Submitted by Marco Antônio de Ramos Chagas (mchagas@ufv.br) on 2016-10-17T16:19:19Z No. of bitstreams: 1 texto completo.pdf: 3803014 bytes, checksum: f60b6cf87d3f9e2554f0dc89ece0a16d (MD5) / Made available in DSpace on 2016-10-17T16:19:19Z (GMT). No. of bitstreams: 1 texto completo.pdf: 3803014 bytes, checksum: f60b6cf87d3f9e2554f0dc89ece0a16d (MD5) Previous issue date: 2005-02-18 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Objetivou-se, neste trabalho, avaliar a sensibilidade à mistura de herbicidas trifloxysulfuron-sodium + ametryn nos cultivares e clones de cana-de-açúcar SP80- 1842, SP79-1011, SP81-3250, SP80-1816, RB855113, RB835486, RB845210, RB867515, RB928064, RB72454, RB855536, RB947643, RB855002, RB957712 e RB957689 e a caracterização anatômica, química e micromorfológica dos cultivares RB855113, SP80-1842, SP80-1816, RB867515 e do clone RB957689 visando melhor entendimento sobre as barreiras que cada material impõe à penetração de herbicidas. O trabalho foi dividido em duas partes. A primeira parte dividida em dois ensaios. No ensaio 1, a mistura herbicida foi aplicada em dose única de 2,00 kg ha -1 sobre todos os cultivares e clones, enquanto no ensaio 2, esta foi aplicada nas doses de 1,00; 2,00; e 6,00 kg ha -1 sobre os cultivares RB855113 e RB867515. Em ambos os ensaios, a partir da aplicação do herbicida, em intervalos semanais, foram feitas avaliações do índice de toxidez (% de fitotoxicidade) e, por ocasião da colheita, realizada aos 45 DAT (dias após a aplicação dos tratamentos), avaliou-se, ainda, a biomassa seca da parte aérea dos 11 cultivares e dos quatro clones. As variáveis altura da planta, número de folhas, área foliar e biomassa seca da parte aérea e do caule foram avaliadas apenas no segundo ensaio. Na segunda parte do trabalho, as primeiras folhas com lígula totalmente visível foram coletadas de cinco plantas de cada cultivar e clone. Das folhas de cada repetição foi obtida uma amostra da região central, com aproximadamente 1cm 2 , as quais foram utilizadas em estudos anatômicos em microscopia de luz e eletrônica de varredura. Foram determinadas as seguintes características anatômicas: área da epiderme adaxial e abaxial; área do mesofilo; área das células buliformes; área dos feixes vasculares; área das células da bainha; área ocupada por fibras, área do xilema e área do floema. Esses dados foram transformados em porcentagem em relação à área total. Determinou-se, também, a densidade de tricomas, a densidade de estômatos, o comprimento do ostíolo, a espessura da epiderme, o perímetro da parede periclinal externa das células buliformes, a espessura da parede periclinal externa das células buliformes e das células epidérmicas comuns nas faces adaxial e abaxial. Também foram coletadas folhas para a avaliação da composição química da cera epicuticular. O cultivar RB855113 apresentou alta sensibilidade à mistura herbicida; os cultivares e clones SP80-1842, SP80-1816, SP79-1011 e RB957689 apresentaram sensibilidade média; e os demais clones mostraram boa tolerância a esse produto. Para o cultivar RB855113, o maior índice de toxidez foi observado no período de 20 a 27 DAT. Nesse período observou-se redução da altura da planta, do número de folhas, da área foliar e da biomassa seca da parte aérea e do caule com o aumento das doses do produto. Considerando as características anatômicas, o cultivar RB855113 apresentou diferenças relevantes em relação aos demais cultivares avaliados, como, maior proporção de células buliformes, chegando a ser o tecido que ocorre em maior quantidade na seção transversal da lâmina foliar desse cultivar, maior densidade de estômatos e de tricomas em ambas as faces, epiderme menos espessa na face adaxial e estômatos com maiores ostíolos nas duas faces. O cultivar RB855113 apresentou composição química da cera epicuticular com polaridade intermediária, porém menos polar que a cera do cultivar RB867515. Na análise micromorfológica não foi observada diferença relevante entre os materiais. De todas as características avaliadas, as anatômicas definiram as principais diferenças entre os cultivares. / This work was carried out in order to evaluate the susceptibility to Trifloxysulfuron-sodium + ametryn herbicide on the cultivars and clones of sugarcane SP80-1842, SP79-1011, SP81-3250, SP80-1816, RB855113, RB835486, RB845210, RB867515, RB928064, RB72454, RB855536, RB947643, RB855002, RB957712 and RB957689, and the anatomical, chemical and Micromorphological characterization of RB855113, SP80-1842, SP80-1816, RB867515 cultivars and RB957689 clone, to understand better the barriers each material imposes upon herbicide penetration. The first part of the work was divided in two assays. In assay 1, the herbicide was applied in one rate 2.00 kg ha -1 in all cultivars and clones, while in assay 2, the rates were 1.00, 2.00 and 6.00 kg ha -1 on the cultivars RB855113 and RB867515. In both assays, it was evaluated the toxicity index from weekly herbicide application and the dry biomass of shoot from harvest, after 45 days after treatment application. Plant height, leaf number, foliar area and dry biomass of shoot and stem were evaluated only in the second assay. On the second part of the work, the first leaves with ligule totally visible were taken from five plants of each cultivar and clone. One sample, with approximately 1 cm 2 , was taken at central portion of the leaves in each replication, to do anatomical studies by using light and scanning-electron microscopes. It was determined the areas of: adaxial and abaxial epiderm, mesophyll, bulliform cells, vascular bundles, sheath cells, occupied by fibers, xylem and phloem, and the data obtained was transformed in percentage related to total area. It was also determined trichome density, stomatic density, ostiole length, epiderm thickness, perimeter of external wall of bulliform cells, periclinal external wall thickness upon bulliform and epidermic cells on abaxial and adaxial sides. Leaves were also used to evaluate chemical content of epicuticular wax. The cultivar RB855113 showed high susceptibility to herbicide; the cultivars and clones SP80-1842, SP80-1816, SP79-1011 and RB957689 showed medium susceptibility and the other clones showed good tolerance to the product. At 20 t0 27 days after treatment application, it was observed to the cultivar RB855113, the highest toxicity index, and reduction on plant height, leaf number, foliar area and dry biomass of shoot and stem, with an increase on the rate of the product. This cultivar showed relevant differences in relation to the other cultivars like higher proportion of bulliform cells, higher density of trichomes and stomata in both sides, smaller thickness of epiderm on adaxial side and stomata with larger ostioles in the two sides. The chemical content of epicuticular wax with intermediate polarity appeared in the cultivar RB855113, however, the polarity was higher in the cultivar RB867515. In the micromorphological analysis, there was not difference among cultivars. The anatomical characteristics defined the main differences among them. / Dissertação importada do Alexandria

Made available in DSpace on 2016-08-29T15:36:24Z (GMT). No. of bitstreams: 1 tese_3379_Sandro Dan tatagiba.pdf: 1080979 bytes, checksum: a308088f8642f1dbfd2416390a5e383f (MD5) Previous issue date: 2006-02-24 / O objetivo desse estudo foi avaliar o efeito do déficit hídrico sobre as características de crescimento: diâmetro do coleto; altura das plantas; área foliar; relação raiz/parte aérea; matéria seca de: folhas, haste e ramos, raízes e total; e as características fisiológicas das trocas gasosas (fotossíntese líquida, condutância estomática, transpiração) e potencial hídrico foliar, em seis clones de eucalipto (1, 2, 3, 4, 5 e 6) crescendo em vasos plásticos de diâmetro de 42 cm e altura de 72 cm, com aproximadamente 100 dm3, de modo a subsidiar a seleção desses materiais genéticos para estabelecimento em áreas com disponibilidade diferenciada de água no solo. Foram avaliadas algumas variáveis climáticas durante todo o período experimental, a fim determinar a condição no ambiente em estudo. As plantas cresceram nos vasos com teor de água próximo a capacidade de campo por um período de 150 dias, quando, então, foram iniciados os manejos hídricos diferenciados até o final do experimento que durou cerca de 250 dias. Os manejos hídricos aplicados foram: sem déficit - manutenção dos vasos próximo à capacidade de campo ao longo de todo o período experimental, ou seja, 250 dias; déficit 1 - corte da irrigação aos 150 dias de experimentação, durante 45 dias e posterior retomada da irrigação por mais 55 dias; déficit 2 - corte da irrigação aos 150 dias de experimentação, prolongando até o final do experimento (100 dias de déficit hídrico). O clone mais afetado pela deficiência hídrica foi o 3, apresentando menor acúmulo de matéria seca total e maior redução em área foliar, provocada principalmente pela abscisão foliar. Contrariamente, os clones 1, 2, 4 foram os mais tolerantes aos efeitos do déficit hídrico, em razão de terem apresentado maior acúmulo de matéria seca total. De modo geral, o déficit hídrico promoveu a redução dos valores da fotossíntese, condutância estomática, e transpiração foliar de todos os clones. Os clones 1, 2 e 4 apresentaram os valores mais altos de potencial hídrico foliar nas plantas submetidas ao déficit hídrico ao longo das campanhas realizadas, mostrando obter mecanismos eficientes no controle de perda de água, já que apresentaram as menores taxas transpiratórias e menores valores de condutância estomática. Pressupõe-se que o aparato fisiológico destes clones permita alcançar um menor consumo de água, favorecendo a planta a suportar um período mais longo de déficit hídrico, e possibilitando uma maior conversão de CO2 atmosférico em carboidrato por período maior de tempo. O clone 3 apresentou menores valores de fotossíntese, condutância estomática e transpiração quando submetido ao déficit hídrico nas duas campanhas realizadas. O clone 6 apresentou maior transpiração no nível de manejo sem déficit e sob déficit em relação aos demais clones, sugerindo elevado consumo de água pelas plantas. Após a retomada da irrigação, todos os clones recuperaram suas taxas fotossintéticas similares às registradas pelos clones mantidos sem déficit, contribuindo para acréscimos na matéria seca total das plantas.

Master of Science / Department of Diagnostic Medicine/Pathobiology / Ying Fang / Seneca Valley Virus (SVV) causes vesicular disease in pigs. Vesicular lesions on the snout and coronary band of hoof mostly resemble lesions caused by Foot-and-Mouth Disease Virus (FMDV), which may lead to the foreign animal disease investigation. In 2015, Brazil experienced major outbreaks of SVV; then in July, sporadic cases of SVV were reported in United States and became a concern in swine industry. A reverse-genetic system serves as a major tool to study pathogenesis of the virus. In our study, a full-length cDNA infectious clone, pKS15-01-Clone, was constructed from an emerging Seneca Valley Virus (SVV; strain KS15-01). To explore the potential use as a viral backbone for expressing marker genes, the enhanced green fluorescent protein (EGFP)-tagged reporter virus (vKS15-01-EGFP) was generated using reverse genetics. Compared to the parental virus, the pKS15-01-Clone derived virus (vKS15-01-Clone) replicated efficiently in vitro and in vivo, and induced similar levels of neutralizing antibody and cytokine responses in infected animals. In contrast, the vKS15-01-EGFP virus showed impaired growth ability and induced lower level of immune response in infected animals. Lesions on the dorsal snout and coronary bands were observed in all pigs infected by parental virus KS15-01, but not in pigs infected with vKS15-01-Clone or vKS15-01-EGFP viruses. These results demonstrated that the infectious clone and EGFP reporter virus will be important tools in further elucidating the SVV pathogenesis and development of control measures.

Objectives: Earlier reports suggested that aggressive periodontitis is common in certain African populations and is associated with the JP2 clone of Aggregatibacter actinomycetemcomitans (Aa). There are few studies that investigated the type of microorganisms that colonize the subgingival sites in young subjects inflicted with a subcategory of aggressive periodontitis that is associated with the Aa-JP2 clone. Hence, the objective of this study was to characterize the subgingival microbiome of JP2 clone-associated aggressive periodontitis. Methods: The study subjects were drawn from a large survey among 14-18 years old schoolchildren in Morocco. The sample included 7 JP2-positive aggressive periodontitis subjects and 14 JP2-negative controls. The controls were selected to be either JP2-positive, JP2-negative (but Aa positive), or Aa-negative. Subgingival samples from these subjects were sequenced for the V1-V3 region (16S rRNA gene) on a Miseq platform. High-quality, non-chimeric merged reads were classified with our previously reported BLASTn-algorithm. Downstream analysis was performed with QIIME and LEfSe. Results: There were no significant differences between the groups in species richness. However, aggressive periodontitis subjects showed significantly lower alpha diversity. The microbiomes of aggressive periodontitis clustered distinctively from the controls. However, there was no significant separation between the subgroups of the control group. Species associated with health included Streptococcus spp., Haemophilus spp., Neisseria spp., Gemella spp., Rothia spp., Fusobacterium nucleatum subsp. polymorphum, Porphyromonas oral taxon 279, Veillonella parvula, Granulicatella adiacens and Lautropia mirabilis. Important periodontal pathogens, including Treponema spp., Fretibacterium spp. P. gingivalis and Tannerella forsythia were significantly enriched in aggressive periodontitis subjects. However, the taxa detected in high abundance and showed strongest association with aggressive periodontitis but not the controls were Pseudomonas oral taxon C61 and Enterobacter cloacae. Conclusions: The results suggest that several periodontal pathogens involved in chronic periodontitis also play a role in aggressive periodontitis. Future studies should investigate the role of Pseudomonas and Enterobacter spp. in the pathogenesis of aggressive periodontitis. / Oral Biology

No description available.

A restricted version of the Galois connection between polymorphisms and invariants, called Pol−CInv, is studied, where the invariant relations are restricted to so-called clausal relations. In this context, the relationship of maximal C-clones and maximal clones is investigated. It is shown that, with the exception of one special case occurring for Boolean domains, maximal C-clones are never maximal clones.:1 Introduction 2 Preliminaries 3 Proof of the main theorem 3.1 Principle of proof 3.2 Bounded orders 3.3 Non-trivial congruences 3.4 Selfdual functions 3.5 Quasilinear functions 3.6 Functions preserving central and h-regular relations 4 Concluding remarks References / Wir untersuchen eine eingeschränkte Variante der Galoisverbindung zwischen Polymorphismen und invarianten Relationen, bezeichnet mit Pol−CInv, wobei die invarianten Relationen auf sogenannte klausale Relationen beschränkt werden. In diesem Zusammenhang wird die Beziehung zwischen maximalen C-Klonen und maximalen Klonen betrachtet. Es wird gezeigt, daß, mit Ausnahme eines Spezialfalles für Boolesche Grundmengen, maximale C-Klone niemals maximale Klone sind.:1 Introduction 2 Preliminaries 3 Proof of the main theorem 3.1 Principle of proof 3.2 Bounded orders 3.3 Non-trivial congruences 3.4 Selfdual functions 3.5 Quasilinear functions 3.6 Functions preserving central and h-regular relations 4 Concluding remarks References

StackOverflow (SO) is a widely used question-and-answer (QandA) website for software developers and computer scientists. GitHub is a code hosting platform for collaboration and version control. Popular software libraries are open-source and published in repositories on GitHub. Preliminary observation shows developers cite SO questions in their GitHub repository. This observation inspired us to explore the relationship between SO posts and GitHub repositories; to help software developers better understand the characterization of SO answers that are reused by GitHub projects. For this study, we conducted an empirical study to investigate the SO answers reused by Java code from public GitHub projects. We used a hybrid approach to ensure precise results: code clone detection, keyword-based search, and manual inspection. This approach helped us identify the leveraged answers from developers. Based on the identified answers, we further investigated the topics of the discussion threads; answer characteristics (e.g., scores, ages, code lengths, and text lengths) and developers' reuse practices. We observed both reused and unused answers. Compared with unused answers, We found that the reused answers mostly have higher scores, longer code, and longer plain text explanations. Most reused answers were related to implementing specific coding tasks. In one of our observations, 9% (40/430) of scenarios, developers entirely copied code from one or multiple answers of an SO discussion thread. Furthermore, we observed that in the other 91% (390/430) of scenarios, developers only partially reused code or created brand new code from scratch. We investigated 130 SO discussion threads referred to by Java developers in 356 GitHub projects. We then arranged those into five different categories. Our findings can help the SO community have a better distribution of programming knowledge and skills, as well as inspire future research related to SO and GitHub. / Master of Science / StackOverflow (SO) is a widely used question-and-answer (QandA) website for software developers and computer scientists. GitHub is a code hosting platform for collaboration and version control. Popular software libraries are open-source and published in repositories on GitHub. Preliminary observation shows developers cite SO questions in their GitHub repository. This observation inspired us to explore the relationship between SO posts and GitHub repositories; to help software developers better understand the characterization of SO answers that are reused by GitHub projects. Our objectives are to guide SO answerers to help developers better; help tool builders understand how SO answers shape software products. Thus, we conducted an empirical study to investigate the SO answers reused by Java code from public GitHub projects. We used a hybrid approach to refine our dataset and to ensure precise results. Our hybrid approach includes three steps. The first step is code clone detection. We compared two code snippets with a code clone detection tool to find the similarity. The second step is a keyword-based search. We created multiple keywords to search within GitHub code to find the referenced answers missed by step one. Lastly, we manually inspected the outputs of both step one and two to ensure zero false positives in our data. This approach helped us identify the leveraged answers from developers. Based on the identified answers, we further investigated the topics of the discussion threads, answer characteristics, and developers' reuse practices. We observed both reused and unused answers. Compared with unused answers, We found that the reused answers mostly have higher scores, longer code, and longer plain text explanations. Most reused answers were related to implementing specific coding tasks. In one of our observations, 9% of scenarios, developers entirely copied code from one or multiple answers of an SO discussion thread. Furthermore, we observed that in the other 91% of scenarios, developers only partially reused code or created brand new code from scratch. Our findings can help the SO community have a better distribution of programming knowledge and skills, as well as inspire future research related to SO and GitHub.

Perdas embrionárias e alterações gestacionais são frequentemente observadas em prenhezes de embriões bovinos manipulados in vitro. Sabe-se que tais anormalidades são resultantes de alterações epigenéticas ocasionadas pela manipulação dos gametas e/ou do embrião durante as técnicas de reprodução assistida (ARTs), com destaque para as técnicas de fecundação in vitro (FIV) e da clonagem por transferência nuclear de células somáticas (SCNT). Tais alterações resultam em distúrbios no desenvolvimento do concepto em algum momento crítico entre a fertilização e o parto, fornecendo bons modelos de estudos sobre a fisiopatologia de perdas embrionárias e dos distúrbios de desenvolvimento. Caracterizam-se como momentos críticos após a transferência do embrião (TE) o desenvolvimento embrionário no útero, o reconhecimento materno da gestação, a placentação e o desenvolvimento da placenta e do feto, os quais tem de ser transpassados sem nenhuma falha, permitindo um desenvolvimento normal do concepto até o termo. Sendo assim, o presente trabalho abordou três fases distintas do amplo período gestacional em prenhezes por ARTs. O Estudo 1 foi realizado durante o período peri-reconhecimento materno da gestação e objetivou descrever a abundância de expressão de genes estimulados pelo interferon tau (ISGs) de células mononucleares do sangue periférico (PBMCs) maternas em gestações oriundas de ARTs no primeiro mês de gestação; o Estudo 2 compreendeu os primeiros 35 dias de gestação e objetivou descrever as mudanças morfológicas e vasculares do complexo útero-concepto-ovário e o estímulo à expressão de ISGs em PBMCs em gestações de conceptos clonados por SCNT com diferentes fenótipos de desenvolvimento, sendo esses denominados gestação anembrionada e CL persistente; e o Estudo 3 foi conduzido durante o período pré-parto e objetivou descrever as alterações na produção de esteroides sexuais e corticosteroides em gestações produzidas por ARTs. Três hipóteses foram testadas: (1) Gestações de conceptos clonados por SCNT apresentam uma baixa e mais tardia estimulação de ISGs em PBMCs maternas quando comparadas com gestações de conceptos produzidos por FIV e IA; (2) O concepto clonado por SCNT apresenta um menor estímulo sobre mudanças morfológicas e vasculares do complexo útero-ovário e ISGs em PBMCs maternas durante os primeiros 35 dias de gestação, quando comparado com conceptos oriundos de IA; e (3) Gestações de embriões oriundos de ARTs apresentam alterações na dinâmica esteroidogênica no pré-parto quando comparados com gestações de IA. No estudo 1 foram coletadas amostras de sangue de gestações produzidas por inseminação artificial (IA), FIV e clonagem por SCNT, nos dias 15, 18, 20, 22, 24, 26, 28 e 31 pós-ovulação e foi realizada mensuração da abundância de transcritos de ISGs (OAS1 e ISG15) em PBMCs maternas. No estudo 2, gestações produzidas por IA e clonagem por SCNT, foram submetidas a escaneamentos ultrassonográficos dos ovários, útero e concepto a cada 3 dias do dia 14 ao 35 (dia 0 = ovulação) e amostras de sangue foram coletadas nos dias 15, 18, 20, 22, 24, 26, 28 e 31 para mensuração da abundância de transcritos de ISGs (OAS1 e ISG15) em PBMCs maternas. No estudo 3, foram coletadas amostras de sangue no último mês em gestações naturais, oriundas de FIV e de clonagem por SCNT para análise hormonal de 10 esteroides utilizando o método de espectrometria de massas multi-hormonal de alta resolução LC-MS/MS. O primeiro estudo mostrou semelhanças na expressão de genes estimulados pelo IFNT em gestações oriundas de ARTs e produzidas por IA. Entretanto, a estimulação nas gestações oriundas de ARTs aparentou ser quatro dias mais prolongada, sugerindo uma maior funcionalidade do trofectoderma em conceptos oriundos de ARTs. O segundo estudo demonstrou um aumento na expressão de ISGs em PBMCs maternas tanto em gestações de conceptos normais quanto em anormais, justificando a manutenção da função luteal mesmo na ausência de detecção do concepto por ultrasonografia. No terceiro estudo, demonstrou-se alterações na esteroidogênese nas gestações de embriões FIV e clonados no último mês de gestação, sendo essas compatíveis com a hiperativação da enzima aromatase durante todo o último mês de gestações oriundas de FIV e hiperativação das enzimas P450C11 e P450C21 trinta dias antes do parto em gestações oriundas de clonagem por SCNT. O presente estudo concluiu que conceptos oriundos de FIV e clonagem por SCNT apresentam um prolongamento no estímulo de ISGs pelo IFNT, conceptos clonados anormalos apresentam estímulo de ISGs, o que justifica a manutenção da função luteal, e, por fim, a cascata esteroidonênica que culmina com o parto apresenta-se alterada em gestações oriundas de FIV e clonagem por SCNT. / Pregnancy losses and gestational abnormalities are frequently observed in pregnancies from in vitro produced embryos in bovines. It is known that these abnormalities are due to epigenetic changes from the manipulation of gametes and/or embryo during the use of assisted reproduction techniques (ARTs), especially for the in vitro fertilization (IFV) and cloning by somatic cells nuclear transfer (SCNT). These changes results in disturbances of conceptus development in any critical stage between the fertilization and parturition, which provides good models for the study of physiopathology of embryo losses and disturbances of development. Critical stages after the embryo transfer (ET) to the uterus are characterized as the maternal recognition of pregnancy, placentation, and fetal-placental development, which needs to be surpassed without failures, in order to develop a normal conceptus until term. Therefore, the present work approached three distinct phases of the wide gestational period in pregnancies from ARTs. The Study 1 was conducted during the maternal peri-recongnition of pregnancy period and aimed to describe the expression of interferon stimulated genes (ISGs) in maternal peripheral blood mononuclear cells (PBMCs) in pregnancies derived of ARTs; the Study 2 comprise the first 35 days of pregnancy and aimed to describe morphological and vascular changes of the complex uterus-ovaries-conceptus, as well as the expression of ISGs in maternal PBMCs in pregnancies of conceptus cloned by SCNT with different phenotypes of development, denominated as anembryonic gestation and persistent CL; the Study 3 was conducted during the pre-partum period and aimed to describe changes in the production of sexual steroids and corticosteroids during the last month of pregnancies derived of ARTs. Three hypothesis were tested: (1) Pregnancies of conceptus cloned by SCNT presented a decrease and delay in the stimulation of ISGs in maternal PBMCs when compared with conceptuses produced by IFV and AI; (2) Stimulus from the conceptus for changes in the morphology and vasculature of the the uterus-ovarian complex, detected by ultrasonography in B and Doppler modes, and the stimulation of ISGs in maternal PBMCs during the first 35 days of pregnancy of conceptus cloned by SCNT are less intense when compared with conceptus derived from AI; and (3) Pregnancies derived of ARTs present changes in the steroidogenic dynamics in the pre-partum, when compared with pregnancies derived from AI. In Study 1 blood samples were collected from pregnancies produced by AI, IVF, and cloning by SCNT, at days 15, 18, 20, 22, 24, 26, 28, and 31 post-ovulation for the measurement of abundance of transcripts of ISGs (OAS1 and ISG15) in maternal PBMCs. In Study 2, pregnancies derived of AI and cloning by SCNT, were submitted to ultrasonographic scans for the evaluation and description of morphological and vascular changes in ovaries, uterus, and conceptus every 3 days from day 14 to 35 (day 0 = ovulation) and blood samples were collected on days 15, 18, 20, 22, 24, 26, 28 e 31 for the measurement of the abundance of transcripts of ISGs (OAS1 and ISG15) in maternal PBMCs. In Study 3, blood samples were collected during the last month of pregnancies naturally conceived, derived of IVF, and cloned by SCNT for the analysis of 10 steroids using the method of mass spectrometry high resolution LC-MS/MS. The first study showed similarities in the ISGs expression stimulation in pregnancies derived of ARTs and AI. However, the stimulation in the ART derived pregnancies was apparently 4 days longer, suggesting a greater placental function in conceptus derived of ARTs. The second study showed an increase in ISG expression in both normal and abnormal conceptus development, which justifies the maintenance of CL in the absence of a conceptus structure detected by ultrasonography. In the third study, was detected changes in the steroidogenesis of pregnancies derived of IFV and cloning by SCNT during the last month of pregnancy, which are compatible with the hyperactivation of the aromatase enzyme during the last month of IFV derived pregnancies, and hyperactivation of the enzymes P450C11 and P450C21 thirty days before parturition in pregnancies derived of cloning by SCNT. The present study concludes that conceptus derived of IFV and cloning by SCNT present a prolonged stimulus of ISGs, cloned conceptus with anomalous development presents a stimulus of ISGs, which justifies the CL function maintenance, and, ultimately, the steroidogenic cascade that culminates with the term is altered in pregnancies derived from IFV and cloning by SCNT.

No Brasil e em diversos países têm sido verificados significativos avanços na silvicultura clonal intensiva com diferenças marcantes existentes entre os clones de espécies e de híbridos de Eucalyptus quanto aos parâmetros de crescimento e de desenvolvimento. Ao mesmo tempo, nos últimos tempos, foi introduzido o conceito de uso múltiplo do lenho das árvores com a utilização da madeira como fonte de celulose e papel, madeira sólida e outras aplicações como estratégia de aumento da rentabilidade do empreendimento florestal. Pelo exposto, o presente trabalho de pesquisa teve como objetivo a caracterização da madeira de árvores de 8 anos de clones de Eucalyptus grandis, E. saligna e de E. grandis x urophylla, visando possibilitar a otimização do seu uso. De plantações clonais localizadas no município de São Miguel de Arcanjo-SP da Cia Suzano de Celulose e Papel, foram selecionados os 5 melhores clones de cada espécie e híbrido pelas suas características silviculturais. Para cada clone foram selecionadas e cortadas as 10 melhores árvores (total de 150 árvores) e foram mensurados os parâmetros dendrométricos (DAP, altura total, altura comercial, espessura da casca). Do tronco dessas árvores foram cortados cilindros de madeira, em 5 porcentagens da altura do tronco e no DAP, em um total de 900 amostras. Em condições de laboratório, as amostras de madeira foram preparadas e analisadas com respeito aos parâmetros de qualidade da madeira anatômicos (caracterização da estrutura anatômica, variabilidade radial das dimensões das fibras, vasos, cerne e alburno) e físicos (densidade básica, densidade aparente através de densitometría de raios X, variabilidade radial da densidade) proporcionando a obtenção do índice de unifomidade da madeira. Os resultados possibilitaram a detalhada caracterização dos clones de cada árvore com respeito às variações ao longo do seu tronco e no sentido radial das amostras do DAP, a saber: (i) o teor médio de casca e alburno foi superior nos clones de E.saligna, o de cerne nos de E. grandis, (ii) os valores médios de densidade básica da madeira foram de 0,46; 0,47 e 0,49 g/cm3 para o E. grandis, E. saligna e E. grandis x urophylla; ocorreram variações longitudinais dentro e entre os clones das espécies de Eucalyptus, (iii) foram observadas variações quali e quantitativas dos elementos anatômicos da madeira entre espécies e híbrido de Eucalyptus, (iv) o valor médio da densidade aparente da madeira foi de 0,46; 0,54 e 0,54 g/cm3 nos clones de E. grandis, E. saligna e E. grandis x urophylla, respectivamente, (v) o índice de uniformidade indicou maior homogeneidade da madeira dos clones de E. grandis e E. saligna. A partir dos resultados obtidos foram preconizados alguns usos potenciais da madeira dos clones das duas espécies e híbrido de Eucalyptus. / In Brazil, as in many other countries, it has been verified significant advances in intensive clone silviculture with remarkable differences between clones of species and hybrids of Eucalyptus based on parameters of growth and development. At the same time, during these last years, it has been introduced the concept of multiple uses of trees log, having the wood as cellulose and paper source, solid wood and other applications as strategies for rising rentability of forestry enterprise. For the above, this research was based on wood of 8-year-old clones of Eucalyptus grandis, E. saligna and E. grandis x urophylla, heading to the optimization of its use. The 5 best clones of each specie and hybrid were selected based on their characteristics silviculturais and were chosen from clone cultures of Cia Suzano de Celulose e Papel, located at the district of São Miguel Arcanjo, in SP. For every clone it was selected and sliced the 10 best trees (over 150) and their dendrometrics parameters were measured (DHP, total highness, commercial highness, thicket bark etc). These trees had their log sliced in cylinders of wood, in 5 percentage of highness of the log and DHP, over a total amount of 900 sample. Under laboratory conditions, these wood samples were prepared and analyzed respecting the parameters about the wood quality related to its aspects anatomic (anatomic structure, radial variability of dimensions fibers, vases, heartwood and sapwood) and physics aspects (basic density, density based on densitometry of x-rays, radial variability of parameters about the wood quality related to its aspects anatomic (anatomic structure, radial variability of dimensions fibers, vases, heartwood and sapwood) and physics aspects (basic density, density based on densitometry of x-rays, radial variability of density) to obtain the uniformity index of wood. The results provided a detailed characterization of the clones of each tree related to the variations all over their log and related to the radial direction of DHP samples, as to know: (i) medium content of bark and sapwood was superior in clones of E.saligna, the heartwood in E.grandis, (ii) the medium values of wood basic density were 0,46; 0,47; 0,49 g/cm3 for the E.grandis, E.saligna and E.grandis x urophylla; it has occurred longitudinal variations inside and between clones of eucalyptus species, (iii) it was observed the variations about quality and quantity of anatomic elements of wood between species and hybrid of eucalyptus, (iv) the medium values of density apparent of wood was 0,46; 0,54; 0,55 g/cm3 in clones of E.grandis, E.saligna and E. grandis x urophylla, (v) the uniformity index indicated more homogeneity of wood in clones of E.grandis and E. saligna. Starting from these results, it had been stated some potencial uses of cloned wood for the 2 species and hibryd of Eucalyptus.

Le virus West Nile (VWN) est un virus neurotrope principalement transmis par piqûre de moustique et dont le réservoir est constitué par la faune aviaire sauvage. Les souches circulant en Europe appartiennent à 4 lignages génétiques différents à l’origine de nombreuses épidémies d’ampleur modérée à faible et limitées géographiquement, contrairement à ce qui a été observé en Amérique du Nord. En 1998 en Israël, une importante épidémie a a été associée pour la première fois à une forte mortalité de la faune aviaire sauvage. Le virus (souche IS-98-ST1, lignage 1a) a été isolé du cerveau d’une cigogne moribonde. L’objet de cette thèse a été de construire un clone infectieux de la souche IS-98-ST1 afin d’en explorer les propriétés de neuroinvasion et de pouvoir mettre en évidence les déterminants moléculaires de sa virulence.Le virus obtenu à partir de la construction clone infectieux s’est révélé posséder les mêmes propriétés biologiques que le virus parental, que ce soit in vitro sur cellules Vero ou in vivo sur souris sensibles ou résistantes ou encore sur l’embryon de poulet. L’embryon de poulet est présenté ici comme un nouveau modèle d’évaluation de la virulence du VWN. Un modèle cellulaire neuronale (lignée de neuroblastomes humains, SK-N-SH) est aussi évalué dans ce manuscrit. En conclusion, un nouvel outil de génétique inverse a été obtenu pour le VWN. Cet outil permettra de travailler sur l’impact de mutations ponctuelles, ou de modifications plus importantes touchant un ou plusieurs gènes viraux sur la virulence du VWN, spécifiquement dans le contexte européen. / West Nile virus (WNV) is a neurotropic virus mainly transmitted through mosquito bites. Wild birds represent the main reservoir hosts. Strains circulating in Europe belong to four lineages and have caused numerous but limited epidemics over the last few years. In 1998, an important outbreak associated to huge bird fatalities caused by a highly neuroinvasive strain (IS-98-ST1) took place in Israel. We aimed at producing a new infectious clone, based on the lineage 1a IS-98-ST1 WNV strain, for the characterization of its neuroinvasion properties as well as the molecular determinants of European WNV virulence. The growth kinetics of recombinant and parental WNV were similar in Vero cells. Moreover, the phenotypes of recombinant and parental WNV were indistinguishable in terms of viremia, viral load in the brain and mortality in susceptible and resistant mice. Finally, the pathobiology of the infectious clone was examined in embryonated chicken eggs, proposed as a new model for the evaluation of WNV virulence. The potential of human neuroblastoma cells (SK-N-SH) to discriminate between highly and mildly virulent WNV strains was assayed. In conclusion: a new molecular tool that is useful for the study of molecular determinants of WNV virulence has been generated. We take advantage of the high genetic stability of our one-piece infectious WNV cDNA clone to produce mutant viruses through the insertion of point mutations or the exchange of genetic fragments between WNV strains into the backbone of the IS-98-ST1 infectious clone.