Assigning human genes to diseases and related phenotypes is an important topic in modern genomics. Human Phenotype Ontology (HPO) is a standardized vocabulary of phenotypic abnormalities that occur in human diseases. Computational methods such as label-propagation and supervised-learning address challenges posed by traditional approaches such as manual curation to link genes to phenotypes in the HPO. It is only in recent years that computational methods have been applied in a network-based approach for predicting genes to disease-related phenotypes. In this thesis, we present an extensive benchmarking of various computational methods for the task of network-based gene classification. These methods are evaluated on multiple protein interaction networks and feature representations. We empirically evaluate the performance of multiple prediction tasks using two evaluation experiments: cross-fold validation and the more stringent temporal holdout. We demonstrate that all of the prediction methods considered in our benchmarking analysis have similar performance, with each of the methods outperforming a random predictor. / Master of Science / For many years biologists have been working towards studying diseases, characterizing dis- ease history and identifying what factors and genetic variants lead to diseases. Such studies are critical to working towards the advanced prognosis of diseases and being able to iden- tify targeted treatment plans to cure diseases. An important characteristic of diseases is that they can be expressed by a set of phenotypes. Phenotypes are defined as observable characteristics or traits of an organism, such as height and the color of the eyes and hair. In the context of diseases, the phenotypes that describe diseases are referred to as clinical phenotypes, with some examples being short stature, abnormal hair pattern, etc. Biologists have identified the importance of deep phenotyping, which is defined as a concise analysis that gathers information about diseases and their observed traits in humans, in finding genetic variants underlying human diseases. We make use of the Human Phenotype Ontology (HPO), a standardized vocabulary of phenotypic abnormalities that occur in human diseases. The HPO provides relationships between phenotypes as well as associations between phenotypes and genes. In our study, we perform a systematic benchmarking to evaluate different types of computational approaches for the task of phenotype-gene prediction, across multiple molecular networks using various feature representations and for multiple evaluation strategies.

Multiple hurdles, such as drug solubility, stability, and physical barriers in the body, hinder bioavailability of many promising therapeutics. Polymeric nanocarriers can encapsulate the therapeutics to protect non-target areas from side effects but also protect the drug from premature degradation for increased circulation and bioavailability. To capitalize on these advantages, the polymer nanoparticle must be properly engineered for increased control in size distribution, therapeutic encapsulation, colloidal stability, and release kinetics. However, each application requires a specific set of characteristics and properties. Being able to tailor these by manipulation of different design parameters is essential to optimize nanoparticles for the application of interest. This study of nanoparticle fabrication and characterization takes us a step closer to building effective delivery systems tailored for specific treatments. Poly(ethylene oxide)-b-poly(D,L-lactic acid) (PEO-b-PDLLA) based nanoparticles were produced to range from 100-200 nm in size. They were fluorescently labeled with a hydrophobic dye 6-13 bis(triisopropylsilylethynyl) pentacene (TIPS pentacene) at an optimal loading of 0.5 wt% with respect to the core. Surfaces were successfully coated with streptavidin to be readily functionalized with various biotinylated compounds such as PD-L1 antibodies or A488 fluorophore. Using the same PEO-b-PDLLA, iron oxide and a conjugated polymer poly(2- methoxy-5-(2-ethylhexyloxy)-1,4-phenylenevinylene) (MEH-PPV) were co-encapsulated to form fluorescently labeled magnetic particles. Using poly(lactic-co-glycolic acid), CRISPR-Cas9 plasmids were encapsulated at 1.6 wt% and most of the payload released within the first 24 hours. The incorporated plasmids were intact enough to have mammalian macrophages successfully express the bacterial protein Cas9. Using similar PLGA based particles, the surface was functionalized with streptavidin and bound to the surface of bacteria as an active carrier for increased penetration of solid tumors averaging ~23 particles per bacterium. PEO-b-PLGA based particles were used in conjunction with a hydrophobic salt former to encapsulate a peptide designed to reduce platelet binding to cancer cells and mitigate extravasation. The peptide encapsulated was increased from < 2 wt% without salt former to 8.5 wt% with the used of hexadecyl phosphonic acid. Although the applications across these projects can be broad, the fundamentals and important design parameters considered contribute to the overarching field of effective carriers for drug delivery. / Ph. D. / There are many reasons why many promising pharmaceutical formulations never make it through regulation and onto market, including low solubility of the drug, low absorbance by the body, and harmful side effects, to name a few. Using polymer drug carriers, these difficulties can be overcome by holding the drug in a more soluble carrier, releasing it on a certain timeline or to a specific location to increase absorbance and decrease side effects. When designing a carrier, the requirements for the product are dependent on the application and the disease of interest. This work looks at the material types and conditions during particle formation to see how it affects the final product to better define and understand how these parameters change the performance. This work shows that the carrier size can be manipulated depending on how much of one material is used versus the other, they can be labeled to fluoresce so they can be tracked during cell and animal studies, and they can be coated with targeting compounds on the surface to increase the specificity of the carrier to localize to a target location of interest. Different particles containing DNA for gene editing, peptides for cancer therapies, and magnetic iron oxides to increase transport across difficult cell barriers have all be fabricated and characterized. The lessons learned through these projects will help guide future work to more effective and efficient delivery of pharmaceuticals to the body.

Functionally low-literate consumers may often struggle to complete everyday adult tasks, such as reading food labels. Food labels are an important source of information for consumers, and assist them to make informed and wise food purchase decisions. However, the manner in which functionally low-literate consumers read, understand and apply this information to their decisionmaking, is different to literate consumers, and minimal research has been conducted regarding functionally low-literate consumers and their use of food labels in South Africa. This study was conducted in a predominantly low-literate and low-income rural area of Valspan, South Africa. Therefore, using food labels to make healthy and financially sound food choices may be considered especially important for this group of consumers. The current study served to conduct an in-depth literature review of functionally low-literate consumers‟ use of food labels in a rural area; explore whether and how functionally low-literate consumers use (read, understand and apply to decision-making) food label information; explore and describe the differences between the demographic characteristics of low-literate consumers and their use (reading, understanding and application) of food labels; make recommendations to educators on how functionally low-literate consumers can improve their use of food labels in a rural area; as well as to make recommendations on how food labels can be adapted to be more user-friendly to functionally low-literate consumers. Data was collected, using 292 interviewer administrated questionnaires, using purposive criterion sampling. The inclusion criteria for respondents were that they: had to be older than 18 years, living in Valspan and must have completed between grades 5 and 8 at school. Data analysis was done, using descriptive statistics, T-tests, ANOVA‟s, Spearman‟s rank order correlations and two-way frequency tables. Effect sizes were taken into consideration for all differences and associations. The results showed that respondents in this study did read food labels. They were also able to understand some simple aspects of the food label, but struggled with other aspects. When respondents struggled to understand food labels, they were selective about who they would ask to assist them, favouring the help of familiar family members and friends. Regarding the respondents‟ ability to apply food label information to their decision-making, respondents were able to identify several store logos, probably due to their tendency to pictorial thinking. Food-related calculations were relatively well completed; however, certain calculation-related terminology was not well understood by respondents. Food label symbols were not well identified, indicating that respondents did not have a good understanding of the meaning of these symbols. Literacy is vital to the use of food labels, as respondents who showed higher literacy levels showed a tendency towards better understanding of food label information, food label symbols and store logos. They also tended to be better equipped to correctly execute product-related calculations. Regarding demographics, respondents who spoke English and Afrikaans and who had a higher income had a tendency to better understand food label information, than respondents who spoke other languages and belonged to lower-income groups. If food labels are adapted, so that even lowliterate consumers are able to efficiently use food labels, they will be able to make informed and wise food product choices. This situation would be beneficial to low-literate and low-income consumers, marketers and retailers, as products with usable labels may consequently become the preferred choice of low-literate consumers and money wary low-income consumers. Additionally, marketers and retailers would be able to capitalise on the mass purchasing power that this target market represents. / M. Consumer Science, North-West University, Potchefstroom Campus, 2014

Functionally low-literate consumers may often struggle to complete everyday adult tasks, such as reading food labels. Food labels are an important source of information for consumers, and assist them to make informed and wise food purchase decisions. However, the manner in which functionally low-literate consumers read, understand and apply this information to their decisionmaking, is different to literate consumers, and minimal research has been conducted regarding functionally low-literate consumers and their use of food labels in South Africa. This study was conducted in a predominantly low-literate and low-income rural area of Valspan, South Africa. Therefore, using food labels to make healthy and financially sound food choices may be considered especially important for this group of consumers. The current study served to conduct an in-depth literature review of functionally low-literate consumers‟ use of food labels in a rural area; explore whether and how functionally low-literate consumers use (read, understand and apply to decision-making) food label information; explore and describe the differences between the demographic characteristics of low-literate consumers and their use (reading, understanding and application) of food labels; make recommendations to educators on how functionally low-literate consumers can improve their use of food labels in a rural area; as well as to make recommendations on how food labels can be adapted to be more user-friendly to functionally low-literate consumers. Data was collected, using 292 interviewer administrated questionnaires, using purposive criterion sampling. The inclusion criteria for respondents were that they: had to be older than 18 years, living in Valspan and must have completed between grades 5 and 8 at school. Data analysis was done, using descriptive statistics, T-tests, ANOVA‟s, Spearman‟s rank order correlations and two-way frequency tables. Effect sizes were taken into consideration for all differences and associations. The results showed that respondents in this study did read food labels. They were also able to understand some simple aspects of the food label, but struggled with other aspects. When respondents struggled to understand food labels, they were selective about who they would ask to assist them, favouring the help of familiar family members and friends. Regarding the respondents‟ ability to apply food label information to their decision-making, respondents were able to identify several store logos, probably due to their tendency to pictorial thinking. Food-related calculations were relatively well completed; however, certain calculation-related terminology was not well understood by respondents. Food label symbols were not well identified, indicating that respondents did not have a good understanding of the meaning of these symbols. Literacy is vital to the use of food labels, as respondents who showed higher literacy levels showed a tendency towards better understanding of food label information, food label symbols and store logos. They also tended to be better equipped to correctly execute product-related calculations. Regarding demographics, respondents who spoke English and Afrikaans and who had a higher income had a tendency to better understand food label information, than respondents who spoke other languages and belonged to lower-income groups. If food labels are adapted, so that even lowliterate consumers are able to efficiently use food labels, they will be able to make informed and wise food product choices. This situation would be beneficial to low-literate and low-income consumers, marketers and retailers, as products with usable labels may consequently become the preferred choice of low-literate consumers and money wary low-income consumers. Additionally, marketers and retailers would be able to capitalise on the mass purchasing power that this target market represents. / M. Consumer Science, North-West University, Potchefstroom Campus, 2014

Introdução: O uso de formas farmacêuticas inadequadas à pediatria, também conhecido como uso off-label, pode levar a uma terapia medicamentosa insegura. No ambiente hospitalar, em especial nas Unidades de Terapia Intensiva Pediátrica, há alta prevalência de prescrições com medicamentos não apropriados para crianças, entre eles os antimicrobianos, devido à gravidade das infecções, o estado crítico dos pacientes internados, a maior realização de procedimentos invasivos e uma maior incidência de bactérias resistentes. Em geral, o uso de antimicrobianos em crianças, bem como dos demais grupos farmacológicos, tem sido baseado principalmente em extrapolações e adaptações do uso em adultos, informações obtidas de raros estudos observacionais e consensos de especialistas na área. Devido ao estado crítico dos pacientes, as complexas terapias envolvendo antimicrobianos nem sempre são feitas de forma adequada, podendo trazer prejuízos à saúde dos pacientes e contribuir, ainda, para a resistência bacteriana. Objetivo: Avaliar a incidência do uso de antimicrobianos em pacientes internados em Unidade de Terapia Intensiva Pediátrica. Método: Estudo do tipo transversal realizado na Unidade de Terapia Intensiva Pediátrica do município de Rio Branco, Acre, no período de agosto de 2014 a julho de 2015. A amostra foi composta por 246 crianças de zero a onze anos de idade, internadas com quaisquer patologias. Foram analisadas as prescrições das primeiras 24 horas. Pesquisou-se o uso off label de antimicrobianos e potenciais interações medicamentosas com esses medicamentos. Resultados: Duzentas e trinta e uma crianças (93,9 por cento ) receberam a prescrição de pelo menos um antimicrobiano. O antimicrobiano mais prescrito foi a ceftriaxona. Em média, cada paciente recebeu dois antimicrobianos. Em 66,8 por cento dos casos, houve uso off-label, sendo a dose prescrita superior ao recomendado em 41 por cento , dos casos. Em 56 por cento das prescrições havia pelo menos uma potencial interação medicamentosa envolvendo antimicrobianos. Ampicilina e gentamicina foram os antibióticos que mais se envolveram em interações. Conclusões: a alta frequência do uso off-label e interações medicamentosas potenciais envolvendo antimicrobianos foi confirmada neste estudo. Tratar crianças como adultos pequenos pode expor esses pacientes a eventos adversos a medicamentos que comprometem a segurança desse grupo específico de pacientes. Isso aponta para a necessidade de equipes multidisciplinares trabalharem em conjunto e serem estimuladas a realizar mais estudos que garantam a segurança do uso de medicamentos na pediatria. / Introduction: The use of inadequate dosage forms for pediatrics, also known as off-label use, may lead to unsafe drug therapy. In the hospital environment, especially in Pediatric Intensive Care Units, there is a high prevalence of prescriptions with drugs not suitable for children, among them antimicrobial agents, due to the severity of the infections, the critical state of the hospitalized patients, more accomplishment of invasive procedures and a higher incidence of resistant bacteria. In general, the use of antimicrobials in children, as well as other pharmacological groups, has been based mainly on extrapolations and adaptations of the use in adults. This information was obtained from rare observational studies and consensus of specialists in the area. Due to the critical condition of patients, the complex therapies involving antimicrobial agents are not always adequately done, which may cause harm to the patients\' health and also contribute to bacterial resistance. Objective: To evaluate the incidence of antimicrobial use in patients hospitalized in a Pediatric Intensive Care Unit. Method: A cross-sectional study conducted at the Pediatric Intensive Care Unit of the municipality of Rio Branco, Acre, from August 2014 to July 2015. The sample consisted of 246 children from zero to eleven years old and hospitalized with any pathologies. The prescriptions of the first 24 hours were analyzed. The use of off-label antimicrobials and potential drug interactions with these drugs was investigated. Results: Two hundred and thirty-one children (93.9 per cent ) received the prescription of at least one antimicrobial. The most commonly prescribed antimicrobial agent was ceftriaxone. On average, each patient received two antimicrobials. In 66.8 per cent of cases, there was an off-label use, with the prescribed dose being higher than recommended in 41 per cent of the cases. In 56 per cent of prescriptions there was at least one potential drug interaction involving antimicrobials. Ampicillin and gentamicin were the antibiotics most involved in interactions. Conclusions: the high frequency of off-label use and potential drug interactions involving antimicrobials was confirmed in this study. Treating children as young adults may expose these patients to adverse drug events that compromise the safety of this particular group of patients. That points to the need for multidisciplinary teams to work together and be encouraged to carry out more studies that ensure the safety of use of medication in pediatrics.

Objetivos: Determinar a frequência de prescrições de medicamentos off label e não licenciados para pediatria na Atenção Primária à Saúde do município de Viamão, no Rio Grande do Sul. Método: Estudo descritivo, analisando prescrições de 323 pacientes emitidas de Agosto a Dezembro de 2012 em duas unidades de atenção primária do Sistema Público de Saúde do município de Viamão, no Rio Grande do Sul. Resultados: Durante o período estudado obteve-se a prescrição de 731 medicamentos e houve frequência de 31,7% de medicamentos prescritos off label, especialmente anti-histamínicos e antiasmáticos (32,3% e 31,5%, respectivamente). O principal tipo de prescrição off-label foi dose (38,8%), seguida de idade (31,5%) e de frequência de administração (29,3%). Com relação à prescrição off label de dose, foi mais frequente a sobredose 93,3% do que a subdose 6,7%. Não foram encontradas prescrições de medicamentos não licenciados. Discussão e Conclusão: As dificuldades relacionadas às pesquisas com crianças propiciam a prescrição de medicamentos off label, prática que, apesar de não ser ilegal, gera insegurança em relação aos possíveis efeitos adversos em uma população com características peculiares como a pediátrica. O estudo mostrou que esta prática é comum na Atenção Primária a Saúde em uma cidade do Rio Grande do Sul, a exemplo de pesquisas em cidades europeias. Este é o primeiro estudo que analisa a frequência de prescrição de medicamentos off label no âmbito ambulatorial do Sistema Único de Saúde no país e espera-se que possa contribuir para a busca de alternativas que promovam o uso racional de medicamentos na pediatria. / Objectives: To determine the frequency of prescriptions for off label and unlicensed medicines for pediatrics at Viamão Primary Health Care in Rio Grande do Sul. Method: Descriptive study analyzing prescriptions of 323 patients, issued from August to December 2012, in two primary care units of Viamão Public Health System in Rio Grande do Sul. Results: During the study period 731 prescription drugs were observed. Off label frequency was 31.7%, especially antihistamines and anti-asthmatic prescriptions (32.3% and 31.5%, respectively). The main type of off-label prescribing was dose (38.8%), followed by age (31.5%) and frequency of administration (29.3%). Regarding the prescription off label of dose, was more frequent overdose (93.3%) than underdose (6.7%). No prescriptions for unlicensed medicines were found. Discussion and Conclusion: The difficulties related to research with children favor prescription drugs off label, a practice that is not illegal, but may produce adverse effects in a population with unique characteristics such as pediatric. The study showed that this practice is common in Primary Health Care in a city in Rio Grande do Sul, as it is in Europe, as reported by other researches. This is the first study that analyzes the frequency of off-label prescribing of drugs in outpatient under the National Health System in Brazil and it is expected to be able to contribute to the search for alternatives that promote rational drug use in pediatrics.

Thesis was carried out to make an estimation of the willingness of consumers to buy the clothes produced under eco-friendly and decent way. Neoclassic demand theory was used and basic idea was to know that either moral effects on a consumer’s individual decision while choosing the products manufactured under friendly environment or not; if yes, then how and how much. Survey method along with choice experiment analysis was used to collect the data. Selected area was Boras Sweden and 50 respondents were considered for the survey. To estimate the degree of willingness of a consumer to pay for a Swan labeled and/or Fairtrade labeled jeans, choice experiments was used. Afterwards, the collected data was used within econometric models. It has been observed from results that consumers were more willing to pay for a Swan labeled and/or Fairtrade labeled jeans than non-labeled jeans. The responsible individuals regarding environment were much more willing to pay for Fairtrade and/or Swan labeled clothes, considering the factor that production does not affect the environment in negative way and it was done under socially acceptable conditions. / Program: Magisterutbildning i Fashion Management

Neste trabalho são estudadas as mioglobínas de Aplysía Brasiliana (MbApB) e da tartaruga marinha \"Dermoche lis Coriacea\" (MbT) focalizando a transição ácida alcalina (TAA), a interação com metais de transição e mudanças conformacionais induzidas - termicamente com objetivo de observar diferenças estruturais destas mioglobinas. A TAA da MbApB possui um pK = 7,2 quando observada Fe3+ em espectro do Fe3+ em g1 = 5,83 e pK = 7,5 quando medida através da absorção óptica (&#955=590nm). O espectro de EPR do Fe+ a pH alcalino revela uma distorção rômbica no campo cristalino do ferro que esta de acordo com a ausência da histidina distantes ta proteína. Além disso, as linhas associadas à configuração de baixo spin são bastante alargadas. Este alargamento de linha PQ de ser explicado por uma flutuação no posicionamento da heme em relação aos eixos de simetria. A MbApB forma complexos com o Cu2+ e Mn2+ . Existe som te um sitio de ligação para esses metais na proteína. Este sítio provavelmente envolve ligantes comuns ao Mn2+ e Cu2+ pois a ligação é competitiva, indicando que o complexo com o Cu2+ é mais estável do que com Mn2+ (KAMn2+ = (11,5-0,8).103 M-1 ). O complexo MbApB: Cu2+ exibe uma transição dependente do pH indicando uma mudança na coordenação do Cu2+ . Esta transição pode ser ajustada teoricamente admitindo-se uma interconversão entre duas formas, fornecendo um pK = 11,0. Observa-se também que a complexação da MbApB com mais de um Cu2+ provoca desnaturação. A TAA da MbT foi estudada por absorção óptica (pK=8,4) e EPR (pK = 8,3). Os espectros de EPR mostram que o Fe3+ esta num campo cristalino com simetria axial e distorção rômbica. As medidas de absorção óptica revelam pontos isosbéstioos praticamente idênticos aos de outras mioglobinas e um espectro na região de Soret típico de hemoproteínas que possuem a histidina distal. Estes dados permitem antecipar que a MbT deve possuir os mesmos resíduos de aminoácidos na região da heme que a Mb(II), porem com a histidina E-7 ligeiramente inclinada em relação ao eixo de simetria da heme. As mudanças conformacionais induzidas pela temperatura foram estudadas para a MbT em várias pH\'s. Esta proteína possui uma cisteína em sua cadeia polipeptídica que permite a ligação de um marcador de spin especifico para os grupos SR. Os resultados obtidos através do espectro de EPR do marcador de spin e da absorção óptica do Fe3+ permitem concluir que esta proteína exibe o fenômeno de pré-desnaturação e a transição da forma nativa para a desnaturada é o resultado de uma seqüência de etapas indicando que existem estados intermediários. / The myoblobins of \"Applysia Brasiliana\" (MbApB) and of the sea turtle \"Dermochelis Coriacea\" (MbT) are studied in this work with special attention devoted to the acid-aIkaIyne transition (AAT) , the interaction with transition metals and temperature induced conformational changes in order to characterize structural differences in these proteins. The AAT of MbApB has a pK = 7.2 obtained from the EPR spectra of Fe3+ at g1 = 5,83 and a pK = 7,5 obtained from optical absorption (&#955=590nm). The EPR Spectrum of Fe3+ at alkaline pH shows a rhombic distortion the ion crystal field which is in agreement with the absence in this protein of the distal histidine residue. The ESR lines associated with the low spin configuration are considerably broadened. This effect can be explained by fluctuations on the heme position relative to the symmetry axis. MbApB forms complexes with both Cu2+ and Mn2+ only one binding site is obtained for both metals in the protein. This site probably has common ligands for Mn2+ and Cu2+ as the binding is competitive, suggesting also that the Cu2+ complex is more stable than the Mn2+ one (KAMn2+ = (11,5-0,8).103 M-1 ). The Cu2+: MbApB complex shows a pH dependent transition probably related to a change in the Cu2+ coordination. This transition can be adjusted assuming an interconversion between two different forms with a pK = 11,0. Addition of more than one Cu2+ ion per MbApB produces the denaturation of the protein. The AAT of MbT was studied through optical absorption (pK =: 8,4) and EPR (pK =: 8.3). The EPR spectra show that the Fe3+ ion is in a crystal field with axial symmetry with rhombic distortion. Measurements of optical absorption show isosbestic points practically identical to other hemoproteins with the distal histidina present. Our data permit to anticipate that MbT probably has the same aminoacid residues in the heme region as Mb (II), and thÉit a slight displacement of the histidine E-7 about the heme symmetry axis is observed. Temperature induced conformational changes were studied in MbT at various pH\'s. This protein has a cysteine residue in the polypeptide chain which allows the binding of a specific spin label, a maleimide derivative. Our results from the EBR spectra of the spin label attached to MbT together with the optical absorption by Fe3+ lead to the conclusion that this protein exhibits a pre-denaturation behaviour and the transition from the native to the denatured forms in a result of a sequence of steps suggesting the existence of intermediate states.

Após setenta anos de sua descoberta, o vírus da zika surgiu no Brasil, espalhou-se rapidamente pelas Américas e trouxe complicações incomuns em doenças causadas por Flavivirus, como a microcefalia. A Organização Mundial da Saúde classifica a zika como a doença viral mais preocupante da atualidade e considera urgente desenvolver novos métodos de diagnóstico para ela e doenças correlatas como a dengue. Embora existam exames para identificar infecções pelos vírus dessas duas doenças, ainda não há um método rápido, específico e de baixo custo para o diagnóstico precoce. Visando preencher essa lacuna, este trabalho teve como objetivo construir dois tipos de biossensores eletroquímicos de DNA para detecção label-free desses dois vírus. Foram fabricados eletrodos descartáveis em substrato de politereftalato de etileno metalizado com filme fino de ouro nas configurações com um e três contatos. As sequências genéticas de iniciadores e sondas de captura foram desenhadas especialmente para este trabalho com base na análise dos genomas dos vírus. O primeiro biossensor utilizou o eletrodo em uma célula eletroquímica e foi capaz de identificar sequências de DNA da zika ou da dengue. As análises por espectroscopia de impedância eletroquímica mostraram que o biossensor é seletivo à sequência alvo com limite de detecção de (9,86 &plusmn; 0,89) nmol L-1. O segundo biossensor utilizou um eletrodo de três contatos para identificação de sequências de DNA em uma gota da amostra. No contato central, usado como eletrodo de trabalho, foi imobilizada a sequência de captura e os contatos laterais funcionaram como eletrodos de referência e auxiliar. Nesse sistema as medidas de impedância indicaram limite de detecção de (25,0 &plusmn; 1,7) nmol L-1. Os biossensores desenvolvidos mostraram seletividade para identificar o material genético dos vírus da zika e da dengue nos ensaios com DNA sintético e, portanto, são promissores para a análise de amostras reais, principalmente de produtos da polimerase da cadeia reversa. / After seventy years of its discovery, zika virus has emerged in Brazil, spread rapidly throughout the Americas, bringing unusual complications in diseases caused by flaviviruses, such as microcephaly. The World Health Organization classifies zika as the most harmful viral disease today and considers urgent the development of new diagnostic methods for zika and related diseases, such as dengue. Although there are tests to identify both infections, no current diagnostic method is rapid, specific and cost-efective. This thesis describes two types of electrochemical DNA biosensors for label-free detection of these zika and dengue. Disposable electrodes were fabricated on polyethylene terephthalate substrates covered with a nanometric gold layer by thermal evaporation, manufactured in one- and three-contact configurations. Genetic sequences of primers and complementary capture probes were designed based on the analysis of the virus genomes. The first biosensor we developed used the new electrode in an electrochemical cell and was able to identify zika or dengue DNA sequences. Analyses by electrochemical impedance spectroscopy showed that these biosensors are selective for zika or dengue with a detection limit of (9.86 &plusmn; 0.89) nmol L-1. A second type of biosensor used a three-contact electrode to identify DNA sequences in a drop of sample. In the central contact, used as a working electrode, the capture sequence was immobilized and the lateral contacts acted as reference and auxiliary electrodes. In this system the impedance measurements indicated a limit of detection of (25.0 &plusmn; 1.7) nmol L-1. The developed biosensors showed selectivity for zika and dengue in the synthetic DNA assays, and therefore are promising for the analysis of real samples, especially the polymerase chain reaction amplicon.

Introdução: O uso de formas farmacêuticas inadequadas à pediatria, também conhecido como uso off-label, pode levar a uma terapia medicamentosa insegura. No ambiente hospitalar, em especial nas Unidades de Terapia Intensiva Pediátrica, há alta prevalência de prescrições com medicamentos não apropriados para crianças, entre eles os antimicrobianos, devido à gravidade das infecções, o estado crítico dos pacientes internados, a maior realização de procedimentos invasivos e uma maior incidência de bactérias resistentes. Em geral, o uso de antimicrobianos em crianças, bem como dos demais grupos farmacológicos, tem sido baseado principalmente em extrapolações e adaptações do uso em adultos, informações obtidas de raros estudos observacionais e consensos de especialistas na área. Devido ao estado crítico dos pacientes, as complexas terapias envolvendo antimicrobianos nem sempre são feitas de forma adequada, podendo trazer prejuízos à saúde dos pacientes e contribuir, ainda, para a resistência bacteriana. Objetivo: Avaliar a incidência do uso de antimicrobianos em pacientes internados em Unidade de Terapia Intensiva Pediátrica. Método: Estudo do tipo transversal realizado na Unidade de Terapia Intensiva Pediátrica do município de Rio Branco, Acre, no período de agosto de 2014 a julho de 2015. A amostra foi composta por 246 crianças de zero a onze anos de idade, internadas com quaisquer patologias. Foram analisadas as prescrições das primeiras 24 horas. Pesquisou-se o uso off label de antimicrobianos e potenciais interações medicamentosas com esses medicamentos. Resultados: Duzentas e trinta e uma crianças (93,9 por cento ) receberam a prescrição de pelo menos um antimicrobiano. O antimicrobiano mais prescrito foi a ceftriaxona. Em média, cada paciente recebeu dois antimicrobianos. Em 66,8 por cento dos casos, houve uso off-label, sendo a dose prescrita superior ao recomendado em 41 por cento , dos casos. Em 56 por cento das prescrições havia pelo menos uma potencial interação medicamentosa envolvendo antimicrobianos. Ampicilina e gentamicina foram os antibióticos que mais se envolveram em interações. Conclusões: a alta frequência do uso off-label e interações medicamentosas potenciais envolvendo antimicrobianos foi confirmada neste estudo. Tratar crianças como adultos pequenos pode expor esses pacientes a eventos adversos a medicamentos que comprometem a segurança desse grupo específico de pacientes. Isso aponta para a necessidade de equipes multidisciplinares trabalharem em conjunto e serem estimuladas a realizar mais estudos que garantam a segurança do uso de medicamentos na pediatria. / Introduction: The use of inadequate dosage forms for pediatrics, also known as off-label use, may lead to unsafe drug therapy. In the hospital environment, especially in Pediatric Intensive Care Units, there is a high prevalence of prescriptions with drugs not suitable for children, among them antimicrobial agents, due to the severity of the infections, the critical state of the hospitalized patients, more accomplishment of invasive procedures and a higher incidence of resistant bacteria. In general, the use of antimicrobials in children, as well as other pharmacological groups, has been based mainly on extrapolations and adaptations of the use in adults. This information was obtained from rare observational studies and consensus of specialists in the area. Due to the critical condition of patients, the complex therapies involving antimicrobial agents are not always adequately done, which may cause harm to the patients\' health and also contribute to bacterial resistance. Objective: To evaluate the incidence of antimicrobial use in patients hospitalized in a Pediatric Intensive Care Unit. Method: A cross-sectional study conducted at the Pediatric Intensive Care Unit of the municipality of Rio Branco, Acre, from August 2014 to July 2015. The sample consisted of 246 children from zero to eleven years old and hospitalized with any pathologies. The prescriptions of the first 24 hours were analyzed. The use of off-label antimicrobials and potential drug interactions with these drugs was investigated. Results: Two hundred and thirty-one children (93.9 per cent ) received the prescription of at least one antimicrobial. The most commonly prescribed antimicrobial agent was ceftriaxone. On average, each patient received two antimicrobials. In 66.8 per cent of cases, there was an off-label use, with the prescribed dose being higher than recommended in 41 per cent of the cases. In 56 per cent of prescriptions there was at least one potential drug interaction involving antimicrobials. Ampicillin and gentamicin were the antibiotics most involved in interactions. Conclusions: the high frequency of off-label use and potential drug interactions involving antimicrobials was confirmed in this study. Treating children as young adults may expose these patients to adverse drug events that compromise the safety of this particular group of patients. That points to the need for multidisciplinary teams to work together and be encouraged to carry out more studies that ensure the safety of use of medication in pediatrics.