Diss. (sammanfattning) Uppsala : Univ., 2001. / Härtill 4 uppsatser.

京都大学 / 新制・課程博士 / 博士(人間健康科学) / 甲第23124号 / 人健博第86号 / 新制||人健||6(附属図書館) / 京都大学大学院医学研究科人間健康科学系専攻 / (主査)教授 岡 昌吾, 教授 藤井 康友, 教授 髙折 晃史 / 学位規則第4条第1項該当 / Doctor of Human Health Sciences / Kyoto University / DFAM

<p> Mammalian sialidases are hydrolytic enzymes that initiate the removal of terminal a2-3, a2-6 and a2-8 sialic acid residues from various sialylated glycoconjugates. Sialidases are reportedly involved in numerous cellular functions involving proliferation, differentiation, antigenic expression, inflammation and the tumorigenicity of malignant cells. Recently, sialidase has been implicated in various immune signaling pathways, involving immune effector cells, such as activated lymphocytes and macrophages. The human lysosomal sialidase gene encodes a 46 kD glycoprotein which exists in a multienzyme complex with β-galactosidase and PPCA. Neurodegenerative diseases such as Tay-Sachs and Sandhoff are characterized by the progressive storage of glycoproteins and sialylated oligosaccharides in the nervous system. The induction of inflammatory mediators is a critical step in the pathogenesis of neurodegeneration that remains largely undefined. As such, an in vitro model of Tay-Sachs disease was used to identify potential mediators involved in disease progression. In addition, we have used the THP-1 monocytic cell line as a model of human macrophages which play a key role in potentiating a variety of immune responses. </p> <p> Translocation of neul from lysosomes to the cell surface and the resulting interaction with signaling molecules suggests neul is involved in the regulation of immune activities. We have investigated the role of sialidase on CD44 expression, an inflammation-associated glycoprotein found on the cell surface. Our data indicate that sialidase interacts with CD44 on the cell surface which may contribute to disease progression in Tay-Sachs disease. We illustrate that overexpression of sialidase stimulates interleukin-6 (IL-6) secretion in both human Tay-Sachs neuroglia and THP-1 derived macrophages. Moreover, the sialidase inhibitor 2-deoxy-2, 3-dehydro-N-acetylneuraminic acid (DANA) was found to attenuate IL-6 secretion and sialidase expression in THP-1 derived macrophages. </p> / Thesis / Master of Science (MSc)

A erliquiose monocítica canina (EMC) é uma doença infecciosa de ocorrência mundial e transmitida pelo carrapato Rhipicephalus sanguineus. As manifestações clínicas são inespecíficas e multissistêmicas. Pode apresentar as fases aguda, assintomática e crônica, as quais podem cursar com alterações hematológicas como trombocitopenia, discreta anemia e leucopenia durante a fase aguda, discreta trombocitopenia na fase assintomática, e pancitopenia nos casos crônicos graves. O estabelecimento de indicadores prognósticos para a doença pode ser de grande valia para o direcionamento do tratamento clínico. O estudo do título de anticorpos em cães infectados poderia auxiliar no diagnóstico da EMC bem como no monitoramento do tratamento. A fim de identificar as principais alterações clínicas, hematológicas e sorológicas de cães infectados por E. canis, foram selecionados 82 animais com diagnóstico etiológico de EMC (positivos a Reação em Cadeia da Polimerase PCR). Os cães foram subdivididos em grupos de animais assintomáticos (n=12), doentes e sobreviventes (n=51) e doentes e não sobreviventes (n=19). Foi realizado ainda acompanhamento clínico, hematológico e sorológico de 28 animais tratados com medicação preconizado, no período de até 180 dias após o tratamento. Na análise clínica, não houve predisposição sexual ou racial, porém cães maiores de 8 anos de idade apresentaram maior frequência. Os sintomas observados mais frequentemente em todos os cães, exceto nos assintomáticos, foram palidez de mucosas, apatia, hiporexia, letargia e gastrenterite. As alterações hematológicas encontradas no grupo dos cães doentes foram anemia e trombocitopenia, que variaram apenas na intensidade quando da comparação dos animais sobreviventes com os não sobreviventes, porém apenas a anemia e a leucopenia apresentaram-se como fatores prognósticos negativos para a doença. Altos títulos de anticorpos (&ge; 2.560) foram observados em grande parte dos animais infectados, porém não puderam ser considerados indicadores de persistência da infecção. Em dois dos 28 animais com monitoramento pós tratamento houve reinfecção, face aos resultados novamente positivos na pesquisa de material genético para E.canis, associados à recidiva de alterações hematológicas em um dos dois animais, e abrupto novo aumento do título de anticorpos. Em outros animais, apesar da rápida ascensão do título concomitante à recidiva de alterações hematológicas, não houve amplificação de DNA erliquial, sugerindo diagnóstico molecular falso negativo. Concluiu-se que alterações clínicas e hematológicas sugestivas da doença foram encontradas nos animais doentes, apenas a anemia e leucopenia podem ser indicadores prognósticos da doença, e que a interpretação do título de anticorpos deve ser feita em consonância com alterações clínicas e hematológicas do cão suspeito. / The canine monocytic ehrlichiosis (CME) is an infectious disease occurring worldwide and is transmitted by the tick Rhipicephalus sanguineus. The clinical manifestations are nonspecific and multisystemic. It may present acute, asymptomatic and chronic phases, which may be presented with haematological changes such as thrombocytopenia, mild anemia and leukopenia during the acute phase, mild thrombocytopenia in asymptomatic phase, and pancytopenia in severe chronic cases. The establishment of prognostic indicators for the disease may be of great value to guide clinical treatment. The study of antibodies titer in infected dogs could help in the diagnosis of CME and in monitoring treatment. In order to identify the main clinical, hematological and serological changes in dogs infected with E. canis, 82 animals were selected with etiologic diagnosis of a CME (positive to Polymerase Chain Reaction - PCR). The dogs were subdivided into groups of asymptomatic animals (n = 12), sick and survivors (n = 51) and sick and non-survivors (n = 19). It was also conducted clinical, hematological and serological monitoring of 28 animals treated with recommended medication for the period until 180 days after treatment. In clinical analysis, there was no racial or sexual predisposition, however dogs older than 8 years of age had a higher frequency. The most frequently reported symptoms in all dogs except the asymptomatic patients were pale mucous membranes, apathy, appetite loss, lethargy and gastroenteritis. Hematological changes found in the group of sick dogs were anemia and thrombocytopenia, which varied only in intensity when comparing the survivors to the nonsurvivors, but only anemia and leukopenia were presented as negative prognostic factors for the disease. High titers of antibodies (2560 &ge;) were observed in most infected animals, but they could not be considered indicators of persistent infection. In two out of the 28 monitored animals after treatment there was reinfection, compared to positive results back on the research of genetic material to E.canis, associated with relapse of hematological changes in one of the two animals, and sudden new increase in antibody titer. In other animals, despite the rapid rise of the concurrent title of relapse hematological changes, there was no erliquial DNA amplification, suggesting molecular diagnostic to be false negative. It was concluded that clinical and hematologic changes suggestive of the disease were found in sick animals, only anemia and leukopenia may be prognostic indicators of the disease, and that the interpretation of antibody titers should be done in line with clinical and hematologic changes of the suspect dog.

Streptococcus pneumoniae[italicized everytime] (S. pneumoniae) causes a spectrum of disease severity, and human host factors likely play a role in this variation. One candidate factor is macrophage migration inhibitory factor (MIF), a pro-inflammatory cytokine and upstream regulator of innate immunity. The MIF[italicized when not in parenthesis] promoter contains two functional polymorphisms, a tetranucleotide (CATT) repeat such that MIF expression increases with repeat number from 5-8 and a single nucleotide polymorphism (SNP) leading to a G-to-C transition, which results in increased MIF expression in cell line reporter assays. Emerging data suggest an association between high-expression MIF alleles and inflammatory disease. This study comprised two parts. For the in vitro portion, we hypothesized that peripheral blood monocytic cells (pBMCs) cultured from healthy individuals with low-expressing MIF genotypes (5-CATT alleles or SNP-GG) would have lower MIF content and release than those from individuals with high-expressing MIF genotypes (7-CATT or SNP-C alleles). For the in vivo study, we hypothesized that individuals with low-expressing MIF genotypes would have less severe systemic inflammatory responses than individuals with high-expressing MIF genotypes in response to S. pneumoniae infection. Blood samples and chart findings were collected prospectively at three Connecticut hospitals from 30 inpatients with documented invasive S. pneumoniae infections. Genomic DNA was isolated from host blood, amplified, and genotyped using fragment analysis (CATT repeat) and allelic discrimination (SNP) methods. Fishers exact tests were used to compare genotypes and disease severity. For the in vitro experiments, there were no differences observed in serum MIF levels or MIF content or release from pBMCs based on MIF genotype. In the cohort of patients infected with S. pneumoniae, serum MIF levels among enrolled subjects were significantly higher than the reported normal values, but levels did not vary with genotype or disease severity. The SNP genotype was not correlated with disease severity or occurrence of meningitis. The CATT genotype did not correlate significantly with disease severity or occurrence of meningitis, although there was a trend suggesting an association between the 7-CATT allele and meningitis (p = 0.1188, 8% without meningitis had a 7-CATT allele vs. 40% with meningitis). More patient samples will need to be analyzed in order to definitively elucidate the role of MIF genetics in infection with S. pneumoniae

Thesis (Ph. D.)--University OF MICHIGAN. / Also issued in print.

Thesis (Ph. D.)--University OF MICHIGAN. / eContent provider-neutral record in process. Description based on print version record.

Thesis (Ph. D.)--University OF MICHIGAN.

Orientador: Katia Denise Saraiva Bresciani / Banca: Caris Maroni Nunes / Banca; Marcos Rogério André / Resumo: A erliquiose canina apresenta elevada ocorrência na rotina da clínica médica de pequenos animais. Esta bactéria multiplica-se nas células epiteliais do intestino, nos hemócitos e nas células das glândulas salivares do carrapato Riphicephalus sanguineus, onde ocorre a transmissão transestadial e provavelmente não ocorre transmissão transovariana. O objetivo do presente estudo foi detectar molecularmente Ehrlichia canis em amostras de linfonodos e medulas ósseas de cães e nos intestinos, ovários e glândulas salivares de seus respectivos carrapatos R. sanguineus. Assim, 720 artrópodes fêmeas (dez de cada animal) foram removidos dos 72 cães examinados. Em seguida, foram efetuadas punções aspirativas de linfonodos e medulas ósseas. Após a dissecação dos carrapatos, seus órgãos (intestinos, ovários e glândulas salivares), bem como as amostras puncionadas de linfonodos e medulas ósseas dos cães foram testadas por meio da Reação em Cadeia da Polimerase (nested-PCR) para amplificação de um fragmento do gene 16S do ácido ribonucleico ribossomal (rRNA) de E. canis e testados também por meio da PCR em Tempo Real Quantitativa (qPCR) para E. canis, baseada em um fragmento do gene dsb. A detecção por meio da nested-PCR foi de 80,5% nos linfonodos e 44,4% nas medulas ósseas, havendo diferença significativa (p<0,05). Já em relação às nested-PCRs dos órgãos dos carrapatos, observamos positividade de 22% nos intestinos, 11% nos ovários e 7% nas glândulas salivares. Na qPCR a detecção foi de 70,... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The canine erlichiosis presents a high level of occurrence in the routine of the medical clinic of small animals. This bacteria multiplies in the epithelial cells of the intestine, in the hemocytes and in the salivary glands of the tick Riphicephalus sanguineus, where there is a transestadial transmission and probably does not occur a transovarial transmission. The objective of our study was to molecularly detect Ehrlichia canis in samples of lymph nodes and bone marrow of dogs and in the intestines, ovaries and salivary glands of their respective ticks R. sanguineus. Therefore, 720 female arthropods (ten of each animal) were removed from the 72 evaluated dogs. Next, aspirative punches were made of lymph nodes e bone marrow. After the dissection of the ticks, their organs (intestines, ovaries and salivary glands), like the punches samples of lymph nodes and bone marrow of the dogs were tested by Polymerase Chain Reaction (nested-PCR) for amplification of the gene 16S rRNA of E. canis and also tested by the Quantitative Real Time PCR (qPCR) to E. canis, based on a fragment of the gene dsb. The detection of the refer pathogen by the nested-PCR was 80,5% in lymph nodes and 44,4% in bone marrows, showing a significant difference (p<0,05). ). In relation to the nested-PCRs of the tick's organs, we observed positivity of 22% on the intestines, 11% on the ovaries and 7% on the salivary glands. In the qPCR the detection was of 70,8% and 44,4% on the lymph nodes e bone marrows, resp... (Complete abstract click electronic access below) / Mestre

A erliquiose monocítica canina (EMC) é uma doença infecciosa de ocorrência mundial e transmitida pelo carrapato Rhipicephalus sanguineus. As manifestações clínicas são inespecíficas e multissistêmicas. Pode apresentar as fases aguda, assintomática e crônica, as quais podem cursar com alterações hematológicas como trombocitopenia, discreta anemia e leucopenia durante a fase aguda, discreta trombocitopenia na fase assintomática, e pancitopenia nos casos crônicos graves. O estabelecimento de indicadores prognósticos para a doença pode ser de grande valia para o direcionamento do tratamento clínico. O estudo do título de anticorpos em cães infectados poderia auxiliar no diagnóstico da EMC bem como no monitoramento do tratamento. A fim de identificar as principais alterações clínicas, hematológicas e sorológicas de cães infectados por E. canis, foram selecionados 82 animais com diagnóstico etiológico de EMC (positivos a Reação em Cadeia da Polimerase PCR). Os cães foram subdivididos em grupos de animais assintomáticos (n=12), doentes e sobreviventes (n=51) e doentes e não sobreviventes (n=19). Foi realizado ainda acompanhamento clínico, hematológico e sorológico de 28 animais tratados com medicação preconizado, no período de até 180 dias após o tratamento. Na análise clínica, não houve predisposição sexual ou racial, porém cães maiores de 8 anos de idade apresentaram maior frequência. Os sintomas observados mais frequentemente em todos os cães, exceto nos assintomáticos, foram palidez de mucosas, apatia, hiporexia, letargia e gastrenterite. As alterações hematológicas encontradas no grupo dos cães doentes foram anemia e trombocitopenia, que variaram apenas na intensidade quando da comparação dos animais sobreviventes com os não sobreviventes, porém apenas a anemia e a leucopenia apresentaram-se como fatores prognósticos negativos para a doença. Altos títulos de anticorpos (&ge; 2.560) foram observados em grande parte dos animais infectados, porém não puderam ser considerados indicadores de persistência da infecção. Em dois dos 28 animais com monitoramento pós tratamento houve reinfecção, face aos resultados novamente positivos na pesquisa de material genético para E.canis, associados à recidiva de alterações hematológicas em um dos dois animais, e abrupto novo aumento do título de anticorpos. Em outros animais, apesar da rápida ascensão do título concomitante à recidiva de alterações hematológicas, não houve amplificação de DNA erliquial, sugerindo diagnóstico molecular falso negativo. Concluiu-se que alterações clínicas e hematológicas sugestivas da doença foram encontradas nos animais doentes, apenas a anemia e leucopenia podem ser indicadores prognósticos da doença, e que a interpretação do título de anticorpos deve ser feita em consonância com alterações clínicas e hematológicas do cão suspeito. / The canine monocytic ehrlichiosis (CME) is an infectious disease occurring worldwide and is transmitted by the tick Rhipicephalus sanguineus. The clinical manifestations are nonspecific and multisystemic. It may present acute, asymptomatic and chronic phases, which may be presented with haematological changes such as thrombocytopenia, mild anemia and leukopenia during the acute phase, mild thrombocytopenia in asymptomatic phase, and pancytopenia in severe chronic cases. The establishment of prognostic indicators for the disease may be of great value to guide clinical treatment. The study of antibodies titer in infected dogs could help in the diagnosis of CME and in monitoring treatment. In order to identify the main clinical, hematological and serological changes in dogs infected with E. canis, 82 animals were selected with etiologic diagnosis of a CME (positive to Polymerase Chain Reaction - PCR). The dogs were subdivided into groups of asymptomatic animals (n = 12), sick and survivors (n = 51) and sick and non-survivors (n = 19). It was also conducted clinical, hematological and serological monitoring of 28 animals treated with recommended medication for the period until 180 days after treatment. In clinical analysis, there was no racial or sexual predisposition, however dogs older than 8 years of age had a higher frequency. The most frequently reported symptoms in all dogs except the asymptomatic patients were pale mucous membranes, apathy, appetite loss, lethargy and gastroenteritis. Hematological changes found in the group of sick dogs were anemia and thrombocytopenia, which varied only in intensity when comparing the survivors to the nonsurvivors, but only anemia and leukopenia were presented as negative prognostic factors for the disease. High titers of antibodies (2560 &ge;) were observed in most infected animals, but they could not be considered indicators of persistent infection. In two out of the 28 monitored animals after treatment there was reinfection, compared to positive results back on the research of genetic material to E.canis, associated with relapse of hematological changes in one of the two animals, and sudden new increase in antibody titer. In other animals, despite the rapid rise of the concurrent title of relapse hematological changes, there was no erliquial DNA amplification, suggesting molecular diagnostic to be false negative. It was concluded that clinical and hematologic changes suggestive of the disease were found in sick animals, only anemia and leukopenia may be prognostic indicators of the disease, and that the interpretation of antibody titers should be done in line with clinical and hematologic changes of the suspect dog.