Submitted by Renorbio (renorbioba@ufba.br) on 2018-04-25T17:41:58Z No. of bitstreams: 1 Tese de Doutorado - Maria da Conceição Aquino de Sá.pdf: 3251110 bytes, checksum: f7580c2d7b59b8cd7c2c1409212927e0 (MD5) / Approved for entry into archive by Delba Rosa (delba@ufba.br) on 2018-05-04T14:16:29Z (GMT) No. of bitstreams: 1 Tese de Doutorado - Maria da Conceição Aquino de Sá.pdf: 3251110 bytes, checksum: f7580c2d7b59b8cd7c2c1409212927e0 (MD5) / Made available in DSpace on 2018-05-04T14:16:30Z (GMT). No. of bitstreams: 1 Tese de Doutorado - Maria da Conceição Aquino de Sá.pdf: 3251110 bytes, checksum: f7580c2d7b59b8cd7c2c1409212927e0 (MD5) / FUNDACAO DE AMPARO A PESQUISA DO ESTADO DA BAHIA - EDITAL 013/2014 Programa De Bolsas De Mestrado e Doutorado / A procura por produtos da caprino-ovinocultura é frequente e contribui para aumentar a economia mundial, por isso é necessário produzir em larga escala e com qualidade para atender à demanda da população. Entretanto, diversos fatores na criação desses animais ainda necessitam melhorar, principalmente na área sanitária, na qual muitas doenças acometem o rebanho, dentre elas, a Linfadenite Caseosa, doença causada por Corynebacterium pseudotuberculosis, uma bactéria que apresenta sua toxicidade através de genes de virulência, assim como pela produção de biofilme, que protege a bactéria de ambientes hostis. Dessa forma, estudos sobre esse patógeno são necessários, a exemplo do sequenciamento genômico, na busca de conhecer genes para análises de biomoléculas e com isso compreender melhor a sua antigenicidade e imunogenicidade com o objetivo de melhorar a especificidade e sensibilidade de testes sorológicos, utilizados como prevenção da doença no rebanho. Além disso, é essencial entender o mecanismo de patogenicidade desta bactéria no hospedeiro e suas formas de defesa no sistema imunológico do animal. Assim, no presente estudo, foram caracterizados os genomas de quatro cepas de C. pseudotuberculosis, comparando as cepas produtoras (OVI2C, CAPJ4) e não produtoras (OVI03, CAP3W) de biofilme. As cepas foram extraídas para produção de antígenos somáticos e de superfícies. A partir de estudos prévios com immunoblotting, foram escolhidas as cepas de origem caprina (CAPJ4 e CAP3W) para o estudo in vivo com dezoito caprinos da raça Canindé, divididos em três grupos experimentais, um grupo controle e dois grupos infectados com cepas de C. pseudotuberculosis, para avaliar a patogenia clinicamente, realizando hemograma, além de outras análises sorológicas através do ensaio de ELISA e Western Blotting. Também foi realizada a cultura de células sanguíneas para o estudo da imunidade celular com IFN- e cultivo de células mononucleares do sangue periférico para observar a modulação da expressão de proteínas do sistema GTPases, Rab 5 e Rab 7. Após esses estudos, observou-se que os genes contidos no genoma das cepas de C. pseudotuberculosis possuem similaridade, mostrando que eles são altamente conservados. A extração das bactérias resultou em antígenos de superfície F3, que apresentaram reatividade no immunoblotting frente aos soros de ovinos experimentalmente infectados. De todos os antígenos, os de origem caprina, apresentaram melhor antigenicidade. No experimento in vivo, após a infecção dos animais, foram realizadas outras colheitas de sangue e foi possível observar que a cepa produtora de biofilme é mais resistente, causando danos maiores aos animais. Para as condições de infecção experimental, os valores do hemograma estão dentro dos parâmetros. Na análise sorológica os caprinos infectados experimentalmente responderam à exposição primária, o que contribui para o aumento da imunoglobulina IgG. A produção de IFN-  também foi observada. Foi verificado que as cepas de C. pseudotuberculosis produtora e não produtora de biofilme podem alterar a expressão de proteínas Rab 5 e Rab 7 em macrófagos derivados de células mononucleares do sangue periférico cultivados in vitro. Esses resultados forneceram informações importantes para a produção de testes diagnósticos, com maior acurácia. / The consumption of sheep and goat products is frequent and increases the values of world economy, so it is necessary to produce large scale and quality to meet the demand of population. However, several factors in the breeding of these animals still have to improve, especially in sanitary area, where many diseases affect the cattle, among them, Caseous Lymphadenitis, caused by Corynebacterium pseudotuberculosis, a bacterium that presents its toxicity through virulence genes, also by production of biofilm that protects the bacteria from hostile environments. Thus, studies of this pathogen are necessary, such as genomic sequencing, in search to know antigenic genes for biomolecule analysis and with this to understand better its antigenicity and immunogenicity, to be possible improve the specificity and sensitivity of serological tests, that are used as prevention of disease in the herd. Besides, it is essential to understand the mechanism of pathogenicity of this bacterium in host and its forms of defense against the animals immune system. At the present study, the genomes of four strains of C. pseudotuberculosis were characterized, comparing the biofilm producing (OVI2C, CAPJ4) and non-producing (OVI03, CAP3W) strains. These strains were extracted for production of somatic and surfaces antigens. From previous studies with immunoblottingting, strains of goat origin (CAPJ4 and CAP3W) were chosen for in vivo study with 18 Canindé goats, divided into three experimental groups, one control group and two groups infected with strains of C. pseudotuberculosis, to evaluate clinically the pathogenesis and through hemogram, in addition to other serological tests as ELISA assay and Western Blotting. Also was performed blood cell culture for the study of cellular immunity with IFN-, as culture of peripheral blood mononuclear cells to observe the modulation of expression of GTPases, Rab 5 and Rab 7 system proteins. After these studies, it was observed that the genes contained in the genome of the strains of C. pseudotuberculosis are genetically homogeneous, showing that they are highly conserved. The extraction of bacteria resulted in F3 surface antigens, that showed reactivity in immunoblottingting against sera from experimentally infected sheep. Of all antigens, those of goat origin had better antigenicity. At the in vivo experiment, after the infection of animals, other blood samples were collected and it was possible to observe that the biofilm producing strain is more resistant, causing greater damages to animals. For the conditions of experimental infection, blood count values are between normal limits. In the serological analysis, the experimentally infected goats responded to primary exposure, which contributes to increase of immunoglobulin IgG. The production of IFN- was also observed. In conclusion, it was observed that strains of C. pseudotuberculosis producing and not producing biofilm can modify the expression of Rab proteins in macrophages derived from mononuclear cells of peripheral blood cultivated in vitro. These results provided important information for production of diagnostic tests, with greater accuracy.

A pluralização dos atendimentos e sua garantia de realização em todos os níveis de atenção conforme os princípios estabelecidos pelo Sistema Único de Saúde (SUS) fizeram com que o atual cenário da saúde pública ganhasse grande complexidade envolvendo o financiamento e suporte para a manutenção desses serviços. Nesse contexto, alguns trabalhos na literatura relatam um excesso no uso desses recursos diagnósticos, que ocorre muitas vezes desnecessariamente, aumentando a chance de riscos à saúde do paciente e sobrecarregando demasiadamente o orçamento financeiro dos serviços de saúde, em particular o hospital. O que ocorre muitas vezes é a falta de critério nas solicitações e problemas de interpretação dos exames, desconhecimento sobre custos dos procedimentos realizados, entre outros fatores que se mostram mais elevados dependendo da complexidade do ambiente do serviço de saúde, do quadro clínico do paciente e da experiência do profissional médico. Dessa forma, é escopo desse projeto avaliar os resultados qualitativos, quantitativos e os custos dos principais parâmetros laboratoriais de exames solicitados nas áreas clínicas de Cardiologia e Endocrinologia da divisão de Clínica Médica do Hospital das Clínicas da Faculdade de Medicina de Ribeirão Preto da Universidade de São Paulo (HCFMRP-USP), e sua correlação com o que está descrito na literatura, objetivando identificar se o uso dos recursos diagnósticos estão sendo utilizados de forma racional e coerente. Para a realização desse estudo, foi realizada uma análise retrospectiva dos prontuários de pacientes pelo sistema HC-Athos em regime de primeira consulta e retorno, e também, de informações e metodologias relacionadas ao custeio de exames laboratoriais junto ao Centro de Custo do HCFMRP-USP, nas áreas clínicas de Endocrinologia, ambulatórios (ENA, END, ETP) e Cardiologia (CAA), no período de Julho de 2012 à Julho de 2013. Observando a análise dos elementos dos exames em ambos amulatórios, os resultados foram próximos: em Cardiologia a taxa média de elementos alterados foi de 20,79%, ao passo que em endocrinologia, o valor médio foi de 22,39%. Dessa forma, com base no que está descrito por alguns autores na literatura, esses achados podem sugerir a possibilidade de haver requisição sem necessidade dos exames laboratoriais. Em relação ao estudo de custos, observamos que existe uma diferença significativa para a relação: valor real gasto (HCFMRP-USP)/valor recebido(SUS). Na cardiologia onde o déficit foi mais elevado, 39,45%, contra os 12,80% observados em endocrinologia. Analisando cada exame separadamente nos dois ambulatórios, observa-se que há situações onde a relação do custo entre real gasto/valor recebido chega a ser bem discrepante, como a variação de 51 à 60% em exames como hemograma, uréia, creatinina, T3 e T4 total. Analisando esses achados, podemos concluir que há de certa forma um subfinanciamento do SUS em relação aos procedimentos laboratoriais realizados pelos hospitais. Com exceção de alguns exames, que isoladamente mostraram relação positiva, o saldo final considerando os exames mais comumente realizados foi negativo, ou seja, nos exames de maior demanda e faturamento, o resultado mostrou se preocupante. / The increasing demand of medical care and its realization warranty on all levels of care according to the principles established by the Unified Health System (SUS) made the current situation of public health gain great complexity involving funding and support to maintain these services. In this context, some studies in the literature report an excess in the use of these diagnostic tools, which is often unnecessarily, increasing the chance of risks to the health of the patient and overly burdening the financial budget of health services, particularly the hospital. What often happens is the lack of criteria in requests and problems of interpretation of the tests, ignorance about costs of the procedures performed, and other factors which show higher depending on the complexity of the health care environment, the patient\'s condition and the doctor\'s professional experience. Thus, is scope of this project, evaluate the qualitative and quantitative results, and the costs of major laboratory parameters tests requested in the clinical areas of Cardiology and Endocrinology of the University Hospital, Faculty of Medicine of RibeirãoPreto, University of São Paulo (HCFMRP-USP), and its correlation with what is described in the literature in order to identify whether the use of diagnostic resources are being used in a rational and coherent manner. To carry out this study, a retrospective analysis of medical records of patients at HC-Athos system in first consultation and return was carried out, and also, information and methodologies related to the cost of laboratory tests by the HCFMRP- USP Cost Center in the clinical areas of Endocrinology, outpatient clinics (ENA, END, ETP) and Cardiology (CAA) in the period July 2012 to July 2013. Noting the analysis of elements in both outpatient examinations, the results were similar: in Cardiology the average rate of change elements was 20.79%, while in endocrinology, the average value was 22.39%. Thus, based on what is described by some authors in the literature, these findings may suggest the possibility of request without laboratory tests. Regarding the study costs, we note that there is a significant difference to the relationship: real amount spent (HCFMRP- USP) / cash received (SUS). In cardiology where the deficit was higher, 39.45%, against 12.80% observed in endocrinology. Analyzing each exam separately in the two clinics, it is observed that there are situations where the ratio of the cost of real expenditure / amount received comes to be well outlier, as the range of 51 to 60% in exams such as blood count, urea, creatinine, T3 and T4 total. Analyzing these findings, we conclude that there is something of a SUS underfunded in relation to laboratory procedures performed by hospitals. Except for some tests, which alone showed a positive relationship, the final balance considering the most commonly performed tests was negative, that means, in higher demand and revenue tests, the results showed worrying.

No description available.

Indiana University-Purdue University Indianapolis (IUPUI) / Background: Ambulatory blood pressure (BP) monitoring (ABPM) remains a reference standard but the number of readings required to make the measurement valid has not been empirically validated. Methods: Among 360 patients with chronic kidney disease and 38 healthy controls, BP was recorded 2 per hour during the night and 3 per hour during the day over 24h using a validated ABPM device; all had at least 90% of the expected readings. From this full set of ABPM recording, a variable number of BP measurements were selected and we compared the performance of the selected readings against that of the full sample using random or sequential selection schemes. To address the question whether random or sequential selection schemes affect the diagnostic performance in diagnosing hypertension control we compared the diagnostic decisions reached with the subsample and the full sample using area under the receiver operating-characteristic curves (AUC ROC). To answer the question regarding the number of readings needed to achieve over 90% coverage of the mean BP of the full ABPM sample we ascertained the point and confidence interval (CI) estimates based on the selected data. Results: To diagnose hypertension control, the number of readings randomly drawn to establish lower bound with 2.5% error of area under the receiver operating-characteristic curve (AUC ROC) of 0.9 was 3, 0.95 was 7, and 0.975 was 13 . In contrast, the corresponding number of readings with serial selections was 18, 30 and 39 respectively. With a random selection scheme, 18 readings provided 80% coverage of the 90th percentile of CI of the true systolic BP mean, for 90% coverage, 26 readings were needed, for 95% coverage 33. With serial selections, the number of readings increased to 42, 47, and 50 respectively. Similar results emerged for diastolic BP. Conclusions: For diagnosing hypertension control 3 random measurements or 18 serial measurements is sufficient. For quantitative analysis, the minimal sufficient number of 24h ambulatory BP is 26 random recordings or 42 serial recordings.

Text in Afrikaans, summary in English / The object of the study was to determine if the use of different types of music would have an influence on the study effectiveness of the high school pupils. The music was used by the educational psychologist together with study methods. An empirical study was done in which three groups of students were used. The first group had no music, the second group had pop music and the last group had baroque music in their study session. The empirical study showed that music and then specifically baroque music had an influence on the study effectiveness of pupils. A program was compiled in which guidelines were given for study, the program is now used in a study program for high school students. Recommendations were made for the use of music with study methods for the Educational Psychologist, teachers and pupils / Educational Studies / M. Ed. (Voorligting)

Bacterial bloodstream infections (BSIs) are a major healthcare problem causing high mortality and economic cost. BSIs require an immediate initiation of antibiotic therapy as any delay is associated with a mortality increase. With the emergence of antimicrobial resistance, the choice of the appropriate antibiotic becomes increasingly difficult, thus creating an urgent need for new diagnostics, ideally to be done at the point of care. The current gold standard is blood culture with subsequent susceptibility testing although several molecular methods have recently entered the market. However, in many instances there is a discrepancy between the in-vitro data provided by the test and the outcome of antimicrobial therapy in-vivo because current diagnostics fail to take into account the impact of the environment in the patient such as the immune system, pharmacokinetics and pharmacodynamics or bacterial fitness. In this thesis, it was hypothesised that the measurement of the bacterial gene expression after the beginning of antibiotic therapy might be a more accurate indicator of the therapy outcome because it reflects the bacterial response under in-vivo conditions. In the first part of the thesis the expression of a set of pre-defined mRNA markers was investigated under various conditions. Experiments conducted with clinical E. coli isolates incubated in human whole blood revealed an excellent correlation between the gene expression, the treatment outcome, the antibiotic susceptibility and the genetic background for three different classes of antimicrobial drugs. The second part of the thesis describes the extraction of bacterial RNA from human whole blood specimen. The effect of different agents for the lysis of human blood cells and the impact of co-purified human RNA were analysed and a method for high yield extraction of undegraded bacterial RNA was established. The third part of the thesis investigates two methods for the sensitive measurement of the bacterial gene expression. This is relevant because the bacterial loads in BSI patients are extremely low. For genes with high gene expression levels both methods yielded reliable results but were unable to quantify the expression of the previously investigated mRNA markers due to their low copy numbers. Other approaches, especially those based on single cell measurements, might be able to overcome the problem in the future and should be explored in greater detail. Overall, the foundations for a future diagnostic test based on the measurement of the bacterial gene expression have been laid in this work. Future work should address the mRNA quantification and further evaluate the connection between gene expression and therapy outcome, e.g. in animal models. A future diagnostic test should also fulfil point-of-care requirements. This will include integrated sample preparation and quantification as well as a time-to-result in the range of a few minutes.

The main purpose of this study was to develop and administer a four-tier test for assessing Turkish pre-service physics teachers&rsquo / misconceptions about geometrical optics. Sixteen pre-service physics teachers, who were selected by maximum variation and convenience sampling methods from three state universities in Ankara, were interviewed in the contexts of plane mirrors, spherical mirrors, and lenses. From these interviews and the studies in the literature, the Open-Ended Geometrical Optics Test was developed. It was administered to 52 pre-service physics teachers from three state universities selected by purposive and convenience sampling. The responses of each subject for each item were categorized in order to determine the alternatives of the multiple-tier multiple-choice misconception test. The initial form of the test, the Three-Tier Geometrical Optics Test (TTGOT), was administered to 53 pre-service physics teachers from three state universities selected by purposive and convenience sampling as a pilot study. The analysis of the results of the TTGOT was used to revise the test items. Finally, the Four-Tier Geometrical Optics Test (FTGOT) was developed and administered to 243 pre-service physics teachers from 12 state universities in Turkey. The validity of the FTGOT scores was established by means of some qualitative and quantitative methods. The Cronbach alpha reliability coefficients were calculated for different test scores. Those for the total correct scores and standardized misconception scores (SUMM4) were found to be .59 and .42, respectively. Some misconceptions, which were held by more than 10 % of the pre-service teachers, were identified and considered to be significant.

The main purpose of this study was to develop a three-tier test for assessing tenthgrade students&rsquo / difficulties about kinematics graphs. In a three-tier test, first tier is classical multiple-choice question, the second tier is also classical multiple-choice question but presents reasons for answers given to the first tier and the third tier asks existence of confidence about the first two tiers. To develop a three-tier test, Turkish translation of the Test of Understanding Graphs in Kinematics (TUG-K) developed by Beichner (1994) was used. One more essay type question of asking students&rsquo / reasons of their answers and blank alternatives to write any suggestion different from choices were added to the each item of Turkish translation of this test translated by Delialioglu (2003). Finally, Kinematics Graphs Test Requesting Reasoning (KGTRR) was developed and administered to 253 students. To determine the distracters of the second-tiers of the Kinematics Graphs Three-Tier Test (KGTTT), answers on each item in KGTRR were categorized according to similarities in their meanings. Considering the frequency of these categories, the KGTTT was developed and administered to 495 students. Both quantitative and qualitative methods were used to determine the validity of the KGTTT. A positive correlation coefficient was calculated between student scores for the first two tiers and confidence levels for the third tiers. Also, percentages of false positives and false negatives were estimated. Cronbach alpha reliability coefficients of correct answers and difficulties of the students for all three tiers together were calculated as 0.84 and 0.69, respectively.

In this study, meta-analysis of diagnostic tests, Summary Receiver Operating Characteristic (SROC) curve, bivariate random effects and Hierarchical Summary Receiver Operating Characteristic (HSROC) curve theories have been discussed and accuracy in literature of Fine Needle Aspiration (FNA) biopsy that is used in the diagnosis of masses in breast cancer (malignant or benign) has been analyzed. FNA Cytological (FNAC) examination in breast tumor is, easy, effective, effortless, and does not require special training for clinicians. Because of the uncertainty related to FNAC&lsquo / s accurate usage in publications, 25 FNAC studies have been gathered in the meta-analysis. In the plotting of the summary ROC curve, the logit difference and sums of the true positive rates and the false positive rates included in the meta-analysis&lsquo / s codes have been generated by SAS. The formula of the bivariate random effects model and hierarchical summary ROC curve is presented in context with the literature. Then bivariate random effects implementation with the new SAS PROC GLIMMIX is generated. Moreover, HSROC implementation is generated by SAS PROC HSROC NLMIXED. Curves are plotted with RevMan Version 5 (2008). It has been stated that the meta-analytic results of bivariate random effects are nearly identical to the results from the HSROC approach. The results achieved through both random effects meta-analytic methods prove that FNA Cytology is a diagnostic test with a high level of distinguish over breast tumor.

Text in Afrikaans, summary in English / The object of the study was to determine if the use of different types of music would have an influence on the study effectiveness of the high school pupils. The music was used by the educational psychologist together with study methods. An empirical study was done in which three groups of students were used. The first group had no music, the second group had pop music and the last group had baroque music in their study session. The empirical study showed that music and then specifically baroque music had an influence on the study effectiveness of pupils. A program was compiled in which guidelines were given for study, the program is now used in a study program for high school students. Recommendations were made for the use of music with study methods for the Educational Psychologist, teachers and pupils / Educational Studies / M. Ed. (Voorligting)