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1.
Acta Cir Bras ; 35(4): e202000406, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32578724

RESUMO

PURPOSE: To investigate the role of Rosmarinic acid (RA) in the prevention of traumatic brain injury and the immunohistochemical analysis of IBA-1 and GFAP expressions. METHODS: Healthy male rats were randomly divided into 3 groups consisting of 10 rats. Groups were as follows; control group, traumatic brain injury (TBI) group, and TBI+RA group. After traumatic brain injury, blood samples were taken from the animals and analyzed with various biochemical markers. And then IBA-1 and GFAP expressions were evaluated immunohistochemically. RESULTS: Significant results were obtained in all biochemical parameters between groups. Immunohistochemical sections showed IBA-1 not only in microglia and macrophage activity but also in degenerative neurons in blood vessel endothelial cells. However, GFAP reaction and post-traumatic rosmarinic acid administration showed positive expression in astrocytes with regular structure around the blood vessel. CONCLUSION: Rosmarinic acid in blood vessel endothelial cells showed that preserving the integrity of astrocytic structure in the blood brain barrier may be an important antioxidant.


Assuntos
Lesões Encefálicas Traumáticas/prevenção & controle , Proteínas de Ligação ao Cálcio/análise , Cinamatos/farmacologia , Craniotomia/métodos , Depsídeos/farmacologia , Proteína Glial Fibrilar Ácida/análise , Proteínas dos Microfilamentos/análise , Fármacos Neuroprotetores/farmacologia , Animais , Astrócitos/efeitos dos fármacos , Lesões Encefálicas Traumáticas/patologia , Lesões Encefálicas Traumáticas/cirurgia , Glutationa Peroxidase/análise , Imuno-Histoquímica , Masculino , Malondialdeído/análise , Distribuição Aleatória , Ratos Sprague-Dawley , Valores de Referência , Reprodutibilidade dos Testes
2.
Mil Med ; 185(Suppl 1): 383-389, 2020 01 07.
Artigo em Inglês | MEDLINE | ID: mdl-32074315

RESUMO

INTRODUCTION: Military and civil aviation have documented physiological episodes among aircrews. Therefore, continued efforts are being made to improve the internal environment. Studies have shown that exposures to many organic compounds present in emissions are known to cause a variety of physiological symptoms. We hypothesize that these compounds may reversibly inhibit acetylcholinesterase, which may disrupt synaptic signaling. As a result, neural proteins leak through the damaged blood-brain barrier into the blood and in some, elicit an autoimmune response. MATERIALS AND METHODS: Neural-specific autoantibodies of immunoglobulin-G (IgG) class were estimated by the Western blotting technique in the sera of 26 aircrew members and compared with the sera of 19 normal healthy nonaircrew members, used as controls. RESULTS: We found significantly elevated levels of circulating IgG-class autoantibodies to neurofilament triplet proteins, tubulin, microtubule-associated tau proteins (Tau), microtubule-associated protein-2, myelin basic protein, and glial fibrillary acidic protein, but not S100 calcium-binding protein B compared to healthy controls. CONCLUSION: Repetitive physiological episodes may initiate cellular injury, leading to neuronal degeneration in selected individuals. Diagnosis and intervention should occur at early postinjury periods. Use of blood-based biomarkers to assess subclinical brain injury would help in both diagnosis and treatment.


Assuntos
Militares/estatística & dados numéricos , Fenômenos Fisiológicos/fisiologia , Medicina Aeroespacial/métodos , Medicina Aeroespacial/estatística & dados numéricos , Aeronaves , Autoanticorpos/análise , Autoanticorpos/sangue , Biomarcadores/análise , Biomarcadores/sangue , Western Blotting/métodos , Proteína Glial Fibrilar Ácida/análise , Proteína Glial Fibrilar Ácida/sangue , Humanos , Imunoglobulina G/análise , Imunoglobulina G/sangue , Proteínas Associadas aos Microtúbulos/análise , Proteínas Associadas aos Microtúbulos/sangue , Proteína Básica da Mielina/análise , Proteína Básica da Mielina/sangue , Proteínas de Neurofilamentos/análise , Proteínas de Neurofilamentos/sangue , Proteínas S100/análise , Proteínas S100/sangue , Tubulina (Proteína)/análise , Tubulina (Proteína)/sangue
3.
Mil Med ; 185(Suppl 1): 279-285, 2020 01 07.
Artigo em Inglês | MEDLINE | ID: mdl-32074333

RESUMO

INTRODUCTION: Posttraumatic stress disorder (PTSD) can develop during the aftermath of traumatic events. Although many are impacted by several stressors, nearly 3.6% suffer from PTSD in the United States with higher incidence reported in military service personnel. Any injury to the blood-brain barrier can ignite an array of biological signaling molecules in the immune-privileged brain parenchyma, which can disrupt the synaptic neural network, resulting in altered behavior. MATERIALS AND METHODS: In this preliminary study, we compared 20 PTSD veterans with age-matched healthy veterans to identify plasma levels of brain-specific protein markers using enzyme-linked immunosorbent assay/immunofluorometric sandwich assay for neurotrophic factors and neuropoietic cytokines, and catalytic activity of matrix metalloproteinase (MMP) by zymography. RESULTS: We observed an increased level of glial fibrillary acidic protein, tumor necrosis factor-alpha, interleukin 6, and MMP2 and MMP9 but decreased level of brain-derived neurotrophic factor, nerve growth factor-beta, and negligible difference in astroglial marker S100 calcium-binding protein B compared to controls. CONCLUSION: Identification of neural biomarkers is essential to understand the subclinical symptoms for the diagnosis PTSD, which may not be visible by magnetic resonance imaging (MRI/fMRI) and may take years to clinically manifest.


Assuntos
Transtornos de Estresse Pós-Traumáticos/etiologia , Adulto , Sintomas Comportamentais/etiologia , Sintomas Comportamentais/psicologia , Fator Neurotrófico Derivado do Encéfalo/análise , Fator Neurotrófico Derivado do Encéfalo/sangue , Feminino , Proteína Glial Fibrilar Ácida/análise , Proteína Glial Fibrilar Ácida/sangue , Humanos , Interleucina-6/análise , Interleucina-6/sangue , Imagem por Ressonância Magnética/métodos , Masculino , Pessoa de Meia-Idade , Fator de Crescimento Neural/análise , Fator de Crescimento Neural/sangue , Subunidade beta da Proteína Ligante de Cálcio S100/análise , Subunidade beta da Proteína Ligante de Cálcio S100/sangue , Transtornos de Estresse Pós-Traumáticos/diagnóstico , Transtornos de Estresse Pós-Traumáticos/psicologia , Fator de Necrose Tumoral alfa/análise , Fator de Necrose Tumoral alfa/sangue , Estados Unidos
4.
Mil Med ; 185(Suppl 1): 197-204, 2020 01 07.
Artigo em Inglês | MEDLINE | ID: mdl-32074362

RESUMO

OBJECTIVES: Serum repositories are foundations for seroepidemiological data, revealing targeted information about morbidities and existing heterogeneity in human populations. With the recent technological advances, we can perform high-throughput screening at an affordable cost using minimal plasma. Monitoring brain health after an injury is critical since mild Traumatic Brain Injury (mTBI) and other neurological symptoms are under-diagnosed. Our objective in this study is to present our preliminary serological data from one of our ongoing studies on mTBI. METHODS: In this retrospective study, we used stored plasma samples to understand biomarkers of mTBI. We compared plasma samples from five patients with mTBI following their first concussive episode to five gender and age-matched healthy controls. We assessed multiple biomarkers to show the importance of biorepositories. RESULTS: Most of the estimated plasma factors in mTBI subjects at baseline were comparable to normal healthy individuals except for the astroglial markers S100B and glial fibrillary acidic protein. Fluctuations of these biomarkers can affect the homeostasis of brain parenchyma by altering the neural network signaling, which in turn may result in intermittent behavioral symptoms. CONCLUSION: Biorepositories are powerful resources for understanding the spectrum of morbidity. Biomarkers serve as a valuable diagnostic and therapeutic tool.


Assuntos
Biomarcadores/análise , Concussão Encefálica/sangue , Guerra , Adulto , Biomarcadores/sangue , Concussão Encefálica/fisiopatologia , Fator Neurotrófico Derivado do Encéfalo/análise , Fator Neurotrófico Derivado do Encéfalo/sangue , Estudos de Coortes , Complemento C3/análise , Proteína 1 de Resposta de Crescimento Precoce/análise , Proteína 1 de Resposta de Crescimento Precoce/sangue , Feminino , Proteína Glial Fibrilar Ácida/análise , Proteína Glial Fibrilar Ácida/sangue , Humanos , Interleucina-6/análise , Interleucina-6/sangue , Estudos Longitudinais , Masculino , Fator de Ativação de Plaquetas/análise , Estudos Retrospectivos , Subunidade beta da Proteína Ligante de Cálcio S100/análise , Subunidade beta da Proteína Ligante de Cálcio S100/sangue , Fator de Necrose Tumoral alfa/análise , Fator de Necrose Tumoral alfa/sangue
5.
Toxicol Mech Methods ; 30(3): 208-218, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31779506

RESUMO

Diallyl sulfide (DAS) is a garlic-derived organosulfur compound. The current study was planned to evaluate the protecting effects of DAS against cyclophosphamide (CP)-induced nephropathic encephalopathy. DAS (100 mg/kg) was orally administered for 4 days, 60 min after the last dose, rats were injected with CP (150 mg/kg). DAS treatment before CP significantly decreased serum urea, creatinine, sodium, potassium, calcium, blood urea nitrogen (BUN), C-reactive protein (CRP), interleukin-6 (IL-6), interleukin-1ß (IL-1ß) and tumor necrosis factor-alpha (TNF-α) compared with CP-treated rats. DAS treatment decreased malondialdehyde (MDA) and increased superoxide dismutase (SOD) and reduced glutathione (GSH) levels in the renal tissues and significantly attenuated the elevated neurotransmitters N-methyl-D-aspartate/adenosine triphosphate (NMDA), γ-aminobutyric acid (GABA) levels and remarkably restored neuronal nitric oxide (NO) level and nitric oxide synthase (nNOS) activity in the brain compared to CP-treated rats. DAS for 4 consecutive days before CP showed moderate positive immunohistochemically expression of the glial fibrillary acidic protein (GFAP) in the brain and kidney tissues comparable to CP-treated rats. DAS afforded renal and neuroprotection against CP-induced nephropathic encephalopathy due to its capacity to ameliorates the afore-mentioned biochemical parameters which were supported by histopathological and immunohistochemically examination.


Assuntos
Compostos Alílicos/farmacologia , Encefalopatias/induzido quimicamente , Ciclofosfamida/toxicidade , Nefropatias/induzido quimicamente , Sulfetos/farmacologia , Animais , Encéfalo/metabolismo , Encéfalo/patologia , Encefalopatias/prevenção & controle , Proteína C-Reativa/análise , Citocinas/sangue , Proteína Glial Fibrilar Ácida/análise , Rim/metabolismo , Rim/patologia , Nefropatias/prevenção & controle , Masculino , Estresse Oxidativo/efeitos dos fármacos , Ratos , Receptores de N-Metil-D-Aspartato/efeitos dos fármacos
6.
Sci Rep ; 9(1): 16161, 2019 11 06.
Artigo em Inglês | MEDLINE | ID: mdl-31695063

RESUMO

The central mechanisms underlying the marked beneficial metabolic effects of bariatric surgery are unclear. Here, we characterized global gene expression in the hypothalamic arcuate nucleus (Arc) in diet-induced obese (DIO) rats following Roux-en-Y gastric bypass (RYGB). 60 days post-RYGB, the Arc was isolated by laser-capture microdissection and global gene expression was assessed by RNA sequencing. RYGB lowered body weight and adiposity as compared to sham-operated DIO rats. Discrete transcriptome changes were observed in the Arc following RYGB, including differential expression of genes associated with inflammation and neuropeptide signaling. RYGB reduced gene expression of glial cell markers, including Gfap, Aif1 and Timp1, confirmed by a lower number of GFAP immunopositive astrocyte profiles in the Arc. Sham-operated weight-matched rats demonstrated a similar glial gene expression signature, suggesting that RYGB and dietary restriction have common effects on hypothalamic gliosis. Considering that RYGB surgery also led to increased orexigenic and decreased anorexigenic gene expression, this may signify increased hunger-associated signaling at the level of the Arc. Hence, induction of counterregulatory molecular mechanisms downstream from the Arc may play an important role in RYGB-induced weight loss.


Assuntos
Núcleo Arqueado do Hipotálamo/metabolismo , Dieta Redutora , Derivação Gástrica , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Gliose/genética , Adiposidade , Animais , Astrócitos/metabolismo , Biomarcadores , Dieta Hiperlipídica , Ingestão de Alimentos , Proteína Glial Fibrilar Ácida/análise , Peptídeo 1 Semelhante ao Glucagon/sangue , Inflamação/genética , Microdissecção e Captura a Laser , Masculino , Neuropeptídeos/biossíntese , Neuropeptídeos/genética , Obesidade/etiologia , Obesidade/cirurgia , Peptídeo YY/sangue , Ratos , Ratos Sprague-Dawley , Análise de Sequência de RNA , Perda de Peso
7.
Arq Neuropsiquiatr ; 77(9): 601-608, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31553389

RESUMO

OBJECTIVE: Hypothalamic inflammation and glial fibrillary acidic protein (GFAP) overexpression in astrocytes are well described in obese animals, as are some cognitive and memory deficits. As the hippocampus plays important roles in the consolidation of information, this investigation aimed to observe the memory function and the astrocyte expression of GFAP in the hippocampus of rats that received either a hypercaloric or a normocaloric diet. METHODS: Adult male Wistar rats received a high-fat (cafeteria) or a standard diet for 60 days. On the 61st day, the rats were submitted to the novel object recognition (NOR) test at three and 24 hours after the first contact with objects, to assess short-term and long-term memory, respectively. Thereafter, the rats were euthanized and their brains were collected for GFAP immunohistochemical investigation in the hippocampus (CA1, CA2, CA3 areas) and hypothalamus (periventricular and arcuate nuclei). Astrocytic reactivity was assessed by morphometry. Different white adipose tissue depots and brown adipose tissue were weighed to calculate the adiposity index. RESULTS: The hypercaloric diet increased body weight gain, adiposity index, white adipose tissue weight (epididymal, subcutaneous and retroperitoneal) and brown adipose tissue weight. Rats fed with the hypercaloric diet showed short-term and long-term memory impairments in the NOR test, as well as increased GFAP expression in astrocytes from all analyzed hypothalamic and hippocampal areas. CONCLUSION: This astrogliosis suggests that the neuroinflammatory response also occurs in the hippocampus and may be involved in the memory losses observed in obese/overweight animals.


Assuntos
Astrócitos/química , Dieta Hiperlipídica/efeitos adversos , Proteína Glial Fibrilar Ácida/análise , Hipocampo/citologia , Transtornos da Memória/etiologia , Obesidade/complicações , Tecido Adiposo/metabolismo , Animais , Proteína Glial Fibrilar Ácida/metabolismo , Imuno-Histoquímica , Masculino , Transtornos da Memória/metabolismo , Obesidade/metabolismo , Ratos Wistar , Valores de Referência , Fatores de Tempo
8.
Arq. neuropsiquiatr ; 77(9): 601-608, Sept. 2019. tab, graf
Artigo em Inglês | LILACS | ID: biblio-1038743

RESUMO

ABSTRACT Objective: Hypothalamic inflammation and glial fibrillary acidic protein (GFAP) overexpression in astrocytes are well described in obese animals, as are some cognitive and memory deficits. As the hippocampus plays important roles in the consolidation of information, this investigation aimed to observe the memory function and the astrocyte expression of GFAP in the hippocampus of rats that received either a hypercaloric or a normocaloric diet. Methods: Adult male Wistar rats received a high-fat (cafeteria) or a standard diet for 60 days. On the 61st day, the rats were submitted to the novel object recognition (NOR) test at three and 24 hours after the first contact with objects, to assess short-term and long-term memory, respectively. Thereafter, the rats were euthanized and their brains were collected for GFAP immunohistochemical investigation in the hippocampus (CA1, CA2, CA3 areas) and hypothalamus (periventricular and arcuate nuclei). Astrocytic reactivity was assessed by morphometry. Different white adipose tissue depots and brown adipose tissue were weighed to calculate the adiposity index. Results: The hypercaloric diet increased body weight gain, adiposity index, white adipose tissue weight (epididymal, subcutaneous and retroperitoneal) and brown adipose tissue weight. Rats fed with the hypercaloric diet showed short-term and long-term memory impairments in the NOR test, as well as increased GFAP expression in astrocytes from all analyzed hypothalamic and hippocampal areas. Conclusion: This astrogliosis suggests that the neuroinflammatory response also occurs in the hippocampus and may be involved in the memory losses observed in obese/overweight animals.


RESUMO Objetivo: A inflamação hipotalâmica e a superexpressão da proteína glial fibrilar ácida (GFAP) em astrócitos são bem descritas em animais obesos, assim como déficits cognitivos e de memória. Como o hipocampo desempenha importante papel na consolidação de informações, esta investigação teve como objetivo observar a função da memória e a expressão astrocitária da GFAP no hipocampo de ratos que receberam dieta hipercalórica ou normocalórica. Métodos: Ratos Wistar machos adultos receberam dieta rica em gordura (cafeteria) ou dieta padrão por 60 dias. No 61º dia, os ratos foram submetidos ao teste de reconhecimento de objetos (NOR) 3 e 24 horas após o primeiro contato com os objetos, para avaliação da memória de curto e de longo prazo, respectivamente. Após, os ratos foram eutanasiados e os encéfalos coletados para pesquisa imuno-histoquímica da expressão astrocitária de GFAP no hipocampo (áreas CA1, CA2 e CA3) e no hipotálamo (núcleos periventricular e arqueado). A reatividade astrocitária foi avaliada por morfometria. Diferentes depósitos de tecido adiposo branco e marrom foram pesados para calcular o índice de adiposidade. Resultados: A dieta hipercalórica aumentou o ganho de peso corporal, o índice de adiposidade, o peso do tecido adiposo branco (epididimal, subcutâneo e retroperitoneal) e marrom. Ratos alimentados com dieta hipercalórica apresentaram prejuízos na memória de curto e longo prazo no teste NOR e aumento da expressão de GFAP em astrócitos de todas as áreas hipotalâmicas e hipocampais analisadas. Conclusão: Esta astrogliose sugere que a resposta neuroinflamatória também ocorre no hipocampo, podendo estar envolvida nas perdas de memória observadas em animais obesos/com sobrepeso.


Assuntos
Animais , Masculino , Astrócitos/química , Dieta Hiperlipídica/efeitos adversos , Proteína Glial Fibrilar Ácida/análise , Hipocampo/citologia , Transtornos da Memória/etiologia , Obesidade/complicações , Valores de Referência , Fatores de Tempo , Imuno-Histoquímica , Tecido Adiposo/metabolismo , Ratos Wistar , Proteína Glial Fibrilar Ácida/metabolismo , Transtornos da Memória/metabolismo , Obesidade/metabolismo
9.
Mol Biol Rep ; 46(6): 5841-5858, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31396803

RESUMO

Parkinson's disease (PD) is a chronic neurodegenerative disease. Unfortunately, the effectiveness of anti-Parkinson treatments gradually diminishes owing to the progressive degeneration of the dopaminergic terminals. The research described here investigated the effect of adipose-derived mesenchymal stem cells (AD-MSC) versus that of an anti-Parkinson drug in a rat model of Parkinsonism. Forty adult rats were divided into four equal groups, each group receiving a different treatment: vehicle, rotenone, rotenone + AD-MSC, or rotenone + carbidopa/levodopa. Behavioral tests were carried out before and at the end of the treatment and specimens harvested from the midbrain were processed for light and electron microscopy. Genetic expression of glial fibrillary acidic protein (GFAP) and Nestin mRNA was assessed. Expression of the Lamin-B1 and Vimentin genes was measured, along with plasma levels of Angiopoietin-2 and dopamine. Treatment with rotenone induced pronounced motor deficits, as well as neuronal and glial alterations. The AD-MSC group showed improvements in motor function in the live animals and in the microscopic picture presented by their tissues. The fold change of both genes (GFAP and Nestin) decreased significantly in the AD-MSC and carbidopa/levodopa groups compared to the group with Parkinson's disease. Plasma levels of Angiopoietin-2 and dopamine were significantly increased after treatment (P < 0.001) compared to levels in the rats with Parkinson's disease. AD-MSC reduced neuronal degeneration more efficiently than did the anti-Parkinson drug in a rat model of Parkinsonism.


Assuntos
Tecido Adiposo/citologia , Transplante de Células-Tronco Mesenquimais , Transtornos Parkinsonianos , Animais , Comportamento Animal/fisiologia , Modelos Animais de Doenças , Proteína Glial Fibrilar Ácida/análise , Proteína Glial Fibrilar Ácida/genética , Proteína Glial Fibrilar Ácida/metabolismo , Masculino , Células-Tronco Mesenquimais/citologia , Nestina/análise , Nestina/genética , Nestina/metabolismo , Transtornos Parkinsonianos/metabolismo , Transtornos Parkinsonianos/patologia , Transtornos Parkinsonianos/fisiopatologia , Transtornos Parkinsonianos/terapia , Ratos , Ratos Wistar , Substância Negra/química , Substância Negra/patologia , Transcriptoma
10.
Sci Rep ; 9(1): 10279, 2019 07 16.
Artigo em Inglês | MEDLINE | ID: mdl-31311947

RESUMO

Extracellular vesicles (EVs) are small, membrane-bound particles released by all cells that have emerged as an attractive biomarker platform. We study the utility of a dielectrophoretic (DEP) micro-chip device for isolation and characterization of EVs derived from plasma specimens from patients with brain tumors. EVs were isolated by DEP chip and subjected to on-chip immunofluorescence (IF) staining to determine the concentration of glial fibrillary acidic protein (GFAP) and Tau. EVs were analyzed from the plasma samples isolated from independent patient cohorts. Glioblastoma cell lines secrete EVs enriched for GFAP and Tau. These EVs can be efficiently isolated using the DEP platform. Application of DEP to clinical plasma samples afforded discrimination of plasma derived from brain tumor patients relative to those derived from patients without history of brain cancer. Sixty-five percent (11/17) of brain tumor patients showed higher EV-GFAP than the maximum observed in controls. Ninety-four percent (16/17) of tumor patients showed higher EV-Tau than the maximum observed in controls. These discrimination thresholds were applied to plasma isolated from a second, independent cohort of 15 glioblastoma patients and 8 controls. For EV-GFAP, we observed 93% sensitivity, 38% specificity, 74% PPV, 75% NPV, and AUC of 0.65; for EV-Tau, we found 67% sensitivity, 75% specificity 83% PPV, 55% NPV, and AUC of 0.71 for glioblastoma diagnosis. This proof-of-principle study provides support for DEP-IF of plasma EVs for diagnosis of glioblastoma.


Assuntos
Biomarcadores Tumorais/análise , Neoplasias Encefálicas/diagnóstico , Vesículas Extracelulares/metabolismo , Proteína Glial Fibrilar Ácida/análise , Glioblastoma/diagnóstico , Proteínas tau/análise , Análise Química do Sangue , Neoplasias Encefálicas/sangue , Estudos de Casos e Controles , Linhagem Celular Tumoral , Eletroforese em Microchip , Imunofluorescência , Regulação Neoplásica da Expressão Gênica , Glioblastoma/sangue , Humanos , Projetos Piloto , Estudo de Prova de Conceito , Análise Serial de Proteínas , Sensibilidade e Especificidade , Regulação para Cima
11.
Methods Enzymol ; 622: 129-151, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31155050

RESUMO

Astrocytes are the most abundant cell type in the brain and are a crucial part of solving its mysteries. Originally assumed to be passive supporting cells, astrocyte's functions are now recognized to include active modulation and information processing at the neural synapse. The full extent of the astrocyte contribution to neural processing remains unknown. This is, in part, due to the lack of methods available for astrocyte identification and analysis. Existing strategies employ genetic tools like the astrocyte specific promoters glial fibrillary acidic protein (GFAP) or Aldh1L1 to create transgenic animals with fluorescently labeled astrocytes. Recently, small molecule targeting moieties have enabled the delivery of bright fluorescent dyes to astrocytes. Here, we review methods for targeting astrocytes, with a focus on a recently developed methylpyridinium targeting moiety's development, chemical synthesis, and elaboration to provide new features like light-based spatiotemporal control of cell labeling.


Assuntos
Astrócitos/citologia , Encéfalo/citologia , Neuroimagem/métodos , Coloração e Rotulagem/métodos , Animais , Astrócitos/ultraestrutura , Encéfalo/ultraestrutura , Corantes Fluorescentes/análise , Proteína Glial Fibrilar Ácida/análise , Proteína Glial Fibrilar Ácida/genética , Humanos , Oxirredutases atuantes sobre Doadores de Grupo CH-NH/análise , Oxirredutases atuantes sobre Doadores de Grupo CH-NH/genética , Regiões Promotoras Genéticas , Compostos de Piridínio/análise , Peixe-Zebra
12.
Cell Physiol Biochem ; 53(1): 76-86, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31192545

RESUMO

BACKGROUND/AIMS: Diabetes causes damage to the enteric nervous system. The enteric nervous system consists of neurons and enteric glial cells (EGCs). The present study evaluated the effects of an ethyl-acetate fraction (EAF) from Trichilia catigua (T. catigua; 200 mg/kg) on the total population of enteric neurons (HuC/D-immunoreactive [IR]) and EGCs (S100-IR and glial fibrillary acidic protein [GFAP]-IR) in the total preparation and jejunal mucosa in diabetic rats. METHODS: The animals were distributed into four groups: normoglycemic rats (N), diabetic rats (D), normoglycemic rats that received the EAF (NC), and diabetic rats that received the EAF (DC). The jejunum was processed for immunohistochemistry to evaluate HuC/D, S100, and GFAP immunoreactivity. The expression of S100 and GFAP proteins was also quantified by Western blot. RESULTS: The D group exhibited a decrease in the number of neurons and EGCs, an increase in the area of cell bodies, an increase in S100 protein expression, a decrease in GFAP protein expression, and a decrease in S100-IR and GFAP-IR EGCs in the jejunal mucosa. The DC group exhibited a decrease in the number of neurons and EGCs, a decrease in the area of cell bodies, a decrease in S100 and GFAP protein expression, and a decrease in S100-IR and GFAP-IR EGCs in the jejunal mucosa. The NC group exhibited maintenance of the number of neurons and EGCs, an increase in the area of cell bodies, and a decrease in S100 and GFAP protein expression. CONCLUSION: The EAF from T. catigua partially conferred protection against diabetic neuropathy in the enteric nervous system.


Assuntos
Diabetes Mellitus Experimental/complicações , Neuropatias Diabéticas/prevenção & controle , Jejuno/inervação , Meliaceae/química , Neuroglia/efeitos dos fármacos , Neurônios/efeitos dos fármacos , Fármacos Neuroprotetores/farmacologia , Extratos Vegetais/farmacologia , Acetatos/química , Animais , Diabetes Mellitus Experimental/patologia , Neuropatias Diabéticas/etiologia , Neuropatias Diabéticas/patologia , Sistema Nervoso Entérico/efeitos dos fármacos , Proteína Glial Fibrilar Ácida/análise , Jejuno/efeitos dos fármacos , Jejuno/patologia , Masculino , Neuroglia/patologia , Neurônios/patologia , Neuroproteção/efeitos dos fármacos , Fármacos Neuroprotetores/química , Fármacos Neuroprotetores/uso terapêutico , Extratos Vegetais/química , Extratos Vegetais/uso terapêutico , Ratos , Ratos Wistar , Proteínas S100/análise
13.
Eur Surg Res ; 60(1-2): 74-85, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31132769

RESUMO

PURPOSE: The involvement of hepatic stellate cells (HSCs) with ischemia-reperfusion (I/R) injury in rat liver was examined using gliotoxin, which is known to induce HSC apoptosis. METHODS: Male Sprague-Dawley rats were used. HSC was represented by a glial fibrillary acidic protein (GFAP)-positive cell. Liver ischemia was produced by cross-clamping the hepatoduodenal ligament. The degree of I/R injury was evaluated by a release of aminotransferases. Sinusoidal diameter and sinusoidal perfusion rates were examined using intravital fluorescence microscopy. RESULTS: Gliotoxin significantly decreased the number of GFAP-positive cells 48 h after dosing (2.50 ± 0.19% [mean ± SD] in the nontreated group vs. 1.91 ± 0.46% in the gliotoxin-treated group). Liver damage was significantly suppressed by the pretreatment with gliotoxin. Sinusoidal diameters in zone 3 were wider in the gliotoxin group (10.25 ± 0.35 µm) than in the nontreated group (8.21 ± 0.50 µm). The sinusoidal perfusion rate was maintained as well in the gliotoxin group as in normal livers, even after I/R. CONCLUSIONS: Pretreatment with gliotoxin significantly reduced the number of HSCs in the liver and further suppressed liver injury following I/R. It is strongly suggested that HSCs play a functional role in exacerbating the degree of I/R injury of the liver.


Assuntos
Células Estreladas do Fígado/fisiologia , Fígado/irrigação sanguínea , Traumatismo por Reperfusão/etiologia , Animais , Proteína Glial Fibrilar Ácida/análise , Gliotoxina/farmacologia , Células Estreladas do Fígado/efeitos dos fármacos , Macrófagos do Fígado/efeitos dos fármacos , Masculino , Microcirculação/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley
14.
Bioelectrochemistry ; 129: 79-89, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31125924

RESUMO

When implantable recording devices for brain or neural electrical activity are designed, the number of available materials for electrodes is quite limited. The material must be biocompatible with respect to ISO10993, its electrochemical properties must remain stable and the response of cells or tissues can be mitigated, especially on the glial scar. This involves electrode characterization pre- implantation and impedance spectroscopy during chronic implantation, in order to evaluate both electrode properties and performance. This study was aimed at a comparison of the long-term behavior of a nanostructured boron-doped diamond (BDD) with a nanostructured Platinum Iridium (PtIr) electrode. Firstly, a batch of cortical grids with bare and modified contacts (2 mm in diameter) was engineered for implantation. Secondly a miniature swine model was developed. This study highlighted the predominant role of electrode surface roughness on the quality of recordings. Rough PtIr contacts and BDD coated ones showed comparable behavior after three-month implantation with a slight increase of the modulus of the impedance and a tissue capsule. Nevertheless, immunohistochemistry analysis did not exhibit either a toxic or irritation reaction. With regard to biocompatibility, promising long term results are shown for both materials.


Assuntos
Materiais Biocompatíveis/química , Boro/química , Diamante/química , Eletrodos Implantados , Nanoestruturas/química , Animais , Materiais Biocompatíveis/efeitos adversos , Boro/efeitos adversos , Encéfalo/ultraestrutura , Diamante/efeitos adversos , Espectroscopia Dielétrica , Técnicas Eletroquímicas , Eletrodos Implantados/efeitos adversos , Proteína Glial Fibrilar Ácida/análise , Nanoestruturas/efeitos adversos , Nanoestruturas/ultraestrutura , Suínos , Porco Miniatura
15.
Brain Behav Immun ; 79: 319-325, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-30953771

RESUMO

Cancer and its treatment are associated with neurotoxic side effects, including cognitive dysfunction, altered functional connectivity in the brain and structural abnormalities in white matter. There is evidence that cancer and its treatment can accelerate aging. Tau is a microtubule associated protein that contributes to microtubule stability thereby playing a key role in neuronal function. Clustering of tau is commonly observed in the aged brain and is related to cognitive decline. We hypothesized that chemotherapy-induced cognitive impairment is associated with accelerated development of tau clustering in the brain as a sign of accelerated aging. We show for the first time that treatment of adult (7-8 month-old) male C57BL/6 mice with cisplatin results in reduced cognitive function and a marked increase in the number of large endogenous tau clusters in the hippocampus when assessed 4 months later. In contrast, we detected only few small tau clusters in the hippocampus of age-matched 11-12 month-old control mice. Astrocyte GFAP expression was increased in close vicinity to the tau clusters in cisplatin-treated mice. We did not detect changes in the microglial marker Iba-1 in the brain of mice treated with cisplatin. The accelerated formation of Tau-1 clusters in cisplatin-treated mice was associated with a decrease in the levels of the post-synaptic marker PSD95 and of the presynaptic marker synaptophysin in the hippocampus. We demonstrate here for the first time that chemotherapy markedly accelerates development of signs of tauopathy and loss of synaptic integrity in the hippocampus. These findings provide a mechanistic link between chemotherapy cognitive decline and accelerated aging in cancer survivors.


Assuntos
Cisplatino/efeitos adversos , Disfunção Cognitiva/metabolismo , Tauopatias/metabolismo , Fatores Etários , Envelhecimento/metabolismo , Animais , Encéfalo/metabolismo , Cognição/efeitos dos fármacos , Disfunção Cognitiva/etiologia , Modelos Animais de Doenças , Tratamento Farmacológico , Efeitos Colaterais e Reações Adversas Relacionados a Medicamentos , Proteína Glial Fibrilar Ácida/análise , Proteína Glial Fibrilar Ácida/efeitos dos fármacos , Hipocampo/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Tauopatias/etiologia , Proteínas tau/metabolismo
16.
Exp Mol Med ; 51(4): 1-9, 2019 04 16.
Artigo em Inglês | MEDLINE | ID: mdl-30992429

RESUMO

Currently, the two primary patient-derived xenograft (PDX) models of glioblastoma are established through intracranial or subcutaneous injection. In this study, a novel PDX model of glioblastoma was developed via intravitreal injection to facilitate tumor formation in a brain-mimicking microenvironment with improved visibility and fast development. Glioblastoma cells were prepared from the primary and recurrent tumor tissues of a 39-year-old female patient. To demonstrate the feasibility of intracranial tumor formation, U-87 MG and patient-derived glioblastoma cells were injected into the brain parenchyma of Balb/c nude mice. Unlike the U-87 MG cells, the patient-derived glioblastoma cells failed to form intracranial tumors until 6 weeks after tumor cell injection. In contrast, the patient-derived cells effectively formed intraocular tumors, progressing from plaques at 2 weeks to masses at 4 weeks after intravitreal injection. The in vivo tumors exhibited the same immunopositivity for human mitochondria, GFAP, vimentin, and nestin as the original tumors in the patient. Furthermore, cells isolated from the in vivo tumors also demonstrated morphology similar to that of their parental cells and immunopositivity for the same markers. Overall, a novel PDX model of glioblastoma was established via the intravitreal injection of tumor cells. This model will be an essential tool to investigate and develop novel therapeutic alternatives for the treatment of glioblastoma.


Assuntos
Glioblastoma/patologia , Injeções Intravítreas/métodos , Adulto , Animais , Linhagem Celular Tumoral , Modelos Animais de Doenças , Feminino , Imunofluorescência , Proteína Glial Fibrilar Ácida/análise , Humanos , Imuno-Histoquímica , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Mitocôndrias/metabolismo , Células Tumorais Cultivadas , Vimentina/análise , Ensaios Antitumorais Modelo de Xenoenxerto
17.
Med Sci Monit ; 25: 525-531, 2019 Jan 18.
Artigo em Inglês | MEDLINE | ID: mdl-30657131

RESUMO

BACKGROUND The incidence of postoperative cognitive dysfunction (POCD) after major joint arthroplasty is high. In the etiology of POCD, many factors have been cited, including thromboembolic complications. The incidence of cerebral embolization after lower extremity arthroplasty may be as high as 40-60%. The potential events of cerebral embolization could lead to a decrease in the regional cerebral oxygenation (rSO2) and increased serum levels of biochemical markers of brain damage. The objective of the study was to test whether there are any changes in the rSO2 values and serum markers of brain damage in patients who underwent total hip arthroplasty. MATERIAL AND METHODS Fifteen patients who underwent primary hip arthroplasty under spinal anesthesia were analyzed. The rSO2 was monitored using infrared spectroscopy. Biochemical analyses of S100 calcium-binding protein B (S100B) protein and fibrillary acidic protein (GFAP) serum concentrations were performed using immunoassay methods. RESULTS The values of rSO2 decreased during the surgery, but this was not related to mean arterial pressure variations or hemoglobin saturation. The concentration of S100B was increased compared to its preoperative values, and there were no changes in GFAP values. The changes in rSO2 readings correlated with the biomarkers' levels just after the surgery. CONCLUSIONS Our results suggest that S100B may be a more specific marker of astroglial damage in patients after primary total hip arthroplasty. The decrease in rSO2 readings may be due to micro-thromboembolic events that occurred during the surgery. However, the results of this study are preliminary, and further studies are needed to establish its clinical efficacy.


Assuntos
Artroplastia de Quadril/efeitos adversos , Cérebro/metabolismo , Oxigênio/metabolismo , Idoso , Idoso de 80 Anos ou mais , Artroplastia/efeitos adversos , Biomarcadores/sangue , Encéfalo/metabolismo , Lesões Encefálicas/sangue , Artérias Cerebrais/metabolismo , Cérebro/irrigação sanguínea , Feminino , Proteína Glial Fibrilar Ácida/análise , Proteína Glial Fibrilar Ácida/sangue , Humanos , Masculino , Pessoa de Meia-Idade , Oximetria/métodos , Oxigênio/análise , Oxigênio/sangue , Período Perioperatório/métodos , Projetos Piloto , Subunidade beta da Proteína Ligante de Cálcio S100/análise , Subunidade beta da Proteína Ligante de Cálcio S100/sangue
18.
J Perinat Med ; 47(2): 222-240, 2019 Feb 25.
Artigo em Inglês | MEDLINE | ID: mdl-30231013

RESUMO

Objectives To investigate mechanisms of in utero death in normally formed fetuses by measuring amniotic fluid (AF) biomarkers for hypoxia (erythropoietin [EPO]), myocardial damage (cardiac troponin I [cTnI]) and brain injury (glial fibrillary acidic protein [GFAP]), correlated with risk factors for fetal death and placental histopathology. Methods This retrospective, observational cohort study included intrauterine deaths with transabdominal amniocentesis prior to induction of labor. Women with a normal pregnancy and an indicated amniocentesis at term were randomly selected as controls. AF was assayed for EPO, cTnI and GFAP using commercial immunoassays. Placental histopathology was reviewed, and CD15-immunohistochemistry was used. Analyte concentrations >90th centile for controls were considered "raised". Raised AF EPO, AF cTnI and AF GFAP concentrations were considered evidence of hypoxia, myocardial and brain injury, respectively. Results There were 60 cases and 60 controls. Hypoxia was present in 88% (53/60), myocardial damage in 70% (42/60) and brain injury in 45% (27/60) of fetal deaths. Hypoxic fetuses had evidence of myocardial injury, brain injury or both in 77% (41/53), 49% (26/53) and 13% (7/53) of cases, respectively. Histopathological evidence for placental dysfunction was found in 74% (43/58) of these cases. Conclusion Hypoxia, secondary to placental dysfunction, was found to be the mechanism of death in the majority of fetal deaths among structurally normal fetuses. Ninety-one percent of hypoxic fetal deaths sustained brain, myocardial or both brain and myocardial injuries in utero. Hypoxic myocardial injury was an attributable mechanism of death in 70% of the cases. Non-hypoxic cases may be caused by cardiac arrhythmia secondary to a cardiac conduction defect.


Assuntos
Eritropoetina/análise , Morte Fetal/etiologia , Doenças Fetais , Hipóxia Fetal , Proteína Glial Fibrilar Ácida/análise , Cardiopatias , Natimorto , Troponina I/análise , Adulto , Amniocentese/métodos , Líquido Amniótico/metabolismo , Lesões Encefálicas/complicações , Lesões Encefálicas/diagnóstico , Causas de Morte , Feminino , Doenças Fetais/diagnóstico , Doenças Fetais/metabolismo , Hipóxia Fetal/complicações , Hipóxia Fetal/diagnóstico , Cardiopatias/complicações , Cardiopatias/diagnóstico , Humanos , Imuno-Histoquímica , Placenta/patologia , Placenta/fisiopatologia , Gravidez , Distribuição Aleatória , Estudos Retrospectivos , Estados Unidos
19.
J Comp Neurol ; 527(7): 1179-1195, 2019 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-30578640

RESUMO

Astrocytes are the main homeostatic cell of the central nervous system. In addition, astrocytes mediate an inflammatory response when reactive to injury or disease known as astrogliosis. Astrogliosis is marked by an increased expression of glial fibrillary acidic protein (GFAP) and cellular hypertrophy. Some degree of astrogliosis is associated with normal aging and degenerative conditions such as Alzheimer's disease (AD) and other dementing illnesses in humans. The recent observation of pathological markers of AD (amyloid plaques and neurofibrillary tangles) in aged chimpanzee brains provided an opportunity to examine the relationships among aging, AD-type pathology, and astrocyte activation in our closest living relatives. Stereologic methods were used to quantify GFAP-immunoreactive astrocyte density and soma volume in layers I, III, and V of the prefrontal and middle temporal cortex, as well as in hippocampal fields CA1 and CA3. We found that the patterns of astrocyte activation in the aged chimpanzee brain are distinct from humans. GFAP expression does not increase with age in chimpanzees, possibly indicative of lower oxidative stress loads. Similar to humans, chimpanzee layer I astrocytes in the prefrontal cortex are susceptible to AD-like changes. Both prefrontal cortex layer I and hippocampal astrocytes exhibit a high degree of astrogliosis that is positively correlated with accumulation of amyloid beta and tau proteins. However, unlike humans, chimpanzees do not display astrogliosis in other cortical layers. These results demonstrate a unique pattern of cortical aging in chimpanzees and suggest that inflammatory processes may differ between humans and chimpanzees in response to pathology.


Assuntos
Envelhecimento/patologia , Doença de Alzheimer/veterinária , Astrócitos/patologia , Encéfalo/patologia , Gliose/veterinária , Pan troglodytes/anatomia & histologia , Doenças dos Primatas/patologia , Doença de Alzheimer/patologia , Peptídeos beta-Amiloides/análise , Animais , Biomarcadores , Química Encefálica , Feminino , Proteína Glial Fibrilar Ácida/análise , Gliose/patologia , Masculino , Especificidade de Órgãos , Placa Amiloide/química , Placa Amiloide/patologia , Proteínas tau/análise
20.
J Histochem Cytochem ; 67(1): 29-39, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-30047826

RESUMO

The immediate alterations following lesions cannot be investigated by using fixed tissues. Here, we employed two-photon microscopy to study the alterations to the permeability of blood-brain barrier and to glio-vascular connections in vivo during the first minutes following cortical lesions in mice. Four models were used: (1) cryogenic lesion, (2) photodisruption using laser pulses, (3) photothrombosis, and (4) bilateral carotid ligation. Sulforhodamine101 was used for supravital labeling of astrocytes and dextran-bound fluorescein isothiocyanate for the assessment of extravasation. Transgenic mice, in which the endothelium and astrocytes expressed a yellow fluorescent protein, were also used. Astrocytic labeling in vivo was verified with postmortem immunostaining against glial fibrillary acidic protein (GFAP). Summary of results: (1) the glio-vascular connections were stable in the intact brain with no sign of spontaneous dynamic attachment/detachment of glial end-feet; (2) only direct vascular damage (photodisruption or cryogenic) resulted in prompt extravasation; (3) even direct damage failed to provoke a prompt astroglial response. In conclusion, the results indicate that a detachment of the astrocytic end-feet does not precede the breakdown of blood-brain barrier following lesions. Whereas vasogenic edema develops immediately after the lesions, this is not the case with cytotoxic edemas. Time-lapse recordings and three-dimensional reconstructions are presented as supplemental materials.


Assuntos
Astrócitos/patologia , Barreira Hematoencefálica/patologia , Encéfalo/patologia , Permeabilidade Capilar , Animais , Barreira Hematoencefálica/diagnóstico por imagem , Barreira Hematoencefálica/fisiopatologia , Encéfalo/irrigação sanguínea , Encéfalo/diagnóstico por imagem , Encéfalo/fisiopatologia , Modelos Animais de Doenças , Feminino , Proteína Glial Fibrilar Ácida/análise , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Microscopia Confocal/métodos , Neuroglia/patologia , Imagem Óptica/métodos , Coloração e Rotulagem/métodos , Fatores de Tempo
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