Freie Universiẗat, Diss., 2004--Berlin. / Dateiformat: zip, Dateien im PDF-Format.- Erscheinungsjahr an der Haupttitelstelle: 2004.

Doctor en Bioquímica / El alcoholismo es una enfermedad de etiología multifactorial que causa un número de problemas sociales, laborales, legales y médicos. El riesgo de un individuo de desarrollar alcoholismo está influenciado por varios genes. Algunos genes permiten el desarrollo del alcoholismo y otros genes protegen a sus portadores de esta condición. La deshidrogenasa aldehídica mitocondrial (ALDH2) metaboliza gran parte del acetaldehído generado en el metabolismo hepático del alcohol. En algunos individuos de la población asiática, una mutación en el gen que codifica la ALDH2, disminuye o elimina la actividad de esta enzima. La inactivación de esta enzima produce una exagerada acumulación del acetaldehído sanguíneo luego del consumo de bebidas alcohólicas, lo que genera efectos disfóricos que producen un rechazo al alcohol. Este fenotipo protector descrito en la población asiática puede ser producido en alcohólicos a los cuales se ha administrado el medicamento aversivo disulfiram (Antabus®). Este fármaco, eficaz en disminuir la actividad ALDH2, es sin embargo poco específico, posee una vida media corta, muestra una gran variabilidad individual y genera una serie de efectos tóxicos por unirse en forma no específica a los grupos sulfhidrilos de varias proteínas. La terapia génica se perfila como una alternativa al actual tratamiento farmacológico del alcoholismo. Un nuevo medicamento aversivo sin los efectos secundarios del disulfiram sería de gran utilidad en el campo del alcoholismo. El tipo de terapia a desarrollar pretende imitar el fenotipo asiático, disminuyendo específicamente la actividad de la ALDH2 mediante la reducción de la traducción del mRNA. La estrategia utilizada para lograr este objetivo, es inhibir la traducción mediante la generación de un RNA antisentido de la deshidrogenasa aldehídica mitocondrial codificado en un vector adenoviral. En estudios en esta tesis, primeramente, el gen antisentido de la ALDH2 (cDNA de Aldh2 en posición 3’ a 5’) contenido en el vector adenoviral bajo el promotor CMV se transdujo a células de hepatoma de rata generando niveles altos de mRNA antisentido. La administración de este vector a células de hepatoma de rata disminuyó de 60 a 70% la actividad ALDH2 y aumentó 8 veces (10 μM a 80 μM) los niveles de acetaldehído acumulado cuando estas células se incubaron con etanol. En estudios in vivo, se administró el adenovirus portador del gen antisentido-Aldh2 a ratas bebedoras UChB (línea de ratas Bebedoras desarrollada en la Universidad de Chile). Las ratas que recibieron una sola dosis del vector (1012 pv/kg) por vía intravenosa, disminuyeron su consumo de alcohol en 50%, y se observó una reducción de 90% en la actividad de la ALDH2. La reducción del consumo de etanol duró 34 días, tiempo al cual el estudio se finalizó. En estudios paralelos de más corta duración (10 días), los niveles plasmáticos de acetaldehído en los animales tratados con el vector anti-Aldh2 aumentó 6 a 8 veces (8 μM a 60 μM) sobre niveles controles (6-14 μM; p<0,01) asociado a un 37,5% de inhibición de la actividad ALDH2 hepática. En conclusión, en animales que reciben el gen Aldh2-antisentido, una reducción específica en la actividad de la ALDH2 permite la elevación de los niveles de acetaldehído sistémico, que genera una aversión al etanol. Los estudios sugieren que es posible desarrollar una terapia génica específica y de larga duración para el alcoholismo / Alcoholism is defined by the compulsive excessive use of alcohol despite of negative consequences perceived by the subject. It is not only one of major public health problems due to the morbidity caused by the alcohol abuse disorder but it is also a major social problem. The risk of developing this disease is genetically influenced, with some genes being permissive and other genes protective. Most of the elimination of ethanol occurs by oxidation to acetaldehyde and further into acetate. The reactions are sequentially catalyzed by alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH). The oxidation of acetaldehyde to acetate is catalyzed primarily by ALDH2, the low Km isozyme of ALDH present in mitochondria. A mutation in the gene coding for ALDH2 present in some Asian populations, lowers or abolishes the activity of this enzyme and results in marked elevations in blood acetaldehyde upon ethanol consumption. The resulting phenotype greatly protects against alcohol abuse and alcoholism. The protective Asian phenotype can be elicited in alcoholics who are administered the aversive medication disulfiram (Antabuse®), affording these individuals the same protection from alcohol abuse. However, disulfiram is a non-specific drug that binds to sulfhydryl groups, has a short half-life, cannot be metabolized by some patients into its active metabolite, and has marked toxicity side effects, as it nonspecifically binds to sulfhydryl groups in other proteins. Gene therapy, as a new approach, opens the possibility of developing a specific aversive medication for alcoholism. This new treatment, based on the reduction of Aldh2 mRNA transduction, provides a means by which gene expression can be specifically inhibited to yield the low-ALDH2 Asian phenotype. The aim of the work presented in this thesis is to reduce Aldh2 mRNA translation in the rat by the intracellular generation of an antisense aldehyde dehydrogenase RNA coded by an antisense gene (Aldh2 cDNA in a 3´ to 5´ orientation) contained in an adenoviral vector. Rat hepatoma cells were transduced with the antisense coding gene against ALDH2 carried by the adenoviral vector, leading to the generation of high levels of antisense mRNA. The cells infected with this vector, reduced their ALDH2 activity by 60 to 70% and, in the presence of ethanol, increased 8-fold the levels of acetaldehyde (10 μM to 80 μM). In studies in vivo, the adenoviral vector coding for the antisense-Aldh2 RNA was administered to UChB rats (University of Chile alcohol dependent rats). The rats infected with a single dose of the vector (1012 vp/kg) by tail vein injection inhibited its ALDH2 activity in 90% and reduced its alcohol consumption in 50%. The reduction in alcohol consumption lasted 34 days, time at which the study was terminated. A parallel study in which rats were infected 10 days earlier with a single dose of the vector carrying the antisense gene showed a 37.5% reduction in hepatic ALDH2 activity, while acetaldehyde plasma levels following the administration of ethanol increased 6-8 fold over controls (5-10 μM to 40-60 μM) (p<0,01). Overall, rats treated with an Aldh2- antisense coding gene show significant reductions in hepatic ALDH2 activity and elevations in systemic acetaldehyde upon ethanol administration, which are accompanied by marked reductions in ethanol voluntary consumption in alcohol dependent animals. These preclinical studies suggest that a long-lasting specific gene therapy against alcoholism may be developed

Submitted by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2016-01-05T13:35:08Z No. of bitstreams: 2 thalita_gomes_miranda.pdf: 848359 bytes, checksum: ea292a0faa794975c2a67fb133b57b6a (MD5) license_rdf: 22974 bytes, checksum: 99c771d9f0b9c46790009b9874d49253 (MD5) / Approved for entry into archive by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2016-01-05T17:58:54Z (GMT) No. of bitstreams: 2 thalita_gomes_miranda.pdf: 848359 bytes, checksum: ea292a0faa794975c2a67fb133b57b6a (MD5) license_rdf: 22974 bytes, checksum: 99c771d9f0b9c46790009b9874d49253 (MD5) / Made available in DSpace on 2016-01-05T17:58:57Z (GMT). No. of bitstreams: 2 thalita_gomes_miranda.pdf: 848359 bytes, checksum: ea292a0faa794975c2a67fb133b57b6a (MD5) license_rdf: 22974 bytes, checksum: 99c771d9f0b9c46790009b9874d49253 (MD5) Previous issue date: 2014 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (Capes) / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico (CNPq) / As pimentas (g?nero Capsicum spp.) s?o mundialmente conhecidas e apreciadas como condimentos al?m de serem excelentes fontes de nutrientes. A crescente demanda faz com que aumente a busca por cultivares que atendam ?s necessidades do mercado para consumo in natura e para fins industriais. A caracteriza??o f?sico-qu?mica dos gen?tipos de pimenta serve de base para o melhoramento gen?tico e lan?amentos de novas cultivares no mercado. Diante disso, o presente trabalho teve como objetivo avaliar gen?tipos de pimenta Capsicum chinense e C. annuum, quanto ?s caracter?sticas f?sico-qu?micas. Foram realizados dois experimentos, um para avalia??es f?sicas e qu?micas e outro para avaliar o teor de capsaicina presente nos frutos. Os experimentos foram conduzidos em casa-de-vegeta??o, em DBC com oito gen?tipos e quatro repeti??es, sendo os gen?tipos BGH-433, BGH-4285, PIM-030, Numex Sweet, Numex Garnet e os h?bridos F1 (Numex Sweet x PIM-030), F1 (Numex Garnet x PIM-030), F1 (BGH-4285 x BGH-433). Para o primeiro experimento foram avaliadas as seguintes caracter?sticas: produtividade total de frutos, peso m?dio do fruto, n?mero de sementes por fruto, altura da planta, produtividade de mat?ria seca total, porcentagem de mat?ria seca do fruto, porcentagem de mat?ria seca da polpa, porcentagem de mat?ria seca da semente, espessura do pericarpo, comprimento do fruto, fen?licos totais, flavonoides, pH, s?lidos sol?veis, vitamina C e acidez titul?vel. No segundo experimento foi avaliado o conte?do de capsaicina presente na semente, na polpa e no fruto inteiro e determinada a pung?ncia dos gen?tipos. Os h?bridos F1 (Numex Sweet x PIM-030), F1 (Numex Garnet x PIM-030) e F1 (BGH-4285 x BGH-433) e as linhagens Numex Garnet e Numex Sweet apresentaram alta produtividade total de frutos e de mat?ria seca total de frutos. O h?brido F1 (BGH-4285 x BGH-433) apresentou altos valores de heterose e heterobeltiose para produtividade total de frutos e produtividade de mat?ria seca total de frutos, e apresentou heterose e heterobeltiose positivos para a porcentagem de mat?ria seca da semente, j? o h?brido F1 (Numex Sweet x PIM-030) apresentou heterose e heterobeltiose negativas para essa caracter?stica. O h?brido F1 (BGH-4285 x BGH-433) apresentou heterobeltiose negativa para a porcentagem de mat?ria seca do fruto. H? correla??es positivas entre caracter?sticas de interesse que podem contribuir significativamente com os programas de melhoramento com C. chinense e C. annuum por meio da sele??o indireta. Os valores observados para os compostos fen?licos, flavonoides, pH, s?lidos sol?veis, vitamina C e acidez total n?o variaram significativamente entre os gen?tipos estudados. O gen?tipo BGH-433 apresentou as maiores concentra??es de capsaicina para semente e polpa. Os h?bridos F1 (BGH-4285 x BGH-433) e BGH-433 apresentaram as maiores concentra??es de capsaicina para fruto inteiro. Os gen?tipos BGH-433, F1 (BGH-4285 x BGH-433) e BGH-4285 foram classificados pela SHU como moderadamente pungentes. Os gen?tipos Numex Sweet, Numex Garnet, PIM-030, F1 (Numex Garnet x PIM-030) e F1 (Numex Sweet x PIM-030) foram classificados como n?o pungentes a levemente pungentes. A temperatura no per?odo de condu??o do experimento pode ter influenciado negativamente na pung?ncia dos frutos, pois alguns gen?tipos apresentaram pung?ncia menor que o esperado. / Disserta??o (Mestrado) ? Programa de P?s-Gradua??o em Produ??o Vegetal, Universidade Federal dos Vales do Jequitinhonha e Mucuri, 2014. / ABSTRACT Peppers (Capsicum spp.) are world known and appreciated as condiments and are excellent sources of nutrients. The increasing demand causes increase of the search for cultivars that meet the needs of the market for fresh consumption and for industrial purposes. The physicochemical characterization of genotypes of pepper is the basis for the breeding and release of new cultivars on the market. Therefore, this study aimed to evaluate genotypes of pepper Capsicum chinense and C. annuum, as the physical and chemical characteristics. Two experiments, one for physical and chemical tests and another to evaluate the content of capsaicin present in the fruits were performed. The experiments were conducted in a greenhouse house in DBC with eight genotypes and four replications, with the genotypes BGH-433, BGH-4285, PIM-030, Numex Sweet, Numex Garnet and F1 hybrids (Numex Sweet x PIM-030), F1 (Numex Garnet x PIM-030), F1 (BGH-4285 x BGH-433). For the first experiment the following characteristics were evaluated: total fruits yield, average fruit weight, number of seeds per fruit, plant height, total dry matter yield, dry matter content of the fruit dry matter content of the pulp, percentage of dry matter of seed, pericarp thickness, fruit length, total phenolics, flavonoids, pH, soluble solids, vitamin C and titratable acidity. In the second experiment evaluated the content of capsaicin present in the seed, in the pulp and whole fruit and given the poignancy of genotypes. The F1 hybrids (Numex Sweet x PIM-030), F1 (Numex Garnet x PIM-030) and F1 (BGH-4285 x BGH-433) and the lines Numex Garnet and Numex Sweet had high total fruit yield and dry matter total fruit. The F1 hybrid (BGH-4285 x BGH-433) showed high levels of heterosis and heterobeltiosis for total fruits yield and productivity of total dry fruits, and showed positive heterosis and heterobeltiosis for dry matter content in the seed, since the hybrid F1 (Numex Sweet x PIM-030) show heterosis and negative heterobeltiosis for this trait. The F1 hybrid (BGH BGH x-4285-433) showed negative for the percentage of dry matter of the fruit heterobeltiosis. There are positive correlations between features of interest that can contribute significantly to the improvement programs with C. chinense and C. annuum by indirect selection. The observed values of the phenolic compounds, flavonoids, pH, soluble solids, total acidity and vitamin C did not differ significantly between genotypes. The BGH-433 genotype showed the highest concentrations of capsaicin for seed and pulp. The F1 hybrids (BGH BGH x-4285-433) and BGH-433 showed the highest concentrations of capsaicin for whole fruit. The BGH-433, F1 genotypes (BGH-4285 x BGH-433) and BGH-4285 were classified as moderately pungent by SHU. Genotypes Numex Sweet, Numex Garnet, PIM-030, F1 (Numex Garnet x PIM-030) and F1 (Numex Sweet x PIM-030) were classified as non-pungent and slightly pungent. The temperature in driving time of the experiment may have negatively influenced the pungency of the fruits, because some genotypes showed less pungency than expected.

The aim of the study was to study the variability of the gene in the population FABP3 pig breed Czech large white. Analyze their association with indicators of pork production. Gene FABP3 belonging to multigene families FABP (fatty acid binding proteins affecting the transport of long chain fatty acids). Analyzed polymorphism was at position X98558:g.1321G>C in the 5'UTR region. The relative frequencies of genotypes were: HH = 0,87, Hh = 0,13. hh genotype was not detected in the monitored individuals. The values of the relative frequencies of alleles were as follows: H = 0,93 h = 0,07. Based on the association analysis showed a statistically significant difference between genotypes for stearic acid and linoleic acid. Highly statistically significant difference was observed with palmitic acid.

Summary Objectives and hypothesis: This thesis concerns the study of plasmids of vancomycin- resistant enterococci isolated from feces of American crows in the years 2012 - 2013 period. The hypothesis is that, in various environments, there is one or more types of epidemiologically significant vanA gene-carrying plasmids that are capable of horizontally spread. Methods: Based on PFGE method the number and size of plasmids were detected in selected isolates of vancomycin-resistant E. faecium. Using PCR method the isolates were subjected to detection of genes of replicases, relaxases and toxin-antitoxin system of plasmid-bound resistance genes. Using 19 primers were characterized types of Tn1546. Results: Of the 12 tested vancomycin-resistant isolates of E. faecium the following number and size of plasmids was proven using PFGE method: 2 isolates contained two plasmids (17%), 3 isolates contained three plasmids (25 %), 5 isolates contained four plasmids (42 %) and 2 isolates contained five plasmids (17 %). All isolates (n = 12) were then subjected to the detection of genes of replicases, relaxases and toxin-antitoxin system for typing of plasmids from each plasmid families. RepA_N family of plasmids: genes characterizing plasmids related to pRUM: rep17 in 11 isolates (92 %), gene Axe-Txe was detected in 5 isolates (42 %) genes characterizing plasmids related to pLG1: rep20 in 7 isolates (58 %) genes characterizing plasmids related to pAD1: relpAD1 gene was detected in one isolate (8 %) Inc18 family of plasmids: genes characterizing plasmids related to pIL501: rep1 gene detected in one case (8 %) genes characterizing plasmids related to pRES25: rep2 gene in 2 isolates (17 %) genes characterizing plasmids related to pEF1: relpEF1 detected in 11 isolates (92 %) pHTB family of plasmids: genes characterizing plasmids related to pHTB: rep22 gene was detected in 4 isolates (33%) and in 2 isolates gene relpHTB was detected (17%) RCR family of plasmids: genes characterizing plasmids related to pRI: positive detection of Rep14 gene in 8 isolates (67%) and in 4 isolates relpRI gene was detected Small theta-replicating plasmids: genes characterizing plasmids related to pEF418 plasmids: rep18a gene in 2 isolates (17%) genes characterizing plasmids related to pB82: rep18b gene was detected in one isolate (8%) genes characterizing plasmids related to pCIZ2: relpCIZ2 gene was detected in 9 isolates tested (75%) Types of transposon Tn1546 Using the PCR method types of Tn1546 were characterized. In 4 isolates (n = 12; 33 %) Tn1546 was characterized as a F3 type. In one isolate (8 %) Tn1546 was characterized as a type F5, in one isolate (8 %) as a type PP-16. In 6 isolates Tn1546 was untypeable. Most likely these are new, yet unknown types. Conclusion: This is the first study of plasmids of vancomycin-resistant isolates E. faecium isolated from feces of American crows. These results emphasize not only a high proportion of plasmids in individual isolates, but also a high proportion of genes with horizontal transfer.

No description available.

No description available.

Over the course of the past few decades, first-generation college students have been analyzed from many angles. With research ranging from quantitative reviews of lower graduation and retention rates, as well as higher attrition rates (Engle & Tinto, 2008; Inman & Mayes, 1999; Terenzini et al., 1996; Tinto, 1975), to qualitative case studies focusing on the psychological aspects of preparation, parental support, and identity formation (Lara, 1992; London, 1989; Rendón, 1992; Rodriguez, 1975 1982; Skinner & Richardson, 1988; Weis, 1985), this population has been well documented across a spectrum of research methodologies. More recently, scholarly attention has shifted toward a more individualized approach, focusing on smaller cohorts within the larger first-generation college student population (Collier & Morgan, 2008; Covarrubias et al., 2019; McCoy, 2014; Phinney & Haas, 2003). The goal of this three-article dissertation is to highlight and prioritize first-generation college students’ voices and narratives by emphasizing their lived experiences, as well as reviewing the support services currently available to them. This goal is addressed using three distinct, yet interconnected articles all utilizing different research methodologies. The first article, a phenomenological case study, addressed the experiences of six female first-generation college student caregivers (Orbe, 2004; Pyne & Means, 2013; Covarrubias et al., 2019) at a large, prestigious, research-driven institution in the Northeast. The second study, a singular, narrative case study, utilized the construct of intersectionality (Crenshaw, 1994; Pyne & Means, 2013) to examine the experiences of a female, first-generation college student caregiver of color as she navigated the higher education system. The last article, a comparative case study, examined the available first-gen support programming at three institutions in the same metropolitan area. This final study also included administrator perspectives about what is required to implement and execute first-generation college student support initiatives. The major implications of this dissertation project include the following: a strong recommendation for increased intersectionality in all first-gen support programming; a discovery of the causational relationship of being a first-gen caregiver and the added difficulty that multi-layered identity creates; a demonstration of the need to motivate and utilize collected student data in order to inform first-gen program creation; and a recognition of the stressors placed on certain campus stakeholders and the need for enhanced cross-campus collaboration to improve first-generation college student support. Future research and specific recommendations for the field of higher education are discussed.

The theoretical part of the diploma thesis deals with tumour diseases and a general description of tumour suppressor genes. Special attention is paid to the TP53 gene and its influence on the functioning of the organism, including its production of the p53 protein. Based on an analysis and synthesis of findings from specialised sources, the current methods of detecting the aforesaid gene, in particular immunological and elec-trochemical, were described. The last chapter of the theoretical part is devoted to bio-sensors and their classification, with a special focus on electrochemical biosensors. The practical part involves the construction and optimisation of the biosensor, from the cleaning of electrodes, the deposition of gold nanoparticles, the ssDNA bond, the blocking of free binding points and a synthesis of the complementary chain to the resulting measurement and quality testing. This part also presents the concept of an innovative biosensor that offers an easier and more reliable alternative for the detec-tion of the TP53 gene. The last chapter of this part analyses and comments on the aforesaid results.

Syfte: Syftet med denna studie är att söka djupare förståelse för Gen-Z konsumenters lojalitet i form av hur deras attityder och beteende skiljer sig mot Fast Fashion handel och hur sambandet mellan dem kan se ut. Vidare syftar denna studie att identifiera konsekvenserna av de olika uppfattningarna i form Word of Mouth. Forskningsfrågor: <ul type="disc">Hur ser konsumenternas bild ut av ett varumärke inom Fast Fashion och vad beror detta på? Hur ser skillnaden ut mellan konsumenters attityd och beteende gentemot ett Fast Fashion varumärke? Hur ser konsumenters lojalitet ut i helhet till ett Fast Fashion varumärke baserat på attitydmässiga samt beteendemässiga faktorer? Metod: Studien är av kvalitativ ansats och datan har samlats in med hjälp av fokusgrupper samt djupintervjuer i form av semistrukturerade intervjuer. Teoretisk referensram: De teorier som används i denna studie är Varumärkesbild, Varumärkesattityd, Varumärkeslojalitet, Lojalitetsmodell samt Lojalitetsindelning av Dick och Basu (1994), Lojalitetspyramiden av Aaker (1991) och slutligen Word of Mouth. Slutsatser: Slutsatserna är att respondenterna har en negativ varumärkesbild av H&amp;M som varumärke. Det identifierades även en motsägelse mellan respondenters attityder och beteenden då de skilde sig åt markant. Respondenternas lojalitet till varumärket visade sig vara väldigt svag. De negativa attityderna visade sig även ha stor påverkan på Word of Mouth som sprids konsumenter emellan. / Aim: The aim of this study is to search for a deeper understanding of Gen-Z’s consumer loyalty in the form of their attitudes and behaviour towards Fast Fashion and how the relationship between these factors can look like. Furthermore this study aims to identify the consequences of the the different perceptions in the form of Word of Mouth. Research questions: <ul type="disc">What does consumers brand image look like when it comes to a Fast Fashion brand and what does it depend on? What are the differences between consumers attitudes and behaviour towards a Fast Fashion brand? What does consumers loyalty look like in its whole to a Fast Fashion brand based off attitudinal and behavioural factors? Method: This study is based on a qualitative approach and the data has been collected through semi-structured interviews. Both focus groups and individual interviews have been conducted. Theoretical framework: The theories that have been applied are Brand Image, Brand Attitude, Brand Loyalty, Loyalty Model and Loyalty Classification by Dick &amp; Basu (1994), Loyaltypyramid (Aaker 1991) and finally Word of Mouth. Conclusions: The conclusions are that the respondents has got a negative image of H&amp;M as a brand. A contradiction was also identified between the respondents attitudes and behaviour considering they were very different. The loyalty towards the brand was not very strong. The negative attitudes of the respondents had an significant influence on the brands Word of Mouth.