Orientador: Sandra Helena Penha de Oliveira / Banca: Valeria Marçal Felix de Lima / Banca: Richardt Gama Landgraf / Resumo: Na inflamação alérgica, a histamina desencadeia papel crucial na indução de sintomas, tais como vasodilatação, broncoconstrição e produção de muco. O objetivo deste estudo foi investigar os efeitos da histamina na inflamação e no remodelamento das vias aéreas, avaliando para tanto a produção de muco e colágeno em modelo murino de inflamação pulmonar. Camundongos Balb/c machos de 20-25g foram estimulados, por via intratraqueal, com histamina em diferentes concentrações (10, 50 e 100 µM) e avaliados após 6, 24 e 48 horas. A partir dos dados resultantes do ensaio dose-resposta, foi realizado tratamento farmacológico, onde animais foram separados em 5 grupos (6 camundongos por grupo): Grupo 1 (PBS), controle não estimulado; Grupo 2 (Histamina), controle estimulado; Grupo 3 (Histamina + Loratadina); Grupo 4 (Histamina + Dexametasona) e Grupo 5 (Histamina + Dexametasona e Loratadina). Analisamos a migração de leucócitos no lavado broncoalveolar (LBA) e no tecido pulmonar. As alterações estruturais no tecido pulmonar e na traqueia foram avaliadas por análise histopatológica, bem como a produção de muco e deposição de colágeno usando Alcian blue/Periodic Acid Shiff e Tricômio de Masson, respectivamente. O sobrenadante do LBA e o tecido pulmonar foram coletados para quantificar os níveis SCF e CCL3 por ELISA de captura. A expressão gênica de Muc5ac, Gob-5, Col1a2 e receptores de histamina foi avaliada por RT-PCR em tempo real. Nossos resultados demonstram que a histamina induz infla... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: In allergic inflammation, histamine exerts a crucial role in the induction of symptoms, such as vasodilation, bronchoconstriction and mucus production. The aim of this study was to investigate the effects of histamine in inflammation and airway remodeling, evaluating the mucus production and collagen in a murine model of pulmonary inflammation. Male Balb/c mice of 20-25g were challenged by intratracheal instillation of histamine at different concentrations (10, 50 and 100 μM) and evaluated after 6, 24 and 48 hours. From the data resulting from the dose-response assay, pharmacological treatment was performed, and animals were separated into 5 groups (6 mice per group): Group 1 (PBS), non-challenged control; Group 2 (Histamine), challenged control; Group 3 (Histamine + Loratadine); Group 4 (Histamine + Dexamethasone) and Group 5 (Histamine + Dexamethasone and Loratadine). We analyzed the recruitment of leukocytes in bronchoalveolar lavage (BAL) and lung tissue. Structural changes in lung tissue and trachea were assessed by histopathological analysis, as well as mucus production and collagen deposition using Alcian blue/Periodic Acid Shiff and Masson's Trichrome, respectively. BAL supernatant and lung tissue were collected for SCF and CCL3 levels quantification by capture ELISA. The gene expression of Muc5ac, Gob-5, Col1a2, and histamine receptors was evaluated by RT-PCR real time. Our results demonstrated that histamine induces airway inflammation, with presence of leukocyte infiltrate in BAL and lung tissue. In addition, histamine-challenged animals demonstrated a significant increase in the gene expression of Muc5ac, Gob-5 and Col1a2 in lung tissue, SCF and CCL3 production in BAL and lung, as well as structural changes in the trachea and lung tissue, such as leukocyte infiltrate, presence of goblet cell, mucus production and collagen... / Mestre

Made available in DSpace on 2016-06-02T20:18:19Z (GMT). No. of bitstreams: 1 4861.pdf: 2511284 bytes, checksum: 341c153935f7b09c5b1226bdc2f7241d (MD5) Previous issue date: 2013-02-19 / Universidade Federal de Minas Gerais / The aim of the first study was to investigate behavioural alterations and Fos expression in gerbils submitted to functional training after unilateral cerebral ischaemic injury. The behavioural procedures that were used to test functional recovery were the vibrissal stimulation test and the pellet-collecting task. Twenty-four hours after surgery, the animals were submitted to the experimental procedure, which lasted three weeks. The animals were distributed into four subgroups: sham lesion, sham lesion and trained, ischaemic injury and ischaemic injury and trained. The motor cortex, hippocampus and dorsal striatum were subjected to immunohistochemistry reaction for Fos expression, and the mean number of cells in each region was calculated. Behavioural data were analysed using the Kruskal Wallis test (p &#8804; 0.05), and the morphological data were subjected to one-way Analysis of Variance, both followed by the Student-Newman-Keuls test (p &#8804; 0.05). The animals in the injury group presented significantly lower averages in the pellet-collecting task and higher percentages of error in the vibrissal stimulation test when compared to animals in the sham lesion group, indicating that the lesion impaired motor performance in both tests. After training, the trained groups showed a significant increase in the pellet-collecting task compared to untrained groups, demonstrating that functional training improves task performance in sham and injured animals. A significant increase in the activation of neurons in the motor cortex and hippocampus of the injury and trained group was observed when compared to the injured group. Functional training increased the motor performance of the trained animals, independently of lesion, in the pellet-collecting task and caused increased neuronal activation in the motor cortex and hippocampus in the injury animals. Moreover, the presented method is useful for the evaluation of motor performance in gerbils. The objective of the second study was to investigate the effects of the histamine precursor L-Histidine and the H1-receptor antagonist Pyrilamine on the functional recovery of gerbils (Meriones unguiculatus) following a unilateral ischemic brain injury. Two experiments were conducted under the same experimental procedures to test the effects of two pharmacological treatments in experiment I, L-Histidine doses of 100 mg/kg and 200 mg/kg and in experiment II, Pyrilamine was administered at doses of 35 mg/kg and 70 mg/kg. Both experiments were accompanied by control groups for the drug (saline), the lesion (sham lesion) and training on the pelletcollecting task (untrained). The behavioural tests that were used to test functional recovery were the vibrissal stimulation test and the pellet-collecting task. The Kruskal-Wallis test (p<0.05), the Student-Newman-Keuls multiple comparison test (p<0.05) and a Bonferroni correction (p<0.025) were used for statistical analysis. It was established that a unilateral cerebral ischemic injury impairs motor performance and that training on the pellet-collecting task improves performance independently of the lesion. L-Histidine impaired functional recovery when administered in combination with motor training, while Pyrilamine in combination with training potentiated functional recovery. With respect to the vibrissal stimulation test, the injured animals that received training in combination with L-Histidine showed a memory recall deficit; however, Pyrilamine had the opposite effect in animals that were injured and trained. These results suggest that the neural histaminergic system participates in the process of functional recovery of gerbils following a unilateral ischemic brain injury. / O Estudo I teve como objetivo testar um modelo experimental de recuperação funcional da pata anterior para gerbilos após lesão isquêmica unilateral encefálica e investigar as alterações comportamentais e expressão de Fos em gerbilos submetidos a treino motor após lesão isquêmica unilateral encefálica. Os testes comportamentais utilizados foram o teste de estimulação da vibrissa e a tarefa de recolher tablete. Os animais foram distribuídos em quatro subgrupos: Lesão fictícia (n= 12), Lesão fictícia treinado (n= 11), Lesão (n= 11) e Lesão treinado (n= 12). Cortes das regiões do córtex motor, hipocampo e estriado dorsal foram expostos à reação de imunohistoquímica para expressão da Fos, em seguida, foi calculado o valor médio do número de neurônios em cada região. Os dados comportamentais foram analisados através do teste de Kruskal-Wallis e os dados morfológicos foram submetidos à análise de variância de uma via, ambos seguidos do teste Student-Newman-Keuls (p&#8804;0,05). Os grupos lesados apresentam médias significativamente menores na tarefa de recolher tablete e maiores porcentagens de erro no teste de estimulação da vibrissa quando comparados aos grupos lesão fictícia, indicando que a lesão prejudicou o desempenho motor em ambos os testes. No ultimo treino, os grupos treinados apresentaram aumento significativo na tarefa de recolher tablete em relação aos grupos não treinados, demonstrando que o treino funcional melhora o desempenho da tarefa em animais intactos e lesados. Houve aumento significativo na marcação dos neurônios do córtex motor e hipocampo do grupo lesado treinado quando comparado ao grupo lesado. O treino funcional aumentou o desempenho motor em animais treinados independente da lesão na tarefa de recolher tablete e provocou aumento da ativação neuronal no córtex motor e hipocampo dos animais lesados. Além disso, o modelo experimental utilizado se mostrou útil para avaliar o desempenho motor da pata anterior de gerbilos. O Estudo II teve como objetivo investigar o efeito do precursor da histamina (Lhistidina) e do antagonista dos receptores H1 (Pirilamina) na recuperação funcional de gerbilos (Meriones unguiculatus) após lesão isquêmica unilateral do encéfalo. Para isso, foram realizados dois experimentos, sendo empregados os mesmos procedimentos experimentais, no entanto, diferem nos tratamentos farmacológicos realizados, que foram: I - L-histidina (doses: 100 mg/kg e 200 mg/kg) e II - Pirilamina (doses: 35 mg/kg e 70 mg/kg), sendo ambos acompanhados de grupos controles da droga (salina), cirurgia (lesão fictícia) e treino motor (não treinado). Os testes comportamentais utilizados foram o teste de estimulação da vibrissa e a tarefa de recolher tablete. Foi utilizado o Teste de Kruskal-Wallis (p<0,05), seguido do Teste de comparações multiplas de Student Newman - Keuls (p<0,05) e correção de Bonferroni (p<0,025). Verificou-se que a lesão prejudica o desempenho motor e o treino da tarefa de recolher tablete melhora o desempenho de animais independente da lesão. A L-histidina associada com o treino motor prejudicou a recuperação funcional, enquanto, a pirilamina combinada com o treino potencializou a recuperação funcional. Com relação ao teste de estimulação da vibrissa, os animais lesados submetidos ao treino e tratados com l-histidina apresentaram déficit de evocação da memória, contudo, a pirilamina possui efeito contrário quando aplicada em animais lesados e treinados. Os resultados sugerem que o sistema histaminérgico neural participa do processo de recuperação funcional de gerbilos submetidos à lesão isquêmica unilateral encefálica.

Made available in DSpace on 2016-06-02T20:19:28Z (GMT). No. of bitstreams: 1 887.pdf: 469528 bytes, checksum: fc5bcdbae3a402aaac1aae4dbf59bcd1 (MD5) Previous issue date: 2006-03-03 / Universidade Federal de Sao Carlos / This work intended to investigate in forebrain ablated Carassius auratus the effect of the H1 receptor blockade in a spatial choice task using a T-maze test with positive reinforcement. Sixty-nine goldfish were submitted to surgery for removal of both telencephalic lobes five days before the beginning of the experiment. A T-shaped glass aquarium, which had two feeders located at the extremities of the long arm, was employed. One of the two feeders was blocked. The experimental trials were performed in nine consecutive days. Each fish was individually placed in the short arm and confined there for thirty seconds, then it was allowed to swim through the aquarium to search for the food for ten minutes (maximum period). Time to find food was analysed in seconds. Animals were injected intraperitoneally with chlorpheniramine (CPA) at 16 mg/kg of body weight or saline after every trial, ten minutes after it had been replaced at house aquarium. Data were analyzed via a three-way analysis of variance (ANOVA) with repeated measures, and post hoc comparisons were analyzed using the Student Newman-Keuls test (p<0.05). All the training latencies of A-SAL group were significantly higher than latencies of S-SAL group (p<0.05). S-SAL group showed a significative decrease in latencies at the second trial and maintained that reduction until the end of the experiment (p<0.05), S-CPA group demonstrated a decrease only after the fourth trial (p<0.01), A-SAL group showed reduced latencies after the fifth trial (p<0.05), and ACPA group did not showed any difference between training latencies. Thus, our results showed that CPA seems to impair the consolidation of memory either on telencephalon ablated animals and in sham-operated ones. Furthermore, this work supports evidence that histamine plays a facilitatory role in learning and memory processes of a spatial task through its action in primitive structures of the brain in teleost fish. / O objetivo deste trabalho foi investigar, em peixes da espécie Carassius auratus submetidos à ablação telencefálica, o efeito do bloqueio do receptor H1 em uma tarefa de escolha espacial utilizando um teste de labirinto em T com reforço positivo. Sessenta e nove peixes foram submetidos à cirurgia para a remoção de ambos os lobos telencefálicos cinco dias antes do início do experimento. Foi empregado um aquário de vidro transparente em forma de T, o qual teve dois comedouros fixados nas extremidades do braço longo, sendo que um dos comedouros foi bloqueado durante as sessões de treino. O experimento foi realizado em nove sessões consecutivas realizadas com intervalos de 24 horas. Individualmente, cada peixe foi confinado no compartimento de saída do aquário experimental durante trinta segundos, e então foi permitido a ele explorar todo o aquário para procurar o alimento durante dez minutos (período máximo). O tempo utilizado para encontrar o alimento foi cronometrado em segundos. Os animais foram injetados intraperitonealmente com clorfeniramina (CPA), na dose de 16 mg/kg de peso corporal, ou salina após cada treino, dez minutos após o peixe ter sido recolocado no aquário de manutenção. Os dados foram analisados através de ANOVA de três vias seguida pelo teste de comparações múltiplas de Student Newman-Keuls. Todas as latências de treino do grupo A-SAL foram significativamente maiores do que as do grupo LFSAL (p<0,05). O grupo LF-SAL demonstrou diminuição significativa das latências apartir do segundo dia de treino (p<0,05), o LF-CPA apresentou esta diminuição após o quarto dia de treino (p<0,01), o A-SAL apresentou redução após o quinto dia de treino (p<0,05), e não foram encontradas diferenças entre as latências no grupo A-CPA. Assim, nossos resultados mostraram que a CPA parece prejudicar a consolidação da memória tanto nos animais que tiveram o telencéfalo removido quanto nos que foram submetidos à lesão fictícia. Além disto, este trabalho dá suporte às evidências de que a histamina desempenha um papel facilitatório nos processos de aprendizagem e memória em uma tarefa de escolha espacial, atuando em estruturas primitivas do cérebro de teleósteos.

Este estudo teve por objetivos: a) caracterizar o influxo de leucócitos (LC) induzido pelo veneno de Bothrops moojeni (VBm); b) avaliar o papel dos mastócitos (MC) e da histamina neste evento; c) analisar a liberação de TXA2, PGD2, LTB4, MCP-1 e KC e d) avaliar a capacidade do VBm desgranular MC in vivo e in vitro. A injeção intraperitoneal de VBm, em camundongos, causou o recrutamento de LC e neutrofilia. O tratamento dos animais com cromoglicato aboliu o influxo de LC enquanto a difenidramina, ranitidina e a tioperamida, antagonistas da histamina, reduziram o influxo de neutrófilos. Ainda, o veneno induziu a liberação de PGD2, TXA2, LTB4, MCP-1 e de KC e causou a desgranulação de MC in vivo e a liberação de -hexosaminidase de MC in vitro. Em conclusão, o VBm induz influxo de LC para o local de sua injeção. Este efeito depende da histamina, via receptores H1, H2 e H4 e da desgranulação de MC, que decorre de ação direta do veneno nestas células. A neutrofilia e o TXA2, LTB4, MCP-1 e KC devem contribuir para o influxo de leucócitos causado pelo VBm. / In this study the effects of Bothrops moojeni venom (BmV) on the cellular component of inflammatory responses and the mechanisms involved in this effect were investigated. The effects of venom on peritoneal and circulating leukocyte numbers and on the release of inflammatory mediators, such as LTB4, TXA2, PGD2, MIP-1 and KC, were assessed. The role of both mast cells and histamine receptors in leukocyte recruitment induced by BmV was assessed by selected pharmacological treatments. BmV caused a marked infiltration of leukocytes (3-24 h) when injected into the peritoneal cavity of mice. Neutrophils (PMN) were the predominant cell type in the early stages of response whereas macrophages (MN) were accumulated from 3 up to 24 h. Moreover, BmV increased blood neutrophil numbers at 3 h after injection. The BmV-induced leukocyte influx was abrogated by cromoglicate and significantly reduced either by difenidramine or ranitidine or tioperamide, histamine H1, H2 and H4 receptor antagonists, respectively, at 6 h after injection. Significant increments in peritoneal levels of LTB4, TXA2, PGD2, MIP-1 and KC were detected at distinct periods of time after venom injection. In addition, BmV induced mast cell degranulation both in vivo and in vitro. In conclusion, obtained data demonstrated the ability of BmV induce leukocyte recruitment into the site of its injection. This effect is dependent on mast cell activation and degranulation, which may be due to a direct effect of venom on these cells, and is mediated at least in part by histamine via H1, H2 and H4 receptors. Moreover, the ability of venom to mobilize leukocytes from bone marrow reserve compartments and to release the chemotactic mediators TXA2, LTB4, MCP-1 and KC may be relevant for leukocyte infiltration.

Histamine is an organic base that develops an important physiological and metabolic function in live organisms. It is formed in normal metabolic processes and through microorganisms enzymatic activity during the fermentation or deterioration. This substance may cause problems when ingested in large amounts, when the eliminating mechanism is inefficient or when potentializers are present. The level of histamine in food varies a lot and it depends on the nature, origin, processing stages and microorganisms present. In wine, histamine has its origin in the grape itself or can be formed by the wine elaborating process, during the alcoholic and/or malolatic fermentation. The objective of this paper was to accompany the histamine level increase since the last maturation month of the grape until the wine storage period for three months. Microvinifications were done with inoculation of selected yeast (Saccharomyce cerevisiae variety bayanus and Saccharomyces cerevisiae) and without inoculation of selected yeast. In the end of the alcoholic fermentation the three treatments were divided in two groups: one was inoculated with a lactic bacteria Leuconostoc oenos and the other was not inoculated with a lactic bacteria. After the malolatic fermentation ending the wine was bottled and maintained for three months in environment temperature. The histamine after chemical derivatization with orthophthaldehyde was quantificated by High Performance Liquid Chromatography with fluorescence detection in grapes harvested during the maturation, after the grapes squashing, in the end of the alcoholic fermentation, after the malolatic fermentation and three months after the bottling. During the grape maturation, the histamine concentration increased in a higher proportion close to the harvest date. In the alcohol fermentation ending there was a reduction on the histamine level in all the three treatments. In the end of the malolatic fermentation it was noticed that the histamine concentration increased in all treatments in spite of fermentation been occurred spontaneously or by induction except to the must that realized both fermentations without the inoculation of selected organisms. In the samples collected three months after the bottling the histamine concentration found in the wine did not present a significant difference in relation to the final concentration of malolatic fermentation except to the wine that realized both fermentations without pure culture where a significant increase in the histamine concentration was found (p<0,05). It was not observed a strong correlation between the histamine levels and the chemical characteristics of pH, total acidity, volatile acidity, alcohol content and free SO2. / A histamina é uma base orgânica que desempenha importante função fisiológica e metabólica nos organismos vivos. É formada durante processos metabólicos normais e através de atividade enzimática de microorganismos durante fermentação ou deterioração. A ingestão desta substância pode causar problemas quando for em grande quantidade, quando o mecanismo de eliminação for deficiente ou ainda quando potencializadores estiverem presentes. A quantidade de histamina em alimentos varia bastante, sendo dependente da natureza, origem, etapas do processamento e microorganismos presentes. Em vinhos, a histamina pode ter sua origem na própria uva ou então ser formada durante o processo de fabricação do vinho, durante a fermentação alcoólica e/ou malolática. O objetivo deste trabalho foi acompanhar a evolução dos níveis de histamina desde o último mês de maturação da uva até um período de estocagem do vinho por três meses. Foram realizadas microvinificações com inoculação de leveduras selecionadas (Saccharomyce cerevisiae variedade bayanus e Saccharomyces cerevisiae) e sem inoculação de levedura selecionada. Ao final da fermentação alcoólica, os três tratamentos foram divididos em dois grupos: em um deles houve inoculação de bactéria lática Leuconostoc oenos e no outro não foi inoculada a bactéria lática. Após o término da fermentação malolática, os vinhos foram engarrafados e mantidos por três meses em temperatura ambiente. A histamina após derivatização química com orto-ftaldeído foi quantificada por Cromatografia Líquida de Alta Eficiência com detecção por Fluorescência em uvas colhidas durante a maturação, após o esmagamento das uvas, no final da fermentação alcoólica, após a fermentação malolática e três meses após o engarrafamento. Durante a maturação da uva houve um aumento mais pronunciado na concentração de histamina próximo à data da colheita. Ao final da fermentação alcoólica houve uma redução dos níveis de histamina nos três tratamentos. Ao término da fermentação malolática observou-se que a concentração de histamina aumentou em todos os tratamentos independente de a fermentação ter ocorrido espontaneamente ou por indução, exceto para o mosto que realizou as duas fermentações sem inoculação de microorganismos selecionados. Em amostras coletadas três meses após o engarrafamento, a concentração de histamina encontrada nos vinhos não apresentou diferença significativa em relação à concentração ao final da fermentação malolática, exceto para o vinho que realizou as duas fermentações sem inoculação de cultura pura, onde houve um aumento significativo na concentração de histamina (p 0,05). Não foi observada uma correlação forte entre os níveis de histamina e as características químicas de pH, acidez total, acidez volátil, teor alcoólico e SO2 livre.

A imunodeficiência comum variável (ICV) é uma síndrome heterogênea caracterizada por hipogamaglobulinemia e infecções bacterianas de repetição. As doenças obstrutivas, como a asma, estão presentes em aproximadamente 50% dos pacientes. Os sintomas decorrentes de infecções respiratórias de repetição podem mascarar os sintomas de alergia respiratória. A asma tem alta prevalência no mundo e é observada em aproximadamente 10% da população brasileira. Embora muitos pacientes com ICV apresentem história clínica sugestiva de rinite e/ou asma alérgicas, a participação da atopia não está bem esclarecida e freqüentemente os níveis de IgE total e/ou IgE específica estão baixos. Muitos autores estudam a produção de IgE local e uma correlação entre a concentração de IgE nos fluidos corporais e no soro existe. Os objetivos deste estudo são avaliar a função pulmonar em pacientes com ICV através de: espirometria, provocação brônquica com histamina e com alérgeno; investigar o diagnóstico de asma em pacientes com ICV e realizar a investigação in vivo e in vitro da IgE em pacientes com ICV. Este estudo incluiu 62 pacientes que estavam em acompanhamento ambulatorial no Serviço de Imunologia Clínica e Alergia do Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo. A função pulmonar foi avaliada pela espirometria e pela provocação brônquica com histamina antes e após uma provocação brônquica com Dermatophagoides pteronyssinus (Der p) e a investigação da IgE específica para aeroalérgenos através de teste epicutâneo e avaliação da IgE sérica específica usando ImmunoCAPTM. Vinte e nove (46,7%) tinham história clínica de sugestiva de asma e em relação à atopia, 27 (43,5%) tinham história sugestiva de atopia. Uma associação de asma e atopia no mesmo paciente foi encontrada em 18 pacientes (29%). Nós comparamos o grupo sugestivo de asma alérgica com os outros pacientes pacientes com rinite alérgica ou não-alérgica, asma não-alérgica e pacientes sem sintomas respiratórios. A maioria dos pacientes apresentou níveis séricos de IgE total indetectáveis. Somente dois pacientes apresentaram resultado positivo para IgE específica pelo teste epicutâneo e in vitro. Sessenta e um pacientes realizaram espirometria. Destes, 25 pacientes (41%) apresentaram resultado normal, 29 (47,5%) apresentaram distúrbio ventilatório obstrutivo e 7 (11,5%) apresentaram resultados sugestivos de distúrbio ventilatório restritivo. As provocações brônquicas foram realizadas em 15 pacientes. A provocação brônquica com histamina foi considerada positiva em 3 pacientes com história positiva para asma. Em relação à provocação brônquica com Der p, nenhum paciente apresentou resposta imediata positiva. Entretanto, quando a segunda provocação brônquica com histamina foi realizada (pós-Der p), quatro dos 5 pacientes com história sugestiva de asma alérgica apresentaram resultado positivo, com diminuição de PC20 em relação à primeira provocação vi brônquica com histamina. Uma diferença estatística foi observada nos resultados entre o grupo sugestivo de asma alérgica e os pacientes sem asma alérgica. Ao final do estudo, a asma foi confirmada em 9 pacientes com ICV (14,5%), a atopia foi confirmada em 6 pacientes com ICV (9,7%) e a asma alérgica foi confirmada em 4 pacientes com ICV (6,5%), que correspondeu a 22,2% dos 18 pacientes com suspeita de asma alérgica / Common variable immunodeficiency (CVID) is a heterogenous immunodeficiency syndrome characterized by hypogammaglobulinemia, and recurrent bacterial infections. Obstructive diseases as asthma are present in approximately 50 % of patients. Symptoms due to recurrent respiratory pyogenic infections may mask respiratory allergic symptoms. Asthma has high worldwide prevalence and is observed in approximately 10 % of Brazilian population. Although a number of patients with CVID report a clinical history suggestive of allergic symptoms, the role of atopy is not well established in these individuals; and frequently levels of total IgE and/or specific IgE are low. Local IgE production has been studied and a correlation between IgE concentration in body fluids and serum exists. The objectives of this study are evaluation of pulmonary function in patients with CVID through: spirometry, bronchial challenge with histamine, and with allergen; investigate asthma diagnosis in patients with CVID; perform in vivo and in vitro investigation of IgE in patients with CVID. This study included sixty-two patients, who were being followed at the Service of Clinical Immunology and Allergy of the Hospital das Clínicas of the University of São Paulo Medical School. Pulmonary function was assessed using spirometry and bronchial challenge with histamine before and after a bronchial challenge with Dermatophagoides pteronyssinus (Der p), and investigation of specific IgE for aeroallergens with skin prick test and serum specific IgE evaluation using ImmunoCAPTM. Twenty-nine (46.7 %) had clinical history suggestive of asthma, and in regards to atopy, twenty-seven patients (43.5%) reported atopy suggestive history. An association of asthma and atopy in the same patient was observed in eighteen (29 %) participants. We compared the group of allergic asthma with the other patients patients with allergic or non-allergic rhinitis, non-allergic asthma, and patients without respiratory. Most patients had undetectable levels of total IgE concentration in serum. Only two patients had positive results for specific IgE by prick test and in vitro investigation. All patients, except one, underwent spirometry test for lung function evaluation. Of the sixty-one patients, twenty-five (41 %) had normal spirometry results, twenty-nine (47.5 %) had Obstructive Ventilatory Defect, and seven (11.5 %) had results suggestive of Restrictive Ventilatory Defect. Bronchial challenges were performed in fifteen patients. Bronchial challenge with histamine was considered positive in three patients with a positive history of asthma. Regarding to bronchial challenge with Der p, none presented immediate positive response. However, when the second nonspecific bronchial provocation with histamine was performed (post-Der p), four of the five patients with a history of allergic asthma had positive test results, with lower PC20 than in the first non-specific bronchial provocation with histamine. A statistical difference was noticed in the test results of the group suggestive for allergic asthma and the patients without allergic asthma. viii At the end of this study, asthma had been confirmed in 9 patients with CVID (14.5%), atopy had been confirmed in 6 patients with CVID (9.7%), and allergic asthma had been confirmed in 4 patients with CVID (6.5%), which corresponded to 22.2% of the 18 patients suspected of allergic asthma

Signal Transducers and Activators of Transcription (STATs) are latent transcription factors that mediate several cellular responses. This protein family consists of seven members, STAT1 – 6 including two closely related molecules, STAT5a and STAT5b, that show 96% amino acid sequence homology and are critical for lymphoid, myeloid and erythroid cell development and function. Activated STAT proteins dimerize and translocate to the nucleus, where they bind to high-affinity DNA motifs to modulate gene expression. We recently identified STAT5b as the critical regulator of IgE-mediated cytokine production in mast cells. STAT5b knockout (KO) cells show decreased sensitivity to IgE-mediated passive systemic anaphylaxis accompanied with decreased production of IL-6 and IL-13 compared to wild type counterparts. Interestingly, STAT5b KO mice demonstrated elevated levels of serum IgE but a normal response to histamine-mediated passive systemic anaphylaxis. The current work demonstrates that STAT5b regulates mast cell function both in vivo and in vitro. Additionally, activated STAT proteins can also form tetramers through an N-terminal domain-mediated oligomerization process when bound to low-affinity tandem motifs. Dr. Warren Leonard’s laboratory generated STAT5a-STAT5b double knock-in (DKI) mice in which STAT5 proteins are phosphorylated and can form dimers but not tetramers. We have now found that bone marrow-derived mast cells from STAT5 DKI mice are defective in IgE-induced cytokine and chemokine production and exhibit defective stem cell factor (SCF)-induced migration and survival responses in vitro. Similarly, IgE-mediated passive systemic anaphylaxis is decreased in STAT5 DKI mice. These data indicate that Stat5 tetramers are critical for some aspects of mast cell function in allergic and inflammatory disease.

Chronic pressure overload (PO) is associated with cardiac hypertrophy and altered autonomic control of cardiac function, in which the latter may involve adaptations in central and/or peripheral cardiac neural control mechanisms. To evaluate the specific remodeling of the intrinsic cardiac nervous system following pressure overload, the descending thoracic aorta artery of the guinea pig was constricted ∼20%, and the animals recovered for 9 wk. Thereafter, atrial neurons of the intrinsic cardiac plexus were isolated for electrophysiological and immunohistochemical analyses. Intracellular voltage recordings from intrinsic cardiac neurons demonstrated no significant changes in passive membrane properties or action potential depolarization compared with age-matched controls and sham-operated animals, but afterhyperpolarization duration was increased in PO animals. Neuronal excitability, as determined by the number of action potentials produced with depolarizing stimuli, was differentially increased in phasic neurons derived from PO animals in response to exogenously applied histamine compared with sham and age-matched controls. Conversely, pituitary adenylate cyclase-activating polypeptide-induced increases in intrinsic cardiac neuron evoked AP frequency were similar between control and PO animals. Immunohistochemical analysis demonstrated a two-fold increase in the percentage of neurons immunoreactive for neuronal nitric oxide synthase in PO animals compared with control. The density of mast cells within the intrinsic cardiac plexus from PO animals was also increased twofold compared with preparations from control animals. These results indicate that congestive heart failure associated with chronic pressure overload induces a differential remodeling of intrinsic cardiac neurons and upregulation of neuronal responsiveness to specific neuromodulators.

To explore a recently established association between histaminergic and dopaminergic neuronal phenotypic systems in brain, we determined the effect of the respective histaminergic H3 receptor agonist and antagonist/inverse agonist, imetit and thioperamide, on L-DOPA - derived tissue and extracellular dopamine (DA) and metabolite levels in the striatum of 6-hydroxydopamine (6-OHDA) - lesioned rats (i.e., parkinsonian rats). We also examined the influence of histamine H3 ligands on L-DOPA evoked behavioral responses (locomotor activity, number of rearings, stereotyped behavior and motor coordination). Using HPLC/ED and in vivo microdialysis technique, imetit (5 mg/kg, i.p.) but not thioperamide (5 mg/kg, i.p.) was shown to attenuate an L-DOPA-evoked (15 mg/ kg, i.p.; carbidopa, 30 min pretreatment) increase in extracellular DA in the neostriatum of 6-OHDA-lesioned rats. However, both imetit and thioperamide increased microdialysate levels of DOPAC and HVA, probably by enhancing intraneuronal DA utilization. As indicated by neurochemical analysis of the striatum imetit produced a decrease in tissue DA content. These findings support the hypothesis that central H3 histaminergic receptors have a modulatory role in the storage, metabolism and release of DA derived from exogenous L-DOPA challenge. Furthermore, evidence from behavioral studies indicate that histamine H3 receptor block markedly improved motor coordination. Conversely, histamine H3 receptor stimulation, being without effect on motor coordination, enhanced vertical activity in rats. From the above we conclude that histamine H3 agonism may augment motor dyskinesia in Parkinson's disease (PD) patients and presumably worsen L-DOPA therapy. Consequently, the histaminergic system represents a viable target for modulating the effectiveness of L-DOPA therapy in Parkinson's disease.

The solubility and dissolution of famotidine from solid glass dispersions of xylitol, prepared by the fusion method, were investigated. Preliminary stability studies revealed that famotidine was stable for 72 hours (< 0.5% decomposition) in water at 37° ± 0.5° C. Both the drug and the carrier were stable and did not decompose during the fusion process. About 4% decomposition of famotidine was observed after 72 hours in an aqueous solution of famotidine:xylitol glass dispersion at 37° ± 0.5° C. Solubility of famotidine from solid glass dispersions and physical mixtures with famotidine:xylitol ratios of 1:1, 1:20 and 1:40 was studied at 37° ± 0.5° C and found to be higher than that of famotidine alone in water. The solubility of famotidine from physical mixtures increased linearly with the increase in xylitol concentration, but the relationship was not linear for glass dispersions. The dispersions were more effective in enhancing the solubility of famotidine as compared to physical mixtures of corresponding drug:carrier ratios. A 1:40 glass dispersion increased the solubility by up to 32% while the solubility increase from a 1:40 physical mixture was 14%. Dissolution studies were carried out on glass dispersions with famotidine:xylitol ratios of 1:1, 1:10 and 1:20 in water at 37° ± 0.5° C. Results revealed a marked increase in the dissolution rate of famotidine from solid glass dispersions when compared to the dissolution rate of plain famotidine powder alone. The increase was greatest at the lowest drug level (1:20 drug:carrier ratio) with 100% of the drug dissolving within one minute. The glass dispersions were subjected to thermal analysis. Thermograms obtained by differential scanning calorimetry showed no evidence of chemical interaction between famotidine and xylitol. Phase diagrams were constructed for famotidine:xylitol solid glass dispersions and physical mixtures from melting temperatures determined by the capacity tube method. The phase diagram of the dispersion system suggested the formation of a eutectic mixture of famotidine and xylitol near a drug:carrier ratio of 1:40.