Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG-geförderten) Allianz- bzw. Nationallizenz frei zugänglich.

Tuberculosis remains a major public health issue globally, with an estimated 9.2 million new cases in 2006. A new threat to TB control is the emergence of drug resistant strains. These strains are harder to cure as standard anti-tuberculosis first line treatments are ineffective. Multi Drug Resistant Tuberculosis (MDR-TB) is defined as Mycobacterium tuberculosis that has developed resistance to at least rifampicin and isoniazid, and these strains now account for greater than 5% of worldwide cases. Mutations within the Rifampicin Resistance Determining Region (RRDR) of the rpoB gene are present in greater than 95% of strains that show rifampicin resistance by conventional drug susceptibility testing. As rifampicin mono resistance is extremely rare, and rifampicin resistance is usually associated with isoniaizd resistance, the RRDR region of the rpoB gene is a very useful surrogate marker for MDR-TB. Many molecular assays have been attempted based on this theory and have had varied levels of success. The three methods evaluated in this study are DNA sequencing of the rpoB, katG and inhA genes, the Genotype MTBDRplus line probe assay (Hain Lifesciences) and a novel method incorporating Real-Time PCR with High Resolution Melt analysis targeted at the RRDR using the Rotorgene 6000 (Corbett Lifesciences). The sensitivity for the detection of rifampicin resistance was far better using DNA sequencing or the commercially available line probe assay than detection by the Real-Time PCR method developed in this study.

Detta examensarbete handlar om lärares syn på lässtrategiundervisning. Då mina egna erfarenheter inom skolans årskurs 1–3 vittnar om att läsfixarna används som metod trots att det inte finns forskning som stödjer metoden, har intresset legat i att undersöka lärares syn på läsfixarna som metod. Ett hett ämne inom skoldebatten har länge varit svenska elevers sjunkande resultat i läsförståelse enligt kunskapsmätningar som PISA. Detta menar Barbro Westlund, lektor i läs- och skrivutveckling och deltagare i flera forskningsprojekt, beror på bristande förmåga att plocka fram de rätta strategierna i mötet med olika texttyper. Westlund betonar vikten av att undervisa om lässtrategier ämnesspecifikt vilket ökade mitt intresse av att även undersöka hur lärare använder läsfixarna i samhälls- och naturorienterande ämnen. Undersökningen bygger på sju intervjuer av verksamma lärare i årskurs 1–3 samt en observation av ett lektionstillfälle. Resultatet visar en positiv syn på läsfixarna som metod som går att knyta till tidigare forskning. Resultatet har även visat på att metoden till största del används i svenskämnet, men att det på flera sätt går att använda strategierna ämnesspecifikt. Den positiva synen på läsfixarna som metod väcker nya frågeställningar om varför det saknas vetenskapliga bevis på att metoden är effektiv för läsförståelseutveckling. Det här är dock något som öppnar nya dörrar för vidare undersökningar i bredare omfattning på huruvida läsfixarna som metod hjälper läsaren att utveckla läsförståelse eller inte. / This master thesis is about the teachers view of reading strategy teaching. As my own experiences within the school grades 1-3 testifies that “läsfixarna”, which is a method of reading comprehension strategies, as method is used despite there existing no research that supports the method. For this reason, thesis aims to investigate the teachers’ view of “läsfixarna” as a method. A hot topic in school debate has been for a long time the decreasing results for reading comprehension for Swedish students in knowledge test like the PISA test. This is a cause according to Barbro Westlund, a senior lecturer in reading and writing development and participant in multiple research projects, of the lack of ability to use the correct strategies when encountering different kinds of text. Westlund emphasizes the importance of teaching about the reading strategies specific to the study which increased the interest to also investigate on how teachers use “läsfixarna” in society and science studies. The present study is based on seven interviews with working teachers in grades 1-3 and an observation of a period. The results are show a positive view of “läsfixarna” as a method which is also connected to earlier research of reading comprehension strategies. The results also suggest that the method is largely used in the Swedish subject, but it is also possible to use the strategies in various subject-specific ways. The positive view of “läsfixarna” as a method raises new issues, as there is no scientific evidence that the method is effective for reading comprehension development. This is something that opens doors for further and new investigations to a much wider extent on whether “läsfixarna” as a method help readers to develop reading comprehension or not.

Diabetes mellitus currently affects over 422 million people globally and over 80% of patients with diabetes will develop diabetic retinopathy. Patients with diabetic retinopathy initially develop background retinopathy, which does not cause significant deterioration of visual function; however, background retinopathy may progress and lead to proliferative diabetic retinopathy and diabetic macular oedema, both of which cause severe visual dysfunction if left untreated. Current therapies for diabetic retinopathy include invasive intravitreal injections and laser photocoagulation; however these treatments only attenuate the progression of proliferative diabetic retinopathy and diabetic macular oedema. Aside from prevention by maintaining good blood glucose and blood pressure control, there are currently no treatments to prevent progression to late-stage diabetic retinopathy and new innovations in the field have not significantly progressed. For this reason, we have used untargeted –omics approaches to identify previously unknown pathological pathways in diabetes. In this thesis, I have analysed a range of trace metals in donor retinas and found that total copper was increased in diabetic retinas compared with non-diabetic. This result was replicated in streptozotocin-induced diabetic rat retinas and further evidenced by upregulation of metallothioneins and caeruloplasmin in diabetic rat retinas compared with non-diabetic. Treatment with the copper chelator triethylenetetramine modulated these changes, the downstream effects of which require further study. This is the first description, to our knowledge, of dysregulated copper homeostasis in the diabetic retina. I have also mapped metabolic changes in streptozotocin-induced diabetic rat retinas and found previously undescribed metabolite changes such as diabetes-induced downregulation of scyllo inositol. This coincided with substantial changes to retinal lipids during diabetes and changes to individual lipids were consistent within their respective class. I have also found a pattern whereby regardless of the extent of change to a lipid class in diabetes, lipids containing docosahexaenoic acid (22:6 carbon chain) were consistently downregulated. This is thought to be the first study to describe this range of metabolite changes in the diabetic retina but also the first study to describe this range of metabolite analysis concomitantly within the same tissue sample. The data from this study provides new insights into metallomic and metabolic dysfunction in diabetic retinopathy and shown that these data are reproducible. We suggest that there is plenty of scope for further research to investigate mechanisms behind copper dysregulation, how this affects pathogenesis of diabetic retinopathy along with new insights into dysregulated metabolic pathways.

The potential for private sector healthcare business in Africa has been forecasted to reach $35 billion by 2016, with South Africa being regarded as the most industrially advanced country on the continent. South Africa’s entry to modern biotechnology is fairly recent, though, with companies in the private sector still in a developmental phase, and most having limited bioproduct ranges. While considerable research has been conducted in the past to attempt to define the biotechnology environment of South Africa, as yet, a concise overview is lacking. In particular, a synopsis of the biomedical or commercial health technology environment has not been forthcoming for entrepreneurs to refer to as a ‘roadmap’. The purpose of this study was to perform a comprehensive study on the attributes that should be met for a successful, sustainable health technology venture (HTV) to be started in South Africa; while identifying the opportunities and threats that have existed in the South African market; thereby, affecting their success and sustainability to date. In this study, two phases of research were conducted. The first was a small-sampled mixed-methods (both qualitative and quantitative) study involving 21 medical devices, biogenerics, diagnostics, and contract services companies. The second was a quantitative study, involving 107 vaccines, biogenerics, therapeutics, nutraceuticals, reagents, diagnostics, medical devices, biotools, contract services and public services companies. Inferential statistical tests were conducted on the data, including Pearson’s Chi-Square, ANOVA, bivariate correlation, linear regression, logistic regression and multinomial logistic regression. From the study, the overall proportion of business sustainability for HTVs was found to be 66.7%, and at least 30% were unsustainable (or not yet at a level of sustainability). Variations were observed in the overall rate of sustainability for companies, based on their core functional classification, location, production type, size and start-up or R&D spending. By converting the observed frequencies of activity level, as an indication of sustainability, into a probability, it was possible to observe the company type that was most, and least likely to succeed in South Africa. Based on the statistical observations in this study, the HTV type most likely to succeed in South Africa, with a 63.7% probability of reaching sustainability, is a ‘vaccines’, ‘biotools’ or ‘public services’ company from Johannesburg with at least 20 employees; that has developed its goods or services internally, but manufactured externally and spent between R20 million–and–R30 million on its R&D or start-up. Conversely, least likely to succeed (3.2% probability) is a nutraceutical company from Cape Town with between six and 20 employees, that has developed and produced internally, and which has spent between R1 million–and–R10million on its start-up. / Life and Consumer Sciences / M. Sc. (Life Sciences)

Project title: Optimalization of the regime of marker's examination of clinically important infections in blood donors Project author: Daniela Dušková, M.D. Project supervisor: prof. Vladimír Tesař, M.D., Dr.Sc., MBA, FASN The aim of this project is to contribute to the discussion about introducing the methods of molecular biology into the routine blood donor testing in the transfusions departments in the Czech Republic. The theoretical part includes a brief history and some turning points in transfusion medicine. The next part within the theoretical section is dedicated to the problems of infectious diseases concerning transfusion and the general examination processes used during the selection of blood donors. The end of the theoretical part concentrates on existing possibilities of markers' examination of clinically important infections in blood donors, including the list of processes performed in the Czech Republic, the European Union and other countries. The practical part describes this study, ie. the routine screening test of blood donors using the CMIA method (a routine method) and using RT-Real Time PCR method (a molecular biology method) for detecting infectious markers (HCV, HBV, HIV). Within this part, the principle of both methods and the process of actual examinations are described in...

Rotavirus diarrhea is still an important cause of mortality in children worldwide, responsible each year for about 500,000 deaths. The introduction of the vaccine against the virus could lead to a reduction in morbidity and mortality associated with rotavirus. In Brazil, the Rotarix ® rotavirus vaccine, which contains the genotype G1P [8], was included in the national programme of immunization since March 2006. This project aims to evaluate clinical and epidemiological data related to infection by RVA, rotavirus genotypes prevalent. The research project is a cross-sectional study in which children with acute diarrhoea were recorded prospectively from 2010 to 2012. The location of collection of fecal samples was the Hospital of urgency of Sergipe in Pediatric Emergency sector, in Aracaju/SE. The clinical and epidemiological information was obtained through a questionnaire. The clinical severity was determined by a scoring system 20, calculated from the survey point. It was verified the episodes of acute diarrhea and stool samples collected for research and genotyping of rotavirus by ELISA and RT-PCR method.Descriptive statistical calculations were carried out to define the epidemiology of rotavirus. EIA positive results were found in 78 of the 790 samples. The most frequent genotype was G2 P [4], followed by the G8 genotype P [4], P [8] G1, G3 P [8], G1 P [6]. Mixed infections were detected G2 P [4] P [8], P [4] and G1G2 G2G8 P [4] and in general there was a cocirculação of different genotypes in the years studied, moreover, shows an alternation between the genotypes every year. The results obtained in this study observed a variability of positive cases distributed, confirming that the seasonality in the region is not remarkable. Therefore, new strains of rotavirus are emerging and associated with severe diarrhea. For this reason, monitoring is required to follow changes in the epidemiology of rotavirus. / A diarreia por rotavírus ainda é uma importante causa de mortalidade infantil em todo o mundo, sendo responsável a cada ano por cerca de 500.000 mortes. A introdução da vacina contra o vírus pode levar a uma redução da morbidade e mortalidade associadas ao rotavírus. No Brasil, a Rotarix®, a vacina contra o rotavírus, que contém o genótipo G1P[8], foi incluído no programa nacional de imunizações em março de 2006. Este projeto teve como objetivo avaliar dados clínicos, epidemiológicos e genótipos predominantes relacionados à infecção por RV-A. O projeto de pesquisa é um estudo transversal em que as crianças com diarreia aguda foram registrados prospectivamente a partir de 2010 a 2012. O local de coleta das amostras fecais foi o Hospital de Urgência de Sergipe no setor de Urgência Pediátrica, em Aracaju/SE. A gravidade clínica foi determinada por um sistema de pontuação 20, calculado a partir do ponto de questionário. Foram verificados os episódios de diarreia aguda e coletadas amostras de fezes para pesquisa e genotipagem de rotavírus pelo método ELISA e RT-PCR. Cálculos estatísticos descritivos foram realizados para definir a epidemiologia do rotavírus. Resultados positivos foram encontrados em 78 das 790 amostras. O genótipo mais frequente foi G2P[4], seguido do genótipo G8P[4], G1P[8], G3P[8], G1P[6]. Foram detectadas infecções mistas G2 P[4] P[8], G1G2 P[4] e G2G8 P[4] e de um modo geral observou-se uma cocirculação de distintos genótipos nos anos estudados, além disso, nota-se uma alternância entre os genótipos a cada ano. O resultado obtido neste estudo observou uma variabilidade dos casos positivos distribuídos, confirmando que a sazonalidade na região não é marcante. Portanto, novas cepas de rotavírus estão surgindo e associados à diarreia grave. Por esta razão, a vigilância contínua é necessária para acompanhar as mudanças na epidemiologia do rotavírus.

INTRODUÇÃO: Poucos estudos têm focado as modificações fisiológicas que ocorrem no trato genital de mulheres menopausadas infectadas pelo HIV e sua associação com a excreção genital do vírus. Nesse estudo de corte transversal, comparou-se a excreção genital do HIV em mulheres menopausadas e em idade fértil em acompanhamento em um centro especializado em São Paulo, Brasil. Investigou-se também a associação entre a excreção genital de RNA de HIV e a viremia em ambos os grupos. Fatores associados com a intensidade da excreção genital de HIV também foram pesquisados, incluindo achados ginecológicos e marcadores de progressão da infecção por HIV. MÉTODOS: 146 mulheres infectadas pelo HIV [73 menopausadas (M)/73 em idade fértil (F)] foram selecionadas em Serviço de Extensão ao Atendimento de Pacientes com HIV/Aids Casa da Aids do Hospital das Clínicas da FMUSP, São Paulo, Brasil. As mulheres menopausadas referiram tempo médio de 8,17 anos (DP=6 anos) de menopausa. A contagem de linfócitos T CD4+ foi obtida por citometria de fluxo e a quantificação do RNA do HIV no plasma e no lavado cervicovaginal (LCV) foi realizada por RT-PCR quantitativo, utilizando-se o kit Cobas Amplicor HIV-1 Monitor Test®, no método ultrasensível. Cloreto de lítio foi introduzido no tampão para obtenção do LCV e quantificado antes e depois da coleta do lavado, a fim de determinar o fator de diluição de cada amostra. A deteção do gene SRY por PCR também foi realizada a fim de eliminar amostras com eventual contaminação espermática. A prevalência de excreção genital foi estimada para ambos os grupos e os fatores associados à intensidade da excreção viral foram investigados, utilizando-se modelo de regressão linear múltipla. As variáveis com p<0,2 na análise bivariada foram incluídas na análise multivariada, assim como o grupo em estudo (M ou F). O modelo final incluiu fatores que se mostraram independentemente associados com a intensidade da excreção genital de HIV. RESULTADOS: A prevalência de excreção genital de HIV-RNA foi similar em ambos os grupos (M: 17,8%, IC 95% 9,8 28,5; F: 22%, IC 95% 13,1 33,1, p=0,678). Similarmente, a intensidade de excreção genital do HIV também não se mostrou diferente entre os grupos (mediana - M: 1,4log/mL; F: 1,4log/mL, p=0,587). A carga viral plasmática foi detectável em 34,2% das pacientes menopausadas (IC 95% 23,5 46,3) e em 42,5% entre as pacientes em idade fértil (IC 95% 31 54,6, p=0,395). Três pacientes (2 M/1 F) exibiram excreção genital de HIV-RNA na ausência de viremia detectável. Existe evidência de correlação entre a carga viral plasmática e a genital em ambos os grupos (rM: 0,658; rF: 0,684, p<0,01). Adicionalmente, o número de células CD4+ periféricas mostrou-se negativamente correlacionada à excreção genital do HIV em ambos os grupos (rM: -0,250; rF: -0,248, p<0,05). À análise multivariada, a carga viral plasmática mostrou-se independentemente associada à ocorrência de excreção genital do HIV em ambos os grupos (OR 4,03, IC 95% 2,52 6,45, p<0,001). Já a intensidade de excreção genital mostrou-se independentemente associada ao pH vaginal (p<0,001), concentração de TNF- no LCV (p=0,01), e à carga viral plasmática (p=0,001), todos com correlação positiva. CONCLUSÕES: Apesar das modificações significativas que ocorrem na mucosa vaginal da mulher menopausada, a excreção cervicovaginal do HIV parece não ser significativamente influenciada por esse estado. A carga viral plasmática e o número de células CD4+ periféricas estão correlacionadas com a excreção genital do vírus. A frequência de excreção genital mostrouse independentemente associada à intensidade de viremia. Além disso, o aumento do pH vaginal e evidência de inflamação genital, associada à concentração de TNF- no LCV, independentemente aumentam a intensidade de excreção genital nas mulheres estudadas. / BACKGROUND: Few studies have focused on physiological modifications that occur in the genital tract of HIV-infected postmenopausal women and their association with HIV cervicovaginal shedding. In this cross-sectional study we evaluated and compared HIV genital shedding among postmenopausal and fertile-aged women under care at a specialized center in Sao Paulo, Brazil, investigating the association between HIV-RNA shedding and HIV plasma viral loads in both groups. Factors associated with higher HIV shedding were also investigated, including gynaecological features and HIV disease progression markers. METHODS: 146 women living with HIV [73 postmenopausal (PM)/73 in fertile-aged (F)] were enrolled at the HIV Clinic, University of São Paulo Medical School, Brazil. Postmenopausal women referred a mean duration of 8.17y (SD=6y) since menopause. CD4+ cell counts were obtained by flow cytometry and HIV-RNA was quantified in plasma and in cervicovaginal lavages (CVL) by RT-PCR, using Cobas Amplicor HIV-1 Monitor Ultrasensitive Test. Lithium chloride was introduced into the CVL buffer and measured before and after CVL collection in order to determine the dilution factor for each specimen. SRY gene detection by PCR was also performed in all samples in order to rule out sperm contamination. Prevalence of HIV genital shedding was estimated for both groups and factors associated with the intensity of viral shedding were investigated, using a multiple linear regression model. Variables with p<0.2 in bivariate analysis were included in multivariate analysis, as well as the study group (PM and F). The final model included factors shown to be independently associated with intensity of HIV genital shedding. RESULTS: The prevalence of HIV-RNA genital shedding was similar in both groups. (PM: 17.8%, 95%CI 9.8 28.5; F: 22%, 95%CI 13.1 33.1, p=0.678). Likewise, the intensity of HIV shedding was shown not to differ between PM and F women (means - PM: 1.4log/mL; F: 1.4log/mL, p=0.587). Plasma viral loads were detectable in 34.2% of PM patients (95%CI 23.5 46.3), as compared to 42.5% among F women (95%CI 31 54.6) (p=0.395). Three patients (2 PM/1 F) exhibited HIV-RNA genital shedding in the absence of detectable viremia. We found evidence of correlation between HIV plasma viral load and HIV cervicovaginal shedding in both groups (rPM: 0.658; rF: 0.684, p<0.01). In addition, CD4+ cell counts were shown negatively correlated to HIV shedding in both groups (rPM: -0.250; rF: -0.248, p<0.05). In multivariate analysis, HIV plasma viral load was shown independently associated with occurrence of HIV genital shedding in both groups (OR 4.03, 95%CI 2.52 6.45, p<0.001). In addition, the intensity of HIV shedding was shown independently associated with vaginal pH (p<0.001), TNF- concentrations in CVL (p=0.01), and with HIV plasma viral loads (p=0.001), all of them with positive correlation. CONCLUSION: Despite the significant changes that occur in the vaginal mucosa of postmenopausal women, HIV cervicovaginal shedding does not seem to be significantly influenced by this state. Plasma viral loads and CD4+ cell counts are correlated to HIV genital shedding. The frequency of HIV genital shedding was shown independently associated with viremia intensity. Moreover, increased vaginal pH and evidence of genital inflammation associated with TNF- concentration independently enhanced the intensity of HIV shedding in postmenopausal and fertile-aged women.

In my workplace, the pupils' shortcomings in reading comprehension were noticed after a teacher at theschool noticed that only a few students actively participated in the whole class teaching and that few pupils could answer questions related to the text that was read in the whole class. The pupils in year sixat the school already have uneven results in reading comprehension and the reading lessons that has been conducted in the two classes has for the most part consisted of whole class tutoring and individual work.  Based on the development needs that I have noticed in my workplace, the purpose of this study is to analyze four student groups' work with text conversations and find out what role support has in text conversations to develop students' reading comprehension. Furthermore, the study intends to investigate whether text conversations in small groups can increase students' participation in reading instruction inthe the swedish subject. Based on this, the following questions have been formulated:  How can text conversations in small groups serve as support for students and promote the use of reading strategies? How can text conversations in small groups increase students' participation in reading lessons?  The analytical tools that has been used to analyse the empirical data is scaffolding, reciprocal teaching and the participation model. Qualitative methods have been used to collect empirical material. These arefocus group interviews and observations. The results show that support, participation and interaction are important concepts that can, to a large extent, effect students' reading comprehension development.

This presenting master’s thesis deals with a problem of digitaly controled room temperature using the software PLC. The result of this work was creating of an algorithm to control temperature on the requested leve ofl setpoint, minimizing operating costs of the building. The PLC softwares from different manufacturers were analyzed and compared with conventional PLC. The testing facility was disigned and used to control the temperature in the room and data visualization. Room temperature was regulated by PS controller or two point regulator.