RESUMO
Background: Amyotrophic lateral sclerosis (ALS) is one of the most frequently occurring neurodegenerative diseases affecting speech and swallowing. This preliminary study aimed to investigate whether an autologous lineage-negative stem/progenitor cell therapy applied to ALS patients affects the level of selected trophic and proinflammatory factors, and subsequently improves the articulation. Methods: We enrolled 12 patients with sporadic ALS, who underwent autologous bone marrow-derived lineage negative (LIN-) cells administration into cerebrospinal fluid (CSF). We evaluated patients' articulation using the Frenchay Dysarthria Assessment on days 0 and 28 following the LIN- cells administration. Concentrations of various factors (BDNF, NGF, ANGP-2, VEGF, PDGF-AA, PEDF, COMP-FH, CRP, C3, C4) in CSF were quantified by multiplex fluorescent bead-based immunoassays in the samples collected on the day of LIN- cells administration and 28 days later. On top of this, we assessed levels of BDNF and NGF in the patients' plasma on the day of the injection, three, seven days and three months after the treatment. Results: Of the 12 patients who received the LIN- cell therapy 8 showed short-termed improvement in articulatory functions (group I), which was particularly noticeable in better phonation time, lips and soft palate performance, swallowing reflex and voice loudness. Four patients (group II) did not show substantial improvement. CSF concentrations of BDNF, ANGP-2 and PDGF-AA in group I decreased significantly 28 days after LIN- cells administration. The highest concentration levels of BDNF in group II and NGF in both groups in blood plasma were observed on day 3 following the injection. Conclusions: The outcomes of the LIN- cell application in ALS treatment of articulatory organs are promising. The procedure proved to be safe and feasible. A short-lasting trophic effect of autologous LIN- administration could encourage repeated cell's application in order to sustain their beneficial effects, however this approach needs further investigation.
Assuntos
Esclerose Lateral Amiotrófica/terapia , Terapia Baseada em Transplante de Células e Tecidos/métodos , Transplante de Células-Tronco Mesenquimais , Fatores de Crescimento Neural/líquido cefalorraquidiano , Adulto , Esclerose Lateral Amiotrófica/líquido cefalorraquidiano , Esclerose Lateral Amiotrófica/genética , Esclerose Lateral Amiotrófica/patologia , Linhagem da Célula/genética , Feminino , Humanos , Masculino , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Pessoa de Meia-Idade , Fatores de Crescimento Neural/genéticaRESUMO
OBJECTIVE: To determine the safety and efficacy of mesenchymal stem cell (MSC)-neurotrophic factor (NTF) cells (NurOwn®, autologous bone marrow-derived MSCs, induced to secrete NTFs) delivered by combined intrathecal and intramuscular administration to participants with amyotrophic lateral sclerosis (ALS) in a phase 2 randomized controlled trial. METHODS: The study enrolled 48 participants randomized 3:1 (treatment: placebo). After a 3-month pretransplant period, participants received 1 dose of MSC-NTF cells (n = 36) or placebo (n = 12) and were followed for 6 months. CSF was collected before and 2 weeks after transplantation. RESULTS: The study met its primary safety endpoint. The rate of disease progression (Revised ALS Functional Rating Scale [ALSFRS-R] slope change) in the overall study population was similar in treated and placebo participants. In a prespecified rapid progressor subgroup (n = 21), rate of disease progression was improved at early time points (p < 0.05). To address heterogeneity, a responder analysis showed that a higher proportion of treated participants experienced ≥1.5 points/month ALSFRS-R slope improvement compared to placebo at all time points, and was significant in rapid progressors at 4 and 12 weeks (p = 0.004 and 0.046, respectively). CSF neurotrophic factors increased and CSF inflammatory biomarkers decreased in treated participants (p < 0.05) post-transplantation. CSF monocyte chemoattractant protein-1 levels correlated with ALSFRS-R slope improvement up to 24 weeks (p < 0.05). CONCLUSION: A single-dose transplantation of MSC-NTF cells is safe and demonstrated early promising signs of efficacy. This establishes a clear path forward for a multidose randomized clinical trial of intrathecal autologous MSC-NTF cell transplantation in ALS. CLASSIFICATION OF EVIDENCE: This phase II study provides Class I evidence.
Assuntos
Esclerose Lateral Amiotrófica/terapia , Transplante de Células-Tronco Mesenquimais/métodos , Fatores de Crescimento Neural/líquido cefalorraquidiano , Adulto , Idoso , Método Duplo-Cego , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Transplante AutólogoRESUMO
BACKGROUND: Neurotrophins, especially brain-derived neurotrophic factor (BDNF) have gained significant therapeutic interest particularly in neurologic and psychiatric disorders and they have been found in human breast milk of mothers who suffered from adverse outcomes in pregnancy. This study tested the hypothesis that oral administration of BDNF/GDNF (glial cell line-derived neurotrophic factor) can exert a biological effect in a rat model of severe neuropathology induced by olfactory bulbectomy (OBX), which exhibits dysregulation of BDNF signaling and impaired blood-brain barrier. METHODS: Adult male albino Sprague-Dawley rats underwent the OBX surgery and separate groups of OBX and sham-operated controls received one oral dose of vehicle, BDNF (0.005 mg/kg), GDNF (0.03 mg/kg) or their combination. One week after neurotrophin dosing the rats were sacrificed and BDNF level was assessed by ELISA in the blood serum and cerebrospinal fluid. RESULTS: A significant decrease of serum BDNF level was found in the OBX model. This alteration was normalized by all types of treatment BDNF, GDNF, or their combination. No influence of sham surgery or treatment was observed in the control rats. BDNF levels in cerebrospinal fluid were below detection limit. CONCLUSION: This study indicates that oral administration of neurotrophins is able to exert a biological effect in the OBX model. There is a number of potential mechanisms, which remain to be elucidated.
Assuntos
Barreira Hematoencefálica/metabolismo , Encefalopatias/sangue , Fator Neurotrófico Derivado do Encéfalo/sangue , Fatores de Crescimento Neural/sangue , Administração Oral , Animais , Transporte Biológico , Barreira Hematoencefálica/efeitos dos fármacos , Fator Neurotrófico Derivado do Encéfalo/administração & dosagem , Fator Neurotrófico Derivado do Encéfalo/líquido cefalorraquidiano , Modelos Animais de Doenças , Masculino , Fatores de Crescimento Neural/administração & dosagem , Fatores de Crescimento Neural/líquido cefalorraquidiano , Bulbo Olfatório/cirurgia , Estudo de Prova de Conceito , Ratos Sprague-Dawley , Proteínas RecombinantesRESUMO
The central nervous system (CNS) plays a central role in the control of sensory and motor functions, and the disruption of its barriers can result in severe and debilitating neurological disorders. Neurotrophins are promising therapeutic agents for neural regeneration in the damaged CNS. However, their penetration across the blood-brain barrier remains a formidable challenge, representing a bottleneck for brain and spinal cord therapy. Herein, a nanocapsule-based delivery system is reported that enables intravenously injected nerve growth factor (NGF) to enter the CNS in healthy mice and nonhuman primates. Under pathological conditions, the delivery of NGF enables neural regeneration, tissue remodeling, and functional recovery in mice with spinal cord injury. This technology can be utilized to deliver other neurotrophins and growth factors to the CNS, opening a new avenue for tissue engineering and the treatment of CNS disorders and neurodegenerative diseases.
Assuntos
Barreira Hematoencefálica/metabolismo , Nanocápsulas/química , Fatores de Crescimento Neural/farmacologia , Regeneração Nervosa/efeitos dos fármacos , Traumatismos da Medula Espinal/tratamento farmacológico , Resinas Acrílicas/química , Animais , Materiais Biocompatíveis/química , Barreira Hematoencefálica/ultraestrutura , Reagentes de Ligações Cruzadas/química , Liberação Controlada de Fármacos , Injeções Intravenosas , Macaca mulatta , Metacrilatos/química , Camundongos Endogâmicos BALB C , Fatores de Crescimento Neural/administração & dosagem , Fatores de Crescimento Neural/sangue , Fatores de Crescimento Neural/líquido cefalorraquidiano , Células PC12 , Permeabilidade , Fosforilcolina/análogos & derivados , Fosforilcolina/química , Poliésteres/química , Ratos , Traumatismos da Medula Espinal/patologia , Traumatismos da Medula Espinal/fisiopatologiaRESUMO
Over the last decade, finding a reliable biomarker for the early detection of schizophrenia (Scz) has been a topic of interest. The main goal of the current review is to provide a comprehensive view of the brain, blood, cerebrospinal fluid (CSF), and serum biomarkers of Scz disease. Imaging studies have demonstrated that the volumes of the corpus callosum, thalamus, hippocampal formation, subiculum, parahippocampal gyrus, superior temporal gyrus, prefrontal and orbitofrontal cortices, and amygdala-hippocampal complex were reduced in patients diagnosed with Scz. It has been revealed that the levels of interleukin 1ß (IL-1ß), IL-6, IL-8, and TNF-α were increased in patients with Scz. Decreased mRNA levels of brain-derived neurotrophic factor (BDNF), tropomyosin receptor kinase B (TrkB), neurotrophin-3 (NT-3), nerve growth factor (NGF), and vascular endothelial growth factor (VEGF) genes have also been reported in Scz patients. Genes with known strong relationships with this disease include BDNF, catechol-O-methyltransferase (COMT), regulator of G-protein signaling 4 (RGS4), dystrobrevin-binding protein 1 (DTNBP1), neuregulin 1 (NRG1), Reelin (RELN), Selenium-binding protein 1 (SELENBP1), glutamic acid decarboxylase 67 (GAD 67), and disrupted in schizophrenia 1 (DISC1). The levels of dopamine, tyrosine hydroxylase (TH), serotonin or 5-hydroxytryptamine (5-HT) receptor 1A and B (5-HTR1A and 5-HTR1B), and 5-HT1B were significantly increased in Scz patients, while the levels of gamma-aminobutyric acid (GABA), 5-HT transporter (5-HTT), and 5-HT receptor 2A (5-HTR2A) were decreased. The increased levels of SELENBP1 and Glycogen synthase kinase 3 subunit α (GSK3α) genes in contrast with reduced levels of B-cell translocation gene 1 (BTG1), human leukocyte antigen DRB1 (HLA-DRB1), heterogeneous nuclear ribonucleoprotein A3 (HNRPA3), and serine/arginine-rich splicing factor 1 (SFRS1) genes have also been reported. This review covers various dysregulation of neurotransmitters and also highlights the strengths and weaknesses of studies attempting to identify candidate biomarkers.
Assuntos
Encéfalo/metabolismo , Esquizofrenia/sangue , Esquizofrenia/líquido cefalorraquidiano , Biomarcadores/sangue , Biomarcadores/líquido cefalorraquidiano , Biomarcadores/metabolismo , Fator Neurotrófico Derivado do Encéfalo/sangue , Fator Neurotrófico Derivado do Encéfalo/líquido cefalorraquidiano , Fator Neurotrófico Derivado do Encéfalo/metabolismo , Catecol O-Metiltransferase/sangue , Catecol O-Metiltransferase/líquido cefalorraquidiano , Catecol O-Metiltransferase/metabolismo , Moléculas de Adesão Celular Neuronais/sangue , Moléculas de Adesão Celular Neuronais/líquido cefalorraquidiano , Moléculas de Adesão Celular Neuronais/metabolismo , Disbindina/sangue , Disbindina/líquido cefalorraquidiano , Disbindina/metabolismo , Proteínas da Matriz Extracelular/sangue , Proteínas da Matriz Extracelular/líquido cefalorraquidiano , Proteínas da Matriz Extracelular/metabolismo , Humanos , Fatores de Crescimento Neural/sangue , Fatores de Crescimento Neural/líquido cefalorraquidiano , Fatores de Crescimento Neural/metabolismo , Proteínas do Tecido Nervoso/sangue , Proteínas do Tecido Nervoso/líquido cefalorraquidiano , Proteínas do Tecido Nervoso/metabolismo , Neurotrofina 3 , Córtex Pré-Frontal/metabolismo , Proteína Reelina , Esquizofrenia/diagnóstico por imagem , Serina Endopeptidases/sangue , Serina Endopeptidases/líquido cefalorraquidiano , Serina Endopeptidases/metabolismo , Fator A de Crescimento do Endotélio VascularRESUMO
SCOPE: The aim of this study was to develop and evaluate a parallel reaction monitoring mass spectrometry (PRM-MS) assay consisting of a panel of potential protein biomarkers in cerebrospinal fluid (CSF). EXPERIMENTAL DESIGN: Thirteen proteins were selected based on their association with neurodegenerative diseases and involvement in synaptic function, secretory vesicle function, or innate immune system. CSF samples were digested and two to three peptides per protein were quantified using stable isotope-labeled peptide standards. RESULTS: Coefficients of variation were generally below 15%. Clinical evaluation was performed on a cohort of 10 patients with Alzheimer's disease (AD) and 15 healthy subjects. Investigated proteins of the granin family exhibited the largest difference between the patient groups. Secretogranin-2 (p<0.005) and neurosecretory protein VGF (p<0.001) concentrations were lowered in AD. For chromogranin A, two of three peptides had significantly lowered AD concentrations (p<0.01). The concentrations of the synaptic proteins neurexin-1 and neuronal pentraxin-1, as well as neurofascin were also significantly lowered in AD (p<0.05). The other investigated proteins, ß2-microglobulin, cystatin C, amyloid precursor protein, lysozyme C, neurexin-2, neurexin-3, and neurocan core protein, were not significantly altered. CONCLUSION AND CLINICAL RELEVANCE: PRM-MS of protein panels is a valuable tool to evaluate biomarker candidates for neurodegenerative disorders.
Assuntos
Doença de Alzheimer/genética , Proteína C-Reativa/genética , Moléculas de Adesão Celular Neuronais/genética , Moléculas de Adesão Celular/genética , Cromogranina A/genética , Fatores de Crescimento Neural/genética , Proteínas do Tecido Nervoso/genética , Secretogranina II/genética , Idoso , Doença de Alzheimer/líquido cefalorraquidiano , Doença de Alzheimer/diagnóstico , Doença de Alzheimer/patologia , Sequência de Aminoácidos , Biomarcadores/líquido cefalorraquidiano , Proteína C-Reativa/líquido cefalorraquidiano , Proteínas de Ligação ao Cálcio , Estudos de Casos e Controles , Moléculas de Adesão Celular/líquido cefalorraquidiano , Moléculas de Adesão Celular Neuronais/líquido cefalorraquidiano , Cromatografia Líquida , Cromogranina A/líquido cefalorraquidiano , Feminino , Expressão Gênica , Humanos , Masculino , Pessoa de Meia-Idade , Fatores de Crescimento Neural/líquido cefalorraquidiano , Proteínas do Tecido Nervoso/líquido cefalorraquidiano , Moléculas de Adesão de Célula Nervosa , Fragmentos de Peptídeos/líquido cefalorraquidiano , Projetos Piloto , Proteólise , Proteômica/métodos , Secretogranina II/líquido cefalorraquidiano , Espectrometria de Massas em Tandem/métodosRESUMO
Pigment-epithelium derived factor (PEDF) is a neurotrophic factor with neuroprotective, anti-tumorigenic, and anti-angiogenic effects. Elevated levels of PEDF have previously been proposed as a cerebrospinal fluid (CSF) biomarker for Alzheimer's disease. However, the origin of PEDF in CSF, i.e. whether it is derived from the brain or from the systemic circulation, and the specificity of this finding hitherto remained unclear. Here, we analyzed levels of PEDF in paired CSF and serum samples by ELISA in patients with Alzheimer's disease (AD, n=12), frontotemporal dementia (FTD, n=6), vascular dementia (n=4), bacterial meningitis (n=8), multiple sclerosis (n=32), pseudotumor cerebri (n=36), and diverse non-inflammatory neurological diseases (n=19). We established CSF/serum quotient diagrams to determine the fraction of intrathecally synthesized PEDF in CSF. We found that PEDF is significantly increased in CSF of patients with AD, FTD, and bacterial meningitis. Remarkably, PEDF concentrations were also significantly elevated in serum of patients with AD. CSF/serum quotient diagrams demonstrated that elevated PEDF concentrations in CSF of patients with AD are mostly due to elevated PEDF concentrations in serum. These findings underscore the importance of relating concentrations of proteins in CSF to their respective concentrations in serum to avoid erroneous interpretations of increased protein concentrations in lumbar CSF.
Assuntos
Doença de Alzheimer/líquido cefalorraquidiano , Proteínas do Olho/líquido cefalorraquidiano , Fatores de Crescimento Neural/líquido cefalorraquidiano , Serpinas/líquido cefalorraquidiano , Adulto , Fatores Etários , Idoso , Doença de Alzheimer/sangue , Análise de Variância , Ensaio de Imunoadsorção Enzimática , Proteínas do Olho/sangue , Feminino , Demência Frontotemporal/líquido cefalorraquidiano , Humanos , Masculino , Pessoa de Meia-Idade , Fatores de Crescimento Neural/sangue , Serpinas/sangueAssuntos
Encefalite/diagnóstico , Encefalite/genética , Pesquisa/tendências , 8-Hidroxi-2'-Desoxiguanosina , Doença Aguda , Biomarcadores/sangue , Biomarcadores/líquido cefalorraquidiano , Criança , Citocromos c/líquido cefalorraquidiano , Desoxiguanosina/análogos & derivados , Desoxiguanosina/líquido cefalorraquidiano , Eletroencefalografia , Encefalite/fisiopatologia , Encefalite/terapia , Humanos , Interleucina-6/líquido cefalorraquidiano , Monitorização Fisiológica , Mutação , Canal de Sódio Disparado por Voltagem NAV1.2/genética , Fatores de Crescimento Neural/sangue , Fatores de Crescimento Neural/líquido cefalorraquidiano , Fosfopiruvato Hidratase/líquido cefalorraquidiano , Prognóstico , Subunidade beta da Proteína Ligante de Cálcio S100/sangue , Subunidade beta da Proteína Ligante de Cálcio S100/líquido cefalorraquidiano , Subunidade beta-1 do Canal de Sódio Disparado por Voltagem/genéticaRESUMO
Several types of animal models have been developed to investigate Alzheimer's disease (AD). Okadaic acid (OA), a potent inhibitor of phosphatases 1 and 2A, induces characteristics that resemble AD-like pathology. Memory impairment induced by intra-hippocampal injection of OA has been reported, accompanied by remarkable neuropathological changes including hippocampal neurodegeneration, a paired helical filament-like phosphorylation of tau protein, and formation of ß-amyloid containing plaque-like structures. Rats were submitted to bilateral intrahippocampal okadaic acid-injection (100 ng) and, 12 days after the surgery, behavioral and biochemical tests were performed. Using this model, we evaluated spatial cognitive deficit and neuroglial alterations, particularly astroglial protein markers such as glial fibrillary acidic protein (GFAP) and S100B, metabolism of glutamate, oxidative parameters and alterations in MAPKs. Our results indicate significant hippocampal changes, including increased GFAP, protein oxidation, and phosphorylation of p38(MAPK); and decreases in glutathione content, transporter EAAT2/GLT-1, and glutamine synthetase activity as well as a decrease in cerebrospinal fluid S100B. No alterations were observed in glutamate uptake activity and S100B content. In conclusion, the OA-induced model of dementia caused spatial cognitive deficit and oxidative stress in this model and, for the first time to our knowledge, specific astroglial alterations. Findings contribute to understanding diseases accompanied by cognitive deficits and the neural damage induced by AO administration.
Assuntos
Demência/metabolismo , Modelos Animais de Doenças , Hipocampo/efeitos dos fármacos , Neuroglia/metabolismo , Animais , Transtornos Cognitivos/líquido cefalorraquidiano , Transtornos Cognitivos/complicações , Transtornos Cognitivos/metabolismo , Demência/líquido cefalorraquidiano , Demência/induzido quimicamente , Demência/complicações , Demência/psicologia , Transportador 2 de Aminoácido Excitatório/metabolismo , Proteína Glial Fibrilar Ácida/metabolismo , Glutamato-Amônia Ligase/metabolismo , Ácido Glutâmico/metabolismo , Glutationa/metabolismo , Hipocampo/metabolismo , Humanos , Masculino , Microinjeções , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Fatores de Crescimento Neural/líquido cefalorraquidiano , Fatores de Crescimento Neural/metabolismo , Ácido Okadáico/administração & dosagem , Oxirredução , Estresse Oxidativo/efeitos dos fármacos , Ratos , Ratos Wistar , Subunidade beta da Proteína Ligante de Cálcio S100 , Proteínas S100/líquido cefalorraquidiano , Proteínas S100/metabolismoRESUMO
BACKGROUND: Inflammatory responses in brain are primarily mediated by microglia, but growing evidence suggests a crucial importance of astrocytes. S100B, a calcium-binding protein secreted by astrocytes, has properties of a neurotrophic or an inflammatory cytokine. However, it is not known whether primary signals occurring during induction of an inflammatory response (e.g. lipopolysaccharide, LPS) directly modulate S100B. METHODS: In this work, we evaluated whether S100B levels in cerebrospinal fluid (CSF) and serum of Wistar rats are affected by LPS administered by intraperitoneal (IP) or intracerebroventricular (ICV) injection, as well as whether primary astrocyte cultures respond directly to lipopolysaccharide. RESULTS: Our data suggest that S100B secretion in brain tissue is stimulated rapidly and persistently (for at least 24 h) by ICV LPS administration. This increase in CSF S100B was transient when LPS was IP administered. In contrast to these S100B results, we observed an increase in in TNFα levels in serum, but not in CSF, after IP administration of LPS. In isolated astrocytes and in acute hippocampal slices, we observed a direct stimulation of S100B secretion by LPS at a concentration of 10 µg/mL. An involvement of TLR4 was confirmed by use of specific inhibitors. However, lower levels of LPS in astrocyte cultures were able to induce a decrease in S100B secretion after 24 h, without significant change in intracellular content of S100B. In addition, after 24 h exposure to LPS, we observed a decrease in astrocytic glutathione and an increase in astrocytic glial fibrillary acidic protein. CONCLUSIONS: Together, these data contribute to the understanding of the effects of LPS on astrocytes, particularly on S100B secretion, and help us to interpret cerebrospinal fluid and serum changes for this protein in neuroinflammatory diseases. Moreover, non-brain S100B-expressing tissues may be differentially regulated, since LPS administration did not lead to increased serum levels of S100B.
Assuntos
Astrócitos/efeitos dos fármacos , Astrócitos/metabolismo , Lipopolissacarídeos/farmacologia , Fatores de Crescimento Neural/líquido cefalorraquidiano , Fatores de Crescimento Neural/metabolismo , Proteínas S100/líquido cefalorraquidiano , Proteínas S100/metabolismo , Animais , Astrócitos/citologia , Células Cultivadas , Córtex Cerebral/citologia , Glutationa/metabolismo , Infusões Intraventriculares , Lipopolissacarídeos/administração & dosagem , Ratos , Ratos Wistar , Subunidade beta da Proteína Ligante de Cálcio S100 , Fator de Necrose Tumoral alfa/sangue , Fator de Necrose Tumoral alfa/líquido cefalorraquidianoRESUMO
Brain damage related to intrathecal methotrexate in children with acute lymphoblastic leukemia (ALL) is still unclear. Neuroinflammatory mechanisms and intracerebral production of specific biomarkers, play a key role in determining neuroprotective mechanisms after brain injury. To determine whether the CSF concentrations of neuron-specific enolase (NSE), neurotrophic factors and doublecortin (DCX) are influenced by repeated intrathecal methotrexate administrations, we prospectively collected CSF samples from 10 children with ALL and 10 controls. Our results showed an increased expression of the liquoral markers. This up-regulation could be interpreted as a neuroprotective response of the brain against the neuronal damages induced by MTX.
Assuntos
Lesões Encefálicas/diagnóstico , Metotrexato/efeitos adversos , Proteínas Associadas aos Microtúbulos/líquido cefalorraquidiano , Fatores de Crescimento Neural/líquido cefalorraquidiano , Neuropeptídeos/líquido cefalorraquidiano , Fármacos Neuroprotetores/líquido cefalorraquidiano , Fosfopiruvato Hidratase/líquido cefalorraquidiano , Leucemia-Linfoma Linfoblástico de Células Precursoras/líquido cefalorraquidiano , Adolescente , Antimetabólitos Antineoplásicos/efeitos adversos , Biomarcadores Tumorais/líquido cefalorraquidiano , Western Blotting , Lesões Encefálicas/líquido cefalorraquidiano , Lesões Encefálicas/induzido quimicamente , Criança , Pré-Escolar , Proteínas do Domínio Duplacortina , Proteína Duplacortina , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Masculino , Leucemia-Linfoma Linfoblástico de Células Precursoras/tratamento farmacológico , Leucemia-Linfoma Linfoblástico de Células Precursoras/patologia , Prognóstico , Estudos Prospectivos , Regulação para CimaRESUMO
BACKGROUND: Central nervous system (CNS) manifestations of hypothyroidism have been associated with cerebrovascular complications. Reports of cerebrospinal fluid (CSF) abnormalities are rare in hypothyroid dogs. OBJECTIVE: The aim of this study was to determine if chronic hypothyroidism causes blood-brain-barrier (BBB) abnormalities that are detectable using indirect CSF biomarkers. METHODS: The study included 18 normal, euthyroid, female mixed-breed dogs. Hypothyroidism was induced by (131) iodine administration in 9 dogs; 9 served as untreated controls. Evaluations included physical and neurologic examination, complete CSF analysis, serum and CSF protein electrophoresis, measurement of plasma vascular endothelial growth factor (VEGF) and serum S-100B concentrations, and calculation of CSF albumin quota (AQ) and were conducted at baseline and 6, 12, and 18 months after induction of hypothyroidism. Data were analyzed using repeated measures ANOVA. RESULTS: At baseline, differences between groups were not detected for any variable. Throughout the study, controls dogs remained free of neurologic disease and had test variables that remained within reference intervals. Two hypothyroid dogs developed CNS signs during the study, and evidence of cerebrovascular disease was found at necropsy. At 12 and 18 months, the CSF total protein, VEGF, S-100B, and fractional albumin concentrations, and AQ were significantly higher (P<.04) in hypothyroid dogs than controls. Among test variables assayed in serum or plasma, the only significant difference was a higher S-100B concentration in hypothyroid dogs (P=.003) at 18 months. CONCLUSIONS: BBB integrity is disrupted in chronic hypothyroidism. Significant increases in CSF concentrations of VEGF and S100-B in hypothyroid dogs indicate dysfunction in both endothelial and glial elements of the BBB.
Assuntos
Barreira Hematoencefálica , Doenças do Cão/líquido cefalorraquidiano , Hipotireoidismo/veterinária , Animais , Cães , Eletroforese em Gel de Ágar/veterinária , Ensaio de Imunoadsorção Enzimática/veterinária , Feminino , Hipotireoidismo/líquido cefalorraquidiano , Hipotireoidismo/complicações , Fatores de Crescimento Neural/sangue , Fatores de Crescimento Neural/líquido cefalorraquidiano , Subunidade beta da Proteína Ligante de Cálcio S100 , Proteínas S100/sangue , Proteínas S100/líquido cefalorraquidiano , Fator A de Crescimento do Endotélio Vascular/sangue , Fator A de Crescimento do Endotélio Vascular/líquido cefalorraquidianoRESUMO
There is an urgent need for both the scientific development and clinical validation of novel therapies for acute spinal cord injury (SCI). The scientific development of novel therapies would be facilitated by a better understanding of the acute pathophysiology of human SCI. Clinical validation of such therapies would be facilitated by the availability of biomarkers with which to stratify injury severity and predict neurological recovery. Cerebrospinal fluid (CSF) samples were obtained over a period of 72 h in 27 patients with complete SCI (ASIA A) or incomplete SCI (ASIA B or C). Cytokines were measured in CSF and serum samples using a multiplex cytokine array system and standard enzyme-linked immunosorbent assay (ELISA) techniques. Neurological recovery was monitored, and patient-reported neuropathic pain was documented. IL-6, IL-8, MCP-1, tau, S100beta, and glial fibrillary acidic protein (GFAP) were elevated in a severity-dependent fashion. A biochemical model was established using S100beta, GFAP, and IL-8 to predict injury severity (ASIA A, B, or C). Using these protein concentrations at 24-h post injury, the model accurately predicted the observed ASIA grade in 89% of patients. Furthermore, segmental motor recovery at 6 months post injury was better predicted by these CSF proteins than with the patients' baseline ASIA grade. The pattern of expression over the first 3 to 4 days post injury of a number of inflammatory cytokines such as IL-6, IL-8, and MCP-1 provides invaluable information about the pathophysiology of human SCI. A prediction model that could use such biological data to stratify injury severity and predict neurological outcome may be extremely useful for facilitating the clinical validation of novel treatments in acute human SCI.
Assuntos
Citocinas/líquido cefalorraquidiano , Mediadores da Inflamação/líquido cefalorraquidiano , Mielite/líquido cefalorraquidiano , Traumatismos da Medula Espinal/líquido cefalorraquidiano , Traumatismos da Medula Espinal/diagnóstico , Medula Espinal/metabolismo , Adulto , Biomarcadores/análise , Biomarcadores/sangue , Biomarcadores/líquido cefalorraquidiano , Quimiocina CCL2/análise , Quimiocina CCL2/sangue , Quimiocina CCL2/líquido cefalorraquidiano , Citocinas/análise , Citocinas/sangue , Ensaio de Imunoadsorção Enzimática , Feminino , Proteína Glial Fibrilar Ácida/análise , Proteína Glial Fibrilar Ácida/sangue , Proteína Glial Fibrilar Ácida/líquido cefalorraquidiano , Humanos , Mediadores da Inflamação/análise , Interleucina-6/análise , Interleucina-6/sangue , Interleucina-6/líquido cefalorraquidiano , Interleucina-8/análise , Interleucina-8/sangue , Interleucina-8/líquido cefalorraquidiano , Masculino , Pessoa de Meia-Idade , Mielite/imunologia , Mielite/fisiopatologia , Fatores de Crescimento Neural/análise , Fatores de Crescimento Neural/sangue , Fatores de Crescimento Neural/líquido cefalorraquidiano , Valor Preditivo dos Testes , Prognóstico , Estudos Prospectivos , Recuperação de Função Fisiológica/fisiologia , Subunidade beta da Proteína Ligante de Cálcio S100 , Proteínas S100/análise , Proteínas S100/sangue , Proteínas S100/líquido cefalorraquidiano , Medula Espinal/imunologia , Medula Espinal/fisiopatologia , Traumatismos da Medula Espinal/fisiopatologia , Índices de Gravidade do Trauma , Adulto Jovem , Proteínas tau/análise , Proteínas tau/sangue , Proteínas tau/líquido cefalorraquidianoRESUMO
Several biomarkers are used in confirming the diagnosis of cognitive disorders. This study evaluates whether the level of these markers after heart surgery correlates with the development of cognitive dysfunction, which is a frequent complication of cardiac interventions. Concentrations of amyloid-ß peptide, tau, and S100ß in the cerebro-spinal fluid were assessed, as well as cognitive functions were evaluated before and after coronary artery bypass grafting, utilizing immuno-assays and psychometric tests, respectively. A drastic rise in the level of S100ß was observed one week after the surgery, a mark of a severe generalized cerebral injury. The level of amyloid-ß peptide significantly decreased, whereas the concentration of tau markedly increased six months postoperatively. Gradual cognitive decline was also present. These findings clearly demonstrate post-surgical cognitive impairment associated with changes in biomarkers similar to that seen in Alzheimer's disease, suggesting a unifying pathognomic factor between the two disorders. A holistic approach to coronary heart disease and Alzheimer's type dementia is proposed.
Assuntos
Doença de Alzheimer/líquido cefalorraquidiano , Ponte de Artéria Coronária/efeitos adversos , Idoso , Doença de Alzheimer/diagnóstico , Peptídeos beta-Amiloides/líquido cefalorraquidiano , Biomarcadores/líquido cefalorraquidiano , Transtornos Cognitivos/líquido cefalorraquidiano , Transtornos Cognitivos/diagnóstico , Doença da Artéria Coronariana/líquido cefalorraquidiano , Doença da Artéria Coronariana/cirurgia , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Fatores de Crescimento Neural/líquido cefalorraquidiano , Testes Neuropsicológicos , Fragmentos de Peptídeos/líquido cefalorraquidiano , Projetos Piloto , Complicações Pós-Operatórias/líquido cefalorraquidiano , Complicações Pós-Operatórias/diagnóstico , Subunidade beta da Proteína Ligante de Cálcio S100 , Proteínas S100/líquido cefalorraquidiano , Proteínas tau/líquido cefalorraquidianoRESUMO
Although the exact cause of Alzheimer's disease remains elusive, many possible risk factors and pathological alterations have been used in the elaboration of in vitro and in vivo models of this disease in rodents, including intracerebral infusion of streptozotocin (STZ). Using this model, we evaluated spatial cognitive deficit and neurochemical hippocampal alterations, particularly astroglial protein markers such as glial fibrillary acidic protein (GFAP) and S100B, glutathione content, nitric oxide production, and cerebrospinal fluid (CSF) S100B. In addition, prevention of these alterations by aminoguanidine administration was evaluated. Results confirm a spatial cognitive deficit and nitrative stress in this dementia model as well as specific astroglial alterations, particularly S100B accumulation in the hippocampus and decreased CSF S100B. The hippocampal astroglial activation occurred independently of the significant alteration in GFAP content. Moreover, all these alterations were completely prevented by aminoguanidine administration, confirming the neuroprotective potential of this compound, but suggesting that nitrative stress and/or glycation may be underlying these alterations. These findings contribute to the understanding of diseases accompanied by cognitive deficits and the STZ-model of dementia.
Assuntos
Demência , Inibidores Enzimáticos/uso terapêutico , Guanidinas/uso terapêutico , Hipocampo/patologia , Estreptozocina , Análise de Variância , Animais , Comportamento Animal/efeitos dos fármacos , Demência/induzido quimicamente , Demência/patologia , Demência/prevenção & controle , Modelos Animais de Doenças , Proteína Glial Fibrilar Ácida/metabolismo , Glutationa/metabolismo , Hipocampo/efeitos dos fármacos , Hipocampo/metabolismo , Masculino , Aprendizagem em Labirinto/efeitos dos fármacos , Fatores de Crescimento Neural/líquido cefalorraquidiano , Óxido Nítrico/metabolismo , Ratos , Ratos Wistar , Tempo de Reação/efeitos dos fármacos , Subunidade beta da Proteína Ligante de Cálcio S100 , Proteínas S100/líquido cefalorraquidianoRESUMO
OBJECTIVE: Thoracic and thoracoabdominal aortic surgery is sometimes complicated by subacute or delayed paraplegia. Pro-inflammatory cytokine interleukin-1 (IL-1) beta has been implicated in extensive inflammation and progressive neurodegeneration after ischemia. Using a rabbit model, we investigated the neuroprotective effects of IL-1 receptor antagonist (IL-1ra) in a temporal fashion. METHODS: Spinal cord ischemia was induced by aortic cross-clamping in New Zealand White rabbits. The animals were assigned to three groups. Group C (n = 20) received saline (0.2-mL) and Group I (n = 20) received IL-1ra (200-microg/0.2-mL) intrathecally just after reperfusion. Group S (n = 3) underwent sham operation without aortic occlusion. We assessed the neuroprotective effects of IL-1ra by evaluating neurological function, histopathological changes, and in-situ terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL staining). We also measured the levels of Nitric Oxide (NO) and S100beta in cerebrospinal fluid (CSF). Each evaluation was performed sequentially within 120 hours after reperfusion. RESULTS: Group C showed progressive deterioration of motor function which became statistically significant from 48 hours after the onset of reperfusion (P < .05, P < .01, P < .001, P < .001 at 48, 72, 96, and 120 hours, respectively). Compared to Group C, a higher number of viable neurons was observed with less severe spinal cord injury in Group I (P < .01, .05 and .05 at 24, 72, and 120 hours, respectively). TUNEL-positive neurons were also significantly reduced by the administration of IL-1ra (P <.01 and .05 at 24, and 120 hours, respectively). The difference between Group C and Group I with regard to NO was significant at 72 and 120 hours (P < .05), while that in terms of S100beta was significant only at 24 hours (P < .05). CONCLUSIONS: Administration of IL-1ra attenuates spinal cord ischemic-reperfusion injury as evidenced by reducing both neuronal necrosis and apoptosis.
Assuntos
Proteína Antagonista do Receptor de Interleucina 1/farmacologia , Fármacos Neuroprotetores/farmacologia , Isquemia do Cordão Espinal/tratamento farmacológico , Animais , Apoptose/efeitos dos fármacos , Modelos Animais de Doenças , Humanos , Marcação In Situ das Extremidades Cortadas , Injeções Espinhais , Proteína Antagonista do Receptor de Interleucina 1/administração & dosagem , Masculino , Atividade Motora/efeitos dos fármacos , Necrose , Fatores de Crescimento Neural/líquido cefalorraquidiano , Neurônios/efeitos dos fármacos , Neurônios/patologia , Fármacos Neuroprotetores/administração & dosagem , Óxido Nítrico/líquido cefalorraquidiano , Coelhos , Proteínas Recombinantes/farmacologia , Subunidade beta da Proteína Ligante de Cálcio S100 , Proteínas S100/líquido cefalorraquidiano , Índice de Gravidade de Doença , Isquemia do Cordão Espinal/líquido cefalorraquidiano , Isquemia do Cordão Espinal/patologia , Isquemia do Cordão Espinal/fisiopatologia , Fatores de TempoRESUMO
S100B expression, particularly extracellular S100B, is used as a parameter of glial activation and/or death in several situations of brain injury. Several immunoassays for S100B measurement are available, which differ with regard to specificity, sensitivity, sample application, and, of course, economic costs. We standardized two protocols for S100B measurement (range between 1.9pg and 10ng/mL) in human and rat samples from brain and adipose tissues, blood serum, cerebrospinal fluid, urine and cell culture. Abundance and secretion of this protein in adipose tissue reinforces the caution about its origin in blood serum. Interestingly, S100B recognition was affected by the redox status of the protein. This aspect should be considered in S100B measurement, assuming that oxidized and reduced forms possibly coexist in vivo and the equilibrium can be modified by oxidative stress of physiological or pathological conditions or even by obtaining sample conditions.
Assuntos
Ensaio de Imunoadsorção Enzimática/métodos , Fatores de Crescimento Neural/análise , Neuroquímica/métodos , Neuroglia/química , Proteínas S100/análise , Tecido Adiposo/metabolismo , Adulto , Animais , Astrócitos/metabolismo , Encéfalo/metabolismo , Química Encefálica/fisiologia , Linhagem Celular Tumoral , Células Cultivadas , Feminino , Humanos , Recém-Nascido , Masculino , Fatores de Crescimento Neural/sangue , Fatores de Crescimento Neural/líquido cefalorraquidiano , Neuroglia/metabolismo , Oxirredução , Estresse Oxidativo , Ratos , Ratos Wistar , Subunidade beta da Proteína Ligante de Cálcio S100 , Proteínas S100/sangue , Proteínas S100/líquido cefalorraquidianoRESUMO
BACKGROUND: Neurotrophic factors play a crucial role in the stimulation of sprouting, synaptic plasticity and reorganization after spinal cord damage. The aim of this study was to investigate the expression of some neurotrophic factors [brain derived neurotrophic factor (BDNF), glial derived neurotrophic factor (GDNF), and nerve growth factor (NGF)] in the cerebrospinal fluid (CSF) of newborns with myelomeningocele (MMC) and to determine their correlations with this malformation. METHODS: To measure the expression of BDNF, GDNF, and NGF, we collected CSF samples of six newborns during the neurosurgical operation to correct the open MMC and of 10 matched controls. Endogenous neurotrophic factor levels were quantified using a two-site immuno-enzymatic assay. The statistical analysis was performed using the Mann-Whitney two-tailed two-sample test. FINDINGS: In the CSF of patients analysis of neurotrophic factor expression showed a significant increase of BDNF, GDNF, and NGF compared to the mean level of the control group (445.8+/-82.3, 86.5+/-2.6, and 59.9+/-6.2 pg/mL, respectively, respect to 10.2+/-5.9, 19.9+/-11.3, and 15.3+/-2.6 pg/mL) (p<0.001). INTERPRETATION: Our study shows an over-expression of neurotrophic factors in the CSF of newborns with MMC. This neurotrophin up-regulation may stimulate axonal sprouting and synaptic reorganization of the damaged neural cells at the site of spinal cord lesion. The neurotrophic factor up-regulation may represent a particularly important biochemical markers of spinal cord damage and might be associated with the severity of spine injury in MMC patients.
Assuntos
Recém-Nascido/metabolismo , Meningomielocele/metabolismo , Fatores de Crescimento Neural/biossíntese , Axônios/fisiologia , Fator Neurotrófico Derivado do Encéfalo/biossíntese , Fator Neurotrófico Derivado do Encéfalo/líquido cefalorraquidiano , Feminino , Fator Neurotrófico Derivado de Linhagem de Célula Glial/biossíntese , Fator Neurotrófico Derivado de Linhagem de Célula Glial/líquido cefalorraquidiano , Humanos , Técnicas Imunoenzimáticas , Masculino , Fatores de Crescimento Neural/líquido cefalorraquidianoRESUMO
UNLABELLED: All data obtained in experimental animal pain models support the role of nerve growth factor (NGF) as a putative candidate intervening in the pathogenesis of chronic pain, including chronic daily headache (CDH). Few studies have been carried out to establish its role in maintaining pain states in humans. The present study was aimed at investigating cerebrospinal fluid (CSF) levels of NGF and brain-derived neurotrophic factor (BDNF), both measured by sensitive immunoassay, in 20 chronic migraine (CM) patients and 20 patients affected by primary fibromyalgia syndrome (PFMS), compared with those of 20 age-matched control subjects. Significantly higher levels of both neurotrophins and glutamate were found. A significantly positive correlation emerged between CSF values of BDNF and those of NGF (r = .61, P < .001; r = .53, P < .01) and glutamate (r = .44, P < .02; r = .51, P < .01) in CM and PFMS patients, respectively. These findings suggest the possibility of a NGF-mediated up-regulation of BDNF involved in the pathophysiological events underlying long-term neuroplastic changes in persistent chronic painful conditions, such as CM and fibromyalgia. NGF might indirectly exert its effect through enhancing glutamatergic transmission via BDNF. The above mechanisms could account for sustained central sensitization in both chronic pain states. PERSPECTIVE: This article presents findings of higher NGF and BDNF levels correlated to increased glutamate levels in the CSF of both chronic migraine and fibromyalgia patients. This opens new insights into the pathogenic mechanisms of chronic pain and offers clinicians new therapeutic perspectives targeting the above mechanisms in both painful disorders.
Assuntos
Fibromialgia/líquido cefalorraquidiano , Fibromialgia/fisiopatologia , Ácido Glutâmico/líquido cefalorraquidiano , Transtornos de Enxaqueca/líquido cefalorraquidiano , Transtornos de Enxaqueca/fisiopatologia , Fatores de Crescimento Neural/líquido cefalorraquidiano , Adulto , Encéfalo/metabolismo , Encéfalo/fisiopatologia , Química Encefálica/fisiologia , Fator Neurotrófico Derivado do Encéfalo/análise , Fator Neurotrófico Derivado do Encéfalo/líquido cefalorraquidiano , Doença Crônica , Feminino , Ácido Glutâmico/análise , Humanos , Imunoensaio , Masculino , Fator de Crescimento Neural/análise , Fator de Crescimento Neural/líquido cefalorraquidiano , Fatores de Crescimento Neural/análise , Valor Preditivo dos Testes , Transmissão Sináptica/fisiologia , Regulação para Cima/fisiologiaRESUMO
OBJECTIVES: Thoracic endovascular aortic repair is associated with postoperative spinal cord ischemia in approximately 1 to 12.5% of all cases. S100beta is a protein that is released during acute damage of the central nervous system. This study was performed to determine the concentration of S100beta in cerebrospinal fluid during and after stenting of the thoracic aorta in patients at high risk for spinal cord ischemia. DESIGN: Prospective clinical study. MATERIALS AND METHODS: Eight patients who underwent elective thoracic aortic stent grafting underwent lumbar spinal fluid drainage. These patients were at high risk to develop spinal cord ischemia. METHODS: CSF samples for analysis of S100beta protein were drawn after induction of anesthesia, during stenting, once every hour the following six hours and 20 hours after repair. RESULTS: No significant increase in S100beta protein could be detected in CSF and no neurological deficits were detected postoperatively. CONCLUSIONS: The results of this study show us that there is no significant release of S100beta protein in CSF during stenting of the thoracic aorta in this subgroup of patients at high risk for spinal cord ischemia, consistent with clinical exam that there was no significant damage to the central nervous system.