Resumo: O Linfoma de Hodgkin (LH) é uma neoplasia linfóide histologicamente caracterizada pela presença de escassas células neoplásicas peculiares, denominadas células de Hodgkin/Reed-Sternberg (H-RS), em meio a infiltrado inflamatório não-neoplásico. Seu curso clínico está diretamente relacionado ao estadiamento, subtipo histológico e presença de alguns fatores prognósticos adversos já estabelecidos. Sintomas constitucionais como febre, perda de peso e sudoreses noturna, associados a um aparente desbalanço da resposta imune celular, são características comuns ao diagnóstico e denotam anormalidade da resposta imunológica desses pacientes, provavelmente causada pela produção de diferentes citocinas, tanto pelas células H-RS como pelo infiltrado inflamatório de permeio. Na maior parte dos casos de LH, há produção aumentada de citocinas de padrão Th2. No presente estudo, foram determinadas, por ELISA, as concentrações séricas de IL-6, IL-10 e IL-13 de 28 pacientes com LH clássico, antes e depois do tratamento anti-neoplásico, e de 26 voluntários saudáveis que compuseram grupo-controle. Dosagens de IL-6 e IL-10 pré-tratamento foram significativamente maiores nos pacientes em relação ao grupo-controle. Todos os pacientes apresentaram redução do nível sérico de IL-6 e IL-10 após o tratamento antineoplásico. Ao diagnóstico, níveis elevados de IL-6 foram associados à presença de linfonodomegalia abdominal, hepatomegalia, sintomas B e anemia. O estudo da regressão linear múltipla mostrou que sintomas B e linfocitopenia são bons preditores de níveis séricos pré-tratamento de IL-6. Níveis elevados de IL-10 foram relacionados à hipoalbuminemia e hepatomegalia. Nenhum paciente apresentou dosagem Resumo 52 sérica detectável de IL-13. Os resultados demonstram que níveis séricos elevados de IL-6 e IL-10 ao diagnóstico estão... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Hodgkin's lymphoma (HL) is a malignant lymphoid neoplasia characterized by abnormal immune response. Part of this disturbance is attributable to the activity of cytokines produced by the malignant Hodgkin/Reed-Sternbergh cells and reactive inflammatory cells. IL-6, IL-10 and IL-13 seem to play an important role in the pathogenesis of HL. Although some features of the disease have been associated with the production of these interleukins, there is insufficient data about changes in its serum levels at diagnosis and after the treatment, as well as its potencial use as biomarkers of HL course. Design and Methods. Serum levels pre and post treatment of IL-6, IL-10 and IL-13 in 28 patients with HL were determined by ELISA. Results were evaluated against clinical and laboratory parameters, as well as response to treatment and presence of infection by the Epstein-Barr virus (EBV), assessed by in situ hybridization with biotinylated probe directed to the viral transcript EBER-1. Serum samples from 26 healthy blood-donors volunteers were evaluated as a control group. Results. IL-6 and IL-10 serum levels before treatment of HL were significantly higher in patients compared to healthy individuals (p<0,001), and a significant reduction after treatment of the disease was observed (p<0,001). Serum levels of IL-13 were indetectable both in patients (before and after treatment) and controls. Serum levels of IL-6 before treatment were higher in patients with abdominal involvement by HL (p=0,03), hepatomegaly (p=0,04), B-symptoms (p=0,01), and anemia (p=0,02). On the other hand, IL-10 pre-treatment levels were higher in patients with hypoalbuminemia (p=0,04) and hepatomegaly (p=0,01). Multivariate analysis Abstract 54 revealed that lymphocytopenia and B-symptoms were good predictors of IL-6 levels in serum before treatment of patients... (Complete abstract click electronic access below) / Orientador: Deilson Elgui de Oliveira / Coorientador: Lígia Niéro-Melo / Banca: José Salvador Rodrigues de Oliveira / Banca: Maria Claudia Nogueira Zerbini / Mestre

Trauma of diverse origins is a common reason for presentation of pets for treatment. It is often difficult clinically to objectively measure the severity of any trauma to an animal. One approach is to measure the changes in the various serum parameters which are known to alter in response to trauma or inflammation. If the changes over time of relevant and easily measurable parameters can be established under two controlled but different conditions of surgical trauma, it may provide the foundation for evaluating their future use in establishing the severity of trauma in a patient. A prospective study was performed on animals presented to the Onderstepoort Veterinary Academic Hospital for either thoracolumbar disc disease or for elective ovariohysterectomy. The two surgical procedures chosen for the study involved significant surgical trauma, particularly to muscle, in the case of thoracolumbar decompression and relatively minor surgical trauma in the case of ovariohysterectomy. Serial evaluation of creatine kinase (CK) and C-reactive protein (CRP) were performed both pre- and post-operatively on two sets of patients derived from the two surgical categories. CK is an enzyme found predominantly in skeletal muscle and significantly elevated serum activity is largely associated with muscle damage. CRP is an acute phase protein which shows elevated serum concentration in response to a broad range of inflammatory stimuli. Analysis of the data showed a very wide range of results at each time point for both CK and CRP. There were no significant differences between the two surgical groups for either analyte preoperatively. Thereafter CK results were markedly and significantly different between the two groups. CRP results were very similar in the two groups with no statistical difference at any time point. The results of this study suggest that the evaluation of CK and CRP at any one time point in a traumatized animal is of limited value. However the evaluation of the trend of these two analytes, even over a relatively short time period, may allow for useful prognostication in clinical cases. / Dissertation (MMedVet)--University of Pretoria, 2008. / Companion Animal Clinical Studies / unrestricted

Made available in DSpace on 2014-06-11T19:27:57Z (GMT). No. of bitstreams: 0 Previous issue date: 2008-08-21Bitstream added on 2014-06-13T18:56:52Z : No. of bitstreams: 1 gaiolla_rd_me_botfm.pdf: 867698 bytes, checksum: ec290cbafd254a98351e5ef8cea0e6de (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / O Linfoma de Hodgkin (LH) é uma neoplasia linfóide histologicamente caracterizada pela presença de escassas células neoplásicas peculiares, denominadas células de Hodgkin/Reed-Sternberg (H-RS), em meio a infiltrado inflamatório não-neoplásico. Seu curso clínico está diretamente relacionado ao estadiamento, subtipo histológico e presença de alguns fatores prognósticos adversos já estabelecidos. Sintomas constitucionais como febre, perda de peso e sudoreses noturna, associados a um aparente desbalanço da resposta imune celular, são características comuns ao diagnóstico e denotam anormalidade da resposta imunológica desses pacientes, provavelmente causada pela produção de diferentes citocinas, tanto pelas células H-RS como pelo infiltrado inflamatório de permeio. Na maior parte dos casos de LH, há produção aumentada de citocinas de padrão Th2. No presente estudo, foram determinadas, por ELISA, as concentrações séricas de IL-6, IL-10 e IL-13 de 28 pacientes com LH clássico, antes e depois do tratamento anti-neoplásico, e de 26 voluntários saudáveis que compuseram grupo-controle. Dosagens de IL-6 e IL-10 pré-tratamento foram significativamente maiores nos pacientes em relação ao grupo-controle. Todos os pacientes apresentaram redução do nível sérico de IL-6 e IL-10 após o tratamento antineoplásico. Ao diagnóstico, níveis elevados de IL-6 foram associados à presença de linfonodomegalia abdominal, hepatomegalia, sintomas B e anemia. O estudo da regressão linear múltipla mostrou que sintomas B e linfocitopenia são bons preditores de níveis séricos pré-tratamento de IL-6. Níveis elevados de IL-10 foram relacionados à hipoalbuminemia e hepatomegalia. Nenhum paciente apresentou dosagem Resumo 52 sérica detectável de IL-13. Os resultados demonstram que níveis séricos elevados de IL-6 e IL-10 ao diagnóstico estão... / Hodgkin’s lymphoma (HL) is a malignant lymphoid neoplasia characterized by abnormal immune response. Part of this disturbance is attributable to the activity of cytokines produced by the malignant Hodgkin/Reed-Sternbergh cells and reactive inflammatory cells. IL-6, IL-10 and IL-13 seem to play an important role in the pathogenesis of HL. Although some features of the disease have been associated with the production of these interleukins, there is insufficient data about changes in its serum levels at diagnosis and after the treatment, as well as its potencial use as biomarkers of HL course. Design and Methods. Serum levels pre and post treatment of IL-6, IL-10 and IL-13 in 28 patients with HL were determined by ELISA. Results were evaluated against clinical and laboratory parameters, as well as response to treatment and presence of infection by the Epstein-Barr virus (EBV), assessed by in situ hybridization with biotinylated probe directed to the viral transcript EBER-1. Serum samples from 26 healthy blood-donors volunteers were evaluated as a control group. Results. IL-6 and IL-10 serum levels before treatment of HL were significantly higher in patients compared to healthy individuals (p<0,001), and a significant reduction after treatment of the disease was observed (p<0,001). Serum levels of IL-13 were indetectable both in patients (before and after treatment) and controls. Serum levels of IL-6 before treatment were higher in patients with abdominal involvement by HL (p=0,03), hepatomegaly (p=0,04), B-symptoms (p=0,01), and anemia (p=0,02). On the other hand, IL-10 pre-treatment levels were higher in patients with hypoalbuminemia (p=0,04) and hepatomegaly (p=0,01). Multivariate analysis Abstract 54 revealed that lymphocytopenia and B-symptoms were good predictors of IL-6 levels in serum before treatment of patients... (Complete abstract click electronic access below)

Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico / A hansenÃase continua sendo um problema de saÃde mundial, sendo o Brasil o segundo paÃs em maior em nÃmero de casos novos. No Cearà a doenÃa à considerada endÃmica e em 2009 foram diagnosticados 1.952 casos novos, alcanÃando um coeficiente de detecÃÃo geral de 22,8/100.00 habitantes. A doenÃa à causada pelo M. leprae, manifesta-se atravÃs de sinais e sintomas dermatoneurolÃgicos e à transmitida de pessoa a pessoa atravÃs do convÃvio de indivÃduos suscetÃveis com doentes bacilÃferos sem tratamento. A interaÃÃo do M. leprae com os subtipos de cÃlulas T produz citocinas do tipo Th1 e Th2, responsÃveis pelas diferentes formas clÃnicas da doenÃa. PadrÃes de citocinas Th1 (IL-2, IFN-&#947; e TNF-&#945;) foram encontrados em lesÃes de pele das formas tuberculÃides e padrÃes de citocinas Th2 (IL-4, IL-5 e IL-10) foram encontrados em lesÃes das formas virchovianas. A hansenÃase à influenciada por vÃrios fatores, sendo os genÃticos os mais estudados no momento. Os genes das citocinas aparecem como fortes candidatos capazes de influenciar a interaÃÃo patÃgeno- hospedeiro e favorecer ou nÃo o desenvolvimento da doenÃa. A IL-10 à uma citocina anti-inflamatÃria e imunomoduladora que possui regiÃes promotoras bastantes polimÃrficas, contendo regiÃes de microssatÃlites e SNPs que formam vÃrios haplÃtipos que estÃo associados a diferentes nÃveis de produÃÃo de citocina in vitro. VÃrios estudos tentam reportar associaÃÃes entre os polimorfismos de IL-10 e o risco ou proteÃÃo para diversas doenÃas. Contudo, os dados relatados tem sido contraditÃrios e a maioria das associaÃÃes entre os polimorfismos e a produÃÃo dessa citocina sÃo baseados em estudos in vitro. Dessa forma o presente estudo teve o objetivo de definir como os polimorfismos da regiÃo promotora da citocina afetam a produÃÃo in vivo frente à infecÃÃo pelo M. leprae. Foram quantificadas as concentraÃÃes sÃricas de IL-10 de 181 indivÃduos, sendo 77 casos Ãndices de hansenÃase, 74 indivÃduos contactantes e 30 controles saudÃveis. Sendo que destes, 31 possuÃam anÃlise genotÃpica de IL-10 no grupo caso, 33 no grupo contactante e 29 no grupo controle. Os pacientes com anÃlise genotÃpica foram estratificados em baixo, mÃdio e alto produtor da citocina. As diferenÃas nos nÃveis da citocina foram comparadas entre os grupos dentro do espectro da hansenÃase (paucibacilar e multibacilar), dos controles externos, dos controles internos, dos genÃtipos e alelos encontrados nos grupos. Foram realizados testes de Kruskall-Wallis e Mann-Whitney para anÃlise das medianas de IL-10 em pg/mL. NÃo foi encontrada diferenÃa significante entre os grupos caso, contactante e controle (p=0,7450), entre os indivÃduos pauci e multibacilar (p=0,7898), entre os fenÃtipos de nÃvel de produÃÃo de citocina (baixo, mÃdio e alto) (p=0,4355). Foi encontrada diferenÃa significante na produÃÃo de IL-10 entre os alelos -1082A,-819C e -592C do grupo caso em relaÃÃo aos controles (p<0,05) e dos alelos -819C e -592C do grupo caso em relaÃÃo aos contactantes (p<0,05). NÃo foi encontrada diferenÃa significante entre os grupos contactante e controle (p>0,05). Em conclusÃo, os genÃtipos de IL-10, -1082G>A, -819C>T e -592G>C nÃo influenciaram a produÃÃo e/ou o desfecho da infecÃÃo pelo M. leprae. Por outro lado, os alelos -1082A,-819C e -592C determinaram menor produÃÃo de IL-10 em indivÃduos com hansenÃase. / Leprosy is a world problem of health and Brazil has the second higher rates of new cases in the world. It is considered an endemic disease at Cearà and a total of 1,952 new cases were detected, reaching a detection rate of 22.8/100,000 inhabitants. The interaction between the M. leprae and different T cells stimulates the production of a Th1 or a Th2 pattern of cytokines, which are responsible for the different clinical forms of leprosy. Th1 cytokines (IL-2, IFN-&#947; and TNF-&#945;) were found in tuberculoid skin lesions while Th2 cytokines (IL-4, IL-5 and IL-10) were found in lepromatoid lesions. Leprosy is influenced by many distinct factors, among them, genetic factors are the most studied at this moment. Cytokine genes seem to influence the interaction between the pathogen and the host and contribute to the development or not of the disease. The IL-10 is an antiinflammatory and immunomodulatory cytokine which has too much polymorphic promoter regions with microsatellites and SNPs. Different haplotypes are associated to distinct levels of cytokine production in vitro. Many studies reported associations between IL-10 polymorphisms and the risk or the protection against many diseases. However, the data reported have been contradictory and most of the associations between these polymorphisms and the production of IL-10 are showed in in vitro studies. The aim of this study was to evaluate how the promoter region polymorphisms of IL-10 influence the cytokine levels production in vivo, during the M. leprae infection. Serum levels of IL-10 were analyzed by ELISA in 181 individuals, 77 of them were leprosy cases, 74 household contacts and 30 healthy controls. Among them, 31 had IL-10 polymorphism typed in the case group, 33 in household contact group and 29 in healthy controls. Cases with polymorphism typed were stratified in low, medium and high levels of cytokine production. Differences in the IL-10 production were compared among leprosy cases (pauci and multibacillary), household contacts, healthy controls, genotypes and alleles distribution found in the groups. Kruskall-Wallis and Mann-Whitney tests were used to analyze mean values of IL-10 levels. No differences were observed between cases, contacts and controls (p=0.7450), pauci and multibacillary (p=0.7898), phenotypes of IL-10 production (low, medium or high) (p=0.4355). A significant difference in the IL-10 levels between cases and controls was found associated to the alleles -1082A,-819C and -592C (p<0.05) and between cases and contacts associated to the alleles -819C and -592C (p<0.05). No differences were found between contacts and controls (p>0.05). In conclusion, the -1082G>A, -819C>T and -592G>C IL-10 genotypes did not influence the IL-10 production or the M. leprae infection outcome. On the other hand, -1082A,-819C e -592C alleles determined a lower production of IL-10 in cases of leprosy. Keywords: Leprosy; Polymorphisms; IL-10; Serum levels; Contacts

<p>Serum levels of eosinophil granule proteins namely ECP, EPO and EPX, which are stored in the matrix of specific granules, were shown to correlate with the course of disease in disorders involving eosinophils. The concentration of eosinophil proteins in serum is the result of their release <i>in vivo</i> and <i>ex vivo</i> during the sampling procedure. Generally, eosinophils release the content of their specific granules in three ways: exocytosis, piecemeal degranulation (PM) or cytolysis. Which of them is operating in circulating eosinophils has not yet been defined. The aim of this thesis was to study the mechanisms of granule protein release from blood eosinophils in respect of protein subcellular localization and cell ultrastructure.</p><p>In patients with bacterial infections, serum levels of ECP but not EPO increased, while in patients with viral infections both proteins remained within the range of healthy controls. G-CSF is a cytokine involved in the response mechanism to bacterial but not viral infections. Administration of G-CSF to healthy subjects induced an elevation of eosinophil numbers and a preferential increase of serum EPX and ECP in comparison to EPO.</p><p>The model of PM consists of the stepwise transportation of specific granule contents from the granules towards the plasma membrane. We observed that administration of G-CSF to healthy subjects and the allergen exposure of allergic subjects during the pollen season, caused changes in the ultrastructure of eosinophil specific granules such as loosening of the matrix, granule matrix lucency and ragged losses of their core. Similar alterations of morphology had been previously described for eosinophils undergoing PM.</p><p>ECP, EPX and EPO were localized not only in the specific granules but also in extra-granular compartments as shown both by immuno electron microscopy and subcelular fractionations, An extra-granular EPX compartment was present in healthy as well as in allergic and in hypereosinophilic subjects, and there were no significant differences in its size between the groups. The size of the extra-granular compartments of ECP and EPO was increased in allergics during the season, and these compartments were clearly separate from that of EPX. Results of this show the differential mobilization ofgranule proteins in blood stream eosinophils serum and indicates PM as its mechanism.</p>

Serum levels of eosinophil granule proteins namely ECP, EPO and EPX, which are stored in the matrix of specific granules, were shown to correlate with the course of disease in disorders involving eosinophils. The concentration of eosinophil proteins in serum is the result of their release in vivo and ex vivo during the sampling procedure. Generally, eosinophils release the content of their specific granules in three ways: exocytosis, piecemeal degranulation (PM) or cytolysis. Which of them is operating in circulating eosinophils has not yet been defined. The aim of this thesis was to study the mechanisms of granule protein release from blood eosinophils in respect of protein subcellular localization and cell ultrastructure. In patients with bacterial infections, serum levels of ECP but not EPO increased, while in patients with viral infections both proteins remained within the range of healthy controls. G-CSF is a cytokine involved in the response mechanism to bacterial but not viral infections. Administration of G-CSF to healthy subjects induced an elevation of eosinophil numbers and a preferential increase of serum EPX and ECP in comparison to EPO. The model of PM consists of the stepwise transportation of specific granule contents from the granules towards the plasma membrane. We observed that administration of G-CSF to healthy subjects and the allergen exposure of allergic subjects during the pollen season, caused changes in the ultrastructure of eosinophil specific granules such as loosening of the matrix, granule matrix lucency and ragged losses of their core. Similar alterations of morphology had been previously described for eosinophils undergoing PM. ECP, EPX and EPO were localized not only in the specific granules but also in extra-granular compartments as shown both by immuno electron microscopy and subcelular fractionations, An extra-granular EPX compartment was present in healthy as well as in allergic and in hypereosinophilic subjects, and there were no significant differences in its size between the groups. The size of the extra-granular compartments of ECP and EPO was increased in allergics during the season, and these compartments were clearly separate from that of EPX. Results of this show the differential mobilization ofgranule proteins in blood stream eosinophils serum and indicates PM as its mechanism.