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Mast cells (MCs) are located in the meninges of the central nervous system (CNS), where they play key roles in the immune response. MC-deficient mice are advantageous in delineating the role of MCs in the immune response in vivo. In this study, we illustrate that a mutation in microphthalmia-associated transcription factor (Mitf) affects meningeal MC number in a dosage-dependent manner. C57BL/6J Mitf null mice lack meningeal MCs completely, whereas heterozygous mice have on average 25% fewer MCs. Mitf heterozygous mice might be a valuable tool to study the role of MCs in the meninges.
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OBJECTIVE: To test if patients recovering from COVID-19 are at increased risk of mental morbidities and to what extent such risk is exacerbated by illness severity. DESIGN: Population-based cross-sectional study. SETTING: Iceland. PARTICIPANTS: A total of 22 861 individuals were recruited through invitations to existing nationwide cohorts and a social media campaign from 24 April to 22 July 2020, of which 373 were patients recovering from COVID-19. MAIN OUTCOME MEASURES: Symptoms of depression (Patient Health Questionnaire), anxiety (General Anxiety Disorder Scale) and posttraumatic stress disorder (PTSD; modified Primary Care PTSD Screen for DSM-5) above screening thresholds. Adjusting for multiple covariates and comorbidities, multivariable Poisson regression was used to assess the association between COVID-19 severity and mental morbidities. RESULTS: Compared with individuals without a diagnosis of COVID-19, patients recovering from COVID-19 had increased risk of depression (22.1% vs 16.2%; adjusted relative risk (aRR) 1.48, 95% CI 1.20 to 1.82) and PTSD (19.5% vs 15.6%; aRR 1.38, 95% CI 1.09 to 1.75) but not anxiety (13.1% vs 11.3%; aRR 1.24, 95% CI 0.93 to 1.64). Elevated relative risks were limited to patients recovering from COVID-19 that were 40 years or older and were particularly high among individuals with university education. Among patients recovering from COVID-19, symptoms of depression were particularly common among those in the highest, compared with the lowest tertile of influenza-like symptom burden (47.1% vs 5.8%; aRR 6.42, 95% CI 2.77 to 14.87), among patients confined to bed for 7 days or longer compared with those never confined to bed (33.3% vs 10.9%; aRR 3.67, 95% CI 1.97 to 6.86) and among patients hospitalised for COVID-19 compared with those never admitted to hospital (48.1% vs 19.9%; aRR 2.72, 95% CI 1.67 to 4.44). CONCLUSIONS: Severe disease course is associated with increased risk of depression and PTSD among patients recovering from COVID-19.
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COVID-19 , Ansiedad/epidemiología , Estudios Transversales , Depresión/epidemiología , Humanos , Islandia/epidemiología , Morbilidad , SARS-CoV-2RESUMEN
Neuroplasticity forms the basis for neuronal circuit complexity and differences between otherwise similar circuits. We show that the microphthalmia-associated transcription factor (Mitf) plays a central role in intrinsic plasticity of olfactory bulb (OB) projection neurons. Mitral and tufted (M/T) neurons from Mitf mutant mice are hyperexcitable, have a reduced A-type potassium current (IA) and exhibit reduced expression of Kcnd3, which encodes a potassium voltage-gated channel subunit (Kv4.3) important for generating the IA Furthermore, expression of the Mitf and Kcnd3 genes is activity dependent in OB projection neurons and the MITF protein activates expression from Kcnd3 regulatory elements. Moreover, Mitf mutant mice have changes in olfactory habituation and have increased habituation for an odorant following long-term exposure, indicating that regulation of Kcnd3 is pivotal for long-term olfactory adaptation. Our findings show that Mitf acts as a direct regulator of intrinsic homeostatic feedback and links neuronal activity, transcriptional changes and neuronal function.
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Factor de Transcripción Asociado a Microftalmía , Bulbo Olfatorio , Animales , Ratones , Factor de Transcripción Asociado a Microftalmía/genética , Neuronas , Odorantes , OlfatoRESUMEN
Although mast cell distribution has been described in both human and canine hearts, cardiac mast cells in mice have yet to be categorically localized. We therefore sought to describe mast cell distribution within the mouse heart and characterize their dependence on the Microphthalmia-associated transcription factor (Mitf). Cardiac mast cells were visualized using Toluidine Blue and avidin staining, and their distribution within the heart described. Cardiac mast cells were most prevalent in the epicardium (50%) or myocardium (45%). Less frequently, mast cells were noted in the endocardium (5%). Within the myocardium, 31% of the mast cells had perivascular location. By studying two different Mitf mutant strains, Mitfmi-vga9 and MitfMi-wh, we demonstrated that these mutations led to near-complete deficiency of cardiac mast cells. Accordingly, expression of the mMCP-4 and mMCP-5 genes was lost and chymase enzyme activity was severely reduced. Additionally, hearts from mice heterozygous for these Mitf mutations contained significantly fewer mast cells compared to wild-type mice. Our results demonstrated that the distribution of cardiac mast cells in mice is different from humans and dogs. Cardiac mast cells are dependent on Mitf expression, with loss-of-function mutation in the Mitf gene leading to near-complete lack of cardiac mast cells. Loss of a single Mitf allele is sufficient for relative mast cell deficiency.
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Regulación de la Expresión Génica/inmunología , Mastocitos/inmunología , Factor de Transcripción Asociado a Microftalmía/inmunología , Miocardio/inmunología , Pericardio/inmunología , Animales , Perros , Humanos , Mastocitos/citología , Ratones , Ratones Transgénicos , Factor de Transcripción Asociado a Microftalmía/genética , Proteínas Quimioatrayentes de Monocitos/genética , Proteínas Quimioatrayentes de Monocitos/inmunología , Mutación , Serina Endopeptidasas/genética , Serina Endopeptidasas/inmunologíaRESUMEN
PURPOSE: The inflammatory markers chitinase-3-like protein 1 (CHI3L1) and chitotriosidase (CHIT1) have both been associated with cardiovascular complications. The aim of this preliminary observational study was to assess the roles and interaction of obstructive sleep apnea (OSA) severity and body mass index (BMI) with plasma CHI3L1 levels and CHIT1 activity in patients with moderate to severe OSA. The second aim was to assess the roles and interaction of positive airway pressure (PAP) treatment and BMI on the expression of the same proteins. METHODS: The study included 97 OSA patients with an apnea-hypopnea index (AHI) ≥ 15 and full usage of PAP treatment after 4 months. Plasma CHI3L1 levels and CHIT1 activity were measured before and after treatment. RESULTS: Multiple linear regression analysis demonstrated an independent association of BMI on CHI3L1 levels (p < 0.05) but not on CHIT1 activity. The OSA severity markers (AHI and oxygen desaturation index) did not independently or in interaction with BMI levels associate with CHI3L1 levels or with CHIT1 activity (p > 0.05). A two-way repeated measures ANOVA revealed a significant interaction between PAP treatment effect (before vs. after) and BMI groups (< 35 kg/m2 vs. ≥ 35 kg/m2) on CHI3L1 levels (p = 0.03) but not on CHIT1 activity (p = 0.98). CONCLUSIONS: Obesity independently associated with CHI3L1 levels. Association between OSA severity and CHI3L1 levels or CHIT1 activity (independent of or dependent on obesity level) could not be confirmed. However, decrease was observed in CHI3L1 levels after PAP treatment in severely obese OSA patients but not in those less obese.
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Proteína 1 Similar a Quitinasa-3/sangre , Obesidad/sangre , Obesidad/terapia , Apnea Obstructiva del Sueño/sangre , Apnea Obstructiva del Sueño/terapia , Adulto , Biomarcadores/sangre , Índice de Masa Corporal , Comorbilidad , Femenino , Humanos , Modelos Lineales , Masculino , Persona de Mediana Edad , Obesidad/complicaciones , Polisomnografía , Índice de Severidad de la Enfermedad , Apnea Obstructiva del Sueño/complicaciones , Resultado del TratamientoRESUMEN
SUMMARY: Melanocytes are pigment producing cells derived from the neural crest. They are primarily found in the skin and hair follicles, but can also be found in other tissues including the eye, ear and heart. Here, we describe the distribution of pigmented cells in C57BL/6J mouse meninges, the membranes that envelope the brain. These cells contain melanosomes of all four stages of development and they depend on Microphthalmia associated transcription factor (MITF), the master regulator of melanocyte development, suggesting that they are bona-fide melanocytes. The location of these pigmented cells is consistent with the location of meningeal melanomas in humans and animal models. SIGNIFICANCE: Here, we document and define pigmented cells in the meninges of the mouse brain and confirm that they are melanocytes. This is important for understanding the role of this cell type and for understanding primary meningeal melanoma, a rare disease that likely arises from normal meningeal melanocytes.
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INTRODUCTION: Acellular fish skin of the Atlantic cod (Gadus morhua) is being used to treat chronic wounds. The prevalence of diabetes and the comorbidity of chronic wounds is increasing globally. The aim of the study was to assess the biocompatibility and biological characteristics of acellular fish skin, important for tissue repair. MATERIALS AND METHODS: The structure of the acellular fish skin was examined with microscopy. Biocompatibility of the graft was conducted by a specialized certified laboratory. Protein extracts from the material were analyzed using gel electrophoresis. Cytokine levels were measured with an enzyme linked immunosorbent assay (ELISA). Angiogenic properties were assessed with a chick chorioallantoic membrane (chick CAM) assay. RESULTS: The structure of acellular fish skin is porous and the material is biocompatible. Electrophoresis revealed proteins around the size 115-130 kDa, indicative of collagens. The material did not have significant effect on IL-10, IL-12p40, IL-6 or TNF-α secretion from monocytes or macrophages. Acellular fish skin has significant effect on angiogenesis in the chick CAM assay. CONCLUSION: The acellular fish skin is not toxic and is not likely to promote inflammatory responses. The graft contains collagen I, promotes angiogenesis and supports cellular ingrowth. Compared to similar products made from mammalian sources, acellular fish skin does not confer a disease risk and contains more bioactive compounds, due to less severe processing.
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Dermis Acelular/metabolismo , Materiales Biocompatibles , Gadus morhua , Regeneración , Ingeniería de Tejidos/métodos , Dermis Acelular/efectos adversos , Animales , Línea Celular , Embrión de Pollo , Colágeno Tipo I/química , Colágeno Tipo I/metabolismo , Citocinas/metabolismo , Humanos , Macrófagos/metabolismo , Peso Molecular , Monocitos/metabolismo , Neovascularización FisiológicaRESUMEN
Recent evidences indicating that cellular kinase signaling cascades are triggered by oligomers of N-acetylglucosamine (ChOS) and that condrocytes of human osteoarthritic cartilage secrete the inflammation associated chitolectin YKL-40, prompted us to study the binding affinity of partially acetylated ChOS to YKL-40 and their effect on primary chondrocytes in culture. Extensive chitinase digestion and filtration of partially deacetylated chitin yielded a mixture of ChOS (Oligomin™) and further ultrafiltration produced T-ChOS™, with substantially smaller fraction of the smallest sugars. YKL-40 binding affinity was determined for the different sized homologues, revealing micromolar affinities of the larger homologues to YKL-40. The response of osteoarthritic chondrocytes to Oligomin™ and T-ChOS™ was determined, revealing 2- to 3-fold increases in cell number. About 500 µg/ml was needed for Oligomin™ and around five times lower concentration for T-ChOS™, higher concentrations abolished this effect for both products. Addition of chitotriose inhibited cellular responses mediated by larger oligosaccharides. These results, and the fact that the partially acetylated T-ChOS™ homologues should resist hydrolysis, point towards a new therapeutic concept for treating inflammatory joint diseases.
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Acetilglucosamina/metabolismo , Adipoquinas/metabolismo , Quitina/metabolismo , Condrocitos/patología , Lectinas/metabolismo , Acetilación , Adipoquinas/genética , Adipoquinas/farmacología , Recuento de Células , Supervivencia Celular , Células Cultivadas , Proteína 1 Similar a Quitinasa-3 , Condrocitos/efectos de los fármacos , Condrocitos/metabolismo , Cromatografía en Gel , Colágeno Tipo II/genética , Colágeno Tipo II/metabolismo , Medios de Cultivo/metabolismo , Humanos , Lectinas/genética , Lectinas/farmacología , Oligosacáridos/metabolismo , Osteoartritis/metabolismo , Osteoartritis/patología , Polimerizacion , Cultivo Primario de Células , Unión Proteica , Trisacáridos/genética , Trisacáridos/metabolismoRESUMEN
A mutation in the human cystatin C gene leads to familial cerebral amyloid angiopathy. This disease is known as "hereditary cerebral hemorrhage with amyloidosis-Icelandic type" or "hereditary cystatin C amyloid angiopathy." The mutant cystatin C protein forms aggregates and amyloid, within the central nervous system almost exclusively in connection with the vascular system. It was not known whether immune cells could remove mutant cystatin C protein aggregates. Ex vivo mutant cystatin C protein aggregates, both in solution and dried onto a glass surface, induced adhesion to the substrate, differentiated the THP-1 monocyte cell line and led to a proinflammatory response. Aggregates were also taken up by both THP-1 cells and THP-1 derived macrophages. These are the same responses induced by other amyloidogenic protein species, such as amyloid ß protein and amylin, supporting the model of all amyloidogenic proteins being toxic due to common structural motifs. Proinflammatory response induced by the ex vivo mutant cystatin C protein aggregates suggests that vascular inflammation plays an important role in hereditary cerebral hemorrhage with amyloidosis-Icelandic type. Ex vivo protein aggregates of cystatin C might better model cellular behavior than in vitro-generated aggregates or supplement in vitro material.
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Cistatina C/inmunología , Cistatina C/farmacología , Endocitosis/inmunología , Inflamación/inmunología , Leucemia Monocítica Aguda/inmunología , Adhesión Celular , Diferenciación Celular/efectos de los fármacos , Línea Celular Tumoral , Angiopatía Amiloide Cerebral , Cistatina C/genética , Endocitosis/efectos de los fármacos , Variación Genética/genética , HumanosRESUMEN
Metal implants and polymeric devices for the application in the clinical treatment of orthopedic tissue injuries are increasingly coated with bioactive biomaterials derived from natural substances to induce desirable biological effects. Many metals and polymers used in biomaterials research show high affinity for endotoxins, which are abundant in the environment. Endotoxin contamination is indicated in the pathology of periodontitis and aseptic implant loosening, but may also affect the evaluation of a biomaterial's bioactivity by inducing strong inflammatory reactions. In this review, we discuss the high affinity of three commonly used implant biomaterials for endotoxins and how the contamination can affect the outcome of the orthopedic fixation. The chemical nature of bacterial endotoxins and some of the clinical health implications are described, as this knowledge is critically important to tackle the issues associated with the measurement and removal of endotoxins from medical devices. Commonly used methods for endotoxin testing and removal from natural substances are examined and the lack of standard guidelines for the in vitro evaluation of biomaterials is discussed.
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Materiales Biocompatibles , Incrustaciones Biológicas/prevención & control , Endotoxinas/química , Endotoxinas/toxicidad , Animales , Prótesis Dental/efectos adversos , Humanos , Periodontitis/inducido químicamente , Periodontitis/etiología , Periodontitis/patologíaRESUMEN
Key roles of macroglia are inextricably coupled to specialized membrane domains. The perivascular endfoot membrane has drawn particular attention, as this domain contains a unique complement of aquaporin-4 (AQP4) and other channel proteins that distinguishes it from perisynaptic membranes. Recent studies indicate that the polarization of macroglia is lost in a number of diseases, including temporal lobe epilepsy and Alzheimer's disease. A better understanding is required of the molecular underpinning of astroglial polarization, particularly when it comes to the significance of the dystrophin associated protein complex (DAPC). Here, we employ immunofluorescence and immunogold cytochemistry to analyze the molecular scaffolding in perivascular endfeet in macroglia of retina and three regions of brain (cortex, dentate gyrus, and cerebellum), using AQP4 as a marker. Compared with brain astrocytes, Müller cells (a class of retinal macroglia) exhibit lower densities of the scaffold proteins dystrophin and α-syntrophin (a DAPC protein), but higher levels of AQP4. In agreement, depletion of dystrophin or α-syntrophin--while causing a dramatic loss of AQP4 from endfoot membranes of brain astrocytes--had only modest or insignificant effect, respectively, on the AQP4 pool in endfoot membranes of Müller cells. In addition, while polarization of brain macroglia was less affected by dystrophin depletion than by targeted deletion of α-syntrophin, the reverse was true for retinal macroglia. These data indicate that the molecular scaffolding in perivascular endfeet is more complex than previously assumed and that macroglia are heterogeneous with respect to the mechanisms that dictate their polarization.
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Astrocitos/metabolismo , Química Encefálica/genética , Encéfalo/metabolismo , Polaridad Celular/genética , Neuroglía/metabolismo , Retina/metabolismo , Animales , Acuaporina 4/metabolismo , Astrocitos/química , Astrocitos/ultraestructura , Encéfalo/ultraestructura , Proteínas de Unión al Calcio/deficiencia , Proteínas de Unión al Calcio/genética , Proteínas de Unión al Calcio/metabolismo , Distrofina/metabolismo , Proteínas Asociadas a la Distrofina/biosíntesis , Proteínas Asociadas a la Distrofina/deficiencia , Proteínas Asociadas a la Distrofina/genética , Inmunohistoquímica , Masculino , Proteínas de la Membrana/deficiencia , Proteínas de la Membrana/genética , Proteínas de la Membrana/metabolismo , Ratones , Ratones de la Cepa 129 , Ratones Endogámicos C57BL , Ratones Endogámicos mdx , Ratones Noqueados , Ratones Transgénicos , Proteínas Musculares/deficiencia , Proteínas Musculares/genética , Proteínas Musculares/metabolismo , Neuroglía/química , Neuroglía/ultraestructura , Retina/química , Retina/ultraestructuraRESUMEN
Expression of the water channel aquaporin-4 (AQP4) at the blood-brain interface is dependent upon the dystrophin associated protein complex. Here we investigated whether deletion of the Aqp4 gene affects the molecular composition of this protein scaffold and the integrity of the blood-brain barrier. High-resolution immunogold cytochemistry revealed that perivascular expression of α-syntrophin was reduced by 60% in Aqp4(-/-) mice. Additionally, perivascular AQP4 expression was reduced by 88% in α-syn(-/-) mice, in accordance with earlier reports. Immunofluorescence showed that Aqp4 deletion also caused a modest reduction in perivascular dystrophin, whereas ß-dystroglycan labeling was unaltered. Perivascular microglia were devoid of AQP4 immunoreactivity. Deletion of Aqp4 did not alter the ultrastructure of capillary endothelial cells, the expression of tight junction proteins (claudin-5, occludin, and zonula occludens 1), or the vascular permeability to horseradish peroxidase and Evans blue albumin dye. We conclude that Aqp4 deletion reduces the expression of perivascular glial scaffolding proteins without affecting the endothelial barrier. Our data also indicate that AQP4 and α-syntrophin are mutually dependent upon each other for proper perivascular expression.
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Acuaporina 4/deficiencia , Barrera Hematoencefálica/citología , Barrera Hematoencefálica/metabolismo , Endotelio/metabolismo , Proteína Ácida Fibrilar de la Glía/metabolismo , Neuroglía/metabolismo , Animales , Acuaporina 4/genética , Vasos Sanguíneos/metabolismo , Vasos Sanguíneos/ultraestructura , Proteínas de Unión al Calcio/metabolismo , Permeabilidad Capilar/genética , Corteza Cerebral/citología , Endotelio/ultraestructura , Azul de Evans , Regulación de la Expresión Génica/genética , Proteínas de la Membrana/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Proteínas de Microfilamentos/metabolismo , Microscopía Inmunoelectrónica , Proteínas Musculares/metabolismo , Neuroglía/ultraestructura , Molécula-1 de Adhesión Celular Endotelial de Plaqueta/metabolismoRESUMEN
It has long been a common view that neurons in the human central nervous system were not capable of self renewal. But in the mid-1990s scientists discovered that certain areas of the human brain do have the ability generate new neurons, at least under certain circumstances. It was subsecuently confirmed that the human central nervous system contains stem cells similar to the cells which originally give rise to the central nervous sysem during fetal development. The possible use of stem cells in the treatment of various neurological disorders, holds great promise. However, much research needs to be carried out before stem cell therapy can be moved from the bench to the bedside. Now researchers are pursuing two fundamental strategies to exploit the possible application of stem cells. One is to cultivate stem cells in vitro and to design the right differentiation profile of cells suitable for implantation. The other strategy relies on studying endogenous signals that could stimulate the patient s own stem cells and repair mechanisms. Here we give an overview of neural stem cells and their possible future use in the treatment of neural diseases such as Parkinson s disease, motor neuron disease and spinal cord injury.