O HPV é transmitido principalmente pelo contato sexual e as infecções causadas por tipos virais oncogênicos estão etiologicamente associadas com o desenvolvimento de câncer de colo de útero, vulva e ânus nas mulheres, câncer de pênis e ânus nos homens, e câncer de cabeça e pescoço em ambos os sexos. Além disso, as verrugas genitais e a rara, mas séria, papilomatose respiratória estão etiologicamente associadas aos HPVs de baixo risco 6 e 11. Ademais, os HPV-16 e 6 estão entre os tipos mais frequentemente detectados em homens, independentemente da origem da amostra estudada, ressaltando a importância epidemiológica do HPV-6. Até o momento, estudos de associação entre variantes moleculares de HPV e o desenvolvimento das doenças associadas foram realizados para os HPVs de alto-risco oncogênico, como os HPV-16 e -18. Em relação à prevalência dos HPVs de baixorisco oncogênico e as implicações da heterogeneidade viral, os dados existentes até o momento são escassos. Pelo exposto, este projeto tem por objetivo: (1) Determinar a prevalência das diferentes variantes moleculares de HPV-6 em esfregaços genitais e lesões genitais externas (LGE), especificamente em verrugas genitais (VGs), entre os participantes do estudo prospectivo multinacional da Infecção por HPV em homens (estudo HIM); (2) Verificar a associação entre a infecção por diferentes variantes moleculares de HPV-6 e o risco de desenvolvimento de LGE nos participantes do estudo HIM. Para atingir os objetivos propostos foram utilizados esfregaços genitais e amostras de verruga genital dos participantes HPV-6 positivos do estudo HIM. Nestas amostras, as variantes de HPV-6 foram caracterizadas através da amplificação por PCR e sequenciamento de um fragmento do gene L2. Isto permitiu classificar as amostras em todas as linhagens (A, B) e sub-linhagens (B1, B2, B3, B4, B5) de HPV-6 descritas. Neste estudo, as variantes da sub-linhagem B3 foram as mais prevalentes. A distribuição das variantes de HPV-6 diferiu entre os países e entre casos e controles. A prevalência das variantes B1 de HPV-6 estava aumentada em VGs e esfregaços genitais de casos em comparação aos controles. Diferenças entre a detecção de variantes B1 e B3 nas VG e no esfregaço genital precedente à lesão foram observadas. Foi encontrada uma associação significativa entre a detecção de variantes da sub-linhagem B1 de HPV-6 e o desenvolvimento de VGs. Em conclusão, variantes B1 de HPV-6 são mais prevalentes em esfregaços genitais normais que precedem o desenvolvimento de VGs. Ademais as variantes B1 conferem risco aumentado para o desenvolvimento de VGs. Estudos futuros são necessários para compreender o possível envolvimento aumentado de variantes B1 de HPV-6 na progressão para lesões clinicamente relevantes / HPV is primarily transmitted through sexual contact and infections caused by oncogenic viral types are etiologically associated with the development of cervical, vulvar and anal cancer, in women, penile and anal cancer in men, and head and neck cancer in both sexes. Moreover, genital warts and the rare, but serious, respiratory papillomatosis are etiologically associated with low-risk HPV types -6 and -11. Additionally, data obtained from different studies show that HPV types -16 and -6 are among the most frequently detected types in men, independently of the origin of the samples studied, underscoring the epidemiological relevance of HPV-6. To date, studies focusing on the association between HPV molecular variants and disease onset have been conducted on high-risk types such as -16 and -18. Regarding the prevalence of low-risk HPVs and the implications of their viral heterogeneity, date is still scarce. In light of these facts, the objectives of this project are to: (1) Determine the prevalence of HPV-6 molecular variants in genital swabs and external genital lesions (EGL), specifically genital warts (GW), among participants of the prospective and multinational HPV infection in men study (HIM study); (2) To verify the association between HPV-6 molecular variants infection and the risk of developing EGL among HIM study participants. To achieve the proposed objectives, genital swabs and genital wart samples from HPV-6 positive HIM study participants were used. In these samples, HPV-6 variants were characterized by PCR amplification followed by sequencing of an L2 gene fragment. This allowed for the classification of the samples into all described HPV-6 lineages (A, B) and sub-lineages (B1, B2, B3, B4, B5). In this study, variants belonging to B3 sub-lineage were the most prevalent. HPV-6 variants distribution differed between countries and between cases and controls. HPV-6 B1 variants prevalence was increased in GWs and genital swabs of cases compared to controls. Differences among B1 and B3 variants detection in GW and the preceding genital swab were observed. A significant association of HPV-6 B1 variants detection with GW development was found. In conclusion, HPV-6 B1 variants are more prevalent in normal genital swabs that precede GW development. Additionally, B1 variants confer an increased risk for GW development. Further research is needed to understand the possible increased involvement of B1 variants in the progression to clinically relevant lesions

Made available in DSpace on 2011-03-23T21:19:16Z (GMT). No. of bitstreams: 0 / Item created via OAI harvest from source: http://www.bdtd.ufpa.br/tde_oai/oai2.php on 2011-03-23T21:19:16Z (GMT). Item's OAI Record identifier: oai:bdtd.ufpa.br:95 / In tropical regions, intensive banana (Musa accuminata) monocropping has severe agronomic and environmental impacts, and the sustainability of this production is actually highly compromised. Yield reductions are principally due to the decrease of soil fertility and root systems damages by sod fungus and nematodes. Several alternatives actually aim to improve sustainable banana cropping systems. Among them, the cultivation of banana in cover-cropping systems with legumes is a promising one. The development of such systems, however, mainly depends on me knowledge and control of how the mixed component crop competes and affects the production of the principal banana crop. The goal of this study is to estimate the functioning of the banana canavalia ensiformis cover-crop system in field conditions Canavalia is an annual legume that is supposed to have nematode control properties. In this study, however, we focus only on the capacity of banana to perform in such cropping systems, under competition for environmental resources. It is a prerequisite to the development of further investigations uppon bio-control mechanisms that could be managed in integrated protection systems. An analysis of the elaboration of yield components during the vegetative phase of the growing cycle of the banana crop was performed: it is during that phase that the potential yield (number of fruits and fruit filling) is determined, and it is assumed that legume competition can affect yield elaboration. Three treatments were carried out: (i) BPO : banana pure crop, (ii) BA0 simultaneous mixed banana-canavalia, (iii) BA60: -mixed banana-canavalia with banana planted 2 months after canavalia. Growth and development parameter were registered weekly by non destructive methods (leaf area, leaf phenology, stipe height and diameter for banana, and canopy extension for canavalia). Destructive sampling were carried out twice a month to determine above ground dry matter (DM) and mineral composition of banana in the different treatments. In addition to this agronomical evaluation, we developed a banana-intercropped modal adapted from the STICS model for validation and scenario analysis. The analysis of the vegetative cycle of banana over 7 months shows that the plantation date is crucial for the success of the intercrop. There was no significant difference between BP0 and BA0 for DM before flowering (4,5 t MS ha-1 for BP0 against 4,2 t MS ha-1 for BA0). The number of weddings is also significantly reduced for 8 to 3 between BP0 and BA0, respectively. Bananas performances are, however, more affected in BP60. Here, banana DM is reduced lo 2,7 t ha-1, that can be explicated by a higher adverse effect by the canavalia that was dominant at the plantation of banana -- 74 cm height against 29 cm for banana. The performed yield analysis may be considered as an efficient approach for the evaluation of the vegetative phase of the banana, growing sole or intercropped. The analysis of the reproductive phase is actually carried out and should complete this first diagnosis. Model validation under simulated vs. observed data was satisfying. Scenario studies of pure crop situations predicted yield values of 28 ha -1 against 20 t ha -1 (1) in situation with 400 kg nitrogen ha -1 and 133 mm irrigation, (ii) in situations without fertilizers and irrigation, respectively. A simulated yield of 16 t ha-1 was obtained in the case of intercrop without inputs. Test of contrasting scenarios may be used as a decision tool for the definition of adapted intercropping patterns - optimization of plant density and synchronization of the plantation dates of the two species, for instance -, and new agronomical standards may be studied. These investigations, however, must be carried on with the purpose of mean and long term effects evaluation of these cover-cropping systems in relation with the evolution of soil bio-physic-chemical properties - organic matter status and evolution, soil fauna transformation, weeding -, and sustainability. / Nas regiões de clima tropical, o monocultivo da banana vem causando conseqüências ambientais desastrosas e, muitas vezes, impedindo uma exploração continuada de uma mesma área. A redução do rendimento é devido principalmente as limitações físico-químicas do solo e a rápida degradação do sistema radicular, agravada pela ação de parasitas do solo (nematóides, fungos, etc.). Em virtude destas limitações, várias iniciativas vem sendo buscadas para a minimização das perdas agronômicas e ambientais, destacando-se o melhoramento e a modificação genética, e a associação deste cultivo com espécies leguminosas. Porém uma das grandes dificuldades de avaliarmos os novos sistemas de cultivo alternativos concentra-se na falta de referenciais agronômicos relacionados principalmente com o funcionamento de sistemas de cultivos associados, especialmente relacionados aos fatores e condições que interferem diretamente na definição do rendimento da espécie principal. O presente estudo testou , em campo experimental, o uso de plantas de serviço associada a bananeira e seus efeitos na produção de biomassa durante seu o ciclo vegetativo. Isto porque é durante esta fase que a bananeira constrói sua capacidade de reservas de fotoassimilados e, consequentemente, define o potencial de produção e enchimento dos frutos. Além do monocultivo, definiu-se mais duas parcelas associadas com o feijão-de- porco: 1) o plantio simultâneo das duas espécies e; 2) o plantio de feijão-de-porco e, após 2 meses, a introdução da banana. Além de acompanhamento semanal das parcelas, realizou-se, bimensalmente, coletas destrutivas de dados sobre produção de matéria seca, superfície foliar e análise nutricional das plantas. Após a análise agronômica da fase vegetativa, aplicou-se a modelização dos sistemas de cultivo estudados e comparou-se os possíveis cenários sobre o rendimento final da bananeira, além de outros indicadores sobre os fatores de crescimento das plantas. Após o acompanhamento dos 7 primeiros meses do ciclo vegetativo, concluiu-se que a data de estabelecimento da associação foi determinante para o sucesso do cultivo associado. Podemos destacar que a associação entre a bananeira e o feijão-de-porco não causou limitações na produção de biomassa (4,2 ton/ha), quando comparada com o monocultivo (4,5 ton/ha). A redução do número de capinas também foi um indicador animador deste sistema de cultivo alternativo. Por outro lado, quando a bananeira foi plantada 60 dias após a leguminosa, a mesma representou uma séria limitação na produção de biomassa (2,7 ton/ha). Esta limitação deveu-se ao estado de forte competição devido a agressividade com que o feijão-de-porco recobria toda a parcela e alcançando uma altura (74 cm) superior que a muda de banana (29 cm). Em relação a primeira parte da metodologia aplicada - o diagnóstico agronômico -, a mesma foi eficiente para a avaliação do ciclo vegetativo da associação estudada, ficando a necessidade da continuidade do acompanhamento do ciclo reprodutivo, para a confirmação dos resultados em termos de formação e produção de frutos. Na fase de modelização, chegou-se a uma leitura dos resultados próxima dos resultados obtidos no campo. Em termos de rendimento em frutos, o monocultivo com adubação (400 kg/ha de nitrogênio) e irrigação (133 mm) teve um aumento na ordem de 50% no rendimento final (28 ton/ha) Quando comparada com a parcela nas condições reais do experimento (19,6 ton/ha). Já o rendimento em frutos da associação, apresentou o mesmo resultado com e sem adubação e irrigação (16 ton/ha). No tocante a contrução dos cenários, confirmou-se novamente algumas das vantagens da associação, principalmente na redução da adubação nitrogenada aplicada nos sistemas convencionais de cultivo. Finalmente, podemos imaginar a construção de várias formas de testar e otimizar o uso destes sistemas associados (cenários). Porém, confirma-se que a construção de novos referenciais agronômicos sobre sistemas de cultivo mais complexos (os cultivos associados) torna-se ainda muito necessário para a realização de avaliações mais precisas sobre estas alternativas. E, com estes novos referenciais técnicos, podemos imaginar, a médio e longo prazo, alguns dos benefícios das leguminosas sobre as propriedades físico-químicas do solo cultivado (cobertura viva, adubo verde, redução de adventícias, etc) e sobre a manutenção do rendimento dos cultivos (adubação verde). / Dans les régions tropicales, la monoculture de la banane (Musa accuminata) a des impacts agro-environnementaux désastreux, et interdit une exploitation continue de la parcelle en culture. La réduction du rendement de la banane est principalement liée aux limitations physico-chimiques du sol et à la rapide dégradation du système racinaire sous l’action des parasites du sol (nématodes et champignons). Du fait de ces limitations, diverses alternatives sont aujourd’hui proposées pour minimiser les problèmes agronomiques et environnementaux, parmi lesquelles l’association de la culture de la banane avec des légumineuses constitue une voie intéressante. Une des grandes difficultés pour évaluer ces nouveaux systèmes de culture tient à l’absence de références agronomiques sur les systèmes de culture associés, notamment des facteurs et conditions de production pouvant affecter l’élaboration du rendement de l’espèce principale. Cette étude a pour objectif l’évaluation au champ de l’association de la banane avec une plante de service - Canavalia ensiformis – qui en plus d’être une légumineuse fixatrice d’azote, posséderait des propriétés némato-régulatrices. Avant de s’intéresser concrètement à ces propriétés, il importe de mieux comprendre l’aptitude du bananier à supporter la présence d’une plante associée. L’analyse des effets du canavalia sur la production de biomasse de la culture de banane a été suivie durant la phase végétative. C’est en effet durant cette phase que la banane élabore ses réserves en hydrates de carbone et que se définit le potentiel de production et de remplissage des fruits. Trois traitements ont été étudiés sous l’angle de l’élaboration des composantes du rendement de la banane : (i) BP0 : banane en culture pure, (ii) BA0 : banane associée au canavalia en plantation simultanée, (iii) BA60 : banane associée au canavalia en respectant un décalage de 2 mois entre la plantation du canavalia et la plantation de la banane. Un suivi hebdomadaire non destructif des paramètres de croissance et de développement des 2 espèces sur les 3 parcelles a été réalisé (surface foliaire, rythme de développement des feuilles, hauteur et diamètre du pseudo-tronc pour le bananier, extension du couvert du canavalia), et des prélèvements destructifs de biomasse de bananier ont été effectuées 2 fois par mois permettant l’analyse de la nutrition minérale des plantes. En parallèle à cette évaluation agronomique, une modélisation de la culture de la banane a été réalisée sur la base du modèle STICS, et différents scénarios de systèmes de cultures ont été étudiés. Le diagnostic portant sur les 7 premiers mois du cycle végétatif montre que la date de mise en place de l’association est déterminante pour le succès de la culture associée. L’association simultanée de la banane et de la légumineuse (BA0) ne provoque pas de réduction significative de la biomasse produite (4,5 t MS ha-1 pour BP0 contre 4,2 t MS ha-1 pour BA0). La réduction du nombre de sarclages pour BA0 est également très significative (8 sarclages en BP0 contre 3 en BA0). En revanche, dans le traitement BA60, la culture de banane subit une série de limitations. La production de biomasse est très affectée (2,7 t MS ha-1), et s’explique par une compétition plus forte exercée par le canavalia qui est beaucoup plus couvrant et qui atteint une hauteur de 74 cm, supérieure à celle de la banane en début de cycle (29 cm). L’analyse des composantes du rendement s’est révélée une méthode efficace pour évaluer la phase végétative de l’association étudiée ; l’analyse de la phase reproductive est en cours et devrait compléter cette première évaluation. La validation du modèle a été probante en permettant de simuler des résultats proches de ceux obtenus sur le terrain. Des études de scénarios ont permis des prédictions de rendement de 28 t ha-1 contre 20 t ha-1, pour le traitement en banane pure, conduit respectivement (i) avec une fertilisation de 400 kg d’azote ha-1 et une irrigation de 133 mm, (ii) sans apports d’engrais. Le rendement de l’association sans fertilisation et sans irrigation atteint 16 tonnes de fruits par hectare. La construction de différents scénarios peut donc être conduite dans la perspective de tester et d’optimiser les modalités d’association : optimisation des densités de plantation et du calage des cycles, par exemple. La construction de nouveaux référentiels agronomiques sur ces bases est donc possible. Ces travaux se doivent d’être poursuivis em vue d’évaluer les effets à moyen et long terme de ce type d’association sur l’évolution des propriétés bio-physico-chimiques du sol cultivé et de sa fertilité - évolution de la matière organique en relation avec les restitutions de la plante de service, évolution de la faune et de la flore du sol, contrôle des mauvaises herbes – , pour statuer sur la durabilité de ces systèmes de culture.

The co-ordination of Ni in hyperaccumulator plants was investigated using a number of mass spectrometry (MS)-based analytical techniques. Initial field studies on nickeliferous (lateritic) soils in Western Australia failed to identify plants containing elevated metal concentrations. For this reason, Ni-hyperaccumulators were collected from known ultramafic sites in New Caledonia, as well as grown under controlled conditions. Using electrospray ionization MS a Ni-nicotianamine (Ni-NA) complex [NiII(C12H20N3O6)]+ was identified in the South African Ni-hyperaccumulator Berkheya coddii. The association between Ni and NA was examined further in a range of Thlaspi species which accumulate different concentrations of Ni and Zn in their foliar tissues. In order to quantitate the concentration of NA a new liquid chromatography-MS (LC-MS) based analytical protocol was developed which allowed the quantification of NA and free amino acids. From the analysis of the leaf tissue in Thlaspi a strong correlation emerged between Ni and NA but not Zn. This suggested that NA plays a role in the transport of Ni in Ni-hyperaccumulating Thlaspi plants. An inverse proportionality was found between Fe and Ni. Higher NA production could be related to maintenance of Fe homeostasis. A correlation was also found between Zn and asparagine. These results are consistent with the operation of separate transport mechanisms for Ni and Zn. Further extreme examples of Ni-hyperaccumulation were also examined using LC-MS and metabolite profiling based on gas chromatography-MS (GC-MS). (For complete abstract open document)

O HPV é transmitido principalmente pelo contato sexual e as infecções causadas por tipos virais oncogênicos estão etiologicamente associadas com o desenvolvimento de câncer de colo de útero, vulva e ânus nas mulheres, câncer de pênis e ânus nos homens, e câncer de cabeça e pescoço em ambos os sexos. Além disso, as verrugas genitais e a rara, mas séria, papilomatose respiratória estão etiologicamente associadas aos HPVs de baixo risco 6 e 11. Ademais, os HPV-16 e 6 estão entre os tipos mais frequentemente detectados em homens, independentemente da origem da amostra estudada, ressaltando a importância epidemiológica do HPV-6. Até o momento, estudos de associação entre variantes moleculares de HPV e o desenvolvimento das doenças associadas foram realizados para os HPVs de alto-risco oncogênico, como os HPV-16 e -18. Em relação à prevalência dos HPVs de baixorisco oncogênico e as implicações da heterogeneidade viral, os dados existentes até o momento são escassos. Pelo exposto, este projeto tem por objetivo: (1) Determinar a prevalência das diferentes variantes moleculares de HPV-6 em esfregaços genitais e lesões genitais externas (LGE), especificamente em verrugas genitais (VGs), entre os participantes do estudo prospectivo multinacional da Infecção por HPV em homens (estudo HIM); (2) Verificar a associação entre a infecção por diferentes variantes moleculares de HPV-6 e o risco de desenvolvimento de LGE nos participantes do estudo HIM. Para atingir os objetivos propostos foram utilizados esfregaços genitais e amostras de verruga genital dos participantes HPV-6 positivos do estudo HIM. Nestas amostras, as variantes de HPV-6 foram caracterizadas através da amplificação por PCR e sequenciamento de um fragmento do gene L2. Isto permitiu classificar as amostras em todas as linhagens (A, B) e sub-linhagens (B1, B2, B3, B4, B5) de HPV-6 descritas. Neste estudo, as variantes da sub-linhagem B3 foram as mais prevalentes. A distribuição das variantes de HPV-6 diferiu entre os países e entre casos e controles. A prevalência das variantes B1 de HPV-6 estava aumentada em VGs e esfregaços genitais de casos em comparação aos controles. Diferenças entre a detecção de variantes B1 e B3 nas VG e no esfregaço genital precedente à lesão foram observadas. Foi encontrada uma associação significativa entre a detecção de variantes da sub-linhagem B1 de HPV-6 e o desenvolvimento de VGs. Em conclusão, variantes B1 de HPV-6 são mais prevalentes em esfregaços genitais normais que precedem o desenvolvimento de VGs. Ademais as variantes B1 conferem risco aumentado para o desenvolvimento de VGs. Estudos futuros são necessários para compreender o possível envolvimento aumentado de variantes B1 de HPV-6 na progressão para lesões clinicamente relevantes / HPV is primarily transmitted through sexual contact and infections caused by oncogenic viral types are etiologically associated with the development of cervical, vulvar and anal cancer, in women, penile and anal cancer in men, and head and neck cancer in both sexes. Moreover, genital warts and the rare, but serious, respiratory papillomatosis are etiologically associated with low-risk HPV types -6 and -11. Additionally, data obtained from different studies show that HPV types -16 and -6 are among the most frequently detected types in men, independently of the origin of the samples studied, underscoring the epidemiological relevance of HPV-6. To date, studies focusing on the association between HPV molecular variants and disease onset have been conducted on high-risk types such as -16 and -18. Regarding the prevalence of low-risk HPVs and the implications of their viral heterogeneity, date is still scarce. In light of these facts, the objectives of this project are to: (1) Determine the prevalence of HPV-6 molecular variants in genital swabs and external genital lesions (EGL), specifically genital warts (GW), among participants of the prospective and multinational HPV infection in men study (HIM study); (2) To verify the association between HPV-6 molecular variants infection and the risk of developing EGL among HIM study participants. To achieve the proposed objectives, genital swabs and genital wart samples from HPV-6 positive HIM study participants were used. In these samples, HPV-6 variants were characterized by PCR amplification followed by sequencing of an L2 gene fragment. This allowed for the classification of the samples into all described HPV-6 lineages (A, B) and sub-lineages (B1, B2, B3, B4, B5). In this study, variants belonging to B3 sub-lineage were the most prevalent. HPV-6 variants distribution differed between countries and between cases and controls. HPV-6 B1 variants prevalence was increased in GWs and genital swabs of cases compared to controls. Differences among B1 and B3 variants detection in GW and the preceding genital swab were observed. A significant association of HPV-6 B1 variants detection with GW development was found. In conclusion, HPV-6 B1 variants are more prevalent in normal genital swabs that precede GW development. Additionally, B1 variants confer an increased risk for GW development. Further research is needed to understand the possible increased involvement of B1 variants in the progression to clinically relevant lesions

Submitted by Reginaldo Soares de Freitas (reginaldo.freitas@ufv.br) on 2015-11-09T13:01:45Z No. of bitstreams: 1 texto completo.pdf: 1860106 bytes, checksum: 353b8088a255cf3282d4aa52ad5a282a (MD5) / Made available in DSpace on 2015-11-09T13:01:45Z (GMT). No. of bitstreams: 1 texto completo.pdf: 1860106 bytes, checksum: 353b8088a255cf3282d4aa52ad5a282a (MD5) Previous issue date: 2207-07-31 / A utilização de plantas medicinais com fins terapêuticos, para tratamento, cura e prevenção de doenças, é uma das mais antigas formas de prática medicinal da humanidade. É cada vez mais freqüente o uso de plantas medicinais, mas muitas vezes as supostas propriedades farmacológicas anunciadas não possuem a validade científica, por não terem sido investigadas, ou por não tido suas ações farmacológicas comprovadas em testes científicos pré-clínicos e clínicos. O medicamento Kraftol é uma associação de extratos fluidos de Cinhona calisaya (Quina amarela), Cola acuminata (noz de cola), Paullinia cupana (guaraná), Iodo, Ácido tânico, Formato de sódio, Glicerofosfato de cálcio, Iodeto de potássio. É um medicamento indicado para o tratamento de convalescenças e nos estados de desnutrição, nas amgdalites, faringites e adenopatias. O objetivo do presente trabalho foi avaliar a ação antiedematogênica e antiulcerativa do medicamento Kraftol e do flavonóide Ipriflavona (7-isopropoxi-isoflavona), em processos inflamatórios como edema e ulcera gástrica em ratos machos da linhagem Wistar; e avaliar a atividade antiulcerativa dos extratos presentes no medicamento Kraftol de forma isoladamente e associados sem os sintéticos. Realizou-se também um estudo de toxicologia oral aguda e subcrônica (doses repetidas) do medicamento Kraftol, tendo como base a resolução No 90, de 16 de março de 2004 da ANVISA. Para o teste nos processos inflamatórios induziu-se edema na pata em ratos com a injeção de 0,02 ml da substância capsaicina (12,5 mg-1) e ácido araquidônico (2 mg/pata) e receberam o tratamento por via oral. As lesões gástricas foram induzidas com administração oral de ácido acetil salicílico (150mg/Kg em 1,5 ml de 0,2N HCl). Para avaliação da toxicidade pré-clínica os animais foram tratados por via oral com doses de acordo com Guia para realização de estudos de toxicidade pré-clínica de fitoterápicos, conforme a resolução vingente. Foram realizadas também análises bioquímicas complementares (leucograma, colesterol total, triglicerídeos, glicose , fosfatase alcalina, TGP, TGO, creatinina, uréia, potássio,ácido úrico, Gama GT) e análise macroscópica e microscópica dos órgãos quando necessário. / The use of medicinal plants with therapeutics purposes for treatment, cure and prevention of diseases is one of most ancient medical practices of humanity. The use of medicinal plants is increasingly spreading, but many times the allegedly pharmacological properties have no scientific validity either because they have not been investigated or because their pharmacological actions have not been proved in pre-clinical and clinical scientific tests. The medicine Kraftol is an association of fluid extracts of Cinhona calisaya (yellow kina), Cola acuminata (cola nut), Paullinia cupana (guaraná), Iodine, tannic acid, sodium phormate, calcium glycerophosphate, potassium iodate. It is a medicine indicated for the treatment of recovery and malnutrition, amygdalitis, pharingitis and adenophaties. The objective of the present work was to evaluate the antiedematogenic and antiulcer action of the medicine Kraftol and of the flavonoid Ipriflavone (7-isopropoxi-isoflavone), in inflammatory processes such as edema and gastric ulcer in Wistar breed male mice; and evaluate the antiulcer activity of the extracts present in the medicine Kraftol, isolatedly and associated without the synthetics. It was also carried out a study on oral sharp and subchronic toxology (repeated doses) of the medicine Kraftol, based on the ANVISA resolution 90, March 16, 2004. For the test in the inflammatory processes, an edema was induced in mice legs by injecting 0,02 ml of the capsaicin substance (12,5 mg -1) and arachdonic acid (2 mg/leg) as well as oral treatment. The gastric wounds were induced by oral administration of acetylsalicylic acid (150mg/Kg em 1,5 ml de 0,2N Hcl). In order to evaluate the pre-clinical toxicity, the animals were treated with oral doses according to the Guide for the accomplishment of studies on phytotherapic pre-clinical toxicity, following the current resolution. Complementary biochemical analyses were also carried out (leucogram, total cholesterol, triglicerids, glucose, alkaline phosphatase, TGP, TGO, creatinin, urea, potassium, uric acid, Gama GT) and microscopic and macroscopic analyses of the organs were performed as well, when necessary.

A key effect of human papillomavirus (HPV) infection is to disrupt the normal cell cycle in order to subvert the cellular DNA replication machinery. Morphologically, condylomata induced by high and low risk HPV types cannot be distinguished and many studies have shown that the pattern of viral gene expression is similar in condylomata caused by both high risk and low risk HPV types. Detailed morphological study of cell cycle protein expression has not previously been performed on condylomata infected with low risk HPV types. The findings presented suggest that the mechanisms employed by low risk HPV6 or HPV11 to subvert cellular functions in condylomata acuminata are similar to those employed by high risk HPVs, with the exception of cyclin D1 and p53 protein over-expression. The differences in p53 expression and cyclin D1 expression seen between high and low risk HPV infection, reflect the known differences between high and low risk types and are in agreement with the known differences between high risk and low risk E6 and E7 proteins. PHK transduction studies demonstrated HPV E6 and E7 induce changes in cell cycle protein expression and that there are differences in cell cycle abrogation between HPV6 and HPV16. Disruption of the p53-MDM2 interaction can lead to activation of the p53 pathway. HPV infected lesions almost always contain wild-type p53. The binding of HPV E6 to p53, and its subsequent targeting for degradation, prevents activation of the p53 pathway in HPV infected cells. Cells over expressing HPV genes were treated with Nutlin-3, a MDM2-small molecule antagonist. The findings presented suggest treatment with Nutlin-3 induces cell cycle arrest in cells expressing HPV16 E7 and HPV6 E6 and HPV6 E7. This suggests a potential role for Nutlin-3 in the treatment of HPV infected cells.

Banana is the most important fruit crop in the world but ironically one of the crops least studied. This fruit constitutes a major staple food for millions of people in developing countries and also it is considered the highest selling fruit in the world market making this crop a very important export commodity for the producing countries. At the present time, one of the most significant constraints of banana production that causes significant economical losses are fungal diseases. Among these, Panama disease, also known as Fusarium wilt has been the most catastrophic. Panama disease is caused by the soil-borne fungus Fusarium oxysporum formae specialis (f.sp) cubense (FOC), which infects susceptible bananas through the roots causing a lethal vascular wilt. To date, the race 4 of this pathogen represents the most serious threat to banana production worldwide since most of the commercial cultivars are highly susceptible to this pathogen. Introduction of FOC resistance into commercial cultivars by conventional breeding has been difficult because edible bananas are sterile polyploids without seeds. Genetic transformation of banana, which has already been established in various laboratories around the world has the potential to solve this problem by transferring a FOC race 4 resistance gene into susceptible banana cultivars (eg. Cavendish cultivars). However, a FOC resistant (R) gene has not been isolated. Genes that confer resistance to Fusarium oxysporum have been isolated from tomato and melon using a map-based positional cloning approach. The tomato I2 and melon Fom-2 genes belong to the non-Toll/interleukin like receptors (TIR) subclass of nucleotide-binding site and leucine-rich repeat (NBS-LRR) R genes. These genes confer resistance only to certain races of F. oxysporum in their corresponding plant families limiting their use in other plant families. The fact that these two Fusarium resistance genes share the same basic non-TIR-NBS-LRR structure suggests a similar Fusarium resistance mechanism is shared between the families Solanaceae and Cucurbitaceae. This observation opens the possibility to find similar Fusarium resistance genes in other plant families including the Musaceae. A remarkable discovery of a population of the wild banana Musa acuminata subspecies (ssp.) malaccensis segregating for FOC race 4 resistance was made by Dr. Ivan Buddenhagen (University of California, Davis) in Southeast Asia. Research carried out at Queensland Department of Primary Industries (Australia) using this plant material has demonstrated that a single dominant gene is involved in FOC race 4 resistance (Dr. Mike Smith, unpublished results). Tissue-culture plantlets of this FOC race 4 segregating population were kindly provided to the Plant Biotechnology Program (Queensland University of Technology) by Dr. Mike Smith to be used in our research. This population holds the potential to assist in the isolation of a FOC race 4 resistance gene and other potential Fusarium resistance genes. The overall aims of this research were to isolate and characterise resistance gene candidates of the NBS-type from M. acuminata ssp. malaccensis and to identify and characterise potential Fusarium resistance genes using a combination of bioinformatics and gene expression analysis. Chapter 4 describes the isolation by degenerate PCR of five different classes of NBS sequences from banana (Musa acuminata ssp malaccensis) designated as resistance gene candidates (RGCs). Deduced amino acid sequences of the RGCs revealed the typical motifs present in the majority of known plant NBS-LRR resistance genes. Structural and phylogenetic analyses showed that the banana RGCs are related to non-TIR subclass of NBS sequences. The copy number of each class was estimated by Southern hybridisation and each RGC was found to be in low copy number. The expression of the RGCs was assessed by RT-PCR in leaf and root tissues of plants resistant or susceptible to Fusarium oxysporum f. sp. cubense (FOC) race 4. Four classes showed a constitutive expression profile whereas no expression was detected for one class in either tissue. Interestingly, a transcriptional polymorphism was found for RGC2 whose expression correlated with resistance to FOC race 4 suggesting a possible role of this gene in resistance to this devastating FOC race. Moreover, RGC2 along with RGC5 showed significant sequence similarity to the Fusarium resistance gene I2 from tomato and were chosen for further characterisation. The NBS sequences isolated in this study represent a valuable source of information that could be used to assist the cloning of functional R genes in banana. Chapter 5 describes the isolation and characterisation of the full open reading frame (ORF) of RGC2 and RGC5 cDNAs. The ORFs of these two banana RGCs were predicted to encode proteins that showed the typical structure of non-TIR-NBS-LRR resistance proteins. Homology searches using the entire ORF of RGC2 and RGC5 revealed significant sequence similarity to the Fusarium resistance gene I2 from tomato. Interestingly, the phylogenetic analysis showed that RGC2 and RGC5 were grouped within the same phylogenetic clade, along with the Fusarium resistance genes l2 and Fom-2. These findings suggest that the banana RGC2 and RGC5 are potential resistance gene candidates that could be associated with Fusarium resistance. The case of RGC2 is more remarkable because its expression was correlated to FOC race 4 resistance (Chapter 4). As a first step to test whether RGC2 has a role in FOC race 4 resistance, different expression constructs were made with the ORF of this sequence. One of the constructs contains a RGC2 putative promoter region that was successfully cloned in this work. These constructs will be used to transform susceptible banana plants that can then be challenged with FOC race 4 to assess whether resistance has been acquired by genetic complementation. The results of this thesis provide interesting insights about the structure, expression and phylogeny of two potential Fusarium resistance genes in banana, and provide a rational starting point for their functional characterisation. The information generated in this thesis may lead to the identification of a Fusarium resistance gene in banana in further studies and may also assist the cloning of Fusarium resistance genes in other plant species.

Objetivos. Determinar la relación costo-efectividad de la vacunación contra el (virus del papiloma humano) VPH y el tamiz de lesiones cervicales, frente a un programa de tamiz solo. Materiales y métodos. Se realizó una evaluación costo-efectividad y se empleó un modelo de Markov, con un horizonte temporal de 70 años y tres alternativas de prevención para el (cáncer del cuello uterino) CCU (tamiz solo, tamiz + vacuna bivalente, y tamiz + vacuna cuadrivalente), en una cohorte hipotética de niñas de diez años, desde la perspectiva del Ministerio de Salud. Resultados. La vacunación contra el VPH y tamiz es más costo-efectiva que el tamiz solo a partir de una voluntad de pago de S/ 2000 (USD 1 290,32). En el análisis determinístico, la vacuna bivalente es marginalmente más costo-efectiva que la vacuna cuadrivalente (S/ 48 [USD 30,97] frente a S/ 166 [USD 107,10] por AVAC, respectivamente). Sin embargo, en el análisis probabilístico ambas intervenciones generan nubes de puntos superpuestos, con una tendencia de la vacuna cuadrivalente a ser más costo-efectiva. Es decir, ambas son costo-efectivas y, por ende, intercambiables. El modelo fue especialmente sensible a variaciones de la cobertura y en la prevalencia de infección persistente por genotipos oncológicos no incluidos en la vacuna. Conclusiones. A partir de una disponibilidad de pago de S/ 2000 [USD 1 290,32] el tamiz y la vacunación son más costo-efectivos que el tamiz solo. La diferencia de costo-efectividad entre ambas vacunas carece de robustez probabilística y ambas vacunas pueden considerarse intercambiables desde la perspectiva costo-efectividad. / Objectives. To determine the cost-effectiveness of human papillomavirus (HPV) vaccination and cervical lesion screening versus screening alone for the prevention of uterine cervical cancer (UCC). Materials and methods. This cost-effectiveness evaluation from the perspective of the Ministry of Health employed a Markov model with a 70-year time horizon and three alternatives for UCC prevention (screening alone, screening + bivalent vaccine, and screening + quadrivalent vaccine) in a hypothetical cohort of 10-year-old girls. Results. Our model, which was particularly sensitive to variations in coverage and in the prevalence of persistent infection by oncologic genotypes not included in the vaccine, revealed that HPV vaccination and screening is more cost-effective than screening alone, assuming a payment availability from S/ 2 000 (US dollars (USD) 1 290.32) per subject. In the deterministic analysis, the bivalent vaccine was marginally more cost-effective than the quadrivalent vaccine (S/ 48 [USD 30.97] vs. S/ 166 [USD 107.10] per quality-adjusted life-year, respectively). However, in the probabilistic analysis, both interventions generated clouds of overlapping points and were thus cost-effective and interchangeable, although the quadrivalent vaccine tended to be more cost-effective. Conclusions. Assuming a payment availability from S/ 2000 [USD 1,290.32], screening and vaccination were more cost-effective than screening alone. The difference in cost-effectiveness between the two vaccines lacked probabilistic robustness, and therefore the vaccines can be considered interchangeable from a cost-effectiveness perspective.

O Papilomavírus humano (HPV) é provavelmente o agente mais prevalente das doenças sexualmente transmissíveis do trato genital.Este estudo foi realizado para comparar as prevalências de HPV nos 144 raspados penianos de homens HIV positivos e negativos.Utilizou PCR PGMY09/11 e hidridização em pontos. A prevalência de HPV nos indivíduos HIV positivo foi de 59% e no HIV negativo de 67%.A lesão aceto-branca pela peniscopia não demonstrou significativa positividade para HPV.Pacientes HIV positivo mostraram múltiplos tipos de HPV e os tipos oncogênicos (16/18) foram os de maior freqüência. Os HPV tipo 6/11 foram os mais freqüentes nos dois grupos. Observou-se maior prevalência de HPV nos HIV positivos com linfócitos T CD4 menor que 200 células/mm3. A carga viral plasmática do HIV não foi um fator de positividade para HPV / Genital tract human papillomaviruses (HPV) are probably the most prevalent sexually transmitted pathogens. This study is to compare HPV DNA prevalence in 144 penile smears, obtained from HIV positive and negative men. It was used PCR employing the PGMY09/11 generic HPV primers and dot blot hybridization. HPV prevalence was 59% in HIV positive men and 67% in HIV negative. Acetic white lesions by peniscopy did not show significant positive of HPV in neither HIV positive and negative groups. HIV positive men had more often multiple and oncogenic HPV types (16/18). HPV types 6/11 were more frequent in both groups. The HIV positive group with lower 200 T CD4 cell counts load reported more HPV prevalence. HIV load was not a positive factor for HPV

O Papilomavírus humano (HPV) é provavelmente o agente mais prevalente das doenças sexualmente transmissíveis do trato genital.Este estudo foi realizado para comparar as prevalências de HPV nos 144 raspados penianos de homens HIV positivos e negativos.Utilizou PCR PGMY09/11 e hidridização em pontos. A prevalência de HPV nos indivíduos HIV positivo foi de 59% e no HIV negativo de 67%.A lesão aceto-branca pela peniscopia não demonstrou significativa positividade para HPV.Pacientes HIV positivo mostraram múltiplos tipos de HPV e os tipos oncogênicos (16/18) foram os de maior freqüência. Os HPV tipo 6/11 foram os mais freqüentes nos dois grupos. Observou-se maior prevalência de HPV nos HIV positivos com linfócitos T CD4 menor que 200 células/mm3. A carga viral plasmática do HIV não foi um fator de positividade para HPV / Genital tract human papillomaviruses (HPV) are probably the most prevalent sexually transmitted pathogens. This study is to compare HPV DNA prevalence in 144 penile smears, obtained from HIV positive and negative men. It was used PCR employing the PGMY09/11 generic HPV primers and dot blot hybridization. HPV prevalence was 59% in HIV positive men and 67% in HIV negative. Acetic white lesions by peniscopy did not show significant positive of HPV in neither HIV positive and negative groups. HIV positive men had more often multiple and oncogenic HPV types (16/18). HPV types 6/11 were more frequent in both groups. The HIV positive group with lower 200 T CD4 cell counts load reported more HPV prevalence. HIV load was not a positive factor for HPV