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BACKGROUND: Contactless vital signs (VS) measurement with video photoplethysmography (vPPG), motion analysis (MA), and passive infrared thermometry (pIR) has shown promise. OBJECTIVES: To compare conventional (contact-based) and experimental contactless VS measurement approaches for emergency department (ED) walk-in triage in pandemic conditions. METHODS: Patients' heart rates (HR), respiratory rates (RR), and temperatures were measured with cardiorespiratory monitor and vPPG, manual count and MA, and contact thermometers and pIR, respectively. RESULTS: There were 475 walk-in ED patients studied (95% of eligible). Subjects were 35.2 ± 20.8 years old (range 4 daysâ95 years); 52% female, 0.2% transgender; had Fitzpatrick skin type of 2.3 ± 1.4 (range 1â6), Emergency Severity Index of 3.0 ± 0.6 (range 2â5), and contact temperature of 36.83°C (range 35.89-39.4°C) (98.3°F [96.6â103°F]). Pediatric HR and RR data were excluded from analysis due to research challenges associated with pandemic workflow. For a 30-s, unprimed "Triage" window in 377 adult patients, vPPG-MA acquired 377 (100%) HR measurements featuring a mean difference with cardiorespiratory monitor HR of 5.9 ± 12.8 beats/min (R = 0.6833) and 252 (66.8%) RR measurements featuring a mean difference with manual RR of -0.4 ± 2.6 beats/min (R = 0.8128). Subjects' Emergency Severity Index components based on conventional VS and contactless VS matched for 83.8% (HR) and 89.3% (RR). Filtering out vPPG-MA measurements with low algorithmic confidence reduced VS acquired while improving correlation with conventional measurements. The mean difference between contact and pIR temperatures was 0.83 ± 0.67°C (range -1.16-3.5°C) (1.5 ± 1.2°F [range -2.1-6.3°F]); pIR fever detection improved with post hoc adjustment for mean bias. CONCLUSION: Contactless VS acquisition demonstrated good agreement with contact methods during adult walk-in ED patient triage in pandemic conditions; clinical applications will need further study.
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Servicio de Urgencia en Hospital , Pandemias , Fotopletismografía , Termografía , Triaje , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Niño , Preescolar , Femenino , Determinación de la Frecuencia Cardíaca/métodos , Humanos , Lactante , Recién Nacido , Masculino , Persona de Mediana Edad , Fotopletismografía/métodos , Frecuencia Respiratoria , Termografía/métodos , Triaje/métodos , Signos Vitales , Adulto JovenRESUMEN
Objective: To investigate features of Guillain-Barré syndrome (GBS) following SARS-CoV-2 vaccines and evaluate for a causal link between the two. Methods: We captured cases of GBS after SARS-CoV-2 vaccination through a national, open-access, online surveillance system. For each case, the certainty of GBS was graded using the Brighton criteria, and the relationship to the vaccine was examined using modified WHO Causality Assessment criteria. We compared age distribution of cases with that of prepandemic GBS cases and clinical features with the International GBS Outcome Study (IGOS). Results: Between 1 January and 30 June 2021, we received 67 reports of GBS following the ChAdOx1 vaccine (65 first doses) and three reports following the BNT162b2 vaccine (all first doses). The causal association with the vaccine was classified as probable for 56 (80%, all ChAdOx1), possible for 12 (17%, 10 ChAdOx1) and unlikely for two (3%, 1 ChAdOx1). A greater proportion of cases occurred in the 50-59 age group in comparison with prepandemic GBS. Most common clinical variants were sensorimotor GBS (n=55; 79%) and facial diplegia with paraesthesias (n=10; 14%). 10% (n=7/69) of patients reported an antecedent infection, compared with 77% (n=502/652) of the IGOS cohort (p<0.00001). Facial weakness (63% (n=44/70) vs 36% (n=220/620); p<0.00001) and sensory dysfunction (93% (n=63/68) vs 69% (n=408/588); p=0.00005) were more common but disease severity and outcomes were similar to the IGOS study. Interpretation: Most reports of GBS followed the first dose of ChAdOx1 vaccine. While our study cannot confirm or refute causation, this observation, together with the absence of alternative aetiologies, different than expected age distribution and the presence of unusual clinical features support a causal link. Clinicians and surveillance bodies should remain vigilant to the possibility of this very rare adverse event and its atypical variants.
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This retrospective study evaluated the phenotypic and genotypic features of 14 patients with McArdle disease attending the West of Scotland adult muscle clinic. Although all patients experienced exercise-induced cramps, exercise intolerance and hyperCKaemia, only 71% (nâ¯=â¯10) experienced the second wind phenomenon, rhabdomyolysis and/or myoglobinuria. We observed a high rate of fixed muscle weakness (50%; nâ¯=â¯7), coronary artery disease (36%; nâ¯=â¯5), and psychological comorbidity (50%; nâ¯=â¯7). Although 79% had symptom onset in the first decade of life, the mean age at presentation and at genetic diagnosis was 43.8 years and 47.7 years, respectively. 93% had at least one copy of the common PYGM pathogenic variant, c.148C > T, p.(Arg50*), with 50% (nâ¯=â¯7) of the cohort being homozygous. Our cohort highlights the phenotypic variability seen in McArdle disease and underscores the potential for late-onset presentations. It emphasises the need for improved awareness and recognition of this condition amongst neurologists, rheumatologists and general physicians. A history of exercise intolerance and second wind phenomenon may not always be volunteered by the patient, underscoring the need to ask specific questions in clinic to extrapolate the relevant symptoms in this patient cohort.
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Genotipo , Enfermedad del Almacenamiento de Glucógeno Tipo V/genética , Fenotipo , Adulto , Anciano , Estudios de Cohortes , Femenino , Homocigoto , Humanos , Masculino , Persona de Mediana Edad , Debilidad Muscular/patología , Músculo Esquelético/patología , Mutación , Mioglobinuria/genética , Estudios Retrospectivos , Rabdomiólisis/genética , EscociaRESUMEN
Reports of Guillain-Barré syndrome (GBS) have emerged during the Coronavirus disease 2019 (COVID-19) pandemic. This epidemiological and cohort study sought to investigate any causative association between COVID-19 infection and GBS. The epidemiology of GBS cases reported to the UK National Immunoglobulin Database was studied from 2016 to 2019 and compared to cases reported during the COVID-19 pandemic. Data were stratified by hospital trust and region, with numbers of reported cases per month. UK population data for COVID-19 infection were collated from UK public health bodies. In parallel, but separately, members of the British Peripheral Nerve Society prospectively reported incident cases of GBS during the pandemic at their hospitals to a central register. The clinical features, investigation findings and outcomes of COVID-19 (definite or probable) and non-COVID-19 associated GBS cases in this cohort were compared. The incidence of GBS treated in UK hospitals from 2016 to 2019 was 1.65-1.88 per 100 000 individuals per year. GBS incidence fell between March and May 2020 compared to the same months of 2016-19. GBS and COVID-19 incidences during the pandemic also varied between regions and did not correlate with one another (r = 0.06, 95% confidence interval: -0.56 to 0.63, P = 0.86). In the independent cohort study, 47 GBS cases were reported (COVID-19 status: 13 definite, 12 probable, 22 non-COVID-19). There were no significant differences in the pattern of weakness, time to nadir, neurophysiology, CSF findings or outcome between these groups. Intubation was more frequent in the COVID-19 affected cohort (7/13, 54% versus 5/22, 23% in COVID-19-negative) attributed to COVID-19 pulmonary involvement. Although it is not possible to entirely rule out the possibility of a link, this study finds no epidemiological or phenotypic clues of SARS-CoV-2 being causative of GBS. GBS incidence has fallen during the pandemic, which may be the influence of lockdown measures reducing transmission of GBS inducing pathogens such as Campylobacter jejuni and respiratory viruses.
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COVID-19/epidemiología , Síndrome de Guillain-Barré/epidemiología , Adulto , Anciano , Anciano de 80 o más Años , Estudios de Cohortes , Femenino , Humanos , Masculino , Persona de Mediana Edad , Estudios Prospectivos , Estudios Retrospectivos , SARS-CoV-2 , Reino Unido/epidemiología , Adulto JovenRESUMEN
We present the case of a 29-year-old south Asian man born of consanguineous marriage, presenting with ataxia, peripheral neuropathy and cognitive impairment. An initial diagnosis of coeliac disease was thought to explain the pertinent clinical features; however, further investigation led to an additional diagnosis of the rare yet treatable autosomal recessive condition, cerebrotendinous xanthomatosis. With both conditions employing highly diverse and overlapping clinical phenotypes, this contributed to a delay in diagnosis. Our report highlights the importance of paying close attention to both the clinical phenotype and family history.
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Ataxia/genética , Enfermedad Celíaca/diagnóstico , Disfunción Cognitiva/genética , Enfermedades del Sistema Nervioso Periférico/genética , Xantomatosis Cerebrotendinosa/diagnóstico , Administración Oral , Adulto , Ataxia/tratamiento farmacológico , Enfermedad Celíaca/complicaciones , Enfermedad Celíaca/dietoterapia , Ácido Quenodesoxicólico/administración & dosificación , Colestanotriol 26-Monooxigenasa/genética , Consanguinidad , Análisis Mutacional de ADN , Diagnóstico Tardío , Dieta Sin Gluten , Humanos , Masculino , Anamnesis , Mutación Missense , Linaje , Enfermedades del Sistema Nervioso Periférico/tratamiento farmacológico , Resultado del Tratamiento , Xantomatosis Cerebrotendinosa/complicaciones , Xantomatosis Cerebrotendinosa/tratamiento farmacológico , Xantomatosis Cerebrotendinosa/genéticaRESUMEN
OBJECTIVE: To determine the prevalence of MPZ mutations that cause Charcot Marie Tooth neuropathy type 1B (CMT1B) and activate the unfolded protein Response (UPR). BACKGROUND: CMT1B is caused by >200 heterozygous mutations in MPZ, the major protein in peripheral nerve myelin. Mutations Ser63del MPZ and Arg98Cys MPZ cause the mutant protein to be retained in the ER and activate the generally adaptive UPR. Treatments that modulate UPR activation have improved cellular and rodent models of CMT1B raising the possibility that other MPZ mutations that activate the UPR would also respond favorably to similar treatment. The prevalence of MPZ mutations that activate the UPR is unknown. METHODS: We developed a dual luciferase reporter assay of Xbp1 splicing using stably transfected RT4 Schwann cells to assay the ability of cDNA constructs bearing 46 distinct MPZ mutations to activate the UPR. Constructs also carried an HA tag to permit detection of ER retention of mutant proteins. UPR activation and ER retention were correlated with clinical phenotypes. RESULTS: Eighteen mutations demonstrated ER retention and UPR activation to a similar degree as Ser63del and Arg98Cys MPZ. Thirty-five of the mutations activated the UPR > 1.5 fold compared to that of wild-type MPZ. Correlation was high between firefly and Nano-luciferase reporters and between both reporters and ER localization. UPR activity did not correlate with clinical onset or severity. CONCLUSION: Many CMT1B causing mutations activate the UPR and may be susceptible to therapeutic efforts to facilitate UPR function.
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Adult brain tumors establish an immunosuppressive tumor microenvironment as a modality of immune escape, with several immunotherapies designed to overcome this barrier. However, the relationship between tumor cells and immune cells in pediatric brain tumor patients is not as well-defined. In this study, we sought to determine whether the model of immune escape observed in adult brain tumors is reflected in patients with pediatric brain tumors by evaluating NKG2D ligand expression on tissue microarrays created from patients with a variety of childhood brain tumor diagnoses, and infiltration of Natural Killer and myeloid cells. We noted a disparity between mRNA and protein expression for the 8 known NKG2D ligands. Surprisingly, high-grade gliomas did not have increased NKG2D ligand expression compared to normal adjacent brain tissue, nor did they have significant myeloid or NK cell infiltration. These data suggest that pediatric brain tumors have reduced NK cell-mediated immune surveillance, and a less immunosuppressive tumor microenvironment as compared to their adult counterparts. These data indicate that therapies aimed to improve NK cell trafficking and functions in pediatric brain tumors may have a greater impact on anti-tumor immune responses and patient survival, with fewer obstacles to overcome.
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Neoplasias Encefálicas/metabolismo , Antígenos de Histocompatibilidad Clase I/metabolismo , Células Asesinas Naturales/inmunología , Células Mieloides/inmunología , Subfamilia K de Receptores Similares a Lectina de Células NK/metabolismo , Escape del Tumor/inmunología , Microambiente Tumoral/inmunología , Neoplasias Encefálicas/sangre , Neoplasias Encefálicas/patología , Niño , Citotoxicidad Inmunológica/inmunología , Humanos , Inmunohistoquímica , Ligandos , ARN Mensajero/metabolismo , Análisis de Matrices TisularesRESUMEN
PURPOSE: To describe an economic (Ec) model for estimating the impact of screening and treatment for retinopathy of prematurity (ROP). DESIGN: EcROP is a cost-effectiveness, cost-utility, and cost-benefit analysis. METHODS: We surveyed caregivers of 52 children at schools for the blind or pediatric eye clinics in Atlanta, Georgia and 43 in Mexico City. A decision analytic model with sensitivity analysis determined the incremental cost-effectiveness (primary outcome) and incremental monetary benefit (secondary outcome) of an ideal (100% screening) national ROP program as compared to estimates of current practice. Direct costs included screening and treatment expenditures. Indirect costs estimated lost productivity of caretaker(s) and blind individuals as determined by face-to-face surveys. Utility and effectiveness were measured in quality-adjusted life years and benefit in US dollars. EcROP includes a sensitivity analysis to assesses the incremental cost-effectiveness and societal impact of ROP screening and treatment within a country or economic region. Estimates are based on evidence-based clinical data and region-specific economic data acquired from direct field survey. RESULTS: In both Mexico and the United States, an ideal national ROP screening and treatment program was highly cost-saving. The incremental net benefit of an ideal ROP program over current practice is $5556 per child ($206 574 333 annually) and $3628 per child ($205 906 959 annually) in Mexico and the United States, respectively. CONCLUSION: EcROP demonstrates that ROP screening and treatment is highly beneficial for quality of life, cost saving, and cost-effectiveness in the United States and Mexico. EcROP can be applied to any country or region to provide data for informed allocation of limited health care resources.
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Costos de la Atención en Salud , Retinopatía de la Prematuridad/economía , Niño , Análisis Costo-Beneficio , Femenino , Humanos , Recién Nacido , Masculino , México , Modelos Económicos , Tamizaje Neonatal/economía , Años de Vida Ajustados por Calidad de Vida , Retinopatía de la Prematuridad/terapia , Estados UnidosRESUMEN
Using exome sequencing in an individual with Charcot-Marie-Tooth disease (CMT) we have identified a mutation in the X-linked dystrophin-related protein 2 (DRP2) gene. A 60-year-old gentleman presented to our clinic and underwent clinical, electrophysiological and skin biopsy studies. The patient had clinical features of a length dependent sensorimotor neuropathy with an age of onset of 50 years. Neurophysiology revealed prolonged latencies with intermediate conduction velocities but no conduction block or temporal dispersion. A panel of 23 disease causing genes was sequenced and ultimately was uninformative. Whole exome sequencing revealed a stop mutation in DRP2, c.805C>T (Q269*). DRP2 interacts with periaxin and dystroglycan to form the periaxin-DRP2-dystroglycan complex which plays a role in the maintenance of the well-characterized Cajal bands of myelinating Schwann cells. Skin biopsies from our patient revealed a lack of DRP2 in myelinated dermal nerves by immunofluorescence. Furthermore electron microscopy failed to identify Cajal bands in the patient's dermal myelinated axons in keeping with ultrastructural pathology seen in the Drp2 knockout mouse. Both the electrophysiologic and dermal nerve twig pathology support the interpretation that this patient's DRP2 mutation causes characteristic morphological abnormalities recapitulating the Drp2 knockout model and potentially represents a novel genetic cause of CMT.
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Enfermedad de Charcot-Marie-Tooth/genética , Enfermedad de Charcot-Marie-Tooth/patología , Proteínas del Citoesqueleto/genética , Proteínas de la Membrana/genética , Proteínas Musculares/genética , Enfermedad de Charcot-Marie-Tooth/fisiopatología , Dermis/inervación , Dermis/ultraestructura , Distroglicanos/metabolismo , Exoma , Humanos , Masculino , Persona de Mediana Edad , Vaina de Mielina/patología , Conducción Nerviosa , Linaje , Ubiquitina Tiolesterasa/metabolismoRESUMEN
OBJECTIVE: To investigate the effects of NEFL Glu396Lys mutation on the expression and assembly of neurofilaments (NFs) in cutaneous nerve fibers of patients with Charcot-Marie-Tooth disease type 2E (CMT2E). METHODS: A large family with CMT2E underwent clinical, electrophysiologic, and skin biopsy studies. Biopsies were processed by indirect immunofluorescence (IF), electron microscopy (EM), and Western blot analysis. RESULTS: The clinical features demonstrated intrafamilial phenotypic variability, and the electrophysiologic findings revealed nerve conductions that were either slow or in the intermediate range. All patients had reduced or absent compound muscular action potential amplitudes. Skin biopsies showed axons labeled with the axonal markers protein gene product 9.5 and α-tubulin, but not with NFs. The results of Western blot analysis were consistent with those of IF, showing reduced or absent NFs and normal expression of α-tubulin. EM revealed clusters of regenerated fibers, in absence of myelin sheath abnormalities. Both IF and EM failed to show NF aggregates in dermal axons. The morphometric analysis showed a smaller axonal caliber in patients than in controls. The study of the nodal/paranodal architecture demonstrated that sodium channels and Caspr were correctly localized in patients with CMT2E. CONCLUSIONS: Decrease in NF abundance may be a pathologic marker of CMT2E. The lack of NF aggregates, consistent with prior studies, suggests that they occur proximally leading to subsequent alterations in the axonal cytoskeleton. The small axonal caliber, along with the normal molecular architecture of nodes and paranodes, explain the reduced velocities detected in patients with CMT2E. Our results also demonstrate that skin biopsy can provide evidence of pathologic and pathogenic abnormalities in patients with CMT2E.
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Enfermedad de Charcot-Marie-Tooth/diagnóstico , Enfermedad de Charcot-Marie-Tooth/metabolismo , Regulación de la Expresión Génica , Filamentos Intermedios/metabolismo , Fibras Nerviosas/metabolismo , Adulto , Femenino , Humanos , Filamentos Intermedios/patología , Masculino , Persona de Mediana Edad , Fibras Nerviosas/patología , Linaje , Piel/metabolismo , Piel/patologíaRESUMEN
Inherited neuropathies known collectively as Charcot-Marie-Tooth disease are one of the most common inherited neurological conditions affecting â¼1 in 2500 people. A heterogenous disorder, CMT is divided into subtypes based on the pattern of inheritance and also by neurophysiological studies. Despite the clinical similarities among patients with demyelinating CMT, it is recognized that this group of disorders is both genetically and phenotypically heterogenous. Understanding the pathogenesis of these disorders requires an intimate knowledge of normal myelin development and homeostasis. Improvements in genetic testing techniques over the last 20 years have contributed majorly to the identification of specific genes, proteins, and molecular pathways that are providing the basis for understanding the disease processes and developing rational approaches to therapy.
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Enfermedad de Charcot-Marie-Tooth , Enfermedades Desmielinizantes , Enfermedad de Charcot-Marie-Tooth/clasificación , Enfermedad de Charcot-Marie-Tooth/genética , Enfermedad de Charcot-Marie-Tooth/patología , Enfermedades Desmielinizantes/clasificación , Enfermedades Desmielinizantes/genética , Enfermedades Desmielinizantes/patología , Humanos , MutaciónRESUMEN
OBJECTIVE: GNE myopathy is a rare recessive myopathy associated with inclusion bodies on muscle biopsy. The clinical phenotype is associated with distal muscle weakness with quadriceps sparing. Most of the current information on GNE myopathy has been obtained through studies of Jewish and Japanese patient cohorts carrying founder mutations in the GNE gene. However, little is known about GNE myopathy in Europe where the prevalence is thought to be very low. METHODS: Patients were referred through the National Specialist Commissioning Team service for limb-girdle muscular dystrophies at Newcastle (UK). All patients harbouring mutations in the GNE gene were recruited for our study. Detailed clinical and genetic data as well as muscle MRIs and muscle biopsies were reviewed. RESULTS: We identified 26 patients harbouring mutations in the GNE gene. Two previously reported mutations (c.1985C>T, p.Ala662Val and c.1225G>T, p.Asp409Tyr) were prevalent in the Scottish, Northern Irish and Northern English populations; with 90% of these patients carrying at least one of the two mutations. Clinically, we confirmed the homogenous pattern of selective quadriceps sparing but noted additional features like asymmetry of weakness at disease onset. CONCLUSIONS: GNE myopathy is an important diagnosis to consider in patients presenting with distal leg muscle weakness. We report, for the first time, two common mutations in the north of Britain and highlight the broader spectrum of clinical phenotypes. We also propose that the prevalence of GNE myopathy may be underestimated due to the frequent absence of rimmed vacuoles in the muscle biopsy.
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Miopatías Distales/genética , Complejos Multienzimáticos/genética , Mutación/genética , Adolescente , Adulto , Niño , Miopatías Distales/epidemiología , Miopatías Distales/patología , Femenino , Mutación del Sistema de Lectura/genética , Humanos , Imagen por Resonancia Magnética , Masculino , Debilidad Muscular/genética , Debilidad Muscular/patología , Músculo Esquelético/patología , Mutación Missense/genética , Reino Unido/epidemiología , Adulto JovenRESUMEN
INTRODUCTION: We explored the modified Rankin scale (mRS) as a tool to quantitate disability in myasthenia gravis (MG). Our aim was to correlate patients' perception of their disability with that of the care provider and determine its relationship with other MG-related scores. METHODS: We evaluated 107 MG patients at 2 neurological centers. Patients were assessed over the telephone before and after clinic visits using the 15-item Myasthenia Gravis Quality-of-Life index (MG-QOL15) and mRS. At the clinic, patients were assessed using the MG-QOL15, MG Composite (MGC), and mRS. RESULTS: The MG-QOL15 correlated with the MGC, mRS, and assessors' scores of patients. Assessors' perception of disease burden was in line with that of the patients' scoring. MG-QOL15 scores obtained over the telephone were consistent with those obtained in the clinic. Scores were generally higher in patients receiving steroids at >5 mg/day and in those receiving or seeking benefits. CONCLUSION: The MG-QOL15 and mRS are useful for estimating disability in MG.
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Evaluación de la Discapacidad , Miastenia Gravis/diagnóstico , Miastenia Gravis/fisiopatología , Anciano , Anciano de 80 o más Años , Personas con Discapacidad , Femenino , Humanos , Masculino , Persona de Mediana Edad , Miastenia Gravis/psicología , Psicometría , Calidad de Vida , Autoevaluación (Psicología) , Índice de Severidad de la Enfermedad , Estadísticas no Paramétricas , Encuestas y Cuestionarios , TeléfonoRESUMEN
Epidemiological evidence suggests that chronic treatment with simvastatin may protect against the development of Alzheimer's disease (AD), but as yet it is unclear how this effect is mediated. Extensive data also indicates that the amyloid ß-protein (Aß) plays a central role in the disease process, and it has been suggested that the protective effects of simvastatin may be mediated by reducing Aß production or by counteracting the toxic effects of Aß. Accordingly, using the AßPPswe/PS1dE9 mouse model of AD, we investigated the effects of simvastatin on long-term potentiation (LTP), amyloid biology, and two key kinases involved in Aß-mediated toxicity. Since burgeoning data indicate that both fibrillar and non-fibrillar forms of Aß play a prominent role in AD pathogenesis, we were careful to investigate the effects of simvastatin on three biochemically distinct pools of Aß. In untreated AßPPswe/PS1dE9 mice, there was a dramatic and significant increase in the levels of water-soluble Aß between 6 and 8 months, but this remained constant between 8 and 18 months. In contrast, the concentrations of detergent-soluble and formic acid (FA)-soluble Aß species increased across all ages examined, thus demonstrating that while amyloid deposition continued, the levels of water-soluble Aß remained relatively constant. LTP was normal at 6 months, but was significantly impaired at 8 and 18 months. Importantly, a diet supplemented with 0.04% simvastatin for one month (at 7 months) positively affected synaptic plasticity in AßPPswe/PS1dE9 mice and did not significantly alter levels of water-soluble, detergent-soluble, or FA-soluble Aß, but did increase phosphorylation of both Akt and GSK-3, while tau and tau phosphorylation were unaltered. These results indicate that the protective effects of simvastatin may be mediated by maintaining signaling pathways that help to protect and rescue LTP.
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Plasticidad Neuronal/efectos de los fármacos , Nootrópicos/farmacología , Simvastatina/farmacología , Sinapsis/efectos de los fármacos , Factores de Edad , Péptidos beta-Amiloides/metabolismo , Precursor de Proteína beta-Amiloide/genética , Animales , Encéfalo/efectos de los fármacos , Encéfalo/fisiopatología , Femenino , Glucógeno Sintasa Quinasa 3/metabolismo , Humanos , Técnicas In Vitro , Potenciación a Largo Plazo/efectos de los fármacos , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Fosforilación/efectos de los fármacos , Presenilina-1/genética , Proteínas Proto-Oncogénicas c-akt/metabolismo , Sinapsis/fisiología , Proteínas tau/metabolismoRESUMEN
Autoantibodies are infrequently detected in the sera of patients with the demyelinating form of Guillain-Barré syndrome most commonly encountered in the Western world, despite abundant circumstantial evidence suggesting their existence. We hypothesised that antibody specificities reliant on the cis interactions of neighbouring membrane glycolipids could explain this discrepancy, and would not have been detected by traditional serological assays using highly purified preparations of single gangliosides. To assess the frequency of glycolipid complex antibodies in a Western European cohort of patients GBS we used a newly developed combinatorial glycoarray methodology to screen against large range of antigens (11 gangliosides, 8 other single glycolipids and 162 heterodimeric glycolipid complexes). Serum samples of 181 patients from a geographically defined, Western European cohort of GBS cases were analysed, along with 161 control sera. Serum IgG binding to single gangliosides was observed in 80.0% of axonal GBS cases, but in only 11.8% of cases with demyelinating electrophysiology. The inclusion of glycolipid complexes increased the positivity rate in demyelinating disease to 62.4%. There were 40 antigens with statistically significantly increased binding intensities in GBS as compared to healthy control sera. Of these, 7 complex antigens and 1 single ganglioside also produced statistically significantly increased binding intensities in GBS versus neurological disease controls. The detection of antibodies against specific complexes was associated with particular clinical features including disease severity, requirement for mechanical ventilation, and axonal electrophysiology. This study demonstrates that while antibodies against single gangliosides are often found in cases with axonal-type electrophysiology, antibodies against glycolipid complexes predominate in cases with demyelinating electrophysiology, providing a more robust serum biomarker than has ever been previously available for such cases. This work confirms the activation of the humoral immune system in the dysimmune disease process in GBS, and correlates patterns of antigen recognition with different clinical features.
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Autoanticuerpos/sangre , Gangliósidos/inmunología , Glucolípidos/inmunología , Síndrome de Guillain-Barré/inmunología , Inmunoglobulina G/inmunología , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Niño , Femenino , Síndrome de Guillain-Barré/sangre , Humanos , Inmunoglobulina M/inmunología , Masculino , Persona de Mediana Edad , Adulto JovenRESUMEN
Pathological and clinical studies implicate antibody-dependent mechanisms in the immunopathogenesis of multiple sclerosis. We tested this hypothesis directly by investigating the ability of patient-derived immunoglobulins to mediate demyelination and axonal injury in vitro. Using a myelinating culture system, we developed a sensitive and reproducible bioassay to detect and quantify these effects and applied this to investigate the pathogenic potential of immunoglobulin G preparations obtained from patients with multiple sclerosis (n = 37), other neurological diseases (n = 10) and healthy control donors (n = 13). This identified complement-dependent demyelinating immunoglobulin G responses in approximately 30% of patients with multiple sclerosis, which in two cases was accompanied by significant complement-dependent antibody mediated axonal loss. No pathogenic immunoglobulin G responses were detected in patients with other neurological disease or healthy controls, indicating that the presence of these demyelinating/axopathic autoantibodies is specific for a subset of patients with multiple sclerosis. Immunofluorescence microscopy revealed immunoglobulin G preparations with demyelinating activity contained antibodies that specifically decorated the surface of myelinating oligodendrocytes and their contiguous myelin sheaths. No other binding was observed indicating that the response is restricted to autoantigens expressed by terminally differentiated myelinating oligodendrocytes. In conclusion, our study identifies axopathic and/or demyelinating autoantibody responses in a subset of patients with multiple sclerosis. This observation underlines the mechanistic heterogeneity of multiple sclerosis and provides a rational explanation why some patients benefit from antibody depleting treatments.
Asunto(s)
Axones/metabolismo , Inmunoglobulina G/farmacología , Esclerosis Múltiple/diagnóstico , Esclerosis Múltiple/inmunología , Proteínas de la Mielina/metabolismo , Vaina de Mielina/metabolismo , Adulto , Anciano , Animales , Axones/efectos de los fármacos , Moléculas de Adhesión Celular/metabolismo , Enfermedades Desmielinizantes/inducido químicamente , Enfermedades Desmielinizantes/tratamiento farmacológico , Relación Dosis-Respuesta a Droga , Embrión de Mamíferos , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Inmunoglobulina G/sangre , Inmunoglobulina G/uso terapéutico , Estudios Longitudinales , Masculino , Persona de Mediana Edad , Esclerosis Múltiple/terapia , Vaina de Mielina/efectos de los fármacos , Factores de Crecimiento Nervioso/metabolismo , Proteínas de Neurofilamentos/metabolismo , Neuronas/citología , Neuronas/efectos de los fármacos , Intercambio Plasmático , Ratas , Médula Espinal/citología , Adulto JovenRESUMEN
Glycolipid-protein interactions are increasingly recognised as critical to numerous and diverse biological processes, including immune recognition, cell-cell signalling, pathogen adherence, and virulence factor binding. Previously, such carbohydrate-lectin interactions have been assessed in vitro largely by assaying protein binding against purified preparations of single glycolipids. Recent observations show that certain disease-associated autoantibodies and other lectins bind only to complexes formed by two different gangliosides. However, investigating such 1:1 glycolipid complexes can prove technically arduous. To address this problem, we have developed a semi-automated system for assaying lectin binding to large numbers of glycolipid complexes simultaneously. This employs an automated thin-layer chromatography sampler. Single glycolipids and their heterodimeric complexes are prepared in microvials. The autosampler is then used to print reproducible arrays of glycolipid complexes onto polyvinylidene difluoride membranes affixed to glass slides. A printing density of 300 antigen spots per slide is achievable. Following overnight drying, these arrays can then be probed with the lectin(s) of interest. Detection of binding is by way of a horseradish peroxidase-linked secondary antibody driving a chemiluminescent reaction rendered on radiographic film. Image analysis software can then be used to measure signal intensity for quantification.
Asunto(s)
Glucolípidos/metabolismo , Lectinas/metabolismo , Análisis por Micromatrices/métodos , Glucolípidos/química , Lectinas/química , Análisis por Micromatrices/instrumentaciónRESUMEN
The presence of oligoclonal bands of IgG (OCB) in cerebrospinal fluid (CSF) is used to establish a diagnosis of multiple sclerosis (MS), but their specificity has remained an enigma since its first description over forty years ago. We now report that the use of lipid arrays identifies heteromeric complexes of myelin derived lipids as a prominent target for this intrathecal B cell response.
Asunto(s)
Inmunoglobulina G/líquido cefalorraquídeo , Metabolismo de los Lípidos , Esclerosis Múltiple , Vaina de Mielina/metabolismo , Bandas Oligoclonales/inmunología , Análisis de Varianza , Animales , Linfocitos B/inmunología , Linfocitos B/metabolismo , Células Cultivadas , Embrión de Mamíferos , Humanos , Esclerosis Múltiple/líquido cefalorraquídeo , Esclerosis Múltiple/inmunología , Esclerosis Múltiple/patología , Proteína Básica de Mielina/metabolismo , Vaina de Mielina/inmunología , Antígenos O/metabolismo , Ratas , Ratas Sprague-Dawley , Médula Espinal/citologíaRESUMEN
Glycolipids are major components of the plasma membrane, interacting with themselves, other lipids, and proteins to form an array of heterogeneous domains with diverse biological properties. Considerable effort has been focused on identifying protein binding partners for glycolipids and the glycan specificity for these interactions, largely achieved through assessing interactions between proteins and homogenous, single species glycolipid preparations. This approach risks overlooking both the enhancing and attenuating roles of heterogeneous glycolipid complexes in modulating lectin binding. Here we report a simple method for assessing lectin-glycolipid interactions. An automatic thin-layer chromatography sampler is employed to create easily reproducible arrays of glycolipids and their heterodimeric complexes immobilized on a synthetic polyvinyl-difluoride membrane. This array can then be probed with much smaller quantities of reagents than would be required using existing techniques such as ELISA and thin-layer chromatography with immuno-overlay. Using this protocol, we have established that the binding of bacterial toxins, lectins, and antibodies can each be attenuated, enhanced, or unaffected in the presence of glycolipid complexes, as compared with individual, isolated glycolipids. These findings underpin the wide-ranging influence and importance of glycolipid-glycolipid cis interactions when the nature of protein-carbohydrate recognition events is being assessed.
Asunto(s)
Glucolípidos/química , Lectinas/química , Análisis por Micromatrices/métodos , Anticuerpos/química , Toxinas Bacterianas/química , Sitios de Unión , Cromatografía en Capa Delgada , Polisacáridos/químicaRESUMEN
Anti-GM1 ganglioside autoantibodies are used as diagnostic markers for motor axonal peripheral neuropathies and are believed to be the primary mediators of such diseases. However, their ability to bind and exert pathogenic effects at neuronal membranes is highly inconsistent. Using human and mouse monoclonal anti-GM1 antibodies to probe the GM1-rich motor nerve terminal membrane in mice, we here show that the antigenic oligosaccharide of GM1 in the live plasma membrane is cryptic, hidden on surface domains that become buried for a proportion of anti-GM1 antibodies due to a masking effect of neighboring gangliosides. The cryptic GM1 binding domain was exposed by sialidase treatment that liberated sialic acid from masking gangliosides including GD1a or by disruption of the live membrane by freezing or fixation. This cryptic behavior was also recapitulated in solid-phase immunoassays. These data show that certain anti-GM1 antibodies exert potent complement activation-mediated neuropathogenic effects, including morphological damage at living terminal motor axons, leading to a block of synaptic transmission. This occurred only when GM1 was topologically available for antibody binding, but not when GM1 was cryptic. This revised understanding of the complexities in ganglioside membrane topology provides a mechanistic account for wide variations in the neuropathic potential of anti-GM1 antibodies.