As espécies Klebsiella pneumoniae, Escherichia coli e Pseudomonas aeruginosa são responsáveis por infecções neonatais hospitalares. Lipopolissacarídeo (LPS) é o principal indutor de respostas inflamatórias. Os objetivos foram avaliar a transferência placentária de IgG reativa ao LPS de K. pneumoniae, E. coli O111, O26 e O6 e P. aeruginosa empregando ELISA para dosar IgG em soro materno e de cordão de 29 neonatos pré-termos e 32 a termo; analisar IgM total e específica no soro materno; e investigar a influência das patologias apresentadas pelas mães na transferência placentária. Concentrações de IgG total foram reduzidas em pré-termos como esperado, porem índices de transferência placentária de IgG total e IgG anti-LPS foram sistematicamente reduzidos quando comparados aos neonatos a termo. Níveis de IgM total e anti-LPS foram equivalentes em mães de ambos os grupos. As patologias das mães influenciaram os níveis de IgM no grupo de mães de pré-termos. Estes resultados indicam uma imunidade adquirida deficiente pelo grupo pré-termo aumentando os riscos de infecção. / Klebsiella pneumoniae, Escherichia coli and Pseudomonas aeruginosa species are responsible for neonatal nosocomial infections. Bacterial lipopolysaccharide (LPS) is the major inducer of the inflammatory responses. The aims were to evaluate the placental transfer of IgG reactive to LPS present in K. pneumoniae, in E. coli O111, O26 and O6 and in P. aeruginosa employing ELISA to detect IgG in maternal and cord sera from 29 preterm and 32 term neonates; to analyze total and specific IgM on the mothers sera; and to investigate the influence of the pathologies presented by some mothers in the placental transfer. Total IgG concentrations were reduced in preterm neonates as expected, but placental transfer indexes of total and anti-LPS IgG were systematically reduced when compared with term neonates. Total and anti-LPS IgM levels were equivalent on mothers of both groups. The mothers pathologies influenced only the IgM levels in the preterm mothers group. These results indicate a deficient acquired immunity by the preterm group increasing the risk of infection.

Acute lymphoblastic leukemia (ALL) encompasses a spectrum of lymphoid progenitors that have undergone malignant transformation and clonal proliferation at various stages of differentiation. Some cases of ALL have been documented to have prenatal origins and in particular neonatal exposure to various environmental pollutants is associated with increased disease risk, including childhood lymphoma and leukemia. Dibenzo[def,p]chrysene (DBC) is a polycyclic aromatic hydrocarbon (PAH) and in our laboratory has been established as a transplacental carcinogen in mice, producing aggressive T-cell lymphoblastic lymphomas, lung, liver, uterine, ovarian, and testicular lesions, depending on timing and dose of exposure. Investigation of the transplacental and translactational transfer of DBC was warranted following a cross-foster experiment demonstrating the greatest tumorigenic response occurred in offspring both gestating in and nursed by an exposed female. [¹⁴C]-DBC (GD17) dosing was utilized to examine time-dependent alterations of [¹⁴C] in maternal and fetal tissues, excreta, and residual levels at weaning. Fetal tissue levels of [¹⁴C]-DBC equivalents were 10-fold lower than maternal tissue, and after weaning the residual body burden was roughly equivalent in offspring exposed only in utero or only via lactation. Certain bioactive food components, including indole-3-carbinol (I3C), 3,3'-diindolylmethane (DIM), and sulforaphane (SFN) from cruciferous vegetables have been shown to target cellular pathways regulating carcinogenesis. In the above mentioned DBC initiated model of carcinogenesis, I3C is an effective transplacental chemopreventive agent. We sought to extend our chemoprevention studies in mice to a human neoplasm in cell culture, analogous to the observed murine T-cell lymphomas. Treatment of the human T-ALL cell line CCRF-CEM (CEM) with I3C reduced cell proliferation and viability only at supraphysiologic concentrations whereas DIM, the primary acid condensation product of I3C, had a marked effect at low micromolar concentrations in vitro and reduced growth of CEM xenografts in vivo. Additional T-ALL lines, selected to represent the heterogeneity of the disease, (CCRF-HSB2, Jurkat, and SUP-T1) responded similarly in vitro, demonstrating a potential therapeutic value of DIM in T-ALL. Given that epigenetic reprograming is especially active during fetal development and that DNA hypermethylation contributes to the etiology of T-ALL we examined genome-wide DNA methylation in CEM. Differential methylation analysis revealed that DIM and I3C alter CpG methylation in unique, yet overlapping, gene targets. DIM treated cells exhibited a dose-dependent decrease in hypermethylation, an observation consistent with an epigenetic mechanism of cancer suppression. Pyroseqencing and RTPCR technologies were utilized to validate changes in DNA methylation and to compare these patterns with a transcriptional response in both novel targets and candidate genes selected from the literature. Collectively, these studies merited returning to the murine transplacental model for further investigation of genetic and epigenetic changes upon maternal dietary intervention with I3C. More importantly we incorporated whole cruciferous vegetable diets (10% broccoli sprouts or 10% Brussels sprouts), SFN diet, or the combination of SFN and I3C, in order to examine matrix and mixture effects. Preliminary analysis suggests a worse prognosis for those animals exposed in utero to SFN or the whole foods, especially males. As this is the first study to administer SFN or whole cruciferous vegetables in a transplacental model of carcinogenesis, our results warrant further study on the concentration dependent influence of these potent phytochemicals during the perinatal window. / Graduation date: 2012

Made available in DSpace on 2014-07-29T15:07:36Z (GMT). No. of bitstreams: 1 Dissertacao Marcelo Sales Guimaraes.pdf: 3899079 bytes, checksum: 38f856923d6a5606a0da0a5054c4e8f9 (MD5) Previous issue date: 2011-02-24 / Bovine neosporosis is a parasitic disease of cosmopolitan distribution, caused by the obligatory intracellular cyst forming protozoan Neospora caninum (Phylum Apicomplexa, family Sarcocystidae). It has a strong association with bovine abortion, being N. caninum protozoan considered the most important for this species. The main route of transmission is vertical, determining the endemic-type neosporosis in cattle. Brazilian local breeds as Curraleiro and Pantaneiro present hardiness and resistance to adverse conditions, e.g. poor nutrition and microbiological challenges, as predominant characteristics. The conservation program of such breeds has encouraged studies aiming at a greater knowledge of these animals peculiarities. The purpose of this study was to assess the seroprevalence of these breeds for N. caninum and the occurrence of vertical transmission in herds, evaluating the importance of bovine neosporosis for these herds. Five farms in the state of Goiás and one in the state of Mato Grosso do Sul, with Curraleiro and Pantaneiro herds, respectively, were observed. Blood was collected from females at reproductive age and their offspring before colostrum intake, if possible. If it was not possible, blood was periodically collected from the offspring (five collections) until about ten months of age. A total of 358 animals was examined, 249 (198 females and 51 calves) Curraleiros and 109 (62 females and 47 calves) Pantaneiros. Seropositivity to N. caninum was evaluated by using the indirect immunofluorescence (cutoff ≥ 1:200) and ELISA (HerdChek® IDEXX Laboratories) (cutoff S/P ≥ 0.50). The total incidence of seropositives was 47,49% (170/358), 51% (127/249) Curraleiro and 39,45% (43/109) Pantaneiro. One hundred percent of the farms observed presented seropositive animals. The highest titer was 1:25600. The overall vertical transmission rate observed in herds was 51% (95%, CI 37%-64%), being 48% (95%, CI 30%-66%) in Curraleiro and 54% (95%, CI 35%-73%) in Pantaneiro. There was no statistically significant association between breed or titer of the cows and the occurrence of vertical transmission (OR 0,79; CI 0,27-2,34 95%). A low-moderate agreement was found between the two diagnostic techniques used (κ = 0.34). Thus, N. caninum is present in Curraleiro and Pantaneiro cattle breeds, with significant rates of seropositivity and vertical transmission. Although widely distributed, N. caninum does not seem to cause problems in Curraleiro and Pantaneiro herds, as abortion problems are rare and females have good fertility, within the constraints of the breed. / A neosporose bovina é uma enfermidade parasitária causada pelo protozoário intracelular obrigatório Neospora caninum (filo Apicomplexa, família Sarcocystidae), formador de cistos. Enfermidade de distribuição cosmopolita, a neosporose apresenta forte associação com o abortamento bovino, sendo N. caninum considerado o protozoário mais importante para essa espécie. A principal rota de transmissão é a vertical determinando a neosporose do tipo endêmica nos rebanhos. Raças locais brasileiras como o gado Curraleiro e Pantaneiro apresentam como características preponderantes a rusticidade e resistência às condições adversas tais como carência nutricional e desafios microbiológicos. O programa de conservação destas raças incentivou estudos que visam o maior conhecimento das particularidades destes animais. A proposta deste trabalho foi avaliar a soropositividade destas raças para N. caninum e a ocorrência de transmissão vertical nos rebanhos, permitindo determinar a importância da neosporose bovina nestes rebanhos. Cinco propriedades criadoras de Curraleiro (estado de Goiás) e uma criadora de Pantaneiro (estado de Mato Grosso do Sul) foram acompanhadas. Foi colhido sangue das fêmeas em idade reprodutiva e de suas crias antes da ingestão do colostro, quando possível. Quando não, foi colhido sangue das crias periodicamente (cinco colheitas) até em torno dos dez meses de idade. Um total de 358 animais foi analisado, sendo 249 (198 fêmeas e 51 bezerros) Curraleiros e 109 (62 fêmeas e 47 bezerros) Pantaneiros. Avaliou-se a soropositividade para N. caninum por meio das técnicas de imunofluorescência indireta (ponto de corte ≥ 1:200) e ELISA indireto (HerdChek® IDEXX Laboratories) (ponto de corte S/P ≥ 0,50). A ocorrência total de soropositivos foi 47,49% (170/358), com 51% (127/249) do gado Curraleiro e 39,45% (43/109) do gado Pantaneiro soropositivo. Cem por cento das propriedades acompanhadas apresentaram animais positivos. O maior título encontrado foi de 1:25600. A taxa de transmissão vertical geral observada nos rebanhos foi de 51% (95%, IC 37%-64%), sendo de 48% (95%, IC 30%-66%) no gado Curraleiro e de 54% (95%, IC 35%-73%) no Pantaneiro. Não houve associação estatisticamente significativa entre título ou raça das mães e ocorrência de transmissão vertical (OR 0,79; IC 0,27-2,34 95%). Uma concordância moderada-baixa foi verificada entre as duas técnicas de diagnóstico utilizadas (κ = 0,34). Assim, N. caninum está presente nos rebanhos bovinos das raças Curraleiro e Pantaneiro, com consideráveis taxas de soropositividade e transmissão vertical. Embora amplamente distribuído, N. caninum parece não representar problema nos rebanhos Curraleiro e Pantaneiro, visto que problemas de abortamento são raros e as fêmeas apresentam boa fertilidade, dentro das limitações das raças.

This experiment was undertaken to study the following aspects of the A. glazioviana poisoning in sheep: (1) abortifacient properties of the plant and the pathogenesis of the abortions and reproductive failure associated with the toxicosis and (2) the morphology and pathogenesis of cardiac and brain lesions. In the first part of the experiment, 17 pregnant ewes were orally fed variable amounts of either green or dried leaves of A. glazioviana fractioned in 1-24 daily doses. All 17 ewes manifested some form of reproductive failure. Nine (52.9%) aborted their fetuses at 4-36 days after starting being fed the leaves of the plant; one had a stillbirth and in another one intrauterine fetal death was diagnosed. The other six ewes delivered 8 weak lambs seven of which died from few minutes to 48 hours after birth. Gross and histopathological changes observed in lambs, stillborn, and in aborted fetuses were rather similar to those found in the spontaneous poisoning by A. glazioviana in ruminants. It is concluded that abortions caused by A. glazioviana are due to transplacental induced fetal lesions consisting of toxic cardiomyopathy and spongy degeneration of the white matter of the brain. For the second part of the experiment 15 mature sheep were fed varying daily amounts of the fresh green leaves of A. glazioviana for different periods of time (1-24 days). Clinical signs observed in poisoned sheep included depression, anorexia, general weakness, staggering gait and prolonged recumbency. One sheep had signs of congestive heart failure. Necropsy findings included subcutaneous and cavitary edema in two sheep and nutmeg liver in one. Histopathological findings included degeneration, necrosis and interstitial fibrosis in the myocardium of 4 sheep and spongy degeneration of cerebral white matter (status spongiosus) in 10 sheep. The ultrastructure of the brain lesion was morphological consistent with those found in diseases grouped as spongiform myelinopathies in which vacuolation of myelin occur in absence of significant myelin break down or phagocytosis / Este estudo foi realizado para investigar os seguintes aspectos da intoxicação por Ateleia glazioviana em ovinos: (1) as propriedades abortivas da planta e a patogênese dos abortos e da falha reprodutiva associada à toxicose e (2) a morfologia e patogênese das lesões cardíacas e encefálicas. Para a primeira parte do experimento, 17 ovelhas prenhes receberam, por via oral, diferentes quantidades das folhas verdes ou secas de A. glazioviana fracionadas em 1-24 doses diárias. Todas as 17 ovelhas manifestaram alguma forma de falha reprodutiva. Nove ovelhas (52,9%) abortaram seus fetos aos 4- 36 dias após o início da administração da planta; houve um natimorto e uma morte intra-uterina. Seis ovelhas pariram 8 cordeiros fracos, sete dos quais morreram de alguns minutos até 48 horas após o nascimento. Alterações macro e microscópicas observadas em cordeiros, num natimorto e nos fetos abortados eram notavelmente semelhantes às observadas na intoxicação espontânea por A. glazioviana em ruminantes. Foi concluído que os abortos causados por A. glazioviana são devidos às lesões induzidas no feto por via transplacentária e consistem de miocardiopatia tóxica e degeneração esponjosa da substância branca do encéfalo. Para a segunda parte do experimento, 15 ovinos adultos receberam diferentes quantidades diárias das folhas verdes frescas de A. glazioviana por vários períodos de tempo (1-24 dias). Os sinais clínicos observados nos ovinos afetados incluíram depressão, anorexia, fraqueza, incoordenação e decúbito prolongado. Um ovino apresentou sinais de insuficiência cardíaca congestiva. Os achados de necropsia incluíram edema subcutâneo e das cavidades orgânicas em dois ovinos e fígado de noz moscada em um. Os achados histopatológicos incluíram degeneração, necrose e fibrose intersticial no miocárdio em 4 ovinos e degeneração esponjosa da substância branca do encéfalo (status spongiosus) em 10 ovinos. A ultra-estrutura da lesão encefálica foi morfologicamente classificada no grupo das mielinopatias espongiformes em que ocorre vacuolização da mielina sem degradação ou fagocitose significativas