Modelo de resposta gradual para testes com penalização para itens dicotômicos

Dissertação (mestrado)—Universidade de Brasília, Instituto de Ciências Exatas, Departamento de Estatística, Programa de Pós-Graduação em Estatística, 2015.

Sistema de informação para a realização de testes de imagens em crianças

O Grupo de Investigação de Bioacústica e Sistemas (GIBS) da Escola Superior Tecnologia da Saúde de Coimbra proporciona várias actividades entre as quais o desenvolvimento de softwares aplicados na área da saúde. Neste trabalho vai ser abordado este tema, em particular, o teste “Peabody picture vocabular test”, um dos testes desenvolvidos no GIBS que vai ser associado a um sistema de informação, por forma a automatizar e dinamizar o procedimento do teste. O teste consiste em disponibilizar uma série de imagens a uma criança, que vai seleccionar as que conhece. Esta escolha tem em conta a idade da criança, bem como o seu desenvolvimento cognitivo, por isso este conjunto de imagens é amplo. Das imagens que a criança seleccionar, o protótipo vai gerar um conjunto de quatro imagens aleatórias, e reproduzir o som alusivo a uma delas. Dado que a criança seleccionou as imagens, caso não consiga identificar existe um potencial problema e o técnico encontra-se em condições de poder realizar mais testes para o identificar. No caso de identificar, o teste decorre sem problemas sendo que no final, o técnico terá acesso aos dados verificando quantas acertou em cada uma das intensidades apresentadas. Estas informações podem ser para cada um dos ouvidos em separado, caso se utilizem auscultadores, ou para ambos os ouvidos se o exame for realizado em campo livre. Actualmente o teste é realizado, numa sala acústica auxiliada por um audiométrico, que ajuda a regular a intensidade do som reproduzido e para qual dos ouvidos vai ser reproduzido. Essas mesmas condições serão mantidas. No entanto, o audiómetro funcionará em conjunto com um computador, com uma versão do protótipo a “correr”, substituindo os cartões com as imagens utilizados actualmente. Para além de ser um sistema totalmente automático, conta ainda com a vantagem de armazenar todas as variáveis associadas a cada teste, desde os dados do utente, os dados do técnico (utilizador), os dados do teste e os resultados. A fim de validar este protótipo, foi empreendido um grupo de experiências, abrangendo aspectos particulares e distintos em cada um deles, verificando-se a adequação do modelo e a utilidade do recurso, automatizado, à experiência passada. Os resultados obtidos nesta avaliação permitem considerar que o protótipo cumpre os requisitos pré-estabelecidos.

Relação da doença periodontal com o cancro do pâncreas: diagnóstico do cancro do pâncreas com recurso a testes salivares

Dissertação para obtenção do grau de Mestre no Instituto Superior de Ciências da Saúde Egas Moniz

Testes físico-mecânicos e físico-químicos do couro da tilápia vermelha

Realizaram-se testes físico-mecânicos e físico-químicos em couro de tilápia vermelha (Oreochromis spp.) a fim de testar a sua resistência. As amostras foram distribuídas em delineamento inteiramente ao acaso com dois tratamentos: no T1, procedeu-se à retirada do corpo-de-prova no sentido longitudinal e, no T2, à retirada do corpo-de-prova no sentido transversal. Para os testes de determinação da resistência à tração, alongamento e rasgamento progressivo, foi utilizado o dinamômetro EMIC, com velocidade de afastamento entre as cargas de 100 ± 20mm/min, em ambiente climatizado (± 23ºC e UR do ar de 50%), por 24 horas. A espessura do couro variou de 0,61 a 0,75mm, mas não houve diferença entre os sentidos analisados. O couro apresentou maior resistência à tração no sentido transversal, 25,89N/mm², (P<0,01), comparado ao sentido longitudinal, 14,20 N/mm². O alongamento foi significativamente (P<0,05) maior no sentido longitudinal, 80,8%, em relação ao transversal, 62,6%. Não houve diferença para o rasgamento progressivo entre os tratamentos. O couro apresentou teor de óxido de cromo de 3,8%, graxa de 15,1% e pH e cifra diferencial de 3,5 e 0,5, respectivamente. Os valores nos testes de resistência e físico-químicos apresentados pelo couro indicam que ele pode ser utilizado para a confecção de vestuário e artefatos de couro em geral.

Sodium metabisulfite-induced changes on testes, spermatogenesis and epididymal morphometric values in adult rats

Background: Sulphites are widely used as a preservative and antioxidant additives in the food and pharmaceutical industries. Many types of biological and toxicological effects of sulphites in multiple organs of mammals have been shown in previous studies. Objective: The aim of this study was to investigate the effects of sodium metabisulfite (SMB) on testicular function and morphometric values of epididymis in adult male Wistar rats. Materials and Methods: A total of 32 rats were randomly divided into four groups. The experimental groups received SMB at doses of 10 mg/kg (S10), 100mg/kg (S100), and 260 mg/kg (S260) while an equal volume of normal saline was administered to the control group via gavage. The rats were anaesthetized after 28 days and the left testis with the head of epididimis was excised following abdominal incision for histological observation using hematoxylin and eosin staining. Serum samples were collected for assay of testosterone level. The initial epididymis was analyzed for motility, morphology, and the number of sperms. Result: The results of this study showed that normal morphology, count, and motility of sperms and testosterone level were decreased in the SMB treated groups. In comparison with the control group, SMB resulted in a lower total number of spermatogonia, primary spermatocyte, spermatids, and Leydig cells. Conclusion: It is suggested that SMB decreases the sperm production and has the potential to affect the fertility adversely in male rats.

Automatização de testes de software no âmbito da certificação de software para aplicações de engenharia civil

Testar software é uma das atividades que faz parte do desenvolvimento de software, e tem como objetivo apresentar evidências de confiabilidade contribuindo para uma maior qualidade do software. Esta atividade consome uma parte significativa do esforço de um projeto de desenvolvimento de software, sempre com o objetivo de encontrar erros antes da fase de manutenção, pois o custo de correção nesta fase pode ser até 100 vezes superior ao custo de correção na fase de conceção. Para auferir mais qualidade ao software, este pode ser certificado por uma norma de qualidade. As normas fornecem processos consistentes, rigorosos e uniformes para o desenvolvimento de software sempre com o objetivo de garantir qualidade ao software. As normas têm um papel importante na definição dos requisitos de teste, casos de teste e relatórios de teste que contemplam a atividade de testes permitindo elaborar um plano de testes mais rigoroso. Como o processo de testes é complexo no desenvolvimento de software, as ferramentas de automatização de testes de software permitem reduzir tempo, recursos e consequentemente os custos para a organização. A automatização deverá ser capaz de produzir os mesmos resultados obtidos através de um processo de testes manual, evidenciando sempre o resultado do teste. Deve também permitir a realização de testes sistemáticos e paralelos em diferentes ambientes de teste, sem o aumento do tempo e de recursos humanos. Nesta dissertação pretende-se desenvolver uma abordagem automatizada com o software Sikuli para a realização de testes seguindo a norma ISO/IEC 25051 para certificação de software. Depois da criação da abordagem e a respetiva criação de testes, é necessário validar a capacidade desta abordagem em comparação com uma abordagem de testes manuais.

Erro de Tipo I no teste de Friedman e nos testes de comparação múltipla

O teste de Friedman e a alternativa não paramétrica à ANOVA de medidas repetidas. Os testes de comparação múltipla são aplicados após a rejeição da hipótese nula do teste Friedman. Neste trabalho, realizamos um estudo de simulação para analisar a probabilidade de erro de Tipo I tanto no teste de Friedman como também nos testes de comparação múltipla mais usuais. Consideraram-se as distribuições discretas vulgarmente utilizadas para modelar dados de contagens, nas áreas da Biologia e da Ecologia. No teste de Friedman a taxa de erro de Tipo I empírica e menor quando se considera a estatística de teste com aproximação ao qui-quadrado. Nos testes de comparação múltipla, a probabilidade de erro de Tipo I aumenta com o aumento do número de blocos, e no teste LSD de Fisher também com o aumento do número de tratamentos. O teste dos Sinais e o mais conservativo e o teste LSD de Fisher o mais liberal.

Testes de parentesco utilizando polimorfismos de nucleotídeo único em dados simulados

2015

Proposta de validação de testes de biopesticidas e biorremediadores em mamíferos.

2008

NMR measurement of 39K detectability and relaxation constants in rat tissue

Differences in the NMR detectability of 39K in various excised rat tissues (liver, brain, kidney, muscle, and testes) have been observed. The lowest NMR detectability occurs for liver (61 ± 3% of potassium as measured by flame photometry) and highest for erythrocytes (100 ± 7%). These differences in detectability correlate with differences in the measured 39K NMR relaxation constants in the same tissues. 39K detectabilities were also found to correlate inversely with the mitochondrial content of the tissues. Mitochondria prepared from liver showed greatly reduced 39K NMR detectability when compared with the tissue from which it was derived, 31.6 ± 9% of potassium measured by flame photometry compared to 61 ± 3%. The detectability of potassium in mitochondria was too low to enable the measurement of relaxation constants. This study indicates that differences in tissue structure, particularly mitochondrial content are important in determining 39K detectability and measured relaxation rates.

The role of immunoglobulins and their transporters in urogenital Chlamydial infections

Chlamydia trachomatis infections of the male and female reproductive tracts are the world's leading sexually transmitted bacterial disease, and can lead to damaging pathology, scarring and infertility. The resolution of chlamydial infection requires the development of adaptive immune responses to infection, and includes cell-mediated and humoral immunity. Whilst cluster of differentiation (CD)4+ T cells are known to be essential in clearance of infection [1], they are also associated with immune cell infiltration, autoimmunity and infertility in the testes [2-3]. Conversely, antibodies are less associated with inflammation, are readily transported into the reproductive tracts, and can offer lumenal neutralization of chlamydiae prior to infection. Antibodies, or immunoglobulins (Ig), play a supportive role in the resolution of chlamydial infections, and this thesis sought to define the function of IgA and IgG, against a variety of chlamydial antigens expressed during the intracellular and extracellular stages of the chlamydial developmental cycle. Transport of IgA and IgG into the mucosal lumen is facilitated by receptor-mediated transcytosis yet the expression profile (under normal conditions and during urogenital chlamydial infection) of the polymeric immunoglobulin receptor (pIgR) and the neonatal Fc receptor (FcRn) remains unknown. The expression profile of pIgR and FcRn in the murine male reproductive tract was found to be polarized to the lower and upper reproductive tract tissues respectively. This demonstrates that the two receptors have a tissue tropism, which must be considered when targeting pathogens that colonize different sites. In contrast, the expression of pIgR and FcRn in the female mouse was found to be distributed in both the upper and lower reproductive tracts. When urogenitally infected with Chlamydia muridarum, both male and female reproductive tracts up-regulated expression of pIgR and down-regulated expression of FcRn. Unsurprisingly, the up-regulation of pIgR increased the concentration of IgA in the lumen. However, down-regulation of FcRn, prevented IgG uptake and led to an increase or pooling of IgG in lumenal secretions. As previous studies have identified the importance of pIgR-mediated delivery of IgA, as well as the potential of IgA to bind and neutralize intracellular pathogens, IgA against a variety of chlamydial antigens was investigated. The protection afforded by IgA against the extracellular antigen major outer membrane protein (MOMP), was found to be dependent on pIgR expression in vitro and in vivo. It was also found that in the absence of pIgR, no protection was afforded to mice previously immunized with MOMP. The protection afforded from polyclonal IgA against the intracellular chlamydial antigens; inclusion membrane protein A (IncA), inclusion membrane proteins (IncMem) and secreted chlamydial protease-like activity factor (CPAF) were produced and investigated in vitro. Antigen-specific intracellular IgA was found to bind to the respective antigen within the infected cell, but did not significantly reduce inclusion formation (p > 0.05). This suggests that whilst IgA specific for the selected antigens was transported by pIgR to the chlamydial inclusion, it was unable to prevent growth. Similarly, immunization of male mice with intracellular chlamydial antigens (IncA or IncMem), followed by depletion CD4+ T cells, and subsequent urogenital C. muridarum challenge, provided minimal pIgR-mediated protection. Wild type male mice immunized with IncA showed a 57 % reduction (p < 0.05), and mice deficient in pIgR showed a 35 % reduction (p < 0.05) in reproductive tract chlamydial burden compared to control antigen, and in the absence of CD4+ T cells. This suggests that pIgR and secretory IgA (SIgA) were playing a protective role (21 % pIgR-mediated) in unison with another antigen-specific immune mechanism (36 %). Interestingly, IgA generated during a primary respiratory C. muridarum infection did not provide a significant amount of protection to secondary urogenital C. muridarum challenge. Together, these data suggest that IgA specific for an extracellular antigen (MOMP) can play a strong protective role in chlamydial infections, and that IgA targeting intracellular antigens is also effective but dependent on pIgR expression in tissues. However, whilst not investigated here, IgA targeting and blocking other intracellular chlamydial antigens, that are more essential for replication or type III secretion, may be more efficacious in subunit vaccines. Recently, studies have demonstrated that IgG can neutralize influenza virus by trafficking IgG-bound virus to lysosomes [4]. We sought to determine if this process could also traffic chlamydial antigens for degradation by lysosomes, despite Chlamydia spp. actively inhibiting fusion with the host endocytic pathway. As observed in pIgR-mediated delivery of anti-IncA IgA, FcRn similarly transported IgG specific for IncA which bound the inclusion membrane. Interestingly, FcRn-mediated delivery of anti-IncA IgG significantly decreased inclusion formation by 36 % (p < 0.01), and induced aberrant inclusion morphology. This suggests that unlike IgA, IgG can facilitate additional host cellular responses which affect the intracellular niche of chlamydial growth. Fluorescence microscopy revealed that IgG also bound the inclusion, but unlike influenza studies, did not induce the recruitment of lysosomes. Notably, anti-IncA IgG recruited sequestosomes to the inclusion membrane, markers of the ubiquitin/proteasome pathway and major histocompatibility complex (MHC) class I loading. To determine if the protection against C. muridarum infection afforded by IncA IgG in vitro translated in vivo, wild type mice and mice deficient in functional FcRn and MHC-I, were immunized, depleted of CD4+, and urogenitally infected with C. muridarum. Unlike in pIgR-deficient mice, the protection afforded from IncA immunization was completely abrogated in mice lacking functional FcRn and MHC-I/CD8+. Thus, both anti-IncA IgA and IgG can bind the inclusion in a pIgR and FcRn-mediated manner, respectively. However, only IgG mediates a higher reduction in chlamydial infection in vitro and in vivo suggesting more than steric blocking of IncA had occurred. Unlike anti-MOMP IgA, which reduced chlamydial infection of epithelial cells and male mouse tissues, IgG was found to enhance infectivity in vitro, and in vivo. Opsonization of EBs with MOMP-IgG enhanced inclusion formation of epithelial cells in a MOMP-IgG dose-dependent and FcRn-dependent manner. When MOMP-IgG opsonized EBs were inoculated into the vagina of female mice, a small but non-significant (p > 0.05) enhancement of cervicovaginal C. muridarum shedding was observed three days post infection in mice with functional FcRn. Interestingly, infection with opsonized EBs reduced the intensity of the peak of infection (day six) but protracted the duration of infection by 60 % in wild type mice only. Infection with EBs opsonized in IgG also significantly increased (p < 0.05) hydrosalpinx formation in the oviducts and induced lymphocyte infiltration uterine horns. As MOMP is an immunodominant antigen, and is widely used in vaccines, the ability of IgG specific to extracellular chlamydial antigens to enhance infection and induce pathology needs to be considered. Together, these data suggest that immunoglobulins play a dichotomous role in chlamydial infections, and are dependent on antigen specificity, FcRn and pIgR expression. FcRn was found to be highly expressed in upper male reproductive tract, whilst pIgR was dominantly expressed in the lower reproductive tract. Conversely, female mice expressed FcRn and pIgR in both the lower and upper reproductive tracts. In response to a normal chlamydial infection, pIgR is up-regulated increasing secretory IgA release, but FcRn is down-regulated preventing IgG uptake. Similarly to other studies [5-6], we demonstrate that IgA and IgG generated during primary chlamydial infections plays a minor role in recall immunity, and that antigen-specific subunit vaccines can offer more protection. We also show that both IgA and IgG can be used to target intracellular chlamydial antigens, but that IgG is more effective. Finally, IgA against the extracellular antigen MOMP can afford protection, whist IgG plays a deleterious role by increasing infectivity and inducing damaging immunopathology. Further investigations with additional antigens or combination subunit vaccines will enhance our understanding the protection afforded by antibodies against intracellular and extracellular pathogenic antigens, and help improve the development of an efficacious chlamydial vaccine.

Orchids : My Intersex Adventure

Orchids: My Intersex Adventure is a multi-award winning autobiographical documentary film. The film follows documentary filmmaker, Phoebe Hart, as she comes clean on her journey of self-discovery to embrace her future and reconcile the past shame and family secrecy surrounding her intersex condition. Despite her mother’s outright refusal to be in the film, Phoebe decides she must push on with her quest to resolve her life story and connect with other intersex people on camera. With the help of her sister Bonnie and support from her partner James, she hits the open road and reflects on her youth. Phoebe’s happy and carefree childhood came to an abrupt end at puberty when she was told she would never menstruate nor have children. But the reasons why were never discussed and the topic was taboo. At the age of 17, Phoebe’s mother felt she was old enough to understand the true nature of her body and the family secret was finally revealed. Phoebe then faced an orchidectomy, invasive surgery to remove her undescended testes, the emotional scars of which are still raw today. Phoebe’s road trip around Australia exposes her to the stories of other intersex people and holds a mirror to her own experience. She learns valuable lessons in resilience and healing but also sees the pervasive impact her condition has on all her relationships. At home, Phoebe and James want to start a family but dealing with infertility and the stress of the adoption process puts pressure on their marriage. Phoebe also starts to understand the difficult decisions her parents faced and is excited but apprehensive when they eventually agree to be interviewed. Will talking openly with her mother give Phoebe the answers she has been looking for? The film was produced and directed by Phoebe Hart and commissioned by the Australian Broadcasting Commission. The film premiered at the Brisbane International Film Festival in 2010 where it was voted the number one film of the festival by audiences. Orchids was broadcast on ABC1 in Australia in 2012, appeared in more than 50 film festivals internationally and has since been broadcast nationally in Switzerland, Sweden, Israel, Spain, France, Russia, Poland, Germany and the USA.

Molecular basis for enhancement of the meiotic DMC1 recombinase by RAD51 associated protein 1 (RAD51AP1)

Homologous recombination is needed for meiotic chromosome segregation, genome maintenance, and tumor suppression. RAD51AP1 (RAD51 associated protein 1) has been shown to interact with and enhance the recombinase activity of RAD51. Accordingly, genetic ablation of RAD51AP1 leads to enhanced sensitivity to and also chromosome aberrations upon DNA damage, demonstrating a role for RAD51AP1 in mitotic homologous recombination. Here we show physical association of RAD51AP1 with the meiosis-specific recombinase DMC1 and a stimulatory effect of RAD51AP1 on the DMC1-mediated D-loop reaction. Mechanistic studies have revealed that RAD51AP1 enhances the ability of the DMC1 presynaptic filament to capture the duplex-DNA partner and to assemble the synaptic complex, in which the recombining DNA strands are homologously aligned. We also provide evidence that functional cooperation is dependent on complex formation between DMC1 and RAD51AP1 and that distinct epitopes in RAD51AP1 mediate interactions with RAD51 and DMC1. Finally, we show that RAD51AP1 is expressed in mouse testes, and that RAD51AP1 foci colocalize with a subset of DMC1 foci in spermatocytes. These results suggest that RAD51AP1 also serves an important role in meiotic homologous recombination.