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"Mémoire Présenté à la Faculté des Études Supérieures en vue de l'obtention du Grade de Maîtrise En Droit Option Recherche" / The past decade has seen an explosion in the availability of genome sequence data from public and private genome projects. Most notably, the complete human DNA genome sequence has been published and the locations of some of the genes have been mapped to individual chromosomes. Commensurate with the growth in sequence information, the biotechnology industry has become firmly established. Gene patents have played an important part in this industry, and there has been a marked increase in patent applications filed in the field of human genetic resource. However, concerns have been raised over the patentability of human genetic material through public protests and international statements, but to little effect. Discussed here are some of these concerns, the patent office's response to them in different jurisdictions, and ways in which to address these issues and to move the debate forward within current legal structures. / Durant la dernière décennie nous avons assisté à une explosion de données génomiques séquentielles provenant de projets publiques et privés. Le séquence complète du génome humain a été publiée ainsi que la localisation certains gènes a été tracée pour des chromosomes individuels. Avec I'accroissement de I'information séquentielle, l'industrie de la biotechnologie s'est sérieusement établie. Le brevets octroyés en regard des gènes a joué un rôle important dans cette industrie, aussi il y a eu une augmentation dans la demande de brevets dans Ie domaine du matériel génétique humaine. Cependant après plusieurs controverses publiques et la créations de règles internationales certains doutes ont été soulevés à propos de la « brevetabilité »du génome humain, mais impact. II sera question dans notre étude ces inquiétudes, de la réponse a ces inquiétudes par les bureaux des brevets différentes juridictions, ainsi que des divers moyens utilisés pour de ces problèmes tout en avançant le présent débat en utilisant les structures légales existantes.

The past decade has seen an explosion in the availability of genome sequence data from public and private genome projects. Most notably, the complete human DNA genome sequence has been published and the locations of some of the genes have been mapped to individual chromosomes. Commensurate with the growth in sequence information, the biotechnology industry has become firmly established. Gene patents have played an important part in this industry, and there has been a marked increase in patent applications filed in the field of human genetic resource. However, concerns have been raised over the patentability of human genetic material through public protests and international statements, but to little effect. Discussed here are some of these concerns, the patent office's response to them in different jurisdictions, and ways in which to address these issues and to move the debate forward within current legal structures. / Durant la dernière décennie nous avons assisté à une explosion de données génomiques séquentielles provenant de projets publiques et privés. Le séquence complète du génome humain a été publiée ainsi que la localisation certains gènes a été tracée pour des chromosomes individuels. Avec I'accroissement de I'information séquentielle, l'industrie de la biotechnologie s'est sérieusement établie. Le brevets octroyés en regard des gènes a joué un rôle important dans cette industrie, aussi il y a eu une augmentation dans la demande de brevets dans Ie domaine du matériel génétique humaine. Cependant après plusieurs controverses publiques et la créations de règles internationales certains doutes ont été soulevés à propos de la « brevetabilité »du génome humain, mais impact. II sera question dans notre étude ces inquiétudes, de la réponse a ces inquiétudes par les bureaux des brevets différentes juridictions, ainsi que des divers moyens utilisés pour de ces problèmes tout en avançant le présent débat en utilisant les structures légales existantes. / "Mémoire Présenté à la Faculté des Études Supérieures en vue de l'obtention du Grade de Maîtrise En Droit Option Recherche"

Ever since the discovery of DNA, there has been many pathologies identified effecting mankind. With the development in technology, there are many methods to alleviate these pathologies. One such is gene therapy or gene delivery. It is a process of introducing some foreign material into the body to correct the effected cells. In principle, it is a modern method to cure cells or a method to transfer nucleic acid into a cell to treat specific cells in the body. The process of delivering a genetic material is carried out using vectors, namely, viral vectors and non-viral vectors. In viral vectors, viruses are modified to make it efficient for delivery into the host cells. This method has high transduction rate as compared to non-viral method. Non-viral methods include chemical and physical transfection methods, which are used to deliver the gene of interest into the host cell unlike viral methods. In this study, a physical method using high voltage is used to deliver a genetic material into cells. High voltages are used to permeabilize the cell to allow the foreign material into it and to express it in the host cell. This process is termed as Electroporation. In specific, in this research, studying a process of charging a region that mimics skin and trying to localize the presence of electric fields on the surface where the strongest uptake of genetic material is found. In other words, region where the gene expression is strongest at a specific region if performed on skin is studied by localizing electric fields on the surfaces. My work is to characterize and develop where this effect takes place on the surface based on both positive and negative electric fields. A physical method is useful as it is a non-toxic way to get a DNA/protein into someone’s body without making them sick, unless if not using a virus to deliver. This is all done using high voltages up to 8kV and the electric fields produced due to high voltages are localized, visualized and characterized with both positive and negative polarities of voltages. In this study, experiments with high voltages are performed and the spread of charges at specific regions are collected using a needle. This needle goes into corona, which may be called as a secondary corona. It might be called a secondary corona because the flat conductor is being charged by a metal finger but not directly by the power supply. Here, the conductor is charged by a metal finger of high input voltage, which ionizes the air molecules above the flat conductor to form a conductive region. As the input voltage is increased further, electrons escape from the needle to air or from molecules to needle forming negative or positive ions respectively. The outputs at needle were measured on the oscilloscope. In this study, repeated sets of experiments are carried out to collect consistent and reliable data. Visualizing/characterizing these fields are important as maximum delivery takes place at high voltage regions, with a condition that permeability of the cells should be known for proper transfection to occur, otherwise cells would die due to high voltages or no transfection takes place due to poor permeability of cell membrane.

Made available in DSpace on 2015-03-26T13:53:33Z (GMT). No. of bitstreams: 1 texto completo.pdf: 402948 bytes, checksum: da3219eefcfbb4dc22f68e1d969b6c0a (MD5) Previous issue date: 2006-03-06 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / Salt accumulation in soils is a frequent event in arid or semi-arid regions, where the evapotranspiration is high and the low precipitation is insufficient to promote leaching of the salt excess from the soil profile. Under such conditions the crop yields are reduced in response to osmotic effects, to the toxicity of specific ions in the soil solution or to a joint effect of these factors. Three trials with the common bean cultivars Diamante Negro and OPNS 331 were performed to isolate the effects caused by the high salt concentrations in the solution and to verify responses to osmotic pressure, to sodium, chloride, bicarbonate, and to pH. The treatments consisted of NaNO3, NaCl, NaHCO3, KNO3, KCl, or KHCO3 solutions, and a control treatment in which the seeds were germinated in deionized water and the plants grown in normal nutrient solution. In the first trial, germination and vigor were studied in 6 dS/m solutions. In the second, germination and vigor were studied in 60 mmol/L solutions and growth in a nutrient solution enriched with salts at the indicated concentration; the pH of the control and the nitrate and chloride solutions was maintained at 5.5, and that of the bicarbonate solutions at 8.5. In the third, the seeds were germinated in deionized water and the plantlets transplanted to a nutrient solution enriched with 60 mmol/L salts at the indicated pH values. In this trial, a treatment was added in which the pH of the nutrient solution with NaCl was raised to 8.5 to coincide with the bicarbonate solutions. The effect of sodium chloride was thus tested at the two indicated pH values. Based on the results, the conclusion was drawn that the harmful effects caused by high salt concentrations and pH, in both common bean varieties under study, can be ranked in the order: osmotic pressure > alkalinity > bicarbonate ≈ chloride > sodium. / O acúmulo de sais nos solos é um evento freqüente em regiões áridas ou semi-áridas, onde a evapotranspiração é elevada e a reduzida precipitação é insuficiente para promover a lixiviação do excesso de sais do perfil do solo. Nessas condições, a produtividade das culturas é reduzida, em resposta a eventuais efeitos osmóticos, à toxicidade de íons específicos presentes na solução do solo ou a um efeito conjunto desses fatores. Com o objetivo de isolar os efeitos decorrentes da presença de concentrações elevadas de sais na solução, verificando respostas a pressão osmótica, sódio, cloreto, bicarbonato e pH, foram realizados três ensaios, utilizando duas cultivares de feijoeiro: Diamante Negro e OPNS 331. Os tratamentos foram constituídos de soluções de NaNO3, NaCl, NaHCO3, KNO3, KCl ou KHCO3, além de um tratamento controle, em que as sementes foram germinadas em água deionizada e as plantas crescidas em solução nutritiva normal. No primeiro ensaio, estudou-se a germinação e o vigor em soluções de 6 dS/m. No segundo, germinação e vigor em soluções de 60 mmol/L e, a seguir, o crescimento em solução nutritiva acrescida dos sais na concentração indicada; o pH da testemunha e das soluções que incluíram nitratos e cloretos foi mantido em 5,5, e o das soluções com bicarbonato, em 8,5. No terceiro, as sementes foram germinadas em água deionizada e as plântulas transplantadas para a solução nutritiva com acréscimo de 60 mmol/L dos sais, nos valores de pH indicados. Neste ensaio, houve acréscimo de um tratamento adicional, no qual a solução nutritiva com NaCl teve seu pH elevado a 8,5, para coincidir com o das soluções de bicarbonato. Assim, testou-se o efeito do cloreto de sódio nos dois valores de pH indicados. Analisados os resultados, pôde concluir-se que, nos dois feijoeiros estudados, os efeitos prejudiciais, decorrentes de concentrações salinas e pH elevados, podem ser hierarquizados segundo a ordem: pressão osmótica > alcalinidade > bicarbonato ≈ cloreto > sódio.

För att bedöma mäns spermiekvalitet och därmed pars chanser till en graviditet har standardparametrar tidigare använts som fertilitetsindikator, men har visat sig vara en mindre tillförlitlig metod. Därför har en ny metod som bygger på flödescytometri och kallas Sperm Chromatin Structure Assay (SCSA) utvecklats som fokuserar på spermiernas arvsmassa, och som då har visat sig vara en mycket mer tillförlitlig metod. Syftet med denna studie var att jämföra DNA Fragmentations Index (DFI) -värde mellan svenska och danska män och se om en signifikant skillnad finns. För att uppfylla detta späddes och färgades spermaproven in och analyserades med SCSA, som därefter gav hur stor andel av spermiernas DNA som är defekt, vilket presenterades som ett procentuellt DFI-värde. Resultatet visade en signifikant skillnad av medianvärdet mellan svenska och danska män med avseende på DFI, där svenska män hade högre medianvärde än de danska männen. Detta i kombination med tidigare forskning som gjordes mellan svenska och danska män och som visade att svenska män hade bättre kvalitet än danska män med avseende på standardparametrarna, stödjer tidigare forskning som visar på att det ej finns ett samband mellan standardparametrar och DFI. Dock hade ytterligare forskning av skillnaderna mellan svenska och danska män behövts, för att validera fynden i denna studie. / To assess men's sperm quality and thus couples' chances of pregnancy, standard parameters have previously been used as a fertility indicator, but have proven to be a less reliable method. Therefore, a new method based on flow cytometry called Sperm Chromatin Structure Assay (SCSA) has been developed which focuses on the sperm's genetic mass, and which has been shown to be a much more reliable method. The purpose of this study was to compare the DNA Fragmentation Index (DFI) value between Swedish and Danish men and see if a significant difference exists. To fulfill this, the sperm samples were diluted and stained and analyzed with SCSA, which then gave the ratio of the sperm's DNA that is defective, which was presented as a percentage DFI value. The result showed a significant difference in the median value between Swedish and Danish men with regard to DFI, where Swedish men had a higher median value than the Danish men. This, in combination with previous research that has been done between Swedish and Danish men which showed that Swedish men had better quality than Danish men in regards to the standard parameters, supports previous research that shows that there is no connection between standard parameters and DFI. However, further research into the differences between Swedish and Danish men is needed to validate the findings in this study.