Establishment of relationships for predicting the lateral drift demands of near-fault ground motions is one of the major challenges in earthquake engineering. Excessive lateral drifts caused by earthquake ground motions are the major causes of structural damage observed in structures. In this study, some of the fundamental characteristics of near-fault ground motions are examined. Response characteristics of elastic frame structures to near-fault ground motions are investigated. An approximate method for estimating the elastic ground story and interstory drifts for regular frame type structures is presented. Inelastic displacement demands imposed on elasto-plastic single degree of freedom (SDOF) systems subjected to near-fault ground are examined. Three equations for estimating the maximum lateral inelastic displacement demand from the maximum elastic displacement demand are established. Two of these equations relate the inelastic and elastic displacement demands through natural period and strength reduction factor. The third equation relates the inelastic and elastic displacement demands through the ratio of natural period to pulse period and the strength reduction factor. Efficiency of the natural period to pulse period ratio for estimating the inelastic displacement ratio is shown. Error statistics of the proposed equations are presented and compared with similar studies in the literature. According to the results, these equations can be used for quick and rough estimates of displacement demands imposed on regular elastic moment resisting frames and elasto-plastic single degree of systems.

The circular, single-stranded (ss) DNA genomes of plant viruses in the families Geminiviridae and Nanoviridae are replicated within the nucleus of a host cell by a mechanism called rolling circle replication (RCR). Although this process relies almost exclusively on the replication machinery of the host cell, initiation occurs via the interaction of the viral replication initiation protein (Rep) with regulatory DNA sequences within the viral genome. The use of a virus-based episomal amplification technology as a plant bioreactor platform exploits the process of Rep-mediated RCR for the high-level amplification of virus-based episomes in plants and subsequent expression of heterologous proteins; such an approach offers advantages over existing gene expression technologies. This PhD thesis describes research towards the development of a virus-based episomal amplification system for use in sugarcane. Such a crop is ideally suited for a plant bioreactor system due to the efficient high-level production of plant biomass and the existence of established production, harvesting and processing infrastructure. In order to rapidly assess the potential of a virus-based episomal amplification system in sugarcane, a transient assay system was established. Sugarcane callus was identified as the most suitable cell preparation; providing rapid cell regeneration, uniform experimental samples and upon isolation, total DNA suitable for Southern analysis. This assay system once established, proved effective in rapidly identifying virus-based episomes capable of undergoing RCR within sugarcane host cells. This transient assay system was then used to test the functionality of a virus-based episomal amplification system based on the ssDNA virus, Banana bunchy top virus (BBTV) in sugarcane. BBTV-based episomal amplification vectors were constructed with a reporter gene expression cassette flanked by two copies of the BBTV regulatory DNA sequences. The episomal amplification vectors were bombarded into sugarcane and banana host cells in various combinations and evidence of RCR was assessed through Southern blot analysis. RCR products were identified in banana host cells bombarded with the BBTV-based episomal amplification vectors in combination with vectors encoding BBTV Master-Rep (M Rep). RCR products were not identified within sugarcane cells bombarded with the same construct combinations. Integrated InPAct (In Plant Activation) episomal vectors based on BBTV were then employed to confirm the transient results, in addition, the functionality of an InPAct vector based on an alternate virus, Tobacco yellow dwarf virus (TYDV) was also assessed. InPAct vectors based on BBTV were constructed with an untranslatable expression cassette for integration within the sugarcane genome. Transient experiments were performed to assess the ability of BBTV M-Rep and TYDV Rep to initiate RCR of their respective InPAct vectors. Visual observation of GFP expression indicated that BBTV M-Rep was capable of initiating RCR of the BBTVbased InPAct vectors within banana host cells but no evidence was observed in sugarcane host cells. TYDV Rep was capable of initiating RCR of the TYDV-based InPAct vector within sugarcane host cells with a 100-fold increase in the number of fluorescent foci compared to cells bombarded with the TYDV InPAct vector alone. The BBTV-based InPAct vector was stably integrated within the sugarcane genome and the ability for BBTV M-Rep to initiate episome formation and RCR was assessed by Southern blot analysis. Evidence of BBTV M-Rep mediated RCR was not detected within the transgenic sugarcane bombarded with BBTV M-Rep. Transgenic sugarcane containing the TYDV-based InPAct vectors was assessed for the ability to be activated by TYDV Rep and undergo RCR. Southern blot analysis demonstrated that TYDV Rep was capable of recognising the integrated TYDVbased InPAct vector and RCR was detected within the transgenic sugarcane. The observation that episomal vectors based on TYDV were functional within sugarcane host cells and BBTV-based vectors were not, was unexpected. It had been hypothesised that an episomal vector based on a monocot-infecting virus would replicate in an alternate monocot host, while an episomal vector based on a dicot infecting virus would not. Virus replication is thought to be host-specific however most host range studies have been conducted with full length infectious clones and not deconstructed virus-based episomes. The implication that viral Reps may be functional in plant cells of non-host species was then investigated. The ability for viral Reps to recognise their cognate IR and initiate RCR of virus-based episomes in different host cells was assessed through cross-replication experiments. Four ssDNA plant viruses; BBTV, TYDV, Chloris striate mosaic virus (CSMV) and Tomato leaf curl virus - Australia (ToLCV-Au) were assessed via Southern blot analysis for their ability to initiate both autonomous replication of infectious clones and episomal amplification within three different plant hosts; tobacco, sugarcane and banana. Results from cross replication studies indicated a complex interaction between viral and host replication components. BBTV infectious clones and episomal vectors were restricted to replication within banana host cells providing a clear indication that episomal amplification vectors based on BBTV are restricted to Musa spp. BBTV M-Rep was unable to recognise the viral regulatory DNA sequences of the other three ssDNA viruses. TYDV infectious clones and episomal vectors were capable of replicating within all three host cells tested, indicating that TYDV is capable of undergoing RCR within a broad range of plant hosts. TYDV Rep was also capable of recognising the viral regulatory DNA sequences of both CSMV and BBTV given favourable conditions within specific plant hosts. Replication of the CSMV infectious clone was not detected in any of the three host cells, although fidelity of this clone requires further confirmation. CSMV episomal vectors were functional within banana host cells only, indicating that although closely related to TYDV, episomal amplification vectors based on CSMV have a restricted host range. CSMV Rep could not initiate RCR of episomal amplification vectors containing the viral regulatory DNA regions of the other three viruses in any of the plant host cells. ToLCV-Au infectious clones were capable of replicating within banana and tobacco host cells. Episomal amplification vectors based on ToLCV-Au extended the host range to sugarcane. ToLCV-Au Rep was unable to recognise the viral regulatory DNA sequences of the other three viruses in any of the plant host cells. The ability for a viral Rep to recognise its own cognate regulatory DNA sequences within alternate plant host cells is variable. Episomal amplification vectors based on TYDV and ToLCV-Au appear to be the most suitable for the further development of a virusbased bioreactor system in sugarcane. This study details the initial steps taken towards the development of a virus-based episomal amplification system in sugarcane. In doing so, fundamental knowledge into the mechanisms involved in Rep recognition of viral regulatory DNA sequences has been gathered. These research findings will provide a solid foundation for the further development of a sugarcane-based bioreactor.

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abstract: HIV/AIDS is the sixth leading cause of death worldwide and the leading cause of death among women of reproductive age living in low-income countries. Clinicians in industrialized nations monitor the efficacy of antiretroviral drugs and HIV disease progression with the HIV-1 viral load assay, which measures the copy number of HIV-1 RNA in blood. However, viral load assays are not widely available in sub-Saharan Africa and cost between 50-$139 USD per test on average where available. To address this problem, a mixed-methods approach was undertaken to design a novel and inexpensive viral load diagnostic for HIV-1 and to evaluate barriers to its adoption in a developing country. The assay was produced based on loop-mediated isothermal amplification (LAMP). Blood samples from twenty-one individuals were spiked with varying concentrations of HIV-1 RNA to evaluate the sensitivity and specificity of LAMP. Under isothermal conditions, LAMP was performed with an initial reverse-transcription step (RT-LAMP) and primers designed for HIV-1 subtype C. Each reaction generated up to a few billion copies of target DNA within an hour. Presence of target was detected through naked-eye observation of a fluorescent indicator and verified by DNA gel electrophoresis and real-time fluorescence. The assay successfully detected the presence of HIV in samples with a broad range of HIV RNA concentration, from over 120,000 copies/reaction to 120 copies/reaction. In order to better understand barriers to adoption of LAMP in developing countries, a feasibility study was undertaken in Tanzania, a low-income country facing significant problems in healthcare. Medical professionals in Northern Tanzania were surveyed for feedback regarding perspectives of current HIV assays, patient treatment strategies, availability of treatment, treatment priorities, HIV transmission, and barriers to adoption of the HIV-1 LAMP assay. The majority of medical providers surveyed indicated that the proposed LAMP assay is too expensive for their patient populations. Significant gender differences were observed in response to some survey questions. Female medical providers were more likely to cite stigma as a source problem of the HIV epidemic than male medical providers while males were more likely to cite lack of education as a source problem than female medical providers. / Dissertation/Thesis / Ph.D. Molecular and Cellular Biology 2011

[pt] A estabilidade do escoamento estratificado de ar e água, sujeito a perturbações periódicas no nível de líquido, é investigada numericamente em um duto horizontal. Selecionou-se o Modelo de Dois Fluidos unidimensional para a simulação do escoamento. As equações de conservação de massa e de quantidade de movimento linear para as fases gás e líquido são discretizadas de acordo com o método dos Volumes Finitos. O acoplamento entre as equações é resolvido sequencialmente com uma versão modificada do método PRIME. Perturbações no nível de líquido foram introduzidas de maneira controlada na entrada da tubulação. A evolução dessas perturbações, ao longo da tubulação, é analisada e os resultados são comparados com as previsões fornecidas por um modelo baseado na teoria linear de Kelvin-Helmholtz. A velocidade de propagação, a frequência e o número de onda das perturbações apresentam excelente concordância entre a simulação e modelo, indicando que, de fato, ambas abordagens são capazes de prever características fundamentais dessas ondas. As taxas de crescimento previstas pelo modelo e as obtidas na simulação, também, foram comparadas apresentando razoável concordância. Os resultados mostram que a frequência da perturbação tem influência na taxa de amplificação e que ondas com frequências mais altas tendem a serem mais instáveis. Para tubulações longas, efeitos não lineares podem ser observados em regiões afastadas da entrada da tubulação. Nesse estágio é possível observar alterações no mecanismo de crescimento das perturbações. / [en] The stability of stratified air-water flow under the influence of periodic disturbances in the liquid level is investigated numerically for a horizontal pipe. One-dimensional two-fluid model is used for flow simulation. Conservation equations for mass and linear momentum are discretized for both phases using a finite volume method. Coupling between equations is achieved by sequentially solving a modified version of PRIME method. Controlled disturbances are introduced in the flow by oscillating the liquid level at the pipe inlet. The evolution of the generated disturbances along the pipe is analyzed and the results are compared with predictions given by a model based on linear theory of Kelvin-Helmholtz (KH). An excellent agreement is obtained for velocity, frequency and wave number of the perturbations. This indicates that both approaches are capable to predict the fundamental characteristics of the waves. Amplifications rates predicted by simulation and model have been also compared and the results show a reasonable agreement. It is found that the frequency of the perturbations plays a role in the amplification rate. For increasingly higher frequencies the disturbances tends to be more unstable. The analysis is extended for long pipes, in such cases the growth rates changes at locations far from the inlet. It is conjectured that non linear mechanisms are related to observations.

Introdução: A infecção urogenital por Chlamydia trachomatis é a doença sexualmente transmissível bacteriana mais prevalente no mundo e afeta principalmente mulheres jovens sexualmente ativas. Infecções não tratadas podem provocar complicações reprodutivas decorrentes do dano tubáreo. Na gestação, aumenta o risco de parto prematuro, baixo peso ao nascer, morte perinatal, conjuntivite e pneumonia neonatal. Existem poucos dados brasileiros referentes à epidemiologia dessa infecção no nosso meio. O objetivo desse estudo foi determinar a prevalência de C. trachomatis em mulheres inférteis e em gestantes. Método: Foram analisadas transversalmente 77 mulheres inférteis e 60 gestantes assintomáticas. Foram coletadas amostras urinárias para ensaio de PCR e amostras sanguíneas para pesquisa de anticorpos IgG através da técnica de imunofluorescência indireta. Todas as participantes responderam um questionário referente ao seu histórico clínico e ginecológico. Resultados: A prevalência, tanto no ensaio de PCR quanto na imunofluorescência indireta (IgG) para C. trachomatis foi similar entre os grupos. Encontramos anticorpos IgG presentes em 61% das mulheres inférteis e em 56,7% das gestantes. Houve somente 1 PCR positivo no grupo das inférteis (1,3%) e nenhum do grupo das gestantes. Conclusão: Encontramos alta prevalência de anticorpos IgG para C. trachomatis em mulheres inférteis e em gestantes, mas verificamos baixa prevalência de PCR positivo nas participantes. A presença de IgG correlacionou-se com comportamento sexual e tabagismo. / Background: Chlamydia trachomatis (CT) is the most prevalent sexually transmitted bacterial infection and affects mainly young, sexually active, women. Untreated infection may lead to reproductive complications due to tubal damage. Infections during pregnancy may cause preterm labor, low birth weight, perinatal death and neonatal conjunctivitis and pneumonia. There is little data on CT infection in Brazil. The aim of this study was to determine CT prevalence on infertile and pregnant women. Methods: A cross-sectional study included 77 infertile and 60 asymptomatic pregnant women. First void urine was tested to CT using PCR and blood samples were collected for CT IgG antibodies testing using Indirect Immunofluorescence. A questionnaire about medical, gynecological and sexual history was applied to all participants. Results: We found statistically similar prevalence of PCR and IgG antibodies between groups. This study observed a 61% prevalence of CT IgG antibodies in infertile women and 56,7% in pregnant women. PCR was positive in only one (1,3%) infertile woman and in none of the pregnant. Conclusion: A high prevalence of C. trachomatis IgG antibody in Brazilian pregnant and infertile women, but a low prevalence of positive PCR on urine samples were demonstrated. CT antibodies were associated with sexual behavior and smoking.

Submitted by VICTOR BERNARDES BARROSO DA COSTA null (victorbernardes_@hotmail.com) on 2016-03-16T04:08:27Z No. of bitstreams: 1 Dissertação - VICTOR (Versão Final).pdf: 20193346 bytes, checksum: f7da64e79358575f01cf61a5054c4587 (MD5) / Approved for entry into archive by Juliano Benedito Ferreira (julianoferreira@reitoria.unesp.br) on 2016-03-18T12:52:04Z (GMT) No. of bitstreams: 1 costa_vbb_me_sjc.pdf: 20193346 bytes, checksum: f7da64e79358575f01cf61a5054c4587 (MD5) / Made available in DSpace on 2016-03-18T12:52:04Z (GMT). No. of bitstreams: 1 costa_vbb_me_sjc.pdf: 20193346 bytes, checksum: f7da64e79358575f01cf61a5054c4587 (MD5) Previous issue date: 2016-01-21 / Submitted by VICTOR BERNARDES BARROSO DA COSTA null (victorbernardes_@hotmail.com) on 2016-01-26T01:08:29Z No. of bitstreams: 1 Dissertação - VICTOR (Versão Final).pdf: 20193437 bytes, checksum: c041b8e5fa02ed4f74d445c5838128fd (MD5) / Approved for entry into archive by Sandra Manzano de Almeida (smanzano@marilia.unesp.br) on 2016-01-26T13:39:40Z (GMT) No. of bitstreams: 1 costa_vbb_me_sjc.pdf: 20193437 bytes, checksum: c041b8e5fa02ed4f74d445c5838128fd (MD5) / Made available in DSpace on 2016-01-26T13:39:40Z (GMT). No. of bitstreams: 1 costa_vbb_me_sjc.pdf: 20193437 bytes, checksum: c041b8e5fa02ed4f74d445c5838128fd (MD5) Previous issue date: 2016-01-21 / Item merged in doublecheck by Felipe Augusto Arakaki (arakaki@reitoria.unesp.br) on 2016-03-21T13:12:44Z Item was identical to item(s): 135509, 132061 at handle(s): http://hdl.handle.net/11449/136305, http://hdl.handle.net/11449/132945 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Fundação de Amparo à Pesquisa do Estado do Amazonas (FAPEAM) / O carcinoma de células escamosas (CEC) de boca é uma neoplasia incomum em pacientes jovens. Na literatura de língua inglesa não há relatos de estudos que investiguem a amplificação do EGFR e a expressão desta proteína neste grupo etário. O objetivo deste estudo foi investigar a amplificação do EGFR através da técnica de hibridização por fluorescência in situ (FISH) e correlacionar os resultados obtidos através da imunomarcação da proteína EGFR com a epidemiologia e com o prognóstico dos pacientes avaliados. Ao final dos testes de FISH e imuno- histoquímicos, 21 amostras do grupo teste (pacientes ≤ 40 anos) e 39 amostras do grupo controle (pacientes ≥ 50 anos) foram consideradas adequadas para avaliação. As variáveis clínicas e anatomopatológicas foram comparadas pelos testes Qui-Quadrado ou exato de Fisher. A expressão do marcador EGFR e do método de FISH foi comparada entre os grupos por meio do teste não paramétrico de Mann-Whitney. As curvas de sobrevida foram calculadas utilizando o método de Kaplan-Meier e suas curvas foram comparadas através do teste de log-rank. Houve maior número de pacientes do sexo masculino, leucodermas, tabagistas e etilistas. A amplificação do EGFR foi maior no grupo teste (p = 0,018). A amplificação do EGFR associou-se estatisticamente com a variável estadiamento clínico avançado (p = 0,013), independente do grupo. A expressão da proteína EGFR correlacionou-se com tumores bem diferenciados (p = 0,011) e a presença de metástase (p = 0,035), independente da idade. A presença de amplificação foi mais frequente no grupo tese. Alguns casos de pacientes ≥40 anos de idade podem ser adequados ao emprego da terapia anti-EGFR, devido à amplificação do EGFR. / Oral squamous cell carcinoma (OSCC) is uncommon neoplasia in young patients. In the English literature, there are no reports of studies that investigate the amplification of EGFR and expression of this protein in this age group. The aim of this study was to investigate the amplification of EGFR by fluorescence in situ hybridization (FISH) and to correlate the results by immunostaining of EGFR protein with clinicopathological features and prognosis. After FISH and immunohistochemistry, 21 samples of the test group (≤ 40 years) and 39 samples of control group (≥ 50 years) were suitable for evaluating. Categorical variables were compared by the Pearson chi-square test or Fisher's exact test. Associations between protein levels and FISH results with clinicopathological characteristics of the patients were analyzed by Mann-Whitney U test. Survival rates were calculated using the Kaplan-Meier method and the curves were compared by the log-rank test. There was predominance for male patients, leucoderma, smoking and alcohol consumption. The EGFR amplification was higher in the test group (p = 0.018) and it was associated statistically with advanced clinical stage (p = 0.013), independent of the group. The expression of EGFR protein was correlated to well differentiated tumors (p = 0.011) and presence of metastasis (p = 0.035), regardless of age. Presence of EGFR amplification and/or expression. Some cases of patients ≥40 years old might be suitable for anti-EGFR therapy because of EGFR amplification. / FAPEAM: 254/2014

Aeglidae Dana (1852) are anomuran crabs which occur exclusively in freshwater. The genus Aegla consists of 63 species and subspecies restricted to the southern South America. Highly sensitive molecular markers can help to elucidate some evolutionary features of the genus. Microsatellite markers are excellent molecular markers because they can reveal fine details of the genetic structure of a population. However, microsatellite isolation employs a laborious and expensive methodology. An alternative approach to the isolation procedure is the utilization of microsatellite markers previously developed for close species. The closer the phylogenetic relationship between two species, the more probable is the conservation of the loci and the successful cross-amplification. The present study aimed to test the crossamplification of microsatellite loci developed for two species of the family Aeglidae in other decapod crustaceans and to verify their potential for application in further studies on population genetics. Primers developed for microsatellite loci previously isolated from Aegla longirostri (AlCA135 and AlCA138) and Aegla uruguayana (Au05) were tested in seven species of the family Aeglidae, Aegla camargoi, Aegla leptodactyla, Aegla plana, Aegla platensis, Aegla spinipalma, Aegla violacea and Aegla sp.n., besides in Emerita brasiliensis (Anomura: Hippidae), Pachycheles laevidactylus (Anomura: Porcellanidae) and Trichodactylus panoplus (Brachyura: Trichodactylidae). DNA was extracted using traditional procedures. DNA samples were submitted to the Polymerase Chain Reaction (PCR) using the primers cited. The PCR products were electrophoresed in 6% polyacrylamide gels, at 100V for 24 hours. The gels were silver-stained and the band patterns were analyzed. The microsatellite markers could be transferred only within the genus Aegla, and the locus AlCA135 presented the greatest rate of success in cross-species transfer. Cross-amplification between families within the infra-order Anomura, as well as between the infra-order Anomura and Brachyura was not possible, indicating that these microsatellite loci are not conserved in distantly related species. Notwithstanding, the evaluated loci present potential for utilization within the family Aeglidae and new tests, using optimized PCR conditions, should improve the success rate in cross-amplification. / Aeglidae Dana (1852) são caranguejos que pertencem a Infraordem Anomura e são exclusivos de água doce. O gênero Aegla é constituído por aproximadamente 63 espécies, sendo restrito ao sul da América do Sul. O emprego de marcadores moleculares altamente sensíveis pode auxiliar no entendimento de aspectos evolutivos e da diversidade do gênero. Os microssatélites são excelentes marcadores moleculares que podem revelar detalhes da estrutura genética de uma população. Entretanto, o isolamento de microssatélites é uma técnica cara e trabalhosa. A utilização de locos de microssatélites previamente desenvolvidos para espécies próximas é uma alternativa ao isolamento. Quanto mais próxima a relação filogenética entre duas espécies, maior a probabilidade de conservação das seqüências e de amplificação cruzada utilizando primers heterólogos. O objetivo deste estudo foi testar a amplificação cruzada de locos de microssatélites desenvolvidos para duas espécies de Aeglidae em outros crustáceos e verificar o seu potencial para aplicação em estudos de genética de populações. Primers desenvolvidos para locos de microssatélite previamente isolados de Aegla longirostri (AlCA 135 e AlCA 138) e Aegla uruguayana (Au05) foram testados em outras sete espécies da família Aeglidae, Aegla camargoi, Aegla leptodactyla, Aegla plana, Aegla platensis, Aegla spinipalma, Aegla violacea, Aegla sp.n. e também em Emerita brasiliensis (Anomura: Hippidae), Pachycheles laevidactylus (Anomura: Porcellanidae) e Trichodactylus panoplus (Brachyura: Trichodactylidae). O DNA foi extraído utilizando o método tradicional de fenol-clorofórmio. As amostras de DNA foram submetidas a uma Reação em Cadeia de Polimerase (PCR) com os primers citados. Os produtos da PCR foram submetidos à eletroforese em gel de poliacrilamida 6% a 100V por 24 horas. Os géis foram corados com nitrato de prata e os padrões de banda foram analisados. Observou-se transferência dos marcadores microssatélites somente dentro do gênero Aegla, e o loco AlCA135 foi o que apresentou maior índice de sucesso na amplificação cruzada. Não foi observada amplificação cruzada entre famílias dentro da infraordem Anomura e nem entre as infraordens Anomura e Brachyura, indicando que esses locos de microssatélites não se encontram conservados em espécies distantemente relacionadas. Entretanto, os locos avaliados apresentam potencial de utilização dentro da família Aeglidae e novos testes, otimizando as condições de PCR, poderão melhorar os índices de sucesso de amplificação cruzada.

Resumo: A maioria dos casos de fraturas ósseas é utilizada nas cirurgias, implantes ortopédicos de osteossíntese (placa-parafuso e parafusos) confeccionados em aço inoxidável austeníticos, devido à sua baixa relação custo-benefício. As infecções associadas aos implantes de osteossintese estão relacionadas com o crescimento de microrganismos em biofilmes, resultando em um processo infeccioso de difícil erradicação. O objetivo do estudo foi identificar os microrganismos recuperados do conjunto metálico placaparafuso e parafusos após remoção cirúrgica, avaliar a capacidade de aderência dos microrganismos, verificar a viabilidade celular, caracterizar genotipicamente os isolados Gram-positivos e detectar a resistência aos antimicrobianos. Os conjuntos placa-parafuso e parafusos de aço inoxidável austenítico ASTM F138/F139 e ISO NBR 5832-1/9 foram transportados em bolsa de polietileno para o Laboratório de Microbiologia Clínica. Os implantes foram lavados em solução tampão fosfato-salino, armazenados em bolsa contendo solução Ringer e submetidos ao banho ultrassônico em freqüência de 40±2 kHz por 5 minutos. O fluido sonicado foi transferido para tubos Falcon e centrifugado a 3.000g durante 20 minutos. O sedimento foi resuspendido em nova solução Ringer, homogeneizado por vortex e 10μL semeados em ágar Sangue de carneiro 5%, MacConkey e Sabouraud. Os meios foram incubados em diferentes condições de temperatura por 7 a 14 dias para recuperação de microrganismos. O perfil de resistência dos isolados foi obtido de acordo com CLSI 2011. Para análise da viabilidade celular foi utilizado o kit Live/Dead e microscópio de fluorescência... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The majority of cases of bone fractures is used in surgery, internal fixation of orthopedic implants (screw-plate and screws) manufactured in austenitic stainless steel due to its low cost-benefit. Infections associated with implant fixation are related to the growth of microorganisms in biofilms, resulting in an infection difficult to eradicate. The objective of study was to identify the microorganisms recovered from the set screwplate and screws metallic after surgical removal, evaluate the capacity of adherence of microorganisms, check the cell viability and characterize the isolates genotypically Gram-positive and detect antimicrobial resistance. The sets screw-plate and screws of austenitic stainless steel ASTM F138/F139 and ISO NBR 5832-1/9 were transported in polyethylene bag to the Laboratory of Clinical Microbiology. The implants were washed in phosphate buffered saline solution, stored in bags containing Ringer's solution and submitted to ultrasonic bath at a frequency of 40 kHz ± 2 for 5 minutes. The sonicate fluid was transferred to falcon tubes and centrifuged at 3.000g for 20 minutes The new sediment was re-suspended in Ringer's solution, homogenized by vortex and 10μL seeded agar 5% sheep blood, MacConkey and Sabouraud. The media were incubated at different temperatures for up to 7 days for growth of CFU mL-1. The resistance profile of the isolates was obtained according to CLSI 2011. For analysis of cell viability kit was used for Live/Dead and fluorescence microscope. After microbiological analysis the isolation was observed in some samples of polymicrobial implants. The data obtained showed that bacteria were the most isolated coagulase-negative... (Complete abstract click electronic access below) / Orientador: Clarice Queico Fujimura Leite / Coorientador: Elisabeth Loshchagin Pizzolitto / Banca: Eliana Aparecida Varanda / Banca: Beatriz Maria Machado de Medeiros / Banca: Denise Crispin Tavares / Mestre

Gliomas are brain tumors arising from neuroglia. In most cases astrocytic or oligodendroglial component is the main element of the tumor. Non-random chromosomal abberations are found in tumor cells as was revealed previously. The aim of this study was a fluorescence in-situ hybridisation analysis (FISH) of tissue samples obtained during neurosurgical procedures, determine the frequence of selected chromosomal abberations, further correlation with morphological and clinical data and statistical analysis of the results. During six years 264 tissue samples were gained in which FISH with defined probes was performed. The acquired results were compared with histological analysis and selected clinical data (age, Karnofsky score, extent of resection, overall survival). The whole series was divided into 7 groups by tumor type for further statistical analysis. In every group median and mean survival time was calculated, Kaplan-Meier analysis was focused on influence of selected parameters to overall survival. In some categories Cox regression model was created to achieve a hazard ratio of selected parameters. In WHO Grade II and III tumors the risk of malignant progression and tumor upgrading is significantly higher in comparison with samples where specific abberations were not found (EGFR amplification, CDKN2A and...