing organizations will be awarded grants to implement LSV. The primary outcome will be the number of Veterans served by LSV in the 12 months after the awarding of the grant. The evaluation has three Aims. Aim 1 will focus on measuring primary and secondary LSV implementation outcomes aligned to RE-AIM. Aim 2 will apply the mixed quantitative-qualitative Matrixed Multiple Case Study method to identify patterns in implementation barriers, enablers, and other i-PARIHS-aligned factors that relate to observed outcomes. Aim 3 involves a mixed-methods economic evaluation to understand the costs and benefits of LSV implementation.
The LSV initiative is a new program that VA is implementing to help Veterans who need legal assistance. To optimize ongoing and future implementation of this program, it is important to rigorously evaluate LSV's outcomes, barriers and enablers, and costs and benefits. We have outlined the protocol for such an evaluation, which will lead to recommending strategies and resource allocation for VA's LSV implementation.
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