Aplicações podem apresentar laços perfeitamente aninhados que demandam um alto poder de processamento. Diversas aplicações científicas contêm laços aninhados em suas estruturas. Tais laços podem processar computações heterogêneas. Uma solução para reduzir o tempo de execução desta classe de aplicações é a paralelização destes laços. A heterogeneidade dos tempos de execução de computações presentes nas iterações de laços perfeitamente aninhados demanda uma paralelização adequada visando uma distribuição de carga homogênea entre os recursos computacionais para reduzir a ociosidade de tais recursos. Esta heterogeneidade implica em um número ideal de recursos computacionais a partir do qual, o seu aumento não impactaria no ganho de desempenho, uma vez que, o tempo mínimo possível é o tempo de execução da tarefa que consome o maior tempo de processamento. Neste trabalho é proposta uma metodologia e ferramentas para paralelização de laços perfeitamente aninhados sem dependência de dados e com processamento heterogêneo em sistemas paralelos e distribuídos. A implementação da metodologia proposta em aplicações melhora o desempenho da execução e reduz a ociosidade dos recursos de processamento. Na metodologia proposta, alguns procedimentos são apoiados por ferramentas desenvolvidas para auxiliá-los. O sistema de processamento poderá ser: um computador Multicore, um Cluster real ou virtual alocado na nuvem. Resultados experimentais são apresentados neste trabalho. Tais resultados mostram a viabilidade e eficiência da metodologia proposta. / Applications may have nested perfectly loops that require a high processing power. Various scientific applications contain nested loops in their structures. Such loops can process heterogeneous computations. A solution to reduce the execution time of this class of applications is the parallelization of these loops. The heterogeneity of the execution times of computations present in the iterations of nested perfectly loops demands an adequate parallelization aiming at a homogeneous load distribution among the computational resources to reduce the idleness of such resources. This heterogeneity implies an ideal number of computational resources which, its increase would not impact the performance gain, since the minimum possible time is the execution time of the task that consumes the longest processing time. In this work is proposed a methodology and tools for parallelization of loops perfectly nested with heterogeneous processing in parallel and distributed systems. The implementation of proposed methodology in application improves execution performance and reduce idles of the processing resources. In the methodology proposed, some procedures are supported by tools developed to assist them. The processing system can be: a computer multicore, a cluster real or virtual allocated in cloud. Experimental results are presented in this work. These results show the feasibility and efficiency of the proposed methodology.

Este estudo teve como objetivo detectar a presença do vírus da raiva em diferentes órgãos de morcegos do gênero Artibeus empregando as técnicas moleculares como RT-PCR, hnRT-PCR e Real Time RT-PCR. De aproximadamente 4000 espécimes de morcegos recebidas no Instituto Pasteur para o diagnóstico da raiva, foram selecionados 30 morcegos do gênero Artibeus, com resultados positivos para raiva pelas técnicas tradicionais de IFD e inoculação em células N2A utilizando suspensões feitas a partir do SNC. Para as técnicas moleculares, foram retirados glândulas salivares, bexigas urinárias, rins, pulmões e conteúdos fecais e ainda foram lavadas as calotas cranianas dos espécimes. Os órgãos e conteúdos fecais foram diluídos a 1:10 (P/V) e as bexigas urinárias a 1:20 (P/V). As suspensões foram inoculadas em células N2A para o isolamento viral. Foi realizada a extração do RNA total usando o TRIzol®, foram realizadas a transcrição reversa seguida da PCR e hnRT-PCR com utilização de primers específicos para o gene codificante da proteína N. A partir do produto da transcrição reversa foi realizada a técnica de Real Time RT-PCR, utilizando primers e sonda específicos para variante antigênica 3. Das 30 suspensões de lavado cerebral, 28 (93,33%) resultaram positivos, na inoculação em cultura de células, seguido de glândulas salivares (36,67%), bexigas (16,67%) e conteúdos fecais (3,33%). Os resultados encontrados da sensibilidade nas técnicas de RT-PCR, hnRT-PCR e Real Time RT-PCR foram 56,25%, 82,57% e 82,19% quando avaliadas as 180 amostras analisadas. A comparação das técnicas de hnRT-PCR e Real Time RT-PCR feita pelo teste exato de Fisher quanto a proporção de positivos detectados mostrou que para o lavado cerebral, órgãos e conteúdos fecais a proporção foi igual (P>0,05). Em relação à positividade os resultados encontrados nas técnicas de hnRT-PCR e Real Time RT-PCR foram 100% em lavado cerebral; 90% e 93,33% em glândulas salivares; 83,33% e 90% em bexigas; 80% e 93,33% em rins; 76,67% e 50% em pulmões e 43,33% em ambas as técnicas em conteúdos fecais. Esses resultados sugerem que tanto as técnicas de hnRT-PCR como Real Time RT-PCR podem ser utilizadas como métodos complementares para o diagnóstico da raiva e são sensíveis o bastante para o uso em estudos de patogênese. A técnica de Real Time RT-PCR realizada neste estudo se mostrou eficiente em detectar o RABV em diferentes órgãos e tecidos extraneurais com a vantagem de ser uma técnica mais rápida e sensível. / This study was aimed to detect the presence of rabies virus in different organs of the genus Artibeus bats using molecular techniques such as RT-PCR, hnRT-PCR, and the Real Time RT-PCR. From about 4,000 specimens of bats received for rabies diagnosis at the Pasteur Institute, 30 bats of the genus Artibeus were then selected. The selected bats presented positive results by the traditional DFA and N2A-cells inoculation test using brain tissue suspensions. Samples of salivary glands, urinary bladders, kidneys, lungs, and fecal contents and washings of the skulls were collected for the molecular techniques testing. The organs and the fecal contents were diluted at 1:10 (w/v) and the urinary bladder, at 1:20 (w/v) and these suspensions were inoculated into N2A cells for viral isolation. The extraction of the total RNA was performed by using TRIzol® and followed by the reverse transcription and the PCR and the hnRT-PCR were performed by using specific primers for the gene encoding the protein N. The product obtained by the reverse transcription technique was submitted to the Real Time RT-PCR technique, using primers and probe specific for antigenic variant 3 of the rabies virus. Of the 30 suspensions of the brain washings, 28 (93.33%) were positive in N2A cell culture inoculation, followed by the suspensions of the salivary glands (36.67%), bladders (16.67%) and fecal contents (3.33%). For the 180 samples evaluated, the results of sensitivity found for the RT-PCR, hnRT-PCR and Real Time RT-PCR techniques were 56.25%, 82.57%, and 82.19%, respectively. A comparison of hnRT-PCR and Real Time RT-PCR techniques performed by Fisher\'s exact test showed that the proportion of positives detected by the brain washings, organs and of the fecal content was non-significant (P> 0.05). Regarding the results found in hnRT-PCR and Real Time RT-PCR techniques, 100% positives were in brain washing, 90% and 93.33% in salivary glands, 83.33% and 90% in bladders, 80% and 93.33% in kidneys, 76.67% and 50% in lungs and 43.33% for both techniques on fecal contents. These results suggest that both hnRT-PCR and Real-Time PCR techniques can be used as complementary methods for the diagnosis of rabies and are sensitive enough for use in pathogenesis studies. The Real Time RT-PCR technique performed in this study proved to be faster and more sensitive and effective in detecting RABV in different organs and extra neural tissues of bats.

Mestrado em Ciências Forenses / Master Degree Course in Forensic Sciences / A possibilidade de analisar amostras com quantidades exíguas de material genético (amostras Low Copy Number ou LCN), em que estão presentes apenas algumas células, tem alterado a forma de encarar a cena do crime. Alguns vestígios que até agora não eram considerados como susceptíveis de proporcionarem resultados, podem actualmente ser analisados com sucesso. As impressões digitais são um bom exemplo. Estes vestígios apresentam um baixo número de células, permitindo apenas recuperar quantidades de DNA inferiores a 100 pg. Assim, para a análise do DNA nuclear, é necessário implementar sistemas muito sensíveis que consistem, habitualmente, no aumento do número de ciclos da PCR. Contudo, alguns artefactos são produzidos, tornando difícil a interpretação dos electroforectogramas. Neste trabalho pretendeu-se comparar a aplicação do estudo de STR autossómicos, Y-STR e miniSTR na análise genética de impressões digitais, partindo do conceito do aumento do número de ciclos como estratégia para se obter maior sensibilidade. Procedeu-se também à amplificação total do genoma e nested-PCR, como métodos alternativos ao aumento do número de ciclos. Adicionalemente, neste estudo tentou-se perceber a influência dos principais métodos reveladores de impressões digitais (cianoacrilato, pó magnético e pó branco) na análise do DNA. Os resultados mostram que o aumento do número de ciclos é a melhor opção como método para aumentar a sensibilidade. Constata-se também que o DNA extraído de impressões digitais encontra-se parcialmente degradado, obtendo-se diferenças significativas entre loci com fragmentos de amplificação menores e maiores do que 200 pb. Dos diferentes marcadores caracterizados verifica-se que, em termos de percentagem de alelos detectados, os miniSTR proporcionam os melhores resultados. Por outro lado, os Y-STR parecem altamente sensíveis à degradação ou presença de inibidores, pelo que são menos robustos para este tipo de análises. Verifica-se também que os perfis LCN são drasticamente afectados por artefactos, principalmente os derivados de variação estocástica, como o allele dropout e o desequilíbrio heterozigótico. A determinação de perfis de consenso permite reduzir alguns destes artefactos. Dos métodos de revelação estudados, o cianoacrilato é o que apresenta menor influência na análise e, pelo contrário, o pó branco provoca os resultados mais negativos. / The possibility to perform low copy number DNA typing, when just a few cells are available, as changed the way how crime scene investigations is faced. Nowadays it is possible to successfully type some evidence that couldn t be considered until now. Fingerprints are a good example of those. Since that just a few cells are present in this evidence (enabling recovery of low quantities of DNA, fewer than 100pg) just very sensitive systems can detect nuclear DNA. The most used method is definitely increasing the number of PCR cycles. However, increased occurrence of stutters and artifacts that reduced the quality of the DNA profile is normally observed. The present work aimed to compare the application of autosomic STR, Y-STR and miniSTR markers, based on the concept of increased number of PCR cycles as a strategy to achieve more sensitivity. Some other methods, such as whole genome amplification and nested-PCR, were also evaluated as an alternative way to reach the desired sensitivity. Another goal was to determine the influence of several reagents for developing latent fingerprints (cyanoacrylate fuming, magnetic powder and white powder) in DNA typing. The results shows that increasing the number of PCR cycles still is the best way to attain the required sensitivity. Moreover we could realize that DNA was partially degraded, once there were observed significant differences between loci larger and smaller than 200bp. Among all markers miniSTR showed to perform the best results in terms of detected alleles percentage. On the other hand, Y-STR seemed to be highly affected in the presence of degraded DNA and PCR inhibitors, which makes them less robust for these analyses. LCN profiles are significantly affected by artifacts, like allele dropout and heterozygous imbalance, derived from stochastic fluctuation. Reporting consensus profiles reduces artifact inherent errors. Finally, cyanoacrylate proved to have a minimum negative effect on DNA profiling, while white powder was the worst reagent.

Made available in DSpace on 2015-03-26T13:51:52Z (GMT). No. of bitstreams: 1 texto completo.pdf: 577759 bytes, checksum: 8b7270200348b4cbbdc4f40d5e821098 (MD5) Previous issue date: 2010-08-19 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / This work aimed to study the diversity of endophytic bacteria associated to Coffea arabica L. fruit during their development. Seven monthly collections were conducted, following coffee fruit development in homogeneous stand of Catuai Vermelho coffee cultivar plantation. Each sample was processed according to protocol aiming to obtain total DNA from the endophytic bacterial community, associated to each stage of fruit development. The strategy of rDNA16S amplification through the Nested-PCR technique and DGGE discrimination were applied. The total population of endophytic bacteria and the phylos Actinobacteria, Firmicutes and Proteobacteria (alpha, beta and gamma classes) were investigated. The analysis of each gel, representing the evolution of a population along time, was carried out using the program Bionumerics®. The existence of endophytic bacteria associated to coffee fruit since the first month of their development was confirmed, by using the universal primer for Eubacteria. Based on the similarity between the distribution pattern of the OTUs on the rays representing the two final months of fruit development, it was inferred that the populations tend to stabilize. Less similarity between the endophytic populations, present in the different phases of fruit development, was confirmed in the phylo Firmicutes, while the beta-Proteobacteria displayed greater similarity among the different phases of fruit development. Amplification of the rDNA by Nested PCR and DGGE discrimination were found to be adequate to distinguish the alteration dynamics among the phylo populations and the bacterial classes studied. / Neste trabalho foi estudada a diversidade de bactérias endofíticas associadas a frutos de Coffea arabica L. durante o seu desenvolvimento. Foram realizadas sete coletas mensais, acompanhando o desenvolvimento dos frutos em cafeeiros de um talhão homogêneo de lavoura do cultivar Catuaí Vermelho. Cada amostra foi processada seguindo protocolo para obter o DNA total da comunidade bacteriana endofítica, associada a cada estádio de desenvolvimento do fruto. Foi empregada a estratégia de amplificação do rDNA16S pela técnica de Nested-PCR e discriminação em DGGE. A população total de bactérias endofíticas e os filos Actinobacteria, Firmicutes e Proteobacteria (classes alfa, beta e gama) foram investigados. A análise de cada gel, representando a evolução de uma população ao longo do tempo, foi realizada empregando o programa Bionumerics®. Demonstrou-se a existência de populações de bactérias endofíticas associadas aos frutos de café desde o primeiro mês de seu desenvolvimento, com emprego do primer universal para Eubacteria. Pela maior similaridade entre o padrão de distribuição das UTOs nas raias que representam os dois meses finais de desenvolvimento dos frutos inferiu-se que as populações tendem para a estabilização. A menor similaridade entre as populações endofíticas, presentes nas diferentes fases de desenvolvimento dos frutos, foi constatada no filo Firmicutes, enquanto as beta- Proteobacteria exibiram maior similaridade entre as diferentes fases de desenvolvimento dos frutos. A amplificação do rDNA por Nested PCR e discriminação em DGGE mostrou-se adequada para a distinção da dinâmica de alterações nas populações entre os filos e as classes de bactérias estudadas.

Este estudo teve como objetivo detectar a presença do vírus da raiva em diferentes órgãos de morcegos do gênero Artibeus empregando as técnicas moleculares como RT-PCR, hnRT-PCR e Real Time RT-PCR. De aproximadamente 4000 espécimes de morcegos recebidas no Instituto Pasteur para o diagnóstico da raiva, foram selecionados 30 morcegos do gênero Artibeus, com resultados positivos para raiva pelas técnicas tradicionais de IFD e inoculação em células N2A utilizando suspensões feitas a partir do SNC. Para as técnicas moleculares, foram retirados glândulas salivares, bexigas urinárias, rins, pulmões e conteúdos fecais e ainda foram lavadas as calotas cranianas dos espécimes. Os órgãos e conteúdos fecais foram diluídos a 1:10 (P/V) e as bexigas urinárias a 1:20 (P/V). As suspensões foram inoculadas em células N2A para o isolamento viral. Foi realizada a extração do RNA total usando o TRIzol®, foram realizadas a transcrição reversa seguida da PCR e hnRT-PCR com utilização de primers específicos para o gene codificante da proteína N. A partir do produto da transcrição reversa foi realizada a técnica de Real Time RT-PCR, utilizando primers e sonda específicos para variante antigênica 3. Das 30 suspensões de lavado cerebral, 28 (93,33%) resultaram positivos, na inoculação em cultura de células, seguido de glândulas salivares (36,67%), bexigas (16,67%) e conteúdos fecais (3,33%). Os resultados encontrados da sensibilidade nas técnicas de RT-PCR, hnRT-PCR e Real Time RT-PCR foram 56,25%, 82,57% e 82,19% quando avaliadas as 180 amostras analisadas. A comparação das técnicas de hnRT-PCR e Real Time RT-PCR feita pelo teste exato de Fisher quanto a proporção de positivos detectados mostrou que para o lavado cerebral, órgãos e conteúdos fecais a proporção foi igual (P>0,05). Em relação à positividade os resultados encontrados nas técnicas de hnRT-PCR e Real Time RT-PCR foram 100% em lavado cerebral; 90% e 93,33% em glândulas salivares; 83,33% e 90% em bexigas; 80% e 93,33% em rins; 76,67% e 50% em pulmões e 43,33% em ambas as técnicas em conteúdos fecais. Esses resultados sugerem que tanto as técnicas de hnRT-PCR como Real Time RT-PCR podem ser utilizadas como métodos complementares para o diagnóstico da raiva e são sensíveis o bastante para o uso em estudos de patogênese. A técnica de Real Time RT-PCR realizada neste estudo se mostrou eficiente em detectar o RABV em diferentes órgãos e tecidos extraneurais com a vantagem de ser uma técnica mais rápida e sensível. / This study was aimed to detect the presence of rabies virus in different organs of the genus Artibeus bats using molecular techniques such as RT-PCR, hnRT-PCR, and the Real Time RT-PCR. From about 4,000 specimens of bats received for rabies diagnosis at the Pasteur Institute, 30 bats of the genus Artibeus were then selected. The selected bats presented positive results by the traditional DFA and N2A-cells inoculation test using brain tissue suspensions. Samples of salivary glands, urinary bladders, kidneys, lungs, and fecal contents and washings of the skulls were collected for the molecular techniques testing. The organs and the fecal contents were diluted at 1:10 (w/v) and the urinary bladder, at 1:20 (w/v) and these suspensions were inoculated into N2A cells for viral isolation. The extraction of the total RNA was performed by using TRIzol® and followed by the reverse transcription and the PCR and the hnRT-PCR were performed by using specific primers for the gene encoding the protein N. The product obtained by the reverse transcription technique was submitted to the Real Time RT-PCR technique, using primers and probe specific for antigenic variant 3 of the rabies virus. Of the 30 suspensions of the brain washings, 28 (93.33%) were positive in N2A cell culture inoculation, followed by the suspensions of the salivary glands (36.67%), bladders (16.67%) and fecal contents (3.33%). For the 180 samples evaluated, the results of sensitivity found for the RT-PCR, hnRT-PCR and Real Time RT-PCR techniques were 56.25%, 82.57%, and 82.19%, respectively. A comparison of hnRT-PCR and Real Time RT-PCR techniques performed by Fisher\'s exact test showed that the proportion of positives detected by the brain washings, organs and of the fecal content was non-significant (P> 0.05). Regarding the results found in hnRT-PCR and Real Time RT-PCR techniques, 100% positives were in brain washing, 90% and 93.33% in salivary glands, 83.33% and 90% in bladders, 80% and 93.33% in kidneys, 76.67% and 50% in lungs and 43.33% for both techniques on fecal contents. These results suggest that both hnRT-PCR and Real-Time PCR techniques can be used as complementary methods for the diagnosis of rabies and are sensitive enough for use in pathogenesis studies. The Real Time RT-PCR technique performed in this study proved to be faster and more sensitive and effective in detecting RABV in different organs and extra neural tissues of bats.

Submitted by isabela.moljf@hotmail.com (isabela.moljf@hotmail.com) on 2017-05-10T14:43:51Z No. of bitstreams: 1 luisadeoliveira.pdf: 1689764 bytes, checksum: 7af6e626808d1ab5d4fc1de9be514aaa (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2017-05-17T15:08:15Z (GMT) No. of bitstreams: 1 luisadeoliveira.pdf: 1689764 bytes, checksum: 7af6e626808d1ab5d4fc1de9be514aaa (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2017-05-17T15:08:48Z (GMT) No. of bitstreams: 1 luisadeoliveira.pdf: 1689764 bytes, checksum: 7af6e626808d1ab5d4fc1de9be514aaa (MD5) / Made available in DSpace on 2017-05-17T15:08:48Z (GMT). No. of bitstreams: 1 luisadeoliveira.pdf: 1689764 bytes, checksum: 7af6e626808d1ab5d4fc1de9be514aaa (MD5) Previous issue date: 2014-10-10 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Hemosporídeos de aves silvestres são considerados importantes agentes etiológicos e podem causar danos às populações dos hospedeiros. O presente estudo teve como objetivos: determinar a prevalência e a parasitemia para Plasmodium spp. e Haemoproteus spp. de Passeriformes da Mata Atlântica de Minas Gerais, por meio de dois métodos de diagnóstico, microscopia e nested PCR e caracterizar morfologicamente a espécie Plasmodium (Haemamoeba) lutzi Lucena, 1939. Amostras de sangue foram obtidas por punção da veia braquial para preparação de esfregaços que foram fixados em metanol e corados em Giemsa. Foram amostradas 237 aves pertencentes a 18 famílias e 56 espécies da ordem Passeriformes provenientes do IBAMA e capturadas no Jardim Botânico da Universidade Federal de Juiz de Fora (JB-UFJF). A prevalência para Plasmodium spp. foi 63,63% em aves do IBAMA e 26,25% em aves do JB-UFJF, apenas três indivíduos encontraram-se parasitados por Haemoproteus spp., sendo dois (2,5%) provenientes do IBAMA e um (0,62%) do JB-UFJF. A parasitemia média encontrada para Plasmodium spp./Haemoproteus spp. foi de 0,08 (± 0,08) no IBAMA e 0,05 (± 0,04) no JB-UFJF. O maior grau de parasitemia média foi registrado nas espécies Turdus albicollis, Zonotrichia capensis (0,22%), Turdus rufiventris (0,18% ± 0,29), Gnorimopsar chopi (0,1% ± 0,07) no IBAMA e Turdus rufiventris (0,17 % ± 0,13), Arremon semitorquatus (0,11% ± 0,13) no JB-UFJF. Foi identificada em uma das aves (A. semitorquatus), a presença da espécie Plasmodium (H.) lutzi, a qual foi morfologicamente caracterizada. A prevalência registrada é considerada alta, entretanto verificou-se uma baixa parasitemia, o que pode estar relacionado à evolução crônica da infecção. Infecções agudas por Plasmodium spp. levam ao aparecimento de sinais clínicos com expressão rápida dos sintomas, sendo que nos casos mais graves pode ocorrer a morte do hospedeiro. Visto a elevada prevalência, levantamentos acerca de infecções por hemoparasitos em aves silvestres podem ajudar na elaboração de estratégias de profilaxia e tratamento de doenças parasitárias para a conservação das espécies. / Hemosporidian wild birds are considered important etiological agents and can cause damage to populations of hosts. The present study aimed to: ¹determine prevalence and parasitemia for Plasmodium spp. and Haemoproteus spp. Passeriformes of the Atlantic Forest of Minas Gerais, through two diagnostic methods, microscopy and nested PCR; and caracterize morphologically the species Plasmodium (Haemamoeba) lutzi Lucena, 1939. Blood samples were obtained by puncturing the brachial vein for preparation of smears, fixed in methanol and stained with Giemsa. 237 birds belonging to 18 families and 56 species of the order Passeriformes from IBAMA were sampled and captured in the Botanical Garden of the Universidade Federal de Juiz de Fora (BG-UFJF). The prevalence of Plasmodium spp. in birds of IBAMA was 63.63% and 26.25% in birds of BG-FUJF, only three individuals were infected by Haemoproteus spp., two (2.5%) from IBAMA and one (0.62 %) of BG-FUJF. The average parasitemia found for Plasmodium spp./ Haemoproteus spp. was 0.08 (± 0.08) at IBAMA and 0.05 (± 0.04) in BG-UFJF. The highest degree of mean parasitaemia was recorded in the species Turdus albicollis, Zonotrichia capensis (0.22%), Turdus rufiventris (0.18% ± 0.29), Gnorimopsar chopi (0.1% ± 0.07) at IBAMA and Turdus rufiventris (0.17% ± 0.13), Arremon semitorquatus (0.11% ± 0.13) in BG-UFJF. Was identified in one of the birds (A. semitorquatus) by means of morphology, the presence of Plasmodium (H.) lutzi species, which was characterized morphologically. Registered prevalence is considered high, however a low parasitaemia was found, which may be related to chronic course of infection.. Acute infections with Plasmodium spp. lead to the onset of clinical signs with rapid symptom expression, and in severe cases death canoccurat the host. Since the high prevalence surveys about hemoparasites infections in wild birds can help in developing strategies for prophylaxis and treatment of parasitic diseases to species conservation.

Legeleistungseinbußen – vor allem mit verminderter Eischalenqualität – stellen in einem Legehennenbetrieb hohe wirtschaftliche Verluste dar. Impfungen gegen entsprechende Erreger, u.a. gegen das aviäre Metapneumovirus (aMPV), sind daher weit verbreitet. AMPV ist seit den 70er Jahren als Auslöser der Rhinotracheitis der Puten (Turkey Rhinotracheitis; TRT) und des sogenannten Swollen Head Syndroms (SHS) der Hühner bekannt. Jedoch liegen nur wenige epidemiologische Studien zu der Verbreitung des Virus und dessen Subtypen in Legehennenbetrieben vor. Ziel der vorliegenden Studie war es daher, die Verbreitung des aMPV, vor allem der Subtypen A und B, zu unterschiedlichen Zeiten der Legeperiode zu untersuchen, um ein besseres Verständnis über den Zeitpunkt der Erstinfektionen sowie evtl. Re- oder Neuinfektionen zu erhalten. Dafür wurden erstmals 18 Legehennenherden in Sachsen alle drei Monate über die gesamte Legeperiode auf das Vorkommen von aMPV-RNA und aMPV-spezifischer Antikörper untersucht. Verschiedene Haltungssysteme wurden berücksichtigt, um ein unterschiedliches seuchenhygienisches Risiko unter Praxisbedingungen bewerten zu können. Pro Herde wurden von je zehn Hühnern Trachealtupfer und Serumproben entnommen. Die Tupferproben wurden mittels duplex nested RT-PCR untersucht, die Serumproben mittels zweier kommerzieller ELISA-Tests. In jeder Herde gelang der aMPV-RNA-Nachweis mindestens einmal zu unterschiedlichen Zeitpunkten. Bereits bei der Einstallung konnten in 17 Herden aMPV-spezifische Antikörper und/oder aMPV-RNA nachgewiesen werden. Diese Ergebnisse zeigen die hohe Verbreitungsrate des aMPV in Legehennenbetrieben. Bereits in der Aufzucht fand in der Mehrzahl der Herden eine aMPV-Infektion statt; während der Legeperiode kam es zu häufigen Re- oder Neuinfektionen bzw. zu einer langen Persistenz des Virus. Subtyp A kam alleine (51%) mehr als doppelt so häufig vor wie ausschließlich Subtyp B (22%). Doppelinfektionen mit den Subtypen A und B (27%) wurden ungefähr so häufig gefunden wie eine Infektion ausschließlich mit Subtyp B. Ein Wechsel der Subtypen A und B während einer Legeperiode wurde am häufigsten beobachtet: zehn der 18 Herden (56%) zeigten diesen Verlauf. Ausschließlich Subtyp A in allen positiven Entnahmen pro Betrieb wurde in vier von 18 Herden gefunden, ausschließlich Subtyp B in drei von 18 Herden, Subtyp A gemeinsam mit Subtyp B in einer von 18 Herden. Dies verdeutlicht die Dominanz des Subtyps A in Legehennenbetrieben. Obwohl drei Herden während der Aufzucht mit einer Subtyp B-Vakzine geimpft wurden, gelang der aMPV-RNA Nachweis in bis zu vier Probenentnahmen. Der Subtyp A dominierte auch in den geimpften Herden. Neben dem Subtyp B Feldvirus wurde in einer Herde zum Zeitpunkt der Einstallung auch ein Subtyp B ähnlich dem Impfstamm nachgewiesen. Es ist daher davon auszugehen, dass trotz bekannter Kreuzimmunität eine Impfung nicht vor Infektionen schützt, aber die Persistenz von Subtyp B vermindert. Die Analyse der Serumproben mit zwei kommerziellen ELISA-Tests ergab zum Teil konträre Ergebnisse. Da die Diagnose einer aMPV-Infektion häufig nur über diese Methode gestellt wird, ist dies von praktischer Relevanz. Eine Evaluierung des ELISA-Tests mit der höchsten Spezifität und Sensitivität sollte daher folgen.

Product line planning aims at optimal planning of product variety. In addition, the traditional product line planning problem develops new product lines based on product attributes without considering existing product lines. However, in reality, almost all new product lines evolve from existing product lines, which leads to the product line evolution problem. Product line evolution involves trade-offs between the marketing perspective and engineering perspective. The marketing concern focuses on maximizing utility for customers; the engineering concern focuses on minimizing engineering cost. Utility represents satisfaction experienced by the customers of a product. Engineering cost is the total cost involved in the process of the development of a product line. These two goals are in conflict since the high utility requires high-end product attributes which could increase the engineering cost and vice versa. Rather than aggregating both problems as one single level optimization problem, the marketing and engineering concerns entail a non-collaborative game per se. This research investigates a game-theoretic approach to the product line evolution problem. A leader-follower joint optimization model is developed to leverage conflicting goals of marketing and engineering concerns within a coherent framework of game theoretic optimization. To solve the joint optimization model efficiently, a bi-level nested genetic algorithm is developed. A case study of smart watch product line evolution is reported to illustrate the feasibility and potential of the proposed approach.

Thesis (MSc)--University of Stellenbosch, 2010. / ENGLISH ABSTRACT: This study examines the phylogeography of the southern African vlei rat, Otomys irroratus using the mtDNA cyt b gene and chromosomal data derived using G-, and C-banding, Ag-NOR staining and fluorescence in situ hybridization (FISH using flow sorts of Myotomys unisulcatus). A total of N = 102 specimens were used from the Western Cape, Eastern Cape, Northern Cape, Free State, Mpumalanga and KwaZulu-Natal provinces of South Africa. Of the N = 102, N = 55 comprised fresh material while N = 47 comprises museum material obtained from the Durban Natural Science Museum of South Africa. Cytogentic analysis of N = 55 specimens collected from seven localities in South Africa revealed intra-specific variation resulting from two rearrangements, namely pericentric inversions and heterochromatin variation. Of the 55 specimens that were analyzed 47% contained inversions or centromeric shifts on four autosomes (OIR1, OIR4, OIR6 and OIR10) which were present singly in specimens (i.e. none of the specimens contained all four inversions concurrently). These inversions were present in both homozygous and heterozygous state over a wide geographic range suggesting that they are floating polymorphisms. Given the potential role of inversions in post-mating isolation (through production of aneuploid gametes), the prevalence of inversions as floating polymorphisms in the vlei rats suggest that they are probably retained in the population through suppression of recombination in the inverted regions of the chromosomes. In addition, differences between populations is due to the presence or absence of heterochromatic arms (and not inversions), which cause variation in the NFa (40 – 49) and supernumerary B chromosomes, resulting in the variation in diploid number (2n = 28 – 32). Analysis of N = 55 specimens revealed Ag-NORs on 7 autosomal pairs 1, 2, 5, 7, 8 and 9 proximal to the centromere on the short arm of the chromosome. Pair 8 also displayed Ag-NOR at the distal end of the long arm of the chromosome in individuals from the Free State province. Pair 3 showed two Ag- NORs occurring proximal to the centromere on the short arm and on the terminal end of the long arm, respectively. I obtained 953bp of mtDNA cyt b from fresh material and 400bp from museum material. Using maximum parsimony and Bayesian inference two main clades were retrieved. Clade A specimens occur mainly in the Western and Eastern Cape provinces of South Africa. Clade B specimens occur in the Eastern Cape, Free State, KwaZulu-Natal, Northern Cape and Mpumalanga provinces of South Africa. The mean sequence divergence between the main clades (A and B) is 7.0% and between sub-clades comprising clade B is 4.8%, while within clade A the sequence divergence was 1.91%. Nested clade analysis revealed allopatric fragmentation within O. irroratus. Chromosomal characters also support the two evolutionary lineages as clade A has pericentric inversions which occur as floating polymorphisms which are absent in clade B. Clade B in turn is fixed for a complex tandem fusion rearrangement which is absent from clade A. Divergence date estimates indicate that the two clades separated around 1.1 MYA, which coincides with climate changes since the late Pliocene/Pleistocene epochs. Cladogenesis within this species complex could therefore have been influenced by habitat fragmentation. A full taxonomic review of O. irroratus is therefore warranted by this study. / AFRIKAANSE OPSOMMING: Die suider Afrikaanse vlei rot, Otomys irroratus word gekenmerk deur fenotipiese konservatisme regoor die spesie se verspreiding en het groot chromosomale variasie met diploied chromosoom getalle wat reeks vanaf 2n = 23 tot 2n = 32. Hierdie variasie binne O. irroratus het gelei tot die beskrywing van drie chromosomale groupe naamlik die A sitotipe wat gekenmerk word deur 'n akrosentriese komplement. Die tweede groep wat die B sitotipe genoem word besit ten minste agt chromosoom pare met heterokromatiese kort arms, onderwyl die derde group (die C sitotipe) vier chromosoom pare het met heterokromatiese kort arms. Hierdie studie bestudeer die bevolkings genetika struktuur van O. irroratus deur 102 monsters te analiseer wat gekollekteer was regoor die spesie se verspreiding binne Suid-Afrika en die mitochondriale merker sitokroom b sowel as chromosoom fluoressent hibridisasie te gebruik. Ek het 55 monsters van sewe lokaliteite binne Suid-Afrika sitogeneties geanaliseer deur gebruik te maak van G- en C-bandbepaling asook die hibridisasie patrone geproduseer deur die vloeisorteerde chromosoome van Myotomys unisulcatus. Die analise het gewys dat 47% van die monsters perisentromeriese inversies besit het, wat slegs aangetref was of die outosome OIR1, OIR4, OIR6 en OIR10. Hierdie inversies was nooit almal teenwoordig binne dieselfde monster nie en was gevind in beide heterosigotiese en homosigotiese vorm. Die inversies kom ook voor oor 'n wye verspreiding wat daarop aandui dat dit swerwende polymorfisme is. Omdat inversies lei tot die produksie van aneuploiede gamete speel hulle 'n belangrike rol in post-parings reproduktiewe isolasie, die verskyning van swerwende inversies binne vlei rotte dui dus daarop dat hulle onderhou word binne populasie verband deur die onderdrukking van rekombinasie in die gedeeltes van die chromosoom. Verdere verskille tussen populasies behels die voorkoms of afwesigheid van heterochromatiese kort arms wat (nie inversies) wat lei tot die variasies in die Nfa (40 – 49). Die variasie in diploied getal (2n = 28 – 32) is eksklusief as gevolg van B chromosoome. Ag-NOR banding het ook gewys dat daar twee evolusionêre lyne binne O. irroratus voorkom. Verder het filogenetiese analise van al die monsters verkryg deur volgorde-bepaling met behulp van maksimale parsimonie en Bayesian afleiding twee klades geidentifiseer. Klade A diere kom voor in die Wes en Oos-Kaap provinsies van Suid-Afrika terwyl klade B diere voorkom in die Oos-Kaap, Vrystaat, KwaZulu-Natal, Noord-Kaap en Mpumalanga provinsies onderskeidelik van Suid-Afrika. Die gemiddelde volgorde-bepalings verskille beloop 7% tussen die twee hoof klades (A en B) en tussen sub-klades 4.8%, terwyl binne klade A die verskille slegs 1.91% beloop het. Analise van die verwantskap tussen die klades het gewys dat allopatriese fragmentasie heel waarskynlik gelei het tot die populasie genetiese struktuur binne O. irroratus. Chromosoom karakters onderskraag die twee evolusionêre lyne waar klade A slegs perisentriese inversies besit wat swerwend wat ontbreek in klade B. Klade B op sy beurt besit 'n komplekse tandemme fusie wat glad nie voorkom in klade A nie. Molekulêre datering het verder gewys dat die twee klades omtrent 1.1 miljoen jaar gelede versprei het, wat ooreenstem met die klimaats veranderinge wat sedert die Peioceen en Pleistoceen plaasgevind het. Klade vorming binne die spesies komples kan daarom as gevolg van habitat fragmentasie plaasgevind het. Hierdie studie dus noodsaak 'n volle taksonomiese ondersoek van O. irroratus ten einde vas te stel hoeveel spesies binne die komplex voorkom.

Hypertension and nutrition: Fat-soluble vitamins A, D and E Weber J1 , Vlcek J1 , Suarez-Varella MM2 1 Department of Social and Clinical Pharmacy, Faculty of Pharmacy in Hradec Králové, Charles University in Prague, Czech Republic 2 Department of Preventive Medicine and Public Health, Faculty of Pharmacy, University of Valencia, Spain Arterial hypertension (AH) is a disease affecting population globally, and thus considered as a problem of public health and socioeconomic. Studies are trying to identify the connection between diet and the prevalence of arterial hypertension. Objective of the study was to determine possible association between an occurrence of AH and fat-soluble vitamins A, D and E intake. The nested, case-control population study investigation was grounded on database from the Spanish Hortega study, and performed on a random sample of 1,514 people (50.3 % women, 49.7 % men). From this sample we selected those aged ≥ 40 years old and untreated for hypertension and divided them into two groups: non-hypertensive (n = 429; 63.6 %) (controls), and newly diagnosed AH (n = 246; 36.4 %) (cases). Biochemical and anthropometric measurements, data on dietary intakes, education, socioeconomic status, place of residence, health habits, comorbidities, consumption of alcohol and tobacco were used for our...