RESUMO
Water quality has become a severe concern on a global scale, owing mostly to the rapid increase of the nation's development. According to Malaysia's Natural Resources and Environment Ministry, poor water management is the primary cause of the country's water quality problems. Many river systems are polluted by home and industrial pollutants, according to the findings of research in Malaysia and comparable difficulties in a few other nations. Hence, the following are the research's goals: (1) To look into what is causing the infractions. (2) To undertake the inquiry, develop a thorough hypothesis. (3) To detect dangerous germs by sampling the most usually infected regions. (4) To develop a test for Total Coliform violations in chlorine-treated water at the water treatment plant and in water distribution systems. As a result, the most major barrier to ensuring the safe delivery of treated water to consumers and protecting human health from water-related diseases is the drinking water treatment process. As a result, practically all water treatment systems around the world, including those in the USA, use a chlorine-based procedure to disinfect the water system during treatment. According to studies, the ideal way of disinfecting treated water is both safe and beneficial. Any sort of pandemic or biologically caused disease has no societal implications. Many countries began to suffer in 2009 as a result of e-coli and total coliform contamination in their water systems, leading to ambiguity in disinfection methods. Some water from UNMC's coolers was within the guidelines, while some exceed them. Water coolers at Block E (614 m) and Block B (605 m), for example, measured 12 CFU/100 ml and 11 CFU/100 ml, respectively. Water coolers should be cleaned regularly to ensure that they perform correctly. Further, the microbial population was found to be higher at water storage tanks than that is at the water cooler. This demonstrates how a water cooler fulfils its purpose of filtering and trapping germs to provide clean drinking water.
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Heart transplantation remains the gold standard of therapy for patients with end-stage heart failure. Submassive pulmonary embolism in a patient with heart failure is generally considered a contraindication to immediate heart transplantation, given the risk of right heart failure posttransplant. Generally patients must wait for extended periods of time to recover from pulmonary embolism therapies before being listed for transplant. We report a case of successful concomitant pulmonary thromboendarterectomy and heart transplantation.
Assuntos
Endarterectomia , Insuficiência Cardíaca/cirurgia , Transplante de Coração , Embolia Pulmonar/cirurgia , Endarterectomia/métodos , Feminino , Insuficiência Cardíaca/complicações , Transplante de Coração/métodos , Humanos , Pessoa de Meia-Idade , Embolia Pulmonar/complicações , Resultado do TratamentoRESUMO
Increased use of continuous-flow left ventricular assist devices (LVADs) to treat advanced heart failure has heightened concern for right ventricular failure after LVAD implantation, which is associated with increased morbidity and mortality. Biventricular support is required in up to 30% of LVAD recipients. Currently, no durable long-term right ventricular assist device (RVAD) has been approved other than the Syncardia (Tucson, AZ) total artificial heart. A recent publication reported the placement of continuous flow LVAD in the heavily trabeculated right ventricle; however, this orientation may jeopardize both assist device and right ventricle function. We describe three cases of right-sided mechanical circulatory support with durable RVAD implanted in the right atrium, allowing long-term support with fewer anatomic limitations as compared with right ventricular cannulation.
Assuntos
Insuficiência Cardíaca/terapia , Coração Auxiliar , Implantação de Prótese/métodos , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Adulto JovemRESUMO
The combination of mass cytometry and immunohistochemistry (IHC) enables new histopathological imaging methods in which dozens of proteins and protein modifications can be visualized simultaneously in a single tissue section. The power of multiplexing combined with spatial information and quantification was recently illustrated on breast cancer tissue and was described as next-generation IHC. Robust, accurate, and high-throughput cell segmentation is crucial for the analysis of this new generation of IHC data. To this end, we propose a watershed-based cell segmentation, which uses a nuclear marker and multiple membrane markers, the latter automatically selected based on their correlation. In comparison with the state-of-the-art segmentation pipelines, which are only using a single marker for object detection, we could show that the use of multiple markers can significantly increase the segmentation power, and thus, multiplexed information should be used and not ignored during the segmentation. Furthermore, we provide a novel, user-friendly open-source toolbox for the automatic segmentation of multiplexed histopathological images.
Assuntos
Neoplasias da Mama/diagnóstico , Diagnóstico por Imagem/métodos , Citometria de Fluxo/métodos , Análise de Célula Única , Neoplasias da Mama/patologia , Feminino , Humanos , Imuno-HistoquímicaRESUMO
Mass cytometry enables high-dimensional, single-cell analysis of cell type and state. In mass cytometry, rare earth metals are used as reporters on antibodies. Analysis of metal abundances using the mass cytometer allows determination of marker expression in individual cells. Mass cytometry has previously been applied only to cell suspensions. To gain spatial information, we have coupled immunohistochemical and immunocytochemical methods with high-resolution laser ablation to CyTOF mass cytometry. This approach enables the simultaneous imaging of 32 proteins and protein modifications at subcellular resolution; with the availability of additional isotopes, measurement of over 100 markers will be possible. We applied imaging mass cytometry to human breast cancer samples, allowing delineation of cell subpopulations and cell-cell interactions and highlighting tumor heterogeneity. Imaging mass cytometry complements existing imaging approaches. It will enable basic studies of tissue heterogeneity and function and support the transition of medicine toward individualized molecularly targeted diagnosis and therapies.
Assuntos
Neoplasias da Mama/metabolismo , Citometria por Imagem/métodos , Proteínas de Neoplasias/metabolismo , Linhagem Celular , Células Epiteliais/citologia , Células Epiteliais/metabolismo , Feminino , Regulação Neoplásica da Expressão Gênica/fisiologia , Humanos , Proteínas de Neoplasias/genéticaRESUMO
In recent years, chemical imaging was prognosticated to become one of the key analytical applications for laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS). However, moderate spatial resolution and the associated measurement time required for a larger sampling area, have restricted this versatile, high sensitivity technique from being routinely used in two-dimensional chemical imaging. This work describes the development and investigation of a low dispersion sample chamber (tube cell), which allows improvement of the imaging capabilities by reduction of the single LA shot duration to 30 ms (full width at 1% maximum). The new tube cell is based on a constant laminar flow and a well-controlled delivery of the laser-ablated aerosol into the transport system, leading to minimized tailing of the aerosol washout and helping to separate the signals even at repetition rates as high as 20-30 Hz. To demonstrate the improved imaging capabilities, microstructured metallic thin film patterns were analyzed at a spatial resolution of a few micrometers. The LA-ICP-MS results obtained were comparable to Synchrotron-based micro-X-ray fluorescence (SR-microXRF). The suitability of the newly designed cell for multielement acquisitions was demonstrated using a simultaneous ICP-Mattauch-Herzog-MS. Finally, the novel laser ablation cell was applied to image the distribution of a metal-tagged biomarker in a thin section of breast cancer tissue. This application demonstrates that the technique is able to produce subcellular (~1 µm) spatial resolution, which is crucial for morphological assessment in cancer diagnostics.
Assuntos
Terapia a Laser , Espectrometria de Massas/métodos , Neoplasias da Mama/patologia , Feminino , Humanos , LasersRESUMO
Growing evidence has suggested that hydrogen sulfide (H2S) acts as a novel neuro-modulator and neuroprotective agent; however, it remains to be investigated whether H2S has a direct effect on neural stem cells (NSCs). We report here that NSCs expressed cystathionine ß synthase (CBS) and addition of exogenous H2S donor, L-cysteine, stimulated proliferation and increased the differentiation potential of NSCs to neurons and astroglia. Moreover, pre-treatment with aminooxyacetic acid, the inhibitor of CBS or knockdown of CBS in using siRNA, significantly attenuated the effects of L-cysteine on elevated H2S levels and the cell proliferation; it also effectively suppressed L-cysteine-induced neurogenesis and astrocytogenesis. Further analysis revealed that L-cysteine-induced proliferation was associated with phosphorylation of extracellular signal-regulated kinases 1/2 and differentiation with altered expression of differentiation-related genes. Taken together, the present data suggest that L-cysteine can enhance proliferation and differentiation of NSCs via the CBS/H2S pathway, which may serve as a useful inference for elucidating its role in regulating the fate of NSCs in physiological and pathological settings.
Assuntos
Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Cistationina beta-Sintase/metabolismo , Cisteína/farmacologia , Sulfeto de Hidrogênio/metabolismo , Células-Tronco Neurais/efeitos dos fármacos , Análise de Variância , Animais , Bromodesoxiuridina/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Embrião de Mamíferos , Citometria de Fluxo , Regulação da Expressão Gênica/efeitos dos fármacos , Camundongos , Proteínas do Tecido Nervoso/genética , Proteínas do Tecido Nervoso/metabolismo , RNA Mensageiro/metabolismo , RNA Interferente Pequeno/farmacologia , Transdução de Sinais/efeitos dos fármacos , Tacrolimo/análogos & derivados , Tacrolimo/farmacologia , Telencéfalo/citologia , Fatores de Tempo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismoRESUMO
Astrocyte activation plays important roles both in physiological and pathological process in the CNS. In the latter, the process is further aggravated by hyperglycemia, leading to diabetes complications of CNS. We report here that high glucose (HG) treatment stimulated astrocytic morphological alteration coupled with changes in glial fibrillary acidic protein (GFAP) and vimentin expression. Additionally, HG upregulated the expression of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), interleukin-1ß (IL-1ß), interleukin-4 (IL-4), and vascular endothelial growth factor (VEGF); however, its effects on transforming growth factor-ß (TGF-ß) expression were not evident. HG treatment induced increased production of reactive oxygen species (ROS) as well as activation of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) and signal transducer and activator transcription 3 (STAT 3). HG-induced expression of TNF-α, IL-6, IL-1ß, IL-4, and VEGF was blocked by ROS scavenger and inhibitors specific for NF-κB and STAT 3, respectively. The results suggest that the aforementioned multiple inflammatory cytokines and mediators that may be linked to the pathogenesis of the diabetes complications of CNS are induced by HG via the key signaling pathways.
Assuntos
Astrócitos/metabolismo , Citocinas/biossíntese , Glucose/toxicidade , Inflamação/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Animais , Astrócitos/efeitos dos fármacos , Western Blotting , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Sequestradores de Radicais Livres/farmacologia , Imuno-Histoquímica , Marcação In Situ das Extremidades Cortadas , Inflamação/induzido quimicamente , Interleucina-1beta/metabolismo , Interleucina-4/metabolismo , Interleucina-6/metabolismo , Melatonina/farmacologia , Camundongos , Camundongos Endogâmicos BALB C , NF-kappa B/metabolismo , Prolina/análogos & derivados , Prolina/farmacologia , RNA/biossíntese , RNA/isolamento & purificação , Reação em Cadeia da Polimerase em Tempo Real , Fator de Transcrição STAT3/metabolismo , Tiocarbamatos/farmacologia , Fator de Crescimento Transformador beta/metabolismo , Triterpenos/farmacologia , Fator de Necrose Tumoral alfa/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
Circadian rhythms in mammals are generated by a feedback loop in which the three PERIOD (PER) proteins, acting in a large complex, inhibit the transcriptional activity of the CLOCK-BMAL1 dimer, which represses their own expression. Although fundamental, the mechanism of negative feedback in the mammalian clock, or any eukaryotic clock, is unknown. We analyzed protein constituents of PER complexes purified from mouse tissues and identified PSF (polypyrimidine tract-binding protein-associated splicing factor). Our analysis indicates that PSF within the PER complex recruits SIN3A, a scaffold for assembly of transcriptional inhibitory complexes and that the PER complex thereby rhythmically delivers histone deacetylases to the Per1 promoter, which repress Per1 transcription. These findings provide a function for the PER complex and a molecular mechanism for circadian clock negative feedback.
Assuntos
Relógios Circadianos , Ritmo Circadiano , Retroalimentação Fisiológica , Proteínas Circadianas Period/genética , Proteínas de Ligação a RNA/metabolismo , Proteínas Repressoras/metabolismo , Fatores de Transcrição ARNTL/genética , Fatores de Transcrição ARNTL/metabolismo , Animais , Proteínas CLOCK/genética , Proteínas CLOCK/metabolismo , Criptocromos/metabolismo , Histona Desacetilase 1/metabolismo , Histonas/metabolismo , Fígado/metabolismo , Pulmão/metabolismo , Espectrometria de Massas , Camundongos , Fator de Processamento Associado a PTB , Proteínas Circadianas Period/metabolismo , Regiões Promotoras Genéticas , Proteínas de Ligação a RNA/genética , Proteínas Recombinantes de Fusão/metabolismo , Complexo Correpressor Histona Desacetilase e Sin3 , Transcrição GênicaRESUMO
PALB2 is a breast and pancreas cancer susceptibility gene whose protein is closely associated with BRCA2 and is essential for BRCA2 anchorage to nuclear structures. This functional relationship made PALB2 a candidate gene for susceptibility to BRCA2-related cancers such as melanoma. The purpose of this study was to screen for the presence of germline mutations in PALB2 in familial melanoma cases. We sequenced the exons and intron-exon boundaries of PALB2 in probands from 53 families with familial melanoma where CDKN2A mutations were absent. A number of previously reported coding and non-coding variants were observed. However, no truncating mutations were identified. These results indicate that deleterious PALB2 mutations are unlikely to play a significant role in familial melanoma.
Assuntos
Predisposição Genética para Doença , Melanoma/genética , Mutação , Proteínas Nucleares/genética , Proteínas Supressoras de Tumor/genética , Proteína do Grupo de Complementação N da Anemia de Fanconi , Feminino , Humanos , Pessoa de Meia-IdadeRESUMO
Granulocyte-colony stimulating factor (G-CSF) is a growth factor that regulates proliferation, differentiation and survival of hematopoietic progenitor cells. There is growing evidence to suggest that G-CSF exerts a powerful neuroprotective effect in different neurological disorders. However, it has remained to be elucidated if G-CSF has a direct effect on neural stem cells (NSCs). Here, we show that G-CSF could stimulate the proliferation of NSCs and promote their differentiation in vitro. Additionally, we have shown that G-CSF-induced proliferation of NSCs is associated with phosphorylation of STAT3, and the differentiation is linked to altered expression of differentiation-related genes. Remarkably, G-CSF could not initiate the differentiation of NSCs. The added roles of G-CSF in regulating proliferation and differentiation of NSCs as shown in this study would serve as a useful reference in designing new stem cell therapy strategies for promoting brain recovery and repair.
Assuntos
Astrócitos/citologia , Fator Estimulador de Colônias de Granulócitos/fisiologia , Neurônios/citologia , Células-Tronco/citologia , Animais , Astrócitos/efeitos dos fármacos , Astrócitos/metabolismo , Fatores de Transcrição Hélice-Alça-Hélice Básicos/biossíntese , Encéfalo/citologia , Diferenciação Celular , Proliferação de Células , Sobrevivência Celular , Células Cultivadas , Embrião de Mamíferos , Fator Estimulador de Colônias de Granulócitos/farmacologia , Camundongos , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Fosforilação , Receptores de Fator Estimulador de Colônias de Granulócitos/biossíntese , Fator de Transcrição STAT3/metabolismo , Células-Tronco/efeitos dos fármacos , Células-Tronco/metabolismoRESUMO
Local microenvironment plays an important role in determining the fate choice of stem cells in the central nervous system (CNS). Astrocytes, a major component of local microenvironment in the CNS, have been demonstrated to influence the proliferation and neural differentiation of stem cells including neural stem/progenitor cells, embryonic stem cells and bone marrow stromal cells (BMSCs). However, it has remained to be ascertained if inflammation-activated astrocytes can affect the behavior of BMSCs. To this end, astrocyte-conditioned medium (ACM) was prepared in this study for treatment of BMSCs. The ACM derived from Wistar rat astrocytes stimulated by lipopolysaccharide for 12, 36 or 72 h, respectively, served as inflammatory ACM (12 h ACM, 36 h ACM and 72 h ACM), while that from unstimulated astrocytes was used as normal control astrocyte-conditioned medium (N-ACM). The results showed that the proliferation and neural differentiation of BMSCs grown in inflammatory ACM were significantly increased compared with those grown in N-ACM. The efficiency of BMSCs exposed to 36 h ACM was significantly greater than that of those exposed to 12 or 72 h ACM. Following neutralization of interleukin-6 (IL-6) of the ACM, both the proliferation and astrocytic differentiation of BMSCs were decreased; on the other hand, the neuronal differentiation was significantly increased. The present findings suggest that inflammation-activated astrocytes can facilitate the proliferation and neural differentiation of BMSCs and activated astrocytes at different phase after CNS injuries might have distinct effects on BMSCs. Moreover, astrocyte-derived IL-6 participates in the proliferation and neural differentiation of BMSCs.
Assuntos
Astrócitos/fisiologia , Células da Medula Óssea/citologia , Diferenciação Celular/fisiologia , Neurogênese/fisiologia , Células-Tronco/citologia , Animais , Astrócitos/citologia , Fatores Biológicos/fisiologia , Células da Medula Óssea/fisiologia , Proliferação de Células , Técnicas de Cocultura , Meios de Cultivo Condicionados , Interleucina-6/fisiologia , Ratos , Ratos Wistar , Células-Tronco/fisiologia , Células EstromaisRESUMO
In an attempt to understand the molecular basis underlying the neural tube defects induced by the teratogen, cyclophosphamide (CP), cDNA microarray analysis was carried out in neural tubes of embryos derived from normal and CP-treated rats. Genes found to have altered expression levels in CP-treated group were clustered into groups on the basis of their biological functions. The expression profile of different genes involved in transcription of molecules related to cell adhesion, inflammation, metabolism and neurotrophic factors pathways as well as in still undefined processes was differentially affected by the teratogen treatment. The most remarkable change was the up-regulation of genes related to an inflammatory process dominated by the fetal brain macrophages viz. amoeboid microglia. Amoeboid microglia/brain macrophage expansion, based on gene expression and histological analysis, was found to be vigorous at the subventricular region. The present results suggest that a vigorous inflammatory response involving amoeboid microglia/brain macrophages primarily is an important component in CP-induced prenatal development disorder.
Assuntos
Ciclofosfamida/toxicidade , Perfilação da Expressão Gênica , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Análise de Sequência com Séries de Oligonucleotídeos , Prosencéfalo/efeitos dos fármacos , Teratogênicos/toxicidade , Animais , Caspase 1/genética , Caspase 1/metabolismo , Imuno-Histoquímica , Hibridização In Situ , Inflamação/genética , Lectinas/metabolismo , Fator Estimulador de Colônias de Macrófagos/genética , Fator Estimulador de Colônias de Macrófagos/metabolismo , Microscopia Confocal , Neuroglia/efeitos dos fármacos , Neuroglia/metabolismo , Óxido Nítrico Sintase Tipo II/genética , Óxido Nítrico Sintase Tipo II/metabolismo , Prosencéfalo/embriologia , Prosencéfalo/metabolismo , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Receptor de Fator Estimulador de Colônias de Macrófagos/genética , Receptor de Fator Estimulador de Colônias de Macrófagos/metabolismo , Reprodutibilidade dos Testes , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de TempoRESUMO
The degradation kinetics of fluorouracil-acetic-acid-dextran conjugate (FUAC-dextran) was investigated in various buffer solutions with different pH value and physiological saline solution at 60 degrees C and 37 degrees C, respectively. The hydrolytic reaction displayed pseudo-first-order degradation kinetics. Hydrolytic rate constant obtained was the function of pH value and independent of species of buffering agents. The smallest rate constant was observed at pH round 3.00. The activation energy of the hydrolytic reaction was estimated from Arrhenius equation as 88.73 +/- 6.00 kJ.mol-1. The special base catalytic degradation of the conjugate was observed from acidic to slight alkaline condition and the special base catalytic rate constants were calculated. The conjugate was more stable in physiological saline than that in buffer solution at pH 7.00 or 9.00 at 37 degrees C. The results revealed that the conjugate was stable in acidic condition and will degrade in alkaline condition.
Assuntos
Antineoplásicos/química , Dextranos/química , Fluoruracila/análogos & derivados , Soluções , Soluções Tampão , Estabilidade de Medicamentos , Fluoruracila/química , Concentração de Íons de Hidrogênio , Hidrólise , Cinética , Modelos Químicos , Cloreto de Sódio/química , Temperatura , ÁguaAssuntos
Variação Genética , Judeus/genética , Melanoma/etnologia , Melanoma/genética , Neoplasias Cutâneas/genética , Adulto , Alelos , Cromossomos Humanos Par 1/genética , Cromossomos Humanos Par 9/genética , Análise Mutacional de DNA , Europa (Continente)/etnologia , Feminino , Genes p16 , Predisposição Genética para Doença , Testes Genéticos , Humanos , Judeus/etnologia , Masculino , Pessoa de Meia-Idade , Linhagem , Neoplasias Cutâneas/epidemiologiaRESUMO
Microarray analysis of RNA from hepatitis C virus (HCV)-infected cirrhotic livers was performed to identify a gene expression signature of liver disease. The expression levels of approximately 13600 genes were analyzed using surgical material and core biopsy specimens from HCV-infected cirrhotic liver explants in comparison with reference samples of normal nondiseased liver. In addition, normal liver samples were compared with each other to determine normal physiologic variation in gene expression. A set of genes, including some associated with stress, acute-phase immune response, and hepatic stellate cell activation, had variable expression levels in normal livers. These genes were subtracted from the sets of genes differentially expressed in cirrhotic livers. To exclude cancer-related genes from our marker sets, we subtracted genes that also were expressed differentially in hepatocellular carcinomas. The resultant HCV- and liver disease-associated gene set provided a molecular portrait of several processes occurring in the HCV-infected liver. It included (1). genes expressed in activated lymphocytes infiltrating the cirrhotic liver, and activated liver macrophages; (2). genes involved in remodeling of extracellular matrix-cell and cell-cell interactions associated with cytoskeleton rearrangements; (3). genes related to the anti-apoptotic pathway of Bcl-2 signaling; and (4). genes involved with the interferon response and virus-host interactions. In conclusion, our microarray analysis identified several potential gene markers of HCV-associated liver disease and contributed to our rapidly expanding database of experiments describing HCV pathogenesis.
Assuntos
Perfilação da Expressão Gênica , Hepatite C/metabolismo , Cirrose Hepática/metabolismo , Apoptose , Citoesqueleto/metabolismo , Proteínas da Matriz Extracelular/biossíntese , Marcadores Genéticos , Hepatite C/patologia , Humanos , Fígado/metabolismo , Fígado/patologia , Cirrose Hepática/patologia , Análise de Sequência com Séries de Oligonucleotídeos , Proteínas Proto-Oncogênicas c-bcl-2/fisiologia , Proteína cdc42 de Ligação ao GTP/fisiologiaRESUMO
Prenatal exposure to teratogen agents is linked to the pathogenesis of neurodevelopment disorders, but the mechanisms leading to the neurodevelopmental disturbance are poorly understood. To elucidate this, an in vitro model of microglial activation induced by neuronal injury has been characterized. In this connection, exposure of primary microglial cells to the conditioned medium from the neuronal damage induced by teratogen, cyclophosphamide, is accompanied by a reactive microgliosis as assessed by reverse transcription-polymerase chain reaction, enzyme-linked immunosorbent assay, lectin histochemistry, double labeling immunohistochemistry and in situ hybridization. Our results showed that reactive microglia were capable of releasing various cytokines such as tumor necrosis factor-alpha, interleukin-1, interleukin-6, transforming growth factor-beta and nitric oxide. Also, we have shown that macrophage colony-stimulating factor (M-CSF) was in fact produced by the reactive microglia. Concomitant to this was the increased expression of M-CSF receptor in these cells following the teratogen-induced neuronal injury. The up-regulation of M-CSF receptor suggests that the cells are capable of responding to self-derived M-CSF in an autocrine fashion. Results with antibody neutralization further suggest that microglial proinflammatory response, as manifested by cytokine expression in culture, is mediated by M-CSF, which acts as a molecular signal that initiates a microglial reaction. We therefore suggest that microglial activation following cyclophosphamide treatment is not only a response to the neuronal damage, but is also a cause of the damage during pathogenesis of neurodevelopment disorders. To this end, the increased expression of M-CSF and its receptor on microglia would be directly linked to the active cell proliferation and proinflammatory response in the teratogen-induced injury.
Assuntos
Córtex Cerebral/efeitos dos fármacos , Ciclofosfamida/toxicidade , Fator Estimulador de Colônias de Macrófagos/metabolismo , Microglia/metabolismo , Neurônios/efeitos dos fármacos , Receptor de Fator Estimulador de Colônias de Macrófagos/metabolismo , Teratogênicos/toxicidade , Animais , Técnicas de Cultura de Células , Relação Dose-Resposta a Droga , Ensaio de Imunoadsorção Enzimática , Imuno-Histoquímica , Hibridização In Situ , Interleucina-1/metabolismo , Interleucina-6/metabolismo , Fator Estimulador de Colônias de Macrófagos/efeitos dos fármacos , Microglia/efeitos dos fármacos , Microglia/imunologia , Microscopia Confocal , Óxido Nítrico Sintase/metabolismo , Óxido Nítrico Sintase Tipo II , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Receptor de Fator Estimulador de Colônias de Macrófagos/efeitos dos fármacos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de Tempo , Fator de Crescimento Transformador beta/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Regulação para CimaRESUMO
OBJECTIVE: To observe the protective effects of natural antioxidant tea polyphenols(TP) on myocardial free radical metabolic disorder in mice induced by inhalation of hypobaric pure oxygen under 5500 m hypobaric condition. METHOD: Forty-two male Kunming mice were randomly divided into three groups (n = 14 each): group A, normal control; group B, inhalation of pure oxygen (> 96 %) at simulated altitude of 5500 m in an animal altitude chamber; group C (TP protection group), same as group B but 100 mg/kg of TP was given orally before the exposure. The exposure time was 2 h/d, 3 d/wk for a total of 8 wk, and distilled water was given to groups A and B before exposure. After experiment, the mice were decapitated on the next day and the heart was quickly removed. Malondialdehyde (MDA) concentration, superoxide dismutase (SOD) activity and nitric oxide (NO) content were measured. In addition, Cu, Zn-SOD and inducible NO synthase (iNOS) enzymatic contents in myocardial tissue were qualitatively examined by immunohistochemical assaying. RESULT: Compared with the control, MDA concentration, SOD activity and Cu, Zn-SOD enzymatic content in group B were significantly increased (P < 0. 05). But in TP protection group, myocardial MDA formation was significantly decreased (P < 0. 01) and SOD activity and Cu, Zn-SOD expression restored to normal. On the contrary, myocardial NO generation and iNOS expression were significantly reduced after repeated inhalation of hypobaric oxygen at 5500 m. NO metabolism regained to normal after repeated administration of TP. CONCLUSION: Natural antioxidant TP had protective effects on myocardial free radical metabolic disorder induced by inhalation of hypobaric pure oxygen under 5500 m hypobaric condition.
Assuntos
Antioxidantes/farmacologia , Flavonoides , Coração/efeitos dos fármacos , Miocárdio/metabolismo , Oxigênio/farmacologia , Fenóis/farmacologia , Polímeros/farmacologia , Chá/química , Altitude , Animais , Antioxidantes/análise , Relação Dose-Resposta a Droga , Sequestradores de Radicais Livres/farmacologia , Peroxidação de Lipídeos/efeitos dos fármacos , Masculino , Malondialdeído/metabolismo , Camundongos , Miocárdio/enzimologia , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase/metabolismo , Oxigenoterapia/efeitos adversos , Fenóis/análise , Polímeros/análise , Polifenóis , Superóxido Dismutase/metabolismoRESUMO
Using in situ hybridization, RT-PCR, lectin histochemistry and immunohistochemistry, this study examined the time course expression and cellular localization of various cytokines including tumor necrosis factor-alpha (TNF-alpha), interleukins (IL-1, IL-6) and transforming growth factor-beta (TGF-beta) in fetal rat brain after a maternal injection of the teratogen cyclophosphamide (CP). Eight hours after CP injection, there was a marked increase in brain macrophages (BM)/amoeboid microglia (AM) in different areas of the fetal brain as determined by lectin histochemistry. Concomitant to this was the induction in mRNA level of TNF-alpha, which was progressively increased with time. TGF-beta mRNA was undetectable until 24 h had elapsed. Expression of IL-1 and IL-6 was undetectable at all stages. In situ hybridization combined with immunohistochemistry has shown the localization of TNF-alpha in BM/AM and neurons. Present results suggest that both TNF-alpha and TGF-beta are involved in the progression of neural damage in the fetal brain induced by the teratogen.
Assuntos
Antígenos CD , Antígenos de Neoplasias , Antígenos de Superfície , Proteínas Aviárias , Proteínas Sanguíneas , Encéfalo/efeitos dos fármacos , Citocinas/metabolismo , Feto/efeitos dos fármacos , Macrófagos/efeitos dos fármacos , Microglia/efeitos dos fármacos , Efeitos Tardios da Exposição Pré-Natal , Teratogênicos/farmacologia , Animais , Basigina , Encéfalo/embriologia , Encéfalo/imunologia , Movimento Celular/efeitos dos fármacos , Movimento Celular/imunologia , Ciclofosfamida/farmacologia , Citocinas/genética , Citocinas/imunologia , Feminino , Feto/embriologia , Feto/imunologia , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Interleucina-1/genética , Interleucina-1/imunologia , Interleucina-1/metabolismo , Interleucina-6/genética , Interleucina-6/imunologia , Interleucina-6/metabolismo , Lectinas/farmacocinética , Macrófagos/imunologia , Macrófagos/metabolismo , Glicoproteínas de Membrana/metabolismo , Microglia/imunologia , Microglia/metabolismo , Mutagênicos/farmacologia , Gravidez , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de Tempo , Fator de Crescimento Transformador beta/genética , Fator de Crescimento Transformador beta/imunologia , Fator de Crescimento Transformador beta/metabolismo , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/imunologia , Fator de Necrose Tumoral alfa/metabolismoRESUMO
This study examined the time course response of amoeboid microglia/brain macrophages in the rat fetus induced by a single intraperitoneal injection of cyclophosphamide, a teratogen, into the mother rat at 13 days of gestation. Compared to the normal fetal brain, a marked increase in amoeboid microglia was observed in the telencephalon and diencephalon of experimental fetuses, especially in those killed at embryonic day 15. Conglomerations of microglia occurred in the dorsal and superior neuroepithelium of diencephalon, basal telencephalon, cortical neuroepithelium, and hippocampal formation as identified with OX-42, OX-18, and ED-1 by immunohistochemistry. Rhodamine isothiocynate (RhIc) as a tracer was injected via the tail vein into the pregnant rat to assess the phagocytic capability of these cells. Following the tracer injection, none of microglial cells in normal fetal brain was detectable. RhIc, however, was readily taken up by amoeboid microglia in fetal brain with injury insult. Double labeling has shown that the RhIc-labeled cells were immunoreactive with ED-1, OX-42, OX-18, and OX-6, confirming their microglial nature. Microglial proliferation was assessed by immunohistochemistry using bromodeoxyuridine, which showed a marked increase in mitotic activity. Confocal microscopic analysis revealed that a varying number of microglia coexpressed iNOS, macrophage colony-stimulating factor (M-CSF), and ICAM-1. RT-PCR analysis showed increased expression of M-CSF mRNA. Furthermore, colony-stimulating factor-1 receptor mRNA was localized in microglia by in situ hybridization. The present results suggest that NO along with M-CSF and ICAM-1 is involved in microglial proliferation in prenatal brain injury.