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Characterization of Hypotonic Shock Induced Ascorbate Release from Pig Coronary Artery Endothelial Cells / Hypotonic Shock Induced Ascorbate ReleaseGill, Rupinder 09 1900 (has links)
Ascorbate (Asc) is a key antioxidant in preventing cardiovascular dysfunction during diseases exacerbated by altered shear stress. According to the literature endothelial responses to hypotonic shock share some characteristics with those induced by shear stress. Thus to study the physiological responses of endothelium to shear stress, the characterization of the Asc release by pig coronary artery endothelial cells in response to hypotonic shock was performed.
The pig coronary artery endothelial cells that had been loaded with ^14C Asc and ^3H deoxyglucose, were exposed to buffers of varying osmolality for different time periods and the release of ^14C Asc and ^3H deoxyglucose was examined. Based on various parameters like relative release of ^14C Asc and ^3H deoxyglucose, their rate of release and protein loss, it was decided to use buffer of .67 percent osmolality for 2 min for these characterization studies. The Asc release was authentic and not a result of membrane damage. The hypotonic shock induced Asc release was not due to endogenously released ATP. The inhibition of ATP induced release by anion channel inhibitors niflumic acid and NPPB was complete but only partial in case of hypotonic shock induced release. The release was not inhibited under nominally Ca^2+ free conditions. Additive release by hypotonic shock and ATP or hypotonic shock and Ca^2+ ionophore A23187 suggests that there are two independent Asc release pathways. Asc release by two different mechanisms may help endothelial cells deal with stressful conditions efficiently and preserve endothelial function. / Thesis / Master of Science (MS)
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G Protein-Coupled Receptor Regulation of ATP release from AstrocytesBlum, Andrew E. 14 June 2010 (has links)
No description available.
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Objemově regulované aniontové kanály u astrocytů - in vitro and in situ analýza / Volume-regulated anion channels in astrocytes- in vitro and in situ analysisHarantová, Lenka January 2012 (has links)
Astrocytes need to preserve constant volume in the face of osmolarity perturbations to function properly. To regain their original volume after hyposmotically induced swelling, they extrude intracellular electrolytes and organic osmolytes, such as inorganic ions, excitative amino acids or polyols, accompanied by osmotically driven water. This process is termed regulatory volume decrease and is ensured by various ion channels and transporters. Recently, much attention has been focused on the ubiquitous volume-regulated anion channels activated by cell swelling. VRACs are moderately outwardly rectifying with intermediary conductance, permeable to inorganic anions and organic osmolytes and sensitive to broad-spectrum anion channels blockers. Using patch-clamp technique we aimed to characterize VRACs in cultured cortical astrocytes isolated from neonatal Wistar rats and to elucidate the effect of intracellular Na+ on VRAC activity. In addition, we also intended to characterize these channels in situ in brain slices of 10 - 12 days old rats, focusing mainly on hippocampal astrocytes. To induce astrocytic swelling, we exposed astrocytes to hypotonic solution (250 mOsm). In agreement with previous findings, we showed that cultured cortical astrocytes activate VRAC currents upon exposure to hypotonic stress, which...
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Objemové změny astrocytů u alfa-syntrofin deficientních myší / Astrocyte volume changes in alpha-syntrophin deficient miceMikešová, Michaela January 2012 (has links)
(EN) The formation of brain oedema, which accompanies ischemic or traumatic brain injuries, originates from a disruption of ionic/neurotransmitter homeostasis that leads to extracellular K+ elevation and neurotransmitter accumulation in the extracellular space. An increased uptake of these osmotically active substances, predominantly provided by astrocytes, is accompanied by intracellular water accumulation via aquaporin-4 (AQP4). Since it has been shown that the removal of perivascular AQP4 via the deletion of α- syntrophin, which is the protein responsible for anchoring AQP4 on the astrocytic membrane (Neely et al. 2001), delays oedema formation and K+ clearance (Amiry-Moghaddam et al. 2003), we aimed to elucidate how the alpha-syntrophin deletion affects astrocyte volume changes in the cortex during pathological states, such as hypoosmotic stress or oxygen- glucose deprivation (OGD), using three-dimensional (3D) confocal morphometry in situ. In order to visualize individual astrocytes that lack alpha-syntrophin, double transgenic mice (GFAP/EGFP/α-Syn-/- ) were generated by crossbreeding GFAP-EGFP mice with α- syntrophin knockout mice. 3D-confocal morphometry revealed that alpha-syntrophin deletion did not alter astrocyte swelling during hypoosmotic stress or their recovery in isotonic solution;...
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Episódio Hipotônico-hiporresponsivo associado à vacina combinada contradifteria, tétano, pertussis e Haemophilus influenzae tipo b: análise da definição de caso para vigilância / Hypotonic-hyporesponsive episode contradifteria associated with the combined vaccine, tetanus, pertussis and Haemophilus influenzae type b: analysis of the case definition for surveillanceNoronha, Tatiana Guimarães de January 2010 (has links)
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Previous issue date: 2010 / Analisar as vantagens e limitações da definição de caso de episódio hipotônico-hiporresponsivo (EHH) baseada na revisão da literatura, e as implicações para o Sistema de Informação de Eventos Adversos Pós-Vacinação do Programa Nacional de Imunizações. Materias e métodos: Este trabalho é baseado em dados obtidos por um estudo de vigilância ativade eventos adversos pós-imunização com a vacina combinada contra difteria, tétano, coqueluche e Haemophilus influenzae tipo b (DTP/Hib), realizado no município do Rio de Janeiro, no ano de 2004 (VIGAT), cujo objetivo primário era determinar em crianças com menos de 1 ano de idade a incidência do episódio hipotônico-hiporresponsivo (EHH) que se inicia nas primeiras 48 horas após a aplicação da vacina DTP/Hib utilizada pelo Programa Nacional de Imunização. Foram analisados o desempenho das perguntas de triagem para potenciais casos de EHH aplicadas no VIGAT, a reprodutibilidade da sua definição de caso de EHH quando aplicada em diferentes níveis de avaliação, o seu valor preditivo positivo, assim como, o seu desempenho em relação a outras definições de caso presentes na literatura, considerando-se o grau em que esse desempenho afeta a vigilância de eventos adversos associados à vacina DTP/Hib. Foi realizada também uma análise demográfica e dos antecedentes pessoais dos casos de EHH, e da sua possível associação com fatores relacionados à vacina, sua aplicação e outros eventos adversos pós-vacinais (EAPV). Resultados: A freqüência de EHH encontrado no VIGAT foi de 8,1 por 10.000 vacinados (IC95 por cento 4,7 - 13,0 por 10.000). Não houve significância estatística para a associação entre o EHH e o sexo do lactente, baixo peso de nascimento, índice de Apgar abaixo do normal, dose da vacina aplicada e lotes dos componentes vacinais. A febre maior ou igual a 39°C foi considerada como fator de risco para a ocorrência de EHH, encontrando-se associação entre eles com risco relativo de 8,2 (IC 95 por cento = 2,6; 25,7). O valor preditivo positivo da definição de caso do EHH adotada no VIGAT (2004) para o diagnóstico de EHH, quando utilizada pelo coordenador local do estudo foi 0,68 (17/25). Houve 100 por cento de concordância no diagnóstico de EHH entre o Grupo de Revisão de Diagnóstico (GRD) e o Comitê de Monitoramento Externo (CME). O Valor Preditivo Positivo (VPP) das duas perguntas de triagem combinadas foi de 17/47 (36,2 por cento). A concordância entre a definição de caso de EHH do VIGAT e a de Braun et al (1998) foi de 0,84 (kappa = 0,68); de 0,8 (kappa = 0,61) em relação a Brighton (2004); de 0,84 (kappa = 0,66) para Vermeer et al (2006); e de 0,92 (kappa = 0,82) para Brighton (2007). Conclusões: A definição de caso de EHH do VIGAT, norteada por perguntas de triagem de elevada sensibilidade e aplicada por uma equipe capacitada, apresentou bom desempenho na vigilância ativa desse EAPV. Entretanto, observou-se uma dificuldade no processo de diagnóstico de EHH, em função das características inerentes a esse evento, chamando atenção para a necessidade de criterios a avaliação dos dados obtidos por vigilância passiva, principalmente, na comparabilidade entre eles. / Objectives: To analyze the advantages and limitations of the case definition for hypotonichyporesponsive episode (HHE) based on the literature review, and the implications for the Postvaccination Adverse Events (VAE) Information System of the National Immunizations Program. Materials and methods: This work is based on data obtained by an active surveillance study of post-vaccination adverse events with the combined diphtheria-tetanus-whole cell pertussis-
Haemophilus influenzae type b vaccine performed in the Rio de Janeiro municipality in 2004
(VIGAT), whose primary objective was to determine the incidence of hypotonic-hyporesponsive
episode (HHE) that began in the first 48 hours after the application of the DTP/Hib vaccine used
by the National Immunization Program in children under the age of 1 year. The performance of
the screening questions for potentials HHE cases performed in VIGAT were analyzed, the
reproducibility of its definition for HHE cases when performed in different evaluation levels, its
positive predictive value, as well as its performance in relation to other case definitions present in the literature, considering in what degree this performance affects adverse events surveillance associated to the DTP/Hib vaccine. A demographic analysis and of the personal antecedents of HHE cases was also accomplished, and of its possible association with vaccine-related factors, its application and other vaccine adverse events (VAE). Results: The frequency of HHE found in VIGAT was 8.1 for 10,000 vaccinated subjects (CI 95% 4.7 –13.0 for 10,000). There was no statistical significance for the association between HHE and the sex of the infant, low birth weight, below normal Apgar index, dosage of vaccine and lots of vaccinal components. Fever equal to or higher than 39°C was considered as risk factor for the occurrence of HHE, having found an association among them with the relative risk of 8.2 (CI 95% = 2.6; 25.7). The predictive positive value for the HHE case definition adopted in VIGAT (2004) for the HHE diagnosis, when used by the local study coordinator was 0.68 (17/25). There was a 100%
agreement in the HHE diagnosis between the Diagnosis Review Group (DRG) and the External
Monitoring Committee (EMC). The Predictive Positive Value (PPV) of the two combined
screening questions was 17/47 (36.2%). The agreement between the HHE case definition of
VIGAT and of Braun et al (1998) was of 0.84 (kappa = 0.68); 0,8 (kappa = 0.61) in relation to
Brighton (2004); 0.84 (kappa = 0.66) for Vermeer et al (2006); and 0.92 (kappa = 0.82) for
Brighton (2007). Conclusions: The case definition for HHE of VIGAT, orientated by screening
questions of high sensibility and applied by a qualified team, presented good performance in the
active surveillance of that VAE. However, a difficulty was observed in the HHE diagnosis
process, due to the inherent characteristics to that event, highlighting the need for a criterial data evaluation obtained by passive surveillance, mainly, in the comparability among them.
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