São estimadas 90 mil espécies de insetos para o Cerrado, o segundo maior bioma brasileiro, com enorme heterogeneidade de habitats e rica fauna. Dentre esses insetos, os Lepidoptera representam cerca de 10% do total de espécies. Entretanto, estudos sobre seus imaturos (ovo, larva e pupa) ainda são incipientes para o bioma, principalmente sobre as mariposas, cujas histórias naturais de muitas espécies ainda são desconhecidas. A falta desses estudos faz com que sejam ignoradas as interações multitróficas em que esses organismos estão envolvidos e, portanto, inviabiliza futuros estudos relacionados ao seu papel em comunidades e redes ecológicas. Esse é o caso de muitas espécies da família Geometridae, incluindo Oospila pallidaria, que é uma mariposa esmeralda (devido à coloração verde esmeralda das suas asas). Oospila pallidaria é uma espécie herbívora que não possuía até o momento nenhuma informação publicada sobre a sua biologia e fatores que condicionam sua ocorrência, tais como: a fenologia da planta hospedeira, que pode determinar o período de melhores condições para crescimento e reprodução; a qualidade nutricional dos recursos alimentares (e. g. quantidade de água e nitrogênio), que pode determinar quais recursos conferem melhor desenvolvimento/performance a um herbívoro; os inimigos naturais, que podem restringir a ocorrência dos herbívoros e a temperatura e pluviosidade, que podem alterar a qualidade e a distribuição geográfica dos recursos alimentares utilizados pelos herbívoros. Nesse sentido, o objetivo principal deste estudo foi descrever, pela primeira vez, aspectos da biologia e história natural de O. pallidaria (Capítulo 1) e avaliar a sua performance larval, a partir de diferentes dietas (Capítulo 2). Os resultados apresentados no Capítulo 1 mostraram que os ovos de O. pallidaria, verdes e com duração média de sete dias, foram solitários e ovipostos principalmente nos tricomas das folhas maduras. As larvas se alimentaram de folhas maduras predominantemente, mas utilizaram folhas jovens e botões florais oportunisticamente. As larvas tiveram coloração críptica, se camuflaram em meio às folhas de Mimosa setosa (Leguminosae: Mimosoideae), sua única hospedeira, e desenvolveram o comportamento de auto-limpeza. Há cinco ínstares larvais, com coloração que variou de amarelo a verde. As larvas apresentaram também um par de projeções no protórax e linha mediana marrom na região dorsal. O comprimento máximo do corpo das larvas foi de 28 mm. As pupas foram predominantemente verdes, com no máximo 10 mm de comprimento. O desenvolvimento do ovo ao adulto durou cerca de 50 dias ( = 42; S = 6; n = 11). Seu único inimigo natural registrado foi o microhimenóptero Cotesia sp. (Braconidae, Microgastrinae). Fêmeas de O. pallidaria produziram 65 ovos em média (S = 7,07; n = 2). A ocorrência de O. pallidaria foi sazonal e sobreposta ao pico de presença de folhas maduras. Larvas foram negativamente relacionadas à temperatura e pluviosidade, com significância estatística somente para a primeira (r = - 0.5889, P < 0.05). No Capítulo 2, foi mostrado que as folhas maduras de M. setosa foram o único recurso alimentar disponível durante todo o ano. Os botões florais foram o recurso com maior qualidade nutricional (conteúdo relativo de água e nitrogênio total), seguidos por folhas jovens e maduras respectivamente. Entretanto, a sobrevivência de O. pallidaria foi maior com folhas maduras. Os resultados indicam que para o herbívoro especialista O. pallidaria, a fenologia da planta hospedeira é crucial para sua sobrevivência, em especial a presença de folhas maduras de M. setosa. Por outro lado, os botões florais são recursos efêmeros, porém importantes para a sobrevivência das larvas de últimos ínstares no final da estação seca, quando as folhas maduras estão ressecadas e/ou senescentes. / It is estimated 90.000 species of insects for the Cerrado, the second largest Brazilian biome, with great diversity of habitats and rich fauna. Among these insects, Lepidoptera represents about 10% of all species. However, studies on their immature stages (egg, larvae and pupae) are incipient for the biome, mostly on moths, whose natural histories of many species are unknown. The lack of these studies makes the multitrophic interactions in that these organisms are involved ignored and, therefore, prevent future studies related to their role in communities and ecological networks. This is the case of many species of Geometridae, including Oospila pallidaria, which is an emerald moth (due to emerald green color of their wings). Oospila pallidaria is a herbivorous species that had not published any information on their biology and factors that influence their occurrence, such as: the phenology of the host plant, which can determine the time of better conditions for growth and reproduction; the nutritional quality of food resources (e. g. amount of water and nitrogen), which can determine what resources provide better development/performance to a herbivore; natural enemies, that could restrict the occurrence of herbivores and the temperature and rainfall, which can change the quality and geographical distribution of food resources used by herbivores. Accordingly, the aim of this study was to describe, for the first time, aspects of the biology and natural history of O. pallidaria (Chapter 1) and to assess their larval performance from different diets (Chapter 2). The results presented in Chapter 1 showed that eggs of O. pallidaria, green and lasting an average of seven days, were lonely and laid especially in trichomes of mature leaves. The larvae fed on mature leaves predominantly, but used young leaves and flower buds opportunistically. Larvae had cryptic coloration, they camouflaged among the leaves of Mimosa setosa (Leguminosae: Mimosoideae), a single host, and developed the self-cleaning behavior. There are five larval instars, with color ranging from yellow to green. The larvae also had a pair of projections on prothorax and brown midline in the dorsal region. The maximum length of the larvaes body was 28 mm. The pupae were predominantly green, with a maximum of 10 mm in length. The development from egg to adult lasted about 50 days ( = 42, S = 6, n = 11). Their only natural enemy recorded was the microhymenopteran Cotesia sp. (Braconidae, Microgastrinae). Oospila pallidaria females produced 65 eggs on average (S = 7.07; n = 2). The occurrence of O. pallidaria was seasonal and overlapped on the peak presence of mature leaves. Larvae were negatively related to temperature and rainfall, with statistical significance only for the first (r = - 0.5889, P < 0.05). In Chapter 2, it was shown that mature leaves of M. setosa were the only food source available throughout the year. The flower buds were the resource with higher nutritional quality (relative water content and total nitrogen), followed by young and mature leaves respectively. However, the survival of O. pallidaria was greater with mature leaves. The results indicate that for the specialist herbivore O. pallidaria, the host plant phenology is critical for its survival, especially the presence of mature leaves of M. setosa. On the other hand, the flower buds are ephemeral resources, but important for the survival of the last instar larvae in the dry season, when mature leaves are withered and/or senescent.

A tribo Omocerini, proposta por Hincks, inclui atualmente sete gêneros (Canistra Erichson, Carlobruchia Spaeth, Cassidinoma Hincks, Cyclosoma Guérin-Meneville, Discomorpha Chevrolat, Omocerus Chevrolat e Polychalca Chevrolat), 17 subgêneros e 147 espécies. Considerando que os estudos filogenéticos de Cassidinae nunca incluíram todos os gêneros e subgêneros da tribo e que sinonímias e mudanças taxonômicas vêm sendo propostas sem embasamento filogenético, este trabalho teve como objetivo avaliar a monofilia da tribo e propor hipóteses sobre as relações entre seus gêneros e subgêneros e testar a validade dos mesmos. Foram analisadas 46 espécies de Omocerini como grupo-interno e 14 espécies de outras seis tribos de Cassidinae como grupo-externo, utilizando 230 caracteres morfológicos (200 referentes a adultos e 30 a imaturos). A análise filogenética com pesos iguais resultou em nove árvores igualmente parcimoniosas com 1101 passos. Na análise com pesagem implícita dos caracteres (K=3) apenas uma árvore foi encontrada. As árvores resultantes de todas as análises corroboram a monofilia da tribo Omocerini com base em 13 sinapomorfias (seis exclusivas). O grupo-irmão de Omocerini é o clado que representa a tribo Stolaini (Mesomphalia gibbosa (Chelymorpha cribraria + Stolas lacordairei)), hipótese de relacionamento nunca proposta anteriormente. As relações encontradas entre os grandes grupos de Omocerini foram: ((Carlobruchia (Cassidinoma (\"Polychalca\" + \"Omocerus\"))) + (Canistra (\"Cyclosoma\" + \"Discomorpha\"))). Os gêneros Cyclosoma, Discomorpha, Omocerus e Polychalca são parafiléticos. Canistra apresenta-se monofilético e grupo-irmão de (\"Cyclosoma\" + \"Discomorpha\"). Carlobruchia é considerado monofilético baseado na análise com pesagem implícita, sendo grupo-irmão de: (Cassidinoma (\"Polychalca\" + \"Omocerus\")). Polychalca foi considerado sinônimo junior de Omocerus e Discomorpha sinônimo junior de Cyclosoma e apenas os sinônimos senior foram considerados válidos. A monofilia dos subgêneros de Omocerini não é suportada nas análises realizadas, portanto, estes não devem ser utilizados como agrupamentos taxonômicos. / The tribe Omocerini, proposed by Hincks, currently comprises seven genera (Canistra Erichson, Carlobruchia Spaeth, Cassidinoma Hincks, Cyclosoma Guerin- Meneville, Discomorpha Chevrolat, Omocerus Chevrolat e Polychalca Chevrolat), 17 subgenera, and 147 species. Given that phylogenetic studies of Cassidinae have never included data regarding all genera and subgenera of the tribe, and synonymization and taxonomic category changes have been proposed without phylogenetic basis, this study aims to evaluate the monophyly of Omocerini based on morphological characters, propose a relationship hypothesis between its genera and subgenera and test their validity. Forty six species of Omocerini were included as ingroup and 14 species of six tribes of Cassidinae representing the outgroup, were analyzed based in 230 morphological characters 200 from adults and 30 from immature). The phylogenetic analysis with equal weighting of the characters yielded nine fundamental trees with 1101 steps. The analysis under implied weighting (k=3) resulted in a single most fit cladogram. The monophyly of the tribe was corroborated in all analysis, with 13 sinapomorphies (six exclusive) supporting it. The sister-group of Omocerini is the clade representing the Stolaini (Mesomphalia gibbosa (Chelymorpha cribraria + Stolas lacordairei)), a relationship hypothesis never proposed before. The relationships obtained between the subunits of Omocerini were: ((Carlobruchia (Cassidinoma (\"Polychalca\" + \"Omocerus\"))) + (Canistra (\"Cyclosoma\" + \"Discomorpha\"))). The genera Cyclosoma, Discomorpha, Omocerus and Polychalca are paraphyletic. Canistra is monophyletic and as the sistergroup of (\"Cyclosoma\" + \"Discomorpha\"). Carlobruchia is considered monophyletic based on the analysis under implied weighting and is sister-group of (Cassidinoma (\"Polychalca\" + \"Omocerus\")). Polychalca is considered a junior synonym of Omocerus and Discomorpha a junior synonym of Cyclosoma, and only the senior synonyms were considered valid. The monophyly of the subgenera of Omocerini were not supported by the analysis, therefore these should not be used as taxonomic groupings.

Orientador: André Victor Lucci Freitas / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-20T03:27:03Z (GMT). No. of bitstreams: 1 Barbosa_EduardoProenca_M.pdf: 42430863 bytes, checksum: 2e52ceb55e75dfe4450c676669d34918 (MD5) Previous issue date: 2012 / Resumo: Muitos aspectos e detalhes da origem, evolução e padrões de distribuição das borboletas ainda não são totalmente compreendidos, como as relações filogenéticas entre a grande maioria dos grupos de borboletas, que permaneceram desconhecidas até recentemente. Estudos filogenéticos até alguns anos atrás foram baseados principalmente em caracteres morfológicos de adultos muito embora o uso de caracteres de estágios imaturos tenha aumentado em importância atualmente, principalmente por se mostrarem informativos onde os caracteres de adultos foram insuficientes. Ainda assim, existem poucos trabalhos que utilizem esses caracteres tanto para a filogenia quanto para a classificação das borboletas. Além das lacunas nas relações de ancestralidade comum das borboletas, também existem lacunas a serem preenchidas com relação aos padrões biogeográficos, principalmente com relação à biogeografia histórica. Nos neotrópicos, a região mais biodiversa do planeta, o estudo da origem e da biogeografia da maioria dos grupos de organismos incluindo borboletas, é um assunto de grande interesse. Isso porque a maioria desses grupos não possui uma boa compreensão de suas histórias evolutivas, e desvendar alguns desses pontos seria dar um passo em direção à compreensão dessa enorme diversidade Neotropical. Apesar de ser um grupo relativamente bem conhecido, o gênero Actinote (Nymphalidae: Heliconiinae) ainda carece de estudos para uma melhor compreensão das relações internas do grupo, bem como sobre os padrões de distribuição na Região Neotropical. No presente estudo foram realizadas, análises filogenéticas baseadas em caracteres morfológicos com 34, 31 e 29 táxons, respectivamente. Nas três análises a maior parte dos clados é suportado por homoplasias, embora alguns grupos reduzidos de espécies, tenham aparecido suportados por sinapomorfias. Embora muitas discussões apontem para uma grande importância no uso de imaturos para se resolver as relações de ancestralidade comum entre as borboletas, os caracteres de estágios imaturos de Actinote não se mostraram muito informativos para elucidar as relações desse grupo, apesar de terem grande valor taxonômico, ajudando na identificação da maioria das espécies. Isso provavelmente se deve há uma grande variação intra-específica e uma baixa congruência com os caracteres de adultos, o que tornaria difícil o uso desses caracteres para a resolução das relações evolutivas para esse grupo em particular. Para alguns autores essa dificuldade poderia ser resultado de uma rápida diversificação durante a história evolutiva dessas espécies, limitando a diferenciação morfológica. A análise biogeográfica foi efetuada com o programa Reconstructing Ancestral States in Phylogeny (RASP). De acordo com a hipótese biogeográfica ótima, o padrão de distribuição observado poderia ser explicado por 41 dispersões, seis eventos vicariantes e três extinções. Segundo a análise o ancestral de Altinote e Actinote era amplamente distribuído, ocupando os Andes e a Serra do Mar, no sudeste do Brasil. Um evento vicariante isolou então a linhagem ancestral de Altinote nos Andes e a linhagem ancestral de Actinote no sudeste da América do Sul. Os eventos vicariantes poderiam estar relacionados a eventos de mudanças climáticas, o que faz sentido para este grupo em particular, que é típico de ambientes frios de grandes altitudes. De acordo com esta hipótese, durante os períodos glaciais essas espécies de grande altitude poderiam se dispersar e alcançar ambientes em baixas altitudes. Quando as temperaturas aumentavam durante os períodos interglaciais, estas espécies eram mantidas nesses locais de maior altitude, onde as temperaturas permaneciam baixas, e ficavam isoladas. Se o período de isolamento fosse grande o bastante, as espécies poderiam então passar por eventos de especiação, se tornando novas espécies / Abstract: Many aspects and details of the origin, evolution and biogeographical patterns of butterflies are not yet fully comprehended. The phylogenetic relationships among most butterfly groups remained unknown until recently. Phylogenetic studies have been based mainly on morphological characters of adults, although characters of immature stages had increased in use and importance nowadays, specially due to their capacity of been informative where adult characters are not enough. There are few studies that use these characters either to phylogeny or classification of the butterflies. Besides the lack of information about the common ancestry of butterflies, there are also gaps to be filled about the biogeographical patterns, primarily related to the historical biogeography. In the neotropics, the planet's most biodiverse region, the study of origin and biogeography of most groups of organisms, including butterflies, is a subject of great interest. This because most of these groups are poorly understood concerning their evolutionary histories and shed light on some of these histories would lead us toward the comprehension of this huge Neotropical diversity. Despite being one relatively well known group, the genus Actinote (Nymphalidae: Heliconiinae) still lacks studies in phylogeny to solve the internal relationships of this group. It also lacks studies that seek to understand the distributional patterns of the group in the Neotropical region and which factors were responsible for the high diversity of Actinote in the southeast region of South America. In the present study we carried out three phylogenetic analyses based on morphology, with 34, 31 and 29 táxons respectively. In all analyses the majority of the clades were supported by homoplasies, although some groups of few species were maintained apparently the same in all the analyses supported by synapomorphies. Despite the fact that many discussions point out to a great importance in the use of immature characters to help solving the common ancestry relationships of butterflies, in the present work these characters were not very informative to elucidate the relations of this group, although they have a great taxonomic value, being helpful in the identification of the majority of species. This is probably due to a great intraspecific variation and a lower congruency with the adults' characters which would make it harder to use these characters to try and solve the evolutionary relationships of this particular group. Some authors claim that this difficulty could be a result of a rapid diversification during the evolutionary history of these species, restricting the morphological differentiation. The biogeographical analysis was carried out with the program Reconstructing Ancestral States in Phylogeny (RASP). According to the optimal biogeographical hypothesis, the distribution pattern observed could be explained by 41 dispersal, six vicariant and three extinction events. The analysis pointed out to a spread distributed ancestor of Altinote and Actinote, occupying the Andes and the Serra do Mar, in southeast Brazil. A vicariant event had isolated the ancestor lineage of Altinote in the Andes and the ancestor lineage of Actinote in the southeast of South America. The vicariant events could be related to climatic changes, what makes sense to this particular group, which is typical of cold high altitude habitats. According to this hypothesis, during the glacial periods the species of high altitude habitats could disperse and reach low altitude habitats. When the temperatures got higher in the interglacial periods these species were trapped in high altitude places, of lower temperature, and became isolated. If this isolation period lasted long enough, the species could pass through speciation events, turning into new species / Mestrado / Ecologia / Mestre em Ecologia

Syssphinx molina (Cramer) é considerada uma espécie de interesse fitossanitário, pois pode ser praga de algumas plantas cultivadas pelo homem, além disso, em lavouras de monoculturas podem ocorrer acidentes com as larvas dessa espécie, causando dermatites nos trabalhadores. O principal objetivo desse estudo é obter e caracterizar as células embrionárias indiferenciadas de Syssphinx molina. Ovos da espécie serão obtidos através de mariposas fêmeas grávidas, coletadas pela equipe da Coleção Zoológica do Maranhão (CZMA), da Universidade Estadual do Maranhão (UEMA), Campus Caxias. Os ovos foram caracterizados e comparados com ovos de outras espécies filogeneticamente próximas à S. molina. Culturas primárias de células embrionárias de S. molina foram cultivadas por 20 dias, ocorreram duas passagens e procedeu-se com o congelamento. Após descongelamento das células ocorreram 10 passagens. Foram analisadas amostras de células de cada passagem para obter dados do ciclo célular e características das células através de uso de marcadores: Anti-Axons, Anti-Caspase 3 ativa, Anti-CD105, Anti-CD117, Anti-CD24, Anti-CD43, Anti-CD73, Anti-CD90/Thy1, Anti-Ciclina D1, Anti GM 130, Anti-HLA DR, Anti-HSP47, Anti-HSP70,. Anti-KI67, Anti-MCP1, Anti- Oct 3/4, Anti-p53, Anti RAB 5, Anti-SSEA4, Anti-Stro-1, Anti-TGF beta 1, Anti-Vimentina e Schneider L2. Pela primeira vez é determinado um protocolo para resfriamento de ovos de S. molina, assim como protocolo para cultura primária e secundária de células embrionárias dessa espécie. São analisadas também as idades gestacionais dos ovos de S. molina e comparadas com ovos de outras espécies de mariposas da mesma subfamília. A análise de ciclo celular e marcadores confirmam a alta taxa de proliferação das células, no entanto, a análise com os anticorpos Anti-Caspase 3 ativa e Anti-P53 mostrou o percentual de morte celular programada (apoptose) é, geralmente, maior que 25% nas populações de células analisadas. Os marcadores Anti GM130 e Anti RAB 5, que participam, respectivamente, do recrutamento de proteínas pela fase cis do aparelho de Golgi e do processo de maturação do endossoma, marcaram mais de 50% das células das amostras analisadas. Anti-HLA-DR, que revela proteínas da membrana do linfócito T, com um percentual geralmente superior a 30% de marcação. Dentre os marcadores de células multi e pluripotentes, aquele que marcou maior taxa de células foi Anti- CD117, que se liga em células estaminais hematopoiéticas. Todos os anticorpos utilizados para marcar células do sistema hematopoiético (Anti-CD24, Anti-CD43, Anti-CD73, Anti-CD90/Thy1, Anti-HLA DR e Anti-MCP1) foram expressos nas células cultivadas de S. molina. Portanto, entende-se que os insetos, a exemplo de S. molina, são um grupo que possuem atividades metabólicas complexas e o entendimento dessas atividades permitirá, no futuro, delinear novas formas de controle biológico. Além disso, os dados inéditos sobre o sistema hematopoiético dos insetos apresentado nesse trabalho, além constitui um subsídio fundamental para estabelecer futuros modelos importantes para estudos de estratégias de controle biológico, como também poderá auxiliar no desenvolvimento de técnicas para combater doenças transmitidas por insetos. / Syssphinx molina (Cramer) is considered a species of phytosanitary interest, it can be curse of some plants cultivated by man, in addition, in monoculture plantations, accidents may occur with the larvae of this species, causing dermatitis in workers. The main objective of this study is to obtain and characterize the undifferentiated embryonic cells of Syssphinx molina. Eggs of the species will be obtained through pregnant female moths, collected by the team of the Coleção Zoológica do Maranhão (CZMA), Universidade Estadual do Maranhão (UEMA), Campus Caxias-MA. The eggs were characterized and compared with eggs of other phylogenetically close species of S. molina. Primary cultures of S. molina embryonic cells were cultured for 20 days, Two passes occurred and proceeded with freezing. After thawing of the cells, there were 10 passes. Cell samples from each passage were analyzed to obtain cell cycle data and cell characteristics through the use of markers: Anti-Axons, Anti-Caspase 3 ativa, Anti-CD105, Anti-CD117, Anti-CD24, Anti-CD43, Anti-CD73, Anti-CD90/Thy1, Anti-Ciclina D1, Anti GM 130, Anti-HLA DR, Anti-HSP47, Anti-HSP70,. Anti-KI67, Anti-MCP1, Anti- Oct 3/4, Anti-p53, Anti RAB 5, Anti-SSEA4, Anti-Stro-1, Anti-TGF beta 1, Anti-Vimentina e Schneider L2. For the first time a protocol for cooling eggs of S. molina is determined, as well as protocol for primary and secondary culture of embryonic cells of this species. The gestational age of S. molina eggs and compared to eggs of other species of moths of the same subfamily. Cell cycle analysis and markers confirm the high rate of cell proliferation, however, analysis with the active Anti-Caspase 3 and Anti-P53 antibodies showed the percentage of programmed cell death (apoptosis) is generally greater than 25 % in the analyzed cell populations. Markers Anti GM130 and anti RAB 5, which participate respectively, recruitment of proteins by cis phase of the Golgi apparatus and endosome maturation process, scored more than 50% of the cells in the samples analyzed. Anti-HLA-DR, which reveals T lymphocyte membrane proteins, with a percentage generally greater than 30% labeling. Among the multi- and pluripotent cell markers, the one that scored the highest cell rate was Anti-CD117, which binds to hematopoietic stem cells. All antibodies used to label cells from the hematopoietic system (Anti-CD24, Anti-CD43, Anti-CD73, Anti-CD90 / Thy1, Anti-HLA DR and Anti-MCP1) were expressed in the cultured cells of S. molina. Therefore, it is understood that insects, like S. molina, are a group that has complex metabolic activities and the understanding of these activities will, in the future, outline new forms of biological control. In addition, the unpublished data on the hamatopoietic system of insects presented in this work, beyond is a key benefit to establish future important models for studies of biological control strategies, but may also assist in the development of techniques to combat diseases transmitted by insects.

Agrobacterium is widely considered, when suitably modified, to be the most effective vector for gene transfer into plant cells. For a long time, many cereals crops (monocotyledonous plants) were recalcitrant species to genetic modification, mainly as a result of their recalcitrance to in-vitro regeneration and their resistance to Agrobacterium infection. However, recently Agrobacterium-mediated transformation has been used to transform monocot crops such as maize (Zea mays) but with severe restrictions on genotype suitability. This study was carried out to evaluate the transformation amenability of 2 Syrian maize varieties and 2 hybrids in comparison with the hybrid line Hi II by the Agrobacterium tumefaciens-mediated transformation technique using a callus induction based system from immature zygotic embryos IZEs. A. tumefaciens strains EHA101, harbouring the standard binary vector pTF102, and the EHA105 containing the pBINPLUS/ARS:PpCBF1 vector were used. The effects of genotypes and the size of IZEs explants on callus induction and development were investigated. Results showed that callus induction and subsequent callus growth were significantly affected by the initial explant size. Calli induction from IZEs explants sized 1.5-2.00mm was 76%. Callus weight however decreased to 8.2g, compared with 11.7g of callus derived from IZEs >2.00mm. Callus induction ranged between 73.6-78.9% for varieties and hybrids respectively. Calli derived from varieties weighed significantly more than those initiated from the hybrids. Results demonstrated that Syrian maize genotypes were efficiently transformed via the A. tumefaciens strains but there was variation in transformation frequency. A transformation frequency of 3.7-4.2% was achieved for hybrids and varieties respectively confirming that the transformation frequency was genotype-dependent. The transformation frequency averaged between 3.2-5.6% for the EHA105 and EHA101 respectively. Fertile transgenic plants were regenerated from mature somatic embryos with an average regeneration frequency of 59.2 and 17% respectively for varieties and hybrids. Transgenic seeds of R0 and R1 progenies were produced from 74% of the outcrosses attempted and more than 98% of transgenic plants were normal in morphology. Fertile transgenic maize plants carrying the transferred gene CBF were produced using the Agrobacterium EHA105/PpCBF1 and these plants were shown to be more salt tolerant. Transient expression of the GUS gene was confirmed in transgenic calli, shoots, leaves, roots and floral parts of transgenic R0 and R1 progenies using histochemical GUS assays. The presence of the introduced bar and CBF genes in the genomic DNA of the transformants was confirmed by the PCR amplification. Further, the stable expression of the CBF and bar transgenes in the maize genome of transgenic R1 progeny was confirmed by qRT-PCR. The transformation protocol developed using an A. tumefaciens standard binary vector system was an effective and reproducible method to transform Syrian maize with an anti-stress gene in which fertile salt-resistant transgenic plants were routinely produced. This approach has great potential for development of Syrian maize breeding programmes for abiotic stress resistance for application in many areas in Syrian maize production.

1 Abstract Currently there are almost 4000 described species of rose chafers (Scarabaeidae: Cetoniinae) but larvae of less than 200 species are described. In this work we studied larval morphology of 86 species belonging to 58 genera with focus on the tribe Goliathini. The larvae of 21 genera were studied for the first time. 77 morphological characters on head, mouth parts, legs and thoracal and abdominal segments were studied. This resulted into large matrix of morphological characters, which were further tested in separate phylogenetical analyses. A checklist of studied characters and their states and also large databasis of photographs of the characters are also amongst the results of the thesis. Five independent datasets were tested, three were based singly on morphology, one on molecular sequences and one on a combined dataset. The phylogenetic analysis was made by three methods, maximum parsimony, maximum likelihood and bayesian interference. Eleven different phylogenetic trees were obtained as a result, based on which we tried to reconstruct the relationships between the inner groups of Cetoniinae with focus on relationship of subtribes of tribe Goliathini. Monophyly of the tribe Goliathini as well as some other groups could not be confirmed, as well as the alleged basal position of the genus...

Diploma thesis "Interaction in multisystemic problems of bronchopulmonary dysplasia - effects of physiotherapy upon lung functions of immature infants" deals with relevant questions about care of immature newborns both during hospitalization at NICU and after discharge, inclusive of examination at Center for comprehensive care for risk newborns. Especially deals with possibilities of physiotherapy of newborns with bronchopulmonary dysplasia pathology and demonstrates the necessity of tight cooperation among physiotherapists, nurses and doctors during daily care of these children. Powered by TCPDF (www.tcpdf.org)

We describe here for the first time an efficient high yield production method for clinical grade recombinant human Oncofetal Antigen/immature laminin receptor protein (OFA/iLRP). We also demonstrate significant antitumor activity for this protein when administered in liposomal delivery form in a murine model of syngeneic fibrosarcoma. OFA/iLRP is a therapeutically very promising universal tumor antigen that is expressed in all mammalian solid tumors tested so far. We have cloned the human OFA/iLRP cDNA in a bacterial expression plasmid which incorporates a 6x HIS-tag. Large scale cultures of the plasmid transformed Escherichia coli were performed and the crude HIS-tagged OFA/iLRP was isolated as inclusion bodies and solubilized in guanidine chloride. The protein was then purified by successive passage through three column chromatography steps of immobilized metal affinity, anion exchange, and gel filtration. The resulting protein was 94% pure and practically devoid of endotoxin and host cell protein. The purified OFA/iLRP was tested in mice for safety and efficacy in tumor rejection with satisfactory results. This protein will be used for loading onto autologous dendritic cells in an FDA approved phase I/II human cancer vaccine trial in OFA/iLRP-positive breast cancer patients.

京都大学 / 0048 / 新制・課程博士 / 博士(理学) / 甲第20557号 / 理博第4315号 / 新制||理||1620(附属図書館) / 京都大学大学院理学研究科生物科学専攻 / (主査)教授 友永 雅己, 准教授 後藤 幸織, 教授 平井 啓久 / 学位規則第4条第1項該当 / Doctor of Science / Kyoto University / DGAM

In vivo studies of mechanical loading on bone have suggested that load magnitude is one of the parameters that play a vital role in bone adaptation. This study examined how longitudinal growth of immature rat metatarsals is affected by different load magnitudes. The main hypotheses were that the longitudinal growth of immature bone would decrease with increased compressive load magnitude, and that the longitudinal growth would be more decelerated the higher the load mag- nitude. The three middle metatarsal bones in the back paws of 19-20 days old Sprague-Dawley rat fetuses were extracted. Metatarsal bones were loaded with 0.05 N, 0.25 N, 1.25 N and 6.25 N. Loading rate and number of cycles were constant at 0.01 mm/sec and 10 cycles respectively. Length measurements occurred every 2-3 day. Concluded from the study was that a load magni- tude of 0.05 N resulted in an increased longitudinal growth, compared to unloaded bones. For the other load magnitudes the results were insufficient and inconsistent and therefore nothing could be suggested for them. The problem remained as before and further studies are needed.