Uncovering Novel Extracellular Matrix Transcriptome Alterations in Lesions of Multiple Sclerosis.

The extracellular matrix (ECM) of the central nervous system (CNS) is an interconnected network of proteins and sugars with critical roles in both homeostasis and disease. In neurological diseases, excessive ECM deposition and remodeling impact both injury and repair. CNS lesions of multiple sclerosis (MS), a chronic inflammatory and degenerative disease, cause prominent alterations of the ECM. However, there are a lack of data investigating how the multitude of ECM members change in relation to each other and how this affects the MS disease course. Here, we evaluated ECM changes in MS lesions compared to a control brain using databases generated in-house through spatial mRNA-sequencing and through a public resource of single-nucleus RNA sequencing previously published by Absinta and colleagues. These results underline the importance of publicly available datasets to find new targets of interest, such as the ECM. Both spatial and public datasets demonstrated widespread changes in ECM molecules and their interacting proteins, including alterations to proteoglycans and glycoproteins within MS lesions. Some of the altered ECM members have been described in MS, but other highly upregulated members, including the SPARC family of proteins, have not previously been highlighted. SPARC family members are upregulated in other conditions by reactive astrocytes and may influence immune cell activation and MS disease course. The profound changes to the ECM in MS lesions deserve more scrutiny as they impact neuroinflammation, injury, and repair.

Single Cell Analysis of High-Parameter Histology Images Using Histoflow Cytometry.

Immunofluorescence histology is commonly used to study immune cells in tissues where the number of fluorescence parameters is normally limited to four or less. This makes it impossible to interrogate multiple subsets of immune cells in tissue with the same precision as flow cytometry. The latter, however, dissociates tissues and loses spatial information. To bridge the gap between these technologies, we developed a workflow to expand the number of fluorescence parameters that can be imaged on widely available microscopes. We instituted a method for identifying single cells in tissue and exporting the data for flow cytometry-based analysis. This histoflow cytometry technique successfully separates spectrally overlapping dyes and identifies similar numbers of cells in tissue sections as manual cell counts. Populations identified through flow cytometry-like gating strategies are mapped to the original tissue to spatially localize gated subsets. We applied histoflow cytometry to immune cells in the spinal cords of mice with experimental autoimmune encephalomyelitis. We ascertained that B cells, T cells, neutrophils, and phagocytes differed in their frequencies in CNS immune cell infiltrates and were increased relative to healthy controls. Spatial analysis determined that B cells and T cells/phagocytes preferentially localized to CNS barriers and parenchyma, respectively. By spatially mapping these immune cells, we inferred their preferred interacting partners within immune cell clusters. Overall, we demonstrate the ease and utility of histoflow cytometry, which expands the number of fluorescent channels used in conventional immunofluorescence and enables quantitative cytometry and spatial localization of histological analyses.

A cadaveric study of arteriovenous trigone of heart: the triangle of Brocq and Mouchet.

Left coronary artery divides into anterior interventricular branch and circumflex branch. As both the arteries run in their corresponding grooves, an arteriovenous trigone is formed between conus arteriosus and left auricle called triangle of Brocq and Mouchet. The triangle base is formed by great cardiac vein. This study aims to describe the frequency of triangle and its type and relationship between various boundaries and content of triangle and to supplement the existing knowledge of clinicians. This observational and descriptive study was conducted on 40 formalin fixed cadaveric hearts in department of anatomy, Kalpana chawla government medical college. The triangle was found in 92.5% of specimen with most common type being closed (51.3%) which is followed by inferiorly open in 35.1%, superiorly open in 8.1% and completely open in 5.4% hearts. Most frequent content of triangle was median artery followed by diagonal branches of anterior interventricular and circumflex branches. The mean area of the triangle was 246.3 mm2. Relationship of vein with two arterial branches was either superficial or deep. The knowledge of different patterns of existence will be required for angiographic procedures. Further the triangle is a potential epicardial access route to left fibrous ring. Thus detailed knowledge of variations will help cardiologist to achieve better outcome in interventional procedures with minimal complications.

Expression of antioxidant enzymes in lesions of multiple sclerosis and its models.

Oxidative stress promotes tissue injury in the central nervous system in neurological disorders such as multiple sclerosis (MS). To protect against this, antioxidant enzymes including superoxide dismutase-1 (SOD1), heme oxygenase-1 (HO-1), peroxiredoxin-5 (PRDX5) and glutathione peroxidase-4 (GPX4) may be upregulated. However, whether antioxidant enzyme elevation in mouse models of neurodegeneration corresponds to their expression in human diseases such as MS requires investigation. Here, we analyzed and compared the expression of SOD1, HO-1, PRDX5 and GPX4 in the murine spinal cord of three models of MS: focal lesions induced by (1) oxidized phosphatidylcholine or (2) lysophosphatidylcholine (lysolecithin), and (3) diffuse lesions of experimental autoimmune encephalomyelitis. Notably, CD68+ microglia/macrophages were the predominant cellular populations that expressed the highest levels of the detected antioxidant enzymes. Overall, the expression patterns of antioxidant enzymes across the models were similar. The increase of these antioxidant enzymes was corroborated in MS brain tissue using spatial RNA sequencing. Collectively, these results show that antioxidant capacity is relatively conserved between mouse models and MS lesions, and suggest a need to investigate whether the antioxidant elevation in microglia/macrophages is a protective response during oxidative injury, neurodegeneration, and MS.

Versican promotes T helper 17 cytotoxic inflammation and impedes oligodendrocyte precursor cell remyelination.

Remyelination failure in multiple sclerosis (MS) contributes to progression of disability. The deficient repair results from neuroinflammation and deposition of inhibitors including chondroitin sulfate proteoglycans (CSPGs). Which CSPG member is repair-inhibitory or alters local inflammation to exacerbate injury is unknown. Here, we correlate high versican-V1 expression in MS lesions with deficient premyelinating oligodendrocytes, and highlight its selective upregulation amongst CSPG members in experimental autoimmune encephalomyelitis (EAE) lesions modeling MS. In culture, purified versican-V1 inhibits oligodendrocyte precursor cells (OPCs) and promotes T helper 17 (Th17) polarization. Versican-V1-exposed Th17 cells are particularly toxic to OPCs. In NG2CreER:MAPTmGFP mice illuminating newly formed GFP+ oligodendrocytes/myelin, difluorosamine (peracetylated,4,4-difluoro-N-acetylglucosamine) treatment from peak EAE reduces lesional versican-V1 and Th17 frequency, while enhancing GFP+ profiles. We suggest that lesion-elevated versican-V1 directly impedes OPCs while it indirectly inhibits remyelination through elevating local Th17 cytotoxic neuroinflammation. We propose CSPG-lowering drugs as potential dual pronged repair and immunomodulatory therapeutics for MS.

Single-cell and spatial RNA sequencing identify perturbators of microglial functions with aging.

Microglia are the immune sentinels of the central nervous system with protective roles such as the removal of neurotoxic oxidized phosphatidylcholines (OxPCs). As aging alters microglial function and elevates neurological disability in diseases such as multiple sclerosis, defining aging-associated factors that cause microglia to lose their custodial properties or even become injurious can help to restore their homeostasis. We used single-cell and spatial RNA sequencing in the spinal cord of young (6-week-old) and middle-aged (52-week-old) mice to determine aging-driven microglial reprogramming at homeostasis or after OxPC injury. We identified numerous aging-associated microglial transcripts including osteopontin elevated in OxPC-treated 52-week-old mice, which correlated with greater neurodegeneration. Osteopontin delivery into the spinal cords of 6-week-old mice worsened OxPC lesions, while its knockdown in 52-week-old lesions attenuated microglial inflammation and axon loss. Thus, elevation of osteopontin and other transcripts in aging disorders including multiple sclerosis perturbs microglial functions contributing to aging-associated neurodegeneration.

B cells in central nervous system disease: diversity, locations and pathophysiology.

B cells represent a relatively minor cell population within both the healthy and diseased central nervous system (CNS), yet they can have profound effects. This is emphasized in multiple sclerosis, in which B cell-depleting therapies are arguably the most efficacious treatment for the condition. In this Review, we discuss how B cells enter and persist in the CNS and how, in many neurological conditions, B cells concentrate within CNS barriers but are rarely found in the parenchyma. We highlight how B cells can contribute to CNS pathology through antibody secretion, antigen presentation and secretion of neurotoxic molecules, using examples from CNS tumours, CNS infections and autoimmune conditions such as neuromyelitis optica and, in particular, multiple sclerosis. Overall, understanding common and divergent principles of B cell accumulation and their effects within the CNS could offer new insights into treating these devastating neurological conditions.

Prognostic Parameters and Their Relevance in Outcome of Middle Ear Surgeries.

To study various parameters, including Middle ear risk index (MERI) and their correlation with outcome of middle ear surgery. The study was conducted from September 2015 to May 2017 in Department of Otolaryngology at our institute. It included 185 cases of safe type of chronic suppurative otitis media. These patients were admitted and treated surgically and record was kept for at least 3 months follow-up in postoperated period. The study concluded that a good correlation exist between MERI and result of tympanoplasty.

Status of Contralateral Ear in Patients with Unilateral Chronic Otitis Media.

This study was conducted to evaluate the existence of otoscopic abnormality, hearing status and radiological changes in contralateral ear of patients with chronic otitis media. 300 patients having unilateral Chronic Otitis Media attending OPD in the Department of Otorhinolaryngology, Institute of Medical Sciences, Banaras Hindu University, Varanasi during the period of March 2019 to March 2020 were selected. Otoscopy, Pure Tone Audiometry and Bilateral X-ray mastoids (lateral oblique view) and/or HRCT Temporal bone were done. Contralateral ear was affected in more than 30% cases. Out of 188 patients having Mucosal COM, 58 cases (30.9%) had abnormal TM. Out of 112 patients having Squamosal COM, 48 cases (42.9%) had abnormal CLE. Out of 300 cases, 231 (77.0%) of them had normal hearing in contralateral ear. It was followed by 65 cases (21.6%) with conductive hearing loss. Mixed hearing loss and SNHL were seen in 2 patients each. In contralateral ear of Mucosal COM, pneumatic pattern of pneumatisation was seen in 69.1% followed by Diploic pattern (30.9%). In squamosal COM, X-ray mastoid showed pneumatic pattern (64.3%) followed by Diploic pattern (33.9%) in the contralateral ear. Sclerotic pattern was seen in only 1.8% of cases in contralateral ear. Chronic otitis media as a disease is not limited to one ear. The precise and critical evaluation of both ears does not play a role in prognostic evaluation of the patient only, but it can also serve as a guide for early detection of probable evolution of the disease process in a patient in contralateral ear with unilateral chronic otitis media.

Derivation of the Buffalo Concussion Physical Examination risk of delayed recovery (RDR) score to identify children at risk for persistent postconcussive symptoms.

OBJECTIVE: The Buffalo Concussion Physical Examination (BCPE) is a brief, but pertinent physical examination designed for the subacute, outpatient assessment of concussion. The purpose of this study was to perform the BCPE on a larger sample and derive a scoring system to identify children at risk for Persistent Post-Concussive Symptoms (PPCS, recovery ≥30 days). METHODS: This prospective, observational cohort study from September 2016 to March 2019 was performed at three university-affiliated concussion clinics. Male and female children (n=270, 14.92±1.86 years, range 8-18, 38% female) were diagnosed with a concussion within 14 days of injury and followed-up until recovery. Logistic regression was used with history and physical examination variables to predict PPCS and a weighted scoring metric was derived. RESULTS: Out of 15 predictor variables, the main effects of 1 preinjury variable (≥3 previous concussions), 2 injury characteristic variables (days-since-injury and type-of-injury), 3 physical examination variables (orthostatic intolerance (OI), vestibulo-ocular reflex (VOR) and tandem gait) and 2 interaction terms (OI/VOR and tandem gait/type-of-injury) produced a score that was 85% accurate for identifying children with low-risk, medium-risk and high-risk for PPCS on cross-validation. CONCLUSION: The Risk for Delayed Recovery (RDR)-Score allows physicians in an outpatient setting to more accurately predict which children are at greater risk for PPCS early after their injury, and who would benefit most from targeted therapies. The RDR-Score is intended to be used as part of a comprehensive assessment that should include validated symptom checklists, mental health history and adjunct testing (eg, cognitive or physical exertion) where clinically indicated.

Prognostic Value of Neutrophil-Lymphocyte Ratio and Platelet-Lymphocyte Ratio in Head and Neck Malignancies.

To study the prognostic significance of neutrophil-lymphocyte ratio (NLR) and platelet-lymphocyte ratio (PLR) in head and neck cancers. The study included 170 cases of histopathologically diagnosed head and neck cancer patients and 80 control subjects. NLR and PLR of patients with head and neck cancers were compared to the control group. The correlation between NLR and PLR values and factors such as age, gender, duration of symptoms, site of tumour, histological type, histological grading, T-category, N-category and TNM stages in cancer patients were analysed. NLR and PLR were statistically higher in cancer patients compared to control. There was a non-significant increase in both NLR and PLR with advancing degree of differentiation and TNM Stages of the cancer patients. A significant increase in NLR and PLR with increasing T Categories and increasing N Categories of head and neck cancer patients was obtained. NLR and PLR can be used to estimate tumour prognosis in head and neck cancers. Increased NLR and PLR values can be used as a marker for poor prognosis. However further studies with larger study groups including treatment response and surveillance should be carried out to corroborate these results.

Penetrating Neck Injuries: from ER to OR.

Neck contains several vital structures, in a small close space, in complex relationship to each other, and unprotected by any bony framework. Any injury to this crucial region, hence mostly becomes an acute emergency. Appropriately managing the same has always been a point of constant discussion amongst head and neck surgeons. The basic aim of the study was to discuss the management, comorbidities, prognosis and associated complications encountered in a series of patients with penetrating neck trauma (piercing platysma), presenting to the emergency over a period of 1 year. Combat injuries and patients declared as brought dead at the time of first examination were excluded. This was a retrospective study of patients with cut throat injury, managed at a tertiary center of northern India from June 2014 to September 2015. Following management in the ER as per ATLS guidelines, all patients were then operated for specific injuries. Graph pad software was used for statistical analysis. Of the 15 patients studied in total, 11 (73.3%) were males. The mean patient age was 33.67 years. Mean duration of presentation was 20.85 h. 60% patients had homicidal injuries. Tracheostomy and Ryle's tube insertion was done in 8 (53.3%) patients. Exploration and surgical repair was done in all patients without any mortality. 4 patients developed post-operative complications. Mean duration of hospital stay was 9.2 days. Immediate resuscitation followed by exploration and primary repair is a must in all patients of penetrating neck injury.

Autoreactive, Low-Affinity T Cells Preferentially Drive Differentiation of Short-Lived Memory B Cells at the Expense of Germinal Center Maintenance.

B cell fate decisions within a germinal center (GC) are critical to determining the outcome of the immune response to a given antigen. Here, we characterize GC kinetics and B cell fate choices in a response to the autoantigen myelin oligodendrocyte glycoprotein (MOG) and compare the response with a standard model foreign antigen. Both antigens generate productive primary responses, as evidenced by GC development, circulating antigen-specific antibodies, and differentiation of memory B cells. However, in the MOG response, the status of the cognate T cell partner drives preferential B cell differentiation to a memory phenotype at the expense of GC maintenance, resulting in a truncated GC. Reduced plasma cell differentiation is largely independent of T cell influence. Interestingly, memory-phenotype B cells formed in the MOG GC are not long lived, resulting in a failure of the B cell response to secondary challenge.

Chronobiological regulation of psychosocial and physiological outcomes in multiple sclerosis.

There is mounting scientific evidence showing the importance of innate biological rhythms on disease onset and progression. Perhaps the most important of these is the circadian rhythm, a cycle of oscillations lasting approximately 24 h. Recent work has shown that circadian rhythms are intrinsically linked to the immune system in a bidirectional fashion, and that disruption of these cycles can contribute to changes in pathology and quality of life (including fatigue, mood, and disability). This is particularly true in diseases of the nervous and immune systems. We review here the current preclinical and clinical literature to highlight interactions between circadian rhythms and multiple sclerosis, as well as its animal model, experimental autoimmune encephalomyelitis. We highlight potential benefits of chronotherapy (the temporal administration of immunomodulatory drugs) in an effort to increase treatment efficacy and reduce the negative side-effects of the drugs that often burden those suffering from the disease.

ICAM1+ neutrophils promote chronic inflammation via ASPRV1 in B cell-dependent autoimmune encephalomyelitis.

Neutrophils contribute to demyelinating autoimmune diseases, yet their phenotype and functions have been elusive to date. Here, we demonstrate that ICAM1 surface expression distinguishes extra- from intravascular neutrophils in the mouse CNS during experimental autoimmune encephalomyelitis (EAE). Transcriptomic analysis of these 2 subpopulations indicated that neutrophils, once extravasated, acquire macrophage-like properties, including the potential for immunostimulation and MHC class II-mediated antigen presentation. In corroboration, super-resolution (3D stimulated emission-depletion [STED]) microscopy revealed neutrophils forming synapses with T and B cells in situ. Further, neutrophils specifically express the aspartic retroviral-like protease ASPRV1, which increases in the CNS during EAE and severe cases of multiple sclerosis. Without ASPRV1, mice immunized with a new B cell-dependent myelin antigen (but not with the traditional myelin oligodendrocyte glycoprotein peptide) develop a chronic phase of EAE that is less severe and even completely fades in many individuals. Therefore, ICAM1+ macrophage-like neutrophils can play both shared and nonredundant roles in autoimmune demyelination, among them perpetuating inflammation via ASPRV1.

Lasing transition at 1.06 µm emission in Nd3+ -doped borate-based tellurium calcium zinc niobium oxide glasses for high-power solid-state lasers.

The spectroscopic properties of Tellurium Calcium Zinc Niobium oxide Borate (TCZNB) glasses of composition (in mol%) 10TeO2  + 15CaO + 5ZnO + 10 Nb2 O5  + (60 - x)B2 O3  + Nd2 O3 (x = 0.1, 0.5, 1.0 or 1.5 mol%) have been investigated experimentally. The three phenomenological intensity parameters Ω2 , Ω4, Ω6 have been calculated using the Judd-Ofelt theory and in turn radiative properties such as radiative transition probabilities, emission cross-sections, branching ratios and radiative lifetimes have been estimated. The trend found in the JO intensity parameter is Ω2  > Ω6  > Ω4 If Ω6  > Ω4 , the glass system is favourable for the laser emission 4 F3/2  â†’ 4 I11/2 in the infrared (IR) wavelength. The experimental values of branching ratio of 4 F3/2  â†’ 4 I11/2 transition indicate favourable lasing action with low threshold power. The evaluated total radiative transition probabilities (AT ), stimulated emission cross-section (σe ) and gain bandwidth parameters (σe  × Δλp ) were compared with earlier reports. An energy level analysis has been carried out considering the experimental energy positions of the absorption and emission bands.

An Unusual Cause of Headache and Fatigue in a Division 1 Collegiate Athlete.

Variegate porphyria (VP) is an autosomal dominant disorder of porphyrin metabolism. We report a case of a 21-year-old male collegiate athlete who complained of recurrent headache and fatigue. Extensive testing after initial presentation failed to identify a cause. Months later, his grandmother was diagnosed with VP after being hospitalized; hence, he was tested. He was positive for a heterozygous missense mutation, R168H, in one protoporphyrinogen oxidase allele. This case highlights a rare disorder of heme synthesis that should be considered in the differential diagnosis of exertional fatigue and headaches in athletes. When other more common causes of fatigue and/or headache are unable to be identified, a more focused history and examination may lead to a more unusual but crucial diagnosis. To our knowledge, there are no reported cases of this condition in Division I collegiate athletes.

Activated B Cells Participating in the Anti-Myelin Response Are Excluded from the Inflamed Central Nervous System in a Model of Autoimmunity that Allows for B Cell Recognition of Autoantigen.

Once activated, T cells gain the ability to access both healthy and inflamed nonlymphoid tissues. They are then reactivated to remain in the tissue and exert their effector function only if they encounter their specific Ag. In this study, we set out to determine if the same is true for B cells using a mouse model of CNS autoimmunity that incorporates both T and B cell recognition of a myelin autoantigen. Both T and B cells were common infiltrates of spinal cords in diseased mice. However, unlike T cells, anti-myelin B cells were excluded from the inflamed tissue. Further, CNS B cells did not have a phenotype consistent with Ag-specific activation as it occurs in lymphatic tissue. Instead, they expressed elevated levels of CD80, indicating that B cells may contribute to local inflammation through nonantigen-specific mechanisms.

Evaluation of the Veterans Health Administration's Specialty Care Transformational Initiatives to Promote Patient-Centered Delivery of Specialty Care: A Mixed-Methods Approach.

INTRODUCTION: Veteran's Affairs Office of Specialty Care (OSC) launched four national initiatives (Electronic-Consults [e-Consults], Specialty Care Access Networks-Extension for Community Healthcare Outcomes [SCAN-ECHO], Mini-Residencies, and Specialty Care Neighborhood) to improve specialty care delivery and funded a center to evaluate the initiatives. METHODS: The evaluation, guided by two implementation frameworks, provides formative (administrator/provider interviews and surveys) and summative data (quantitative data on patterns of use) about the initiatives to OSC. RESULTS: Evaluation of initiative implementation is assessed through CFIR (Consolidated Framework for Implementation Research)-grounded qualitative interviews to identify barriers/facilitators. Depending on high or low implementation, factors such as receiving workload credit, protected time, existing workflow/systems compatibility, leadership engagement, and access to information/resources were considered implementation barriers or facilitators. Findings were shared with OSC and used to further refine implementation at additional sites. Evaluation of other initiatives is ongoing. CONCLUSIONS: The mixed-methods approach has provided timely information to OSC about initiative effect and impacted OSC policies on implementation at additional sites.

Eight years of decreased methicillin-resistant Staphylococcus aureus health care-associated infections associated with a Veterans Affairs prevention initiative.

BACKGROUND: Declines in methicillin-resistant Staphylococcus aureus (MRSA) health care associated infections (HAIs) were previously reported in Veterans Affairs acute care (2012), spinal cord injury (SCIU) (2011), and long-term-care facilities (LTCFs) (2012). Here we report continuing declines in infection rates in these settings through September 2015. METHODS: Monthly data entered into a national database from 127 acute care facilities, 22 SCIUs, and 133 LTCFs were evaluated for trends using negative binomial regression. RESULTS: There were 23,153,240 intensive care unit (ICU) and non-ICU, and 1,794,234 SCIU patient-days from October 2007-September 2015, and 22,262,605 LTCF resident-days from July 2009-September 2015. Admission nasal swabbing remained >92% in all 3 venues. Admission prevalence changed from 13.2%-13.5% in acute care, from 35.1%-32.0% in SCIUs, and from 23.1%-25.0% in LTCFs during the analysis periods. Monthly HAI rates fell 87.0% in ICUs, 80.1% in non-ICUs, 80.9% in SCIUs, and 49.4% in LTCFs (all P values < .0001 for trend). During September 2015, there were 2 MRSA HAIs reported in ICUs, 20 (with 3 in SCIUs) in non-ICUs, and 31 in LTCFs nationwide. CONCLUSIONS: MRSA HAI rates declined significantly in acute care, SCIUs, and LTCFs over 8 years of the Veterans Affairs MRSA Prevention Initiative.