Dissertation (PhD)--University of Stellenbosch, 2007. / ENGLISH ABSTRACT: This dissertation describes how the isolation and characterisation of ovine liver cytochrome b5 was accomplished by referring to the following goals achieved in this study: - The optimisation of the isolation and purification procedure for ovine liver microsomal cytochrome b5 in order to obtain sufficient material for aggregation and immunological studies. - The removal of the membrane binding domain of cytochrome b5 by means of tryptic digestion to establish the role of the carboxyl terminal in ovine cytochrome b5 aggregation. - The raising of antibodies against both the trypsin truncated and intact forms of cytochrome b5 to study the aggregation of the protein. - The investigation into the influence of purified cytochrome b5 on steroidogenesis in ovine adrenal microsomes. / AFRIKAANSE OPSOMMING: Die isolering en karakterisering van skaaplewersitochroom b5, soos beskryf in hierdie proefskrif, is uitgevoer deur die volgende doelwitte suksesvol af te handel: - Die optimalisering van die prosedure vir die suksesvolle isolering en suiwering van skaaplewersitochroom b5 ten einde genoegsame hoeveelhede van die suiwer proteïen te hê vir die bestudering van die aggregasie van die proteïen sowel as ‘n immunologiese studie. - Die verwydering van die membraanbindingsdomein van sitochroom b5 om die invloed van die karboksielterminaal op die aggregering van die proteïen te bestudeer. - Die gebruik van sowel die tripties gesnyde as die intakte vorms van sitochroom b5 om ‘n immuunrespons in hase op te wek vir die verkryging van sitochroom b5 spesifieke anti-liggame. - Die gebruik van die gesuiwerde proteïene om die invloed van sitochroom b5 op adrenale steroïdogenese te bestudeer.

Orientador: João Aristeu da Rosa / Banca: João Aristeu da Rosa / Banca: Eunice Aparecida Bianchi Galati / Banca:Maria Tercilia Vilela de azeredo Oliveira / Resumo: A doença de Chagas tem como agente etiológico o protozoário Trypanosoma cruzi, cujos vetores pertencem à subfamília Triatominae. Os triatomíneos distribuem-se distribuídos por toda região Neotropical e epidemiologicamente se destacam as espécies dos gêneros Panstrongylus, Rhodnius e Triatoma. O gênero Triatoma é o mais numeroso e foi subdividido em complexos específicos de acordo com semelhanças morfológicas e distribuição geográfica de suas espécies. As sete espécies estudadas podem ser encontradas na região Centro-Oeste do Brasil, das quais seis pertencem ao subcomplexo matogrossensis (T. baratai, T. costalimai, T. guazu, T. matogrossensis, T. vandae e T. williami). A outra espécie, T. sordida pertence ao subcomplexo sordida. O objetivo deste estudo foi determinar a posição filogenética das sete espécies ocorrentes na região Centro-Oeste, por meio de comparação das sequências dos fragmentos citocromo b e 16S do DNA mitocondrial, visto que até o momento não foi determinada a posição de T. baratai e T. vandae com base em dados moleculares. Os exemplares avaliados são oriundos de colônias mantidas junto ao Insetário de Triatominae da Faculdade de Ciências Farmacêuticas/UNESP - Araraquara. Após a extração do DNA genômico e da amplificação dos fragmentos 16S e Cytb, procedeu-se o sequenciamento do DNA em sqüenciador automático, modelo ABI 377. As sequências obtidas juntamente com outras sequências (do mesmo fragmento) já disponíveis no GenBank, foram alinhadas com auxílio do programa Clustal W do BioEdit e as inferências filogenéticas foram conduzidas utilizando-se análises de parcimônia e de distância com o programa MEGA 3.1. De acordo com os resultados encontrados, T. costalimai mostrou-se mais distante das demais espécies e foi posicionada como grupo externo. As outras espécies distribuíram-se em dois grupos:.. (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The protozoan Trypanosoma cruzi, the etiologic agent of Chagas' disease, is transmitted by vectors that belong to the subfamily Triatominae. The triatomines are distributed throughout the Neotropical region and species from Panstrongylus, Rhodnius and Triatoma genus stand out epidemiologically. The Triatoma genus is the largest and was divided in specific complexes according to morphological similarities and geographical distribution of its species. The seven species studied can be found in the Central West Region of Brazil, of which six belong to the matogrossensis subcomplex (T. baratai, T. costalimai, T. guazu, T. jurbergi, T. matogrossensis, T. vandae and T. williami). The other specie, T. sordida belong sordida subcomplex. The aim of this study was to determine the phylogenetic position of the seven species from Central West Region by comparing the 16S and Cytb fragments sequences of the mitochondrial DNA, since so far not been given the position of T. baratai and T. vandae based DNA sequences. The specimens evaluated came from colonies maintained at the Insectarium of Triatominae, Faculdade de Ciências Farmacêuticas / UNESP - Araraquara. After extraction of genomic DNA and amplification of the 16S and Cytb fragments, it was sequenced in an automatic DNA sequencer, model ABI 377. The sequences obtained and other sequences (of the same fragment) already available in GenBank were aligned using the Clustal W program of BioEdit and the phylogenetic inferences were conducted using the analysis of parsimony and distance with the MEGA 3.1 program. According to the results, T. costalimai was more distant from other species and was placed as an outside group. Other species are distributed in two groups, the first comprising the species T. vandae very close to T. matogrossensis and external of these, T. sordida. The second group includes T. guazu strongly related to T. williami... (Complete abstract click electronic access below) / Mestre

Bats of the genus Scotophilus are distributed throughout sub-Saharan Africa, parts of southern and Southeast Asia, a majority of the Indomalayan Islands, Reunion Island, and Madagascar. The genus is composed of 14 recognized species with seven distributed throughout sub-Saharan Africa including: (S. dinganii (A. Smith, 1833), S. leucogaster (Cretzschmar, 1830), S. nigritellus de Winton, 1899, S. nigrita (Schreber, 1774), S. nucella Robbins, 1983, S. nux Thomas, 1904, and S. viridis (Peters, 1852). The remaining species include four from southern and southeast Asia (S. celebensis Sody, 1928; S. collinus Sody 1936; S. heathi (Horsfield, 1831); S. kuhlii Leach, 1821), two on Madagascar (S. sp. nov. Goodman et al., in press; and S. robustus Milne-Edwards, 1881), and one endemic to Reunion Island (S. borbonicus (E. Geoffroy, 1803). The systematics and taxonomy of this genus have been controversial and continue to be confusing. The genus is plagued with problems in species definition and the systematic relationships among members of the genus are poorly understood. The major goal of this study was to use a molecular phylogenetic approach to clarify some of the controversy and confusion surrounding the members of this genus. Nucleotide differences from mtDNA and the Y chromosome were used to examine phylogenetic patterns within Scotophilus. Based on these data two new species of Scotophilus were identified. Phylogenetically, African Scotophilus were found to comprise a monophyletic group with S. nux as the most basal African taxon. Overall, the Asian S. kuhlii was the most basal taxon. A distant relationship was identified between S. kuhlii and S. heathi, the other Asian species examined. The multiple origins of Malagasy Scotophilus are apparent as the two Malagasy taxa in the study do not share a sister-group relationship. The large bodied S. nigrita is closely related to S. dinganii and the S. dinganii-like species all share a close relationship. S. nigrita has a S. dinganii-like mtDNA haplotype and a very distinct zfy haplotype, suggesting a possible hybridization event with a S. dinganii-like ancestor.

Family Pinguipedidae belong to the class Actinopterygii, subclass Neopterygii, order Perciformes, suborder Trachinoidei. Currently the interrelationships of the genera within this family and among the families in the Trachinoidei remain unequivocal. Also, whether the Cheimarrichthys should be included in the family Pinguipedidae has also been a controversial issue. This study aimed to reconstruct phylogenetic hypotheses in order to resolve these questions. Species of the Parapercis and Kochichtys in the family Pinguipedidae occur Taiwan. This study used osteological characters, 16S rRNA and Cyt b sequences to conduct phylogenetic analysis such that hypotheses can be proposed. The results revealed the monophyly of Parapercis, a taxonomic view consistent to the prevailary classification. Summarizing all the results, the 17 Parapercis species analysed can be divided into 4 groups. They are (1) Parapercis aurantiaca¡BP. decemfasciata¡BP. mimaseana¡BP. multifasciata¡BP. muronis¡Btwo morphotypes of P. sexfasciata¡F(2) P. cephalopunctata¡BP. clathrata¡BP. hexophthalma¡BP. kamoharai¡BP. tetracantha¡BP. xanthozona¡F(3) P. cylindrica and P. snyderi¡F(4) P. maculata¡BP. ommatura and P. somaliensis. Two color morphotypes have been shown for Parapercis sexfasciata. Data of the present study revealed that the ¡§ autapomorphic¡¨ osteological character known only in Kochichtys also occurred in three Parapercis species. This result supports a close relationship between these species. However, it also challenges the validity of the generic status of Kochichtys. About the dabate of the phylogenetic position of Cheimarrichthys, it should be put into its own family, Cheimarichthyide, rather than placed in the Pinguipedidae. The hypothesis for the sister group of Pinguipedidae to the Cheimarichthyide is not supported by all the data in this study completely. Morphological and molecular evidences are incongruence for closest phylogenetic relationship. Similar results were also obtained when the molecular sequences were analysed using different methods. More data analyses are needed for complete and reliable results. The present study suggests that the Trachinoidei is not a monophyletic group.

Bats of the genus Scotophilus are distributed throughout sub-Saharan Africa, parts of southern and Southeast Asia, a majority of the Indomalayan Islands, Reunion Island, and Madagascar. The genus is composed of 14 recognized species with seven distributed throughout sub-Saharan Africa including: (S. dinganii (A. Smith, 1833), S. leucogaster (Cretzschmar, 1830), S. nigritellus de Winton, 1899, S. nigrita (Schreber, 1774), S. nucella Robbins, 1983, S. nux Thomas, 1904, and S. viridis (Peters, 1852). The remaining species include four from southern and southeast Asia (S. celebensis Sody, 1928; S. collinus Sody 1936; S. heathi (Horsfield, 1831); S. kuhlii Leach, 1821), two on Madagascar (S. sp. nov. Goodman et al., in press; and S. robustus Milne-Edwards, 1881), and one endemic to Reunion Island (S. borbonicus (E. Geoffroy, 1803). The systematics and taxonomy of this genus have been controversial and continue to be confusing. The genus is plagued with problems in species definition and the systematic relationships among members of the genus are poorly understood. The major goal of this study was to use a molecular phylogenetic approach to clarify some of the controversy and confusion surrounding the members of this genus. Nucleotide differences from mtDNA and the Y chromosome were used to examine phylogenetic patterns within Scotophilus. Based on these data two new species of Scotophilus were identified. Phylogenetically, African Scotophilus were found to comprise a monophyletic group with S. nux as the most basal African taxon. Overall, the Asian S. kuhlii was the most basal taxon. A distant relationship was identified between S. kuhlii and S. heathi, the other Asian species examined. The multiple origins of Malagasy Scotophilus are apparent as the two Malagasy taxa in the study do not share a sister-group relationship. The large bodied S. nigrita is closely related to S. dinganii and the S. dinganii-like species all share a close relationship. S. nigrita has a S. dinganii-like mtDNA haplotype and a very distinct zfy haplotype, suggesting a possible hybridization event with a S. dinganii-like ancestor.

Thesis (M.S.)--Brigham Young University. Dept. of Biology, 2007. / Includes bibliographical references.

Thesis (Ph. D.)--University of California, San Diego, 1997. / Vita. Includes bibliographical references.

Made available in DSpace on 2014-06-11T19:27:55Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-01-27Bitstream added on 2014-06-13T20:36:33Z : No. of bitstreams: 1 gardim_s_me_arafcf.pdf: 1853681 bytes, checksum: 3bd67f0a8ef8c760824f1c5ffaea5e3e (MD5) / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / Universidade Estadual Paulista (UNESP) / A doença de Chagas tem como agente etiológico o protozoário Trypanosoma cruzi, cujos vetores pertencem à subfamília Triatominae. Os triatomíneos distribuem-se distribuídos por toda região Neotropical e epidemiologicamente se destacam as espécies dos gêneros Panstrongylus, Rhodnius e Triatoma. O gênero Triatoma é o mais numeroso e foi subdividido em complexos específicos de acordo com semelhanças morfológicas e distribuição geográfica de suas espécies. As sete espécies estudadas podem ser encontradas na região Centro-Oeste do Brasil, das quais seis pertencem ao subcomplexo matogrossensis (T. baratai, T. costalimai, T. guazu, T. matogrossensis, T. vandae e T. williami). A outra espécie, T. sordida pertence ao subcomplexo sordida. O objetivo deste estudo foi determinar a posição filogenética das sete espécies ocorrentes na região Centro-Oeste, por meio de comparação das sequências dos fragmentos citocromo b e 16S do DNA mitocondrial, visto que até o momento não foi determinada a posição de T. baratai e T. vandae com base em dados moleculares. Os exemplares avaliados são oriundos de colônias mantidas junto ao Insetário de Triatominae da Faculdade de Ciências Farmacêuticas/UNESP - Araraquara. Após a extração do DNA genômico e da amplificação dos fragmentos 16S e Cytb, procedeu-se o sequenciamento do DNA em sqüenciador automático, modelo ABI 377. As sequências obtidas juntamente com outras sequências (do mesmo fragmento) já disponíveis no GenBank, foram alinhadas com auxílio do programa Clustal W do BioEdit e as inferências filogenéticas foram conduzidas utilizando-se análises de parcimônia e de distância com o programa MEGA 3.1. De acordo com os resultados encontrados, T. costalimai mostrou-se mais distante das demais espécies e foi posicionada como grupo externo. As outras espécies distribuíram-se em dois grupos:.. / The protozoan Trypanosoma cruzi, the etiologic agent of Chagas' disease, is transmitted by vectors that belong to the subfamily Triatominae. The triatomines are distributed throughout the Neotropical region and species from Panstrongylus, Rhodnius and Triatoma genus stand out epidemiologically. The Triatoma genus is the largest and was divided in specific complexes according to morphological similarities and geographical distribution of its species. The seven species studied can be found in the Central West Region of Brazil, of which six belong to the matogrossensis subcomplex (T. baratai, T. costalimai, T. guazu, T. jurbergi, T. matogrossensis, T. vandae and T. williami). The other specie, T. sordida belong sordida subcomplex. The aim of this study was to determine the phylogenetic position of the seven species from Central West Region by comparing the 16S and Cytb fragments sequences of the mitochondrial DNA, since so far not been given the position of T. baratai and T. vandae based DNA sequences. The specimens evaluated came from colonies maintained at the Insectarium of Triatominae, Faculdade de Ciências Farmacêuticas / UNESP - Araraquara. After extraction of genomic DNA and amplification of the 16S and Cytb fragments, it was sequenced in an automatic DNA sequencer, model ABI 377. The sequences obtained and other sequences (of the same fragment) already available in GenBank were aligned using the Clustal W program of BioEdit and the phylogenetic inferences were conducted using the analysis of parsimony and distance with the MEGA 3.1 program. According to the results, T. costalimai was more distant from other species and was placed as an outside group. Other species are distributed in two groups, the first comprising the species T. vandae very close to T. matogrossensis and external of these, T. sordida. The second group includes T. guazu strongly related to T. williami... (Complete abstract click electronic access below)

Made available in DSpace on 2014-06-11T19:27:55Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-02-12Bitstream added on 2014-06-13T18:56:49Z : No. of bitstreams: 1 rocha_cs_me_arafcf.pdf: 697628 bytes, checksum: 703b47e347bee1b853ea18538df8a3e2 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Atualmente são admitidas 143 espécies da subfamília Triatominae que estão agrupadas em 18 gêneros e seis tribos. Essa classificação baseia-se principalmente em características morfológicas. Dentre essas espécies temos Triatoma rubrovaria, que pode ser encontrado no Estado do Rio Grande do Sul (Brasil), Uruguai e em algumas regiões da Argentina. Algumas espécies de Triatominae apresentam coloração e características morfológicas semelhantes, o que dificulta a identificação dos exemplares. Ferramentas como a morfometria, citogenética, retrocruzamentos e técnicas de biologia molecular são metodologias importantes para a identificação dessa subfamília. Estudos morfométricos prévios realizados com três populações de T. rubrovaria mantidas no Insetário de Triatomíneos do Laboratório de Parasitologia da Faculdade de Ciências Farmacêuticas revelaram a existência de diferenças morfométricas estatisticamente significativas entre a colônia de Caçapava do Sul (CS) e as duas colônias de Quaraí (QI e QII). Diferenças no padrão de cor do pronoto entre as três populações também foram observadas. A fim de avaliar a variabilidade genética dessas populações, analisaram-se as seqüências nucleotídicas do Citocromo B (Cyt B) e 16S, pertencentes ao DNA mitocondrial e o 28S, ao DNA nuclear. Dentre os marcadores utilizados, o Cyt B apresentou maior variabilidade, seguido do 28S e 16S, respectivamente. Devido ao seu maior polimorfismo o Cyt B mostrou-se um marcador eficaz para estudos de variabilidades... / Currently 143 species of the subfamily Triatominae, grouped into 16 genera and 6 tribes are recognized. This classification is mainly based on morphological characteristics. These species include Triatoma rubrovaria, which can be found in the state of Rio Grande do Sul (Brazil), Uruguay and some regions of Argentina. Some species of Triatominae have similar color and morphological characteristics, which complicates the identification of specimens. Morphometry, cytogenetics, backcrossing and molecular biology techniques are important methods for the identification of this subfamily. Previous morphometric studies carried out on three populations of T. rubrovaria maintained in the Insetário de Triatomíneos do Laboratório de Parasitologia of the Faculdade de Ciências Farmacêuticas have revealed the existence of statistically significant morphometric differences among the colony from Caçapava do Sul (CS) and the two colonies from Quaraí (QI and QII). Differences were also observed in the color patterns of the pronotum among the three populations. The sequences of Cytochrome B (Cyt B) and 16S, belonging to the mitochondrial DNA and 28S, the nuclear DNA, were analyzed to assess the genetic variety of these populations . The Cyt B showed the greatest variability, followed by 28S and 16S. It was concluded that Cyt B is the best marker for effective studies of population variability by virtue of its grater polymorphis ...(Complete abstract click electronic access below)

Orientador: João Aristeu da Rosa / Banca: João Aristeu da Rosa / Banca: Regina Maria Barretto Cicarelli / Banca: Eunice Aparecida Bianchi Galati / Resumo: Atualmente são admitidas 143 espécies da subfamília Triatominae que estão agrupadas em 18 gêneros e seis tribos. Essa classificação baseia-se principalmente em características morfológicas. Dentre essas espécies temos Triatoma rubrovaria, que pode ser encontrado no Estado do Rio Grande do Sul (Brasil), Uruguai e em algumas regiões da Argentina. Algumas espécies de Triatominae apresentam coloração e características morfológicas semelhantes, o que dificulta a identificação dos exemplares. Ferramentas como a morfometria, citogenética, retrocruzamentos e técnicas de biologia molecular são metodologias importantes para a identificação dessa subfamília. Estudos morfométricos prévios realizados com três populações de T. rubrovaria mantidas no Insetário de Triatomíneos do Laboratório de Parasitologia da Faculdade de Ciências Farmacêuticas revelaram a existência de diferenças morfométricas estatisticamente significativas entre a colônia de Caçapava do Sul (CS) e as duas colônias de Quaraí (QI e QII). Diferenças no padrão de cor do pronoto entre as três populações também foram observadas. A fim de avaliar a variabilidade genética dessas populações, analisaram-se as seqüências nucleotídicas do Citocromo B (Cyt B) e 16S, pertencentes ao DNA mitocondrial e o 28S, ao DNA nuclear. Dentre os marcadores utilizados, o Cyt B apresentou maior variabilidade, seguido do 28S e 16S, respectivamente. Devido ao seu maior polimorfismo o Cyt B mostrou-se um marcador eficaz para estudos de variabilidades ...(Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Currently 143 species of the subfamily Triatominae, grouped into 16 genera and 6 tribes are recognized. This classification is mainly based on morphological characteristics. These species include Triatoma rubrovaria, which can be found in the state of Rio Grande do Sul (Brazil), Uruguay and some regions of Argentina. Some species of Triatominae have similar color and morphological characteristics, which complicates the identification of specimens. Morphometry, cytogenetics, backcrossing and molecular biology techniques are important methods for the identification of this subfamily. Previous morphometric studies carried out on three populations of T. rubrovaria maintained in the Insetário de Triatomíneos do Laboratório de Parasitologia of the Faculdade de Ciências Farmacêuticas have revealed the existence of statistically significant morphometric differences among the colony from Caçapava do Sul (CS) and the two colonies from Quaraí (QI and QII). Differences were also observed in the color patterns of the pronotum among the three populations. The sequences of Cytochrome B (Cyt B) and 16S, belonging to the mitochondrial DNA and 28S, the nuclear DNA, were analyzed to assess the genetic variety of these populations . The Cyt B showed the greatest variability, followed by 28S and 16S. It was concluded that Cyt B is the best marker for effective studies of population variability by virtue of its grater polymorphis ...(Complete abstract click electronic access below) / Mestre