Made available in DSpace on 2014-10-09T12:51:25Z (GMT). No. of bitstreams: 0 / Made available in DSpace on 2014-10-09T13:56:25Z (GMT). No. of bitstreams: 1 11303.pdf: 21897794 bytes, checksum: dd08773f11bb9dc32569446f5352b1cf (MD5) / Dissertacao (Mestrado Profissionalizante em Lasers em Odontologia) / IPEN/D-MPLO / Intituto de Pesquisas Energeticas e Nucleares, IPEN/CNEN-SP; Faculdade de Odontologia, Universidade de Sao Paulo

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Made available in DSpace on 2014-10-09T12:52:22Z (GMT). No. of bitstreams: 0 / Made available in DSpace on 2014-10-09T14:00:42Z (GMT). No. of bitstreams: 0 / Freqüentemente as condutas terapêuticas utilizadas no tratamento de patologias bucais são cirúrgicas, resultando em falhas de continuidade da mucosa bucal. A possibilidade de obtenção de epitélios transplantáveis, a partir do cultivo in vitro de células da mucosa bucal, abre novas perspectivas de utilização, não se restringindo somente ao seu local de origem, ou seja, a boca, mas também como material de reconstrução para outras regiões, tais como: uretra, córnea, superfície ocular e epitélio córneo-limbal. Os métodos utilizados para a obtenção dessas células ainda são controversos na literatura. Neste sentido, avaliamos e comparamos a eficiência de dois métodos, enzimático e explante, para a obtenção de queratinócitos de mucosa bucal humana. Os fragmentos utilizados para a obtenção dessas células foram obtidos durante procedimentos cirúrgicos de pacientes voluntários saudáveis. Os queratinócitos foram cultivados sobre uma camada de sustentação, feeder-layer, confeccionada com fibroblastos murinos irradiados (3T3 - Swiss albino). Neste estudo foram comparados: o tempo para a obtenção dos queratinócitos, o rendimento obtido entre os dois métodos, a duração da vida útil em cultura, a capacidade que estas células tiveram em formar um epitélio in vitro e a morfologia dos mesmos. Os resultados obtidos, na avaliação dos dois métodos, comprovaram a possibilidade de obtenção dos queratinócitos, a partir de um pequeno fragmento bucal, porém pode-se verificar que existem vantagens e restrições peculiares a cada um dos métodos estudados. / Dissertacao (Mestrado) / IPEN/D / Instituto de Pesquisas Energeticas e Nucleares - IPEN/CNEN-SP

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Submitted by Luciana Ferreira (lucgeral@gmail.com) on 2016-04-18T14:57:11Z No. of bitstreams: 2 Tese - Clayson Moura Gomes - 2015.pdf: 2080474 bytes, checksum: e53c02c9eac98545d091fb13331c2b9a (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2016-04-18T14:58:58Z (GMT) No. of bitstreams: 2 Tese - Clayson Moura Gomes - 2015.pdf: 2080474 bytes, checksum: e53c02c9eac98545d091fb13331c2b9a (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Made available in DSpace on 2016-04-18T14:58:58Z (GMT). No. of bitstreams: 2 Tese - Clayson Moura Gomes - 2015.pdf: 2080474 bytes, checksum: e53c02c9eac98545d091fb13331c2b9a (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2015-08-05 / Fundação de Amparo à Pesquisa do Estado de Goiás - FAPEG / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Leishmaniasis are zoonoses present in several countries, being Leishmania (Viannia) braziliensis the main parasite that causes localized cutaneous (LCL) and / or mucous leishmaniasis (LM) in Brazil. It is not clear yet which factors favor the development of the mucosal form of the disease. The aim of this study was to evaluate the profile of experimental leishmaniasis in wild type mice and mice lacking the IFN, inos or gp91phox genes, after infection with L. (V.) braziliensis amastigotes isolated from patients with LCL or LM. Mice were inoculated with different amount of amastigote parasites in the foot paw and lesion development was measured weekly. Histopathological analyses were performed in hematoxilin eosin stained slides. The number of parasites present in the infected foot paw, lymph node and spleen during infection was estimated by the limiting dilution assay. It was observed that IFNγ or Phox KO mice infected with amastigotes isolated from LCL patients develop lesion earlier than mice infected with amastigotes from LM patients. iNOS KO mice infected with 1000 amastigotes isolated from LM patients developed greater lesion and more severe inflammation than mice infected with parasites from LCL at the 5th week. The IFNγ, iNOS and gp91phox mice had similar parasite number in the infected foot paw after infection with LM or LCL parasites .The parasite burden in lymph node and spleen was higher after infection with parasites from LM than LCL patients. The low ability of human macrophages to produce sufficient amount of NO can favor survive of LM parasites. Additionally, the increased ability of LM parasite to disseminate will facilitate migration to mucosal tissue to promote the mucosal disease. / As leishmanioses são zoonoses presentes em vários países do mundo, sendo Leishmania (Viania) braziliensis o parasito que causa a leishmaniose cutânea e/ou a leishmaniose mucosa no Brasil. Ainda não são claros os fatores que favorecem o desenvolvimento da forma mucosa da doença, porém é possível que fatores presentes nos parasitos sejam responsáveis pelo aparecimento de lesões mucosas. O objetivo deste estudo foi avaliar o perfil imunopatológico da leishmaniose experimental em camundongos do tipo selvagem e também em deficientes nos genes de IFN, inos ou gp91phox após a infecção com amastigotas de L. (V.) braziliensis isoladas de pacientes com leishmaniose cutânea localizada (LCL) ou leishmaniose mucosa (LM). Foram inoculadas diferentes quantidades de parasitos na pata dos camundongos e a progressão da lesão foi mensurada semanalmente. A análise histopatológica da lesão foi feita em lâminas coradas com hematoxilina e eosina. O número de parasitos presentes na pata infectada, linfonodo e baço dos animais infectados foi estimado pelo método de diluição limitante. Observou-se que os camundongos desprovidos de IFN ou phox infectados com amastigotas isoladas de pacientes com LCL desenvolveram a lesão mais precocemente do que camundongos infectados com amastigotas isoladas de pacientes com LM. A infecção de camundongos desprovidos de iNOS com 1000 amastigotas de pacientes com LM desencadeou uma lesão maior e com uma inflamação mais severa que aquela causada por parasitos de pacientes com LCL a partir da 5a semana. Animais deficientes de IFN, iNOS e gp91phox possuíam um número similar de parasitos na pata infectada, independente dos parasitos serem oriundos de pacientes com LCL ou LM. Nos linfonodos e baço foi observado mais parasitos quando os animais eram infectados com amastigotas de pacientes com LM. A pequena habilidade dos macrófagos humanos de produzir uma quantidade suficiente de NO pode favorecer a sobrevida de parasitos oriundos de LM. Além disso, o aumento da habilidade de parasitos de LM em se disseminar facilitaria a migração para os tecidos mucosos e promoveriam a doença na mucosa.

Os escargots da espécie Achatina fulica são caracóis africanos comestíveis e para essa finalidade foram introduzidos no Brasil em 1988 para substituir o escargot europeu Helix sp. Contudo, o hábito alimentar conservador da população brasileira ocasionou prejuízos aos criadores de escargots no Brasil, desencadeando uma soltura irresponsável e anti-ética desses moluscos no meio ambiente, o que proporcionou uma associação direta dos caracóis à impactos ambientais, sendo objeto de estudos e pesquisas correlatas. Todavia, estudos anteriores descreveram efeitos benéficos, antimicrobianos e cicatrizantes do muco extraído de caracóis Achatina sp, e ainda a potencialização destes efeitos a partir do acréscimo de plantas medicinais à dieta base consumida pelos escargots, constatando a capacidade de retenção em seu organismo das propriedades dos alimentos por eles ingeridos. As atividades antimicrobiana e cicatrizante também são conhecidas na utilização da própolis produzida por abelhas da espécie Apis mellifera. Partindo dessas premissas, neste estudo, foi adicionada própolis à ração base dos escargots, objetivando avaliar o efeito cicatrizante desses zooterápicos e suas aplicações. Os resultados obtidos demonstraram que a utilização de muco nas lesões cirurgicamente induzidas em camundongos acelerou o processo de cicatrização, comparativamente ao grupo controle que recebeu apenas tratamento com soro fisiológico. As análises parasitológica e citotóxica realizadas demonstraram que o muco é apto para a utilização proposta. A dieta acrescida de própolis interferiu nas características do muco. Foi possível observar, a partir das avaliações histológicas e macroscópicas uma discreta vantagem no processo de cicatrização para o grupo tratado com muco extraído de escargots que receberam ração base acrescida de própolis em sua dieta. Estes resultados demonstram a potencialidade desta pesquisa em resultar em um biofármaco com propriedades cicatrizantes à base de muco de escargots e ainda uma possível patente deste muco, através do indicativo de sua importância terapêutica para o reparo tecidual de lesões veterinárias e humanas. / The snail species Achatina fulica, is an edible African giant land snail species that was introduced into Brazil in 1988 as a substitute for the european escargot Helix sp. However, the conservative eating habits of the brazilian population have caused losses to breeders of escargot in Brazil due to the irresponsible, unethical release of the species into the environment. This species is known for its invasive nature and most of the literature focuses on the species disruption to the environment. Thus, the first objective of the study and related research focuses on these environmental impacts. Others studies have described beneficial attributes, antimicrobial and healing properties, of the mucous extracted from the small Achatina species. They have also described the potential benefits of feeding the snails a base diet enriched with medicinal plants to prove the capacity of the organisms to retain the aforementioned properties after ingestion. Lastly, our research will also examine the addition of propolis, to the base ration diet of snails. Propolis is a resinous material, obtained from local plant sources, used as a sealant in bee hives. For the purpose of these studies, the focus will be propolis specifically associated with the bees of the species Apis mellifera. The antimicrobial and healing properties of propolis have been well recognized and described. The objective of this work is thus, to evaluate the healing effects of this zootherapics, and its application in veterinary medicine as a biopharmaceutical. The results showed that the use of mucous in the lesions surgically induced in mice accelerated the healing process, compared to a control group that received treatment only with saline. Parasitological and cytotoxic analysis performed demonstrated that the mucous is suitable for the proposed use. The diet added with propolis influenced the parameters of mucous. It was observed from the macroscopic and histological findings a slight advantage in the healing process for the group treated with mucus extracted from snails fed diets basis with propolis in their diet. This results achieved were encouraging, showing the potenciality of this research results in a medication with scaring properties based on snail mucus and further a possible patent of this mucus, through its expressive therapeutics importance to veterinary and human lesions tissue repair.

In our organism, mucous surfaces are important boundaries against the environmental milieu with defined fluxes of metabolites through these surfaces and specific rules for defense reactions. Major mucous surfaces are formed by epithelia of the respiratory system and the digestive tract. The heme peroxidases lactoperoxidase (LPO), myeloperoxidase (MPO), and eosinophil peroxidase (EPO) contribute to immune protection at epithelial surfaces and in secretions. Whereas LPO is secreted from epithelial cells and maintains microbes in surface linings on low level, MPO and EPO are released from recruited neutrophils and eosinophils, respectively, at inflamed mucous surfaces. Activated heme peroxidases are able to oxidize (pseudo)halides to hypohalous acids and hypothiocyanite. These products are involved in the defense against pathogens, but can also contribute to cell and tissue damage under pathological conditions. This review highlights the beneficial and harmful functions of LPO, MPO, and EPO at unperturbed and inflamed mucous surfaces. Among the disorders, special attention is directed to cystic fibrosis and allergic reactions.

Introduction: Despite advances in prevention and treatment of vaccine-preventable diseases, diarrhoeal and pneumococcal diseases remain a major source of morbidity and mortality among children worldwide. The introduction of vaccines has led to dramatic reductions in the burden of infectious diseases and mortality among children. South Africa was the first country in Africa to introduce rotavirus vaccine (RV) and pneumococcal conjugate vaccine (PCV) in 2008 as part of its national immunisation programme. Performance of immunization programmes is commonly measured by the coverage and uptake of vaccines, hence ensuring that every child is immunized at the earliest or appropriate age is an important public health goal. We therefore assessed proportions and factors associated with uptake of RV and PCV among infants who were followed during the routine demographic surveillance system of the Africa Centre Demographic Surveillance Area (DSA) in a rural South Africa setting. Methods: An open cohort of children resident in the DSA aged 12 months or below was prospectively followed between January 2008 and December 2011. Trained interviewers visited households and administered a standardised questionnaire. Mothers and caregivers were asked to show the interviewers the South African Road-To-Health (RTH) card for all children aged 12-23 months at the time of the visit or through maternal recall for children whose RTH card was not available. The RTH card includes dates of all routine vaccinations a child has received. Rotavirus vaccine doses are given at 6 and 14 weeks of age and PCV doses at 6 and 14 weeks and 9 months. Complete uptake was defined as “complete” if a child received all recommended doses of either RV or PCV and incomplete if a child did not receive any dose or received one dose of RV or PCV. Logistic regression models were used to assess factors associated with uptake of RV and PCV separately. Results: A total of 6,263 children were included in the analysis, of which 3,082 (49%) were females. At birth, 3,823 (61%) children were living in rural areas and about one-sixth of the children were living in households located far from a health facility (≥5km). The overall uptake of RV and PCV vaccines among children aged 12 months or below was 50% and 37% respectively. Infants who ever migrated outside the DSA had reduced odds of complete RV and PCV vaccination compared to infants who did not out migrate (adjusted OR=0.49, 95% CI 0.41-0.57) and (adjusted OR=0.52, 95% CI 0.43-0.63) respectively. Complete uptake of RV was associated with the increase in education levels of mothers compared secondary education (adjusted OR=1.70, 95 % CI 1.02-2.34) or tertiary education (adjusted OR=1.80, 95 % CI 0.97-2.44). Infants whose mothers were employed were less likely than infants whose mothers were not employed to have complete vaccination for RV or PCV (adjusted OR=0.71, 95 % CI 0.60-0.84) and (adjusted OR=0.81, 95% CI 0.68-0.96) respectively. Similarly, infants whose mothers were resident in the DSA were more likely than infants whose mothers were not resident to have complete vaccination for RV or PCV (adjusted OR=1.97, 95 % CI 1.49-2.60) and (adjusted OR=1.55, 95% CI 1.16-2.08) respectively. Conclusion and recommendation: The uptake of complete RV and PCV were generally low among children in rural South Africa within our study period. Child outmigration, maternal employment, maternal education and maternal residency in the DSA at child birth were associated with complete uptake of RV and PCV vaccines. Programmes targeting mothers of lower socio-economic status are required. Such programmes may include vaccine awareness and immunization campaigns at the community level to improve vaccine uptake and more targeted interventions in areas with low RV and PCV uptake.