O objetivo da presente tese foi desenvolver um adesivo com nanocápsulas (NCs), contendo indometacina, e um sistema adesivo com nanocápsulas (NC), contendo indometacina e triclosan, e avaliar suas propriedades. As NCs foram produzidas por meio do método de deposição de polímero, secas e caracterizadas quanto ao tamanho de partículas, à forma, quantidade de fármaco encapsulado e citotoxicidade. Uma resina adesiva foi formulada. Foram adicionadas ao adesivo 1%, 2%, 5% e 10% de NCs em massa, e um grupo permaneceu sem NC. As NCs, contendo indometacina e triclosan, foram também incorporadas no primer comercial a 2% em peso e um grupo permaneceu sem NCs. Os adesivos foram avaliados quanto ao GC imediato e tardio, à degradação em solvente, liberação dos fármacos, difusão de indometacina pela dentina e resistência de união. O adesivo com 10% de NCs, contendo indometacina, foi também avaliado quanto à ação anti-inflamatória em modelo animal. O primer e o adesivo com as diferentes concentrações de NCs, contendo indometacina e triclosan, foram avaliados quanto à liberação dos fármacos, difusão de indometacina pela dentina, ao efeito antimicrobiano, grau de conversão (GC) in situ, ângulo de contato e à resistência de união à microtração. Os dados foram analisados por ANOVA, Tukey e teste t. As NCs apresentaram forma esférica e viabilidade celular acima de 80%. As NCs, contendo indometacina, apresentaram diâmetro médio de 165 nm e as NCs, contendo indometacina e triclosan, 159 nm. O adesivo, contendo 10% de NCs com indometacina, apresentou efeito anti-inflamatório. A incorporação de NCs não alterou o GC, que variou de 63,63 ± 1,01% a 70,50 ± 2,08%. A degradação em solvente não foi alterada com 2% de NCs. Tanto os adesivos quanto o primer apresentaram liberação controlada. A indometacina permeou através da dentina. O adesivo e primer também apresentaram efeito antimicrobiano. A incorporação de NCs no primer e no adesivo não influenciou o GC in situ nem a resistência de união imediata, em comparação aos materiais sem NCs. O uso concomitante do primer e adesivo com NCs aumentou o ângulo de contato e diminuiu a resistência de união longitudinal. Conclui-se que o uso do adesivo com a incorporação de NCs tem potencial para proporcionar ações terapêuticas à adesão dentinária. / The aim of the present thesis was to develop an adhesive with nanocapsules (NCs) containing indomethacin and an adhesive system with nanocapsules containing indomethacin and triclosan and to evaluate their properties. NCs were prepared by the interfacial deposition of preformed polymer technique, dried and characterized regarding particle size, encapsulated drug content and cytotoxicity. Adhesive resin was produced. Concentrations of 1%, 2%, 5% and 10% of NCs were added in the adhesive and a group was maintained with no NCs. Indomethacin and triclosan-loaded NCs were also incorporated into a commercial primer in a concentration of 2% and a group was maintained with no NCs. Adhesives were evaluated regarding immediate and late degree of conversion (DC), degradation in solvent, drug release, indomethacin diffusion through dentin and bond strength. The adhesive with 10% of NCs containing indomethacin was also evaluated regarding the anti-inflammatory effect in an animal model. Primer and adhesive with different concentrations of NCs containing indomethacin and triclosan were evaluated regarding drug release, indomethacin diffusion through dentin, antimicrobial effect, in situ degree of conversion, contact angle and bond strength. Data were analyzed through ANOVA, Tukey post-hoc and t-test.NCs presented a spherical shape and cell viability higher than 80%. NCs containing indomethacin presented an averaged size of 165 nm and NCs containing indomethacin and triclosan, 159 nm. The adhesive with 10% of NCs containing indomethacin presented anti-inflammatory effect. The incorporation of NCs presented no alteration of DC, varying from 63.63 ± 1.01% a 70.50 ± 2.08%. Degradation in solvent suffers no influence of NCs with 2% of NCs. Adhesives and primer presented controlled drug release. Indomethacin diffused through dentin. Adhesive and primer also presented an antimicrobial effect. The incorporation of NCs in adhesive and primer showed no influence on in situ DC and immediate bond strength compared to materials with no NCs. The use in combination of primer and adhesive with NCs resulted in higher contact angle and lower longitudinal bond strength. It is possible to conclude that the use adhesive with incorporation of NCs has potential to provide therapeutic effects on dentin adhesion.

El insight, o conciencia de enfermedad, en los trastornos psicóticos ha despertado recientemente un gran interés en la Psiquiatría y la Psicología Clínica debido a la relación que ha mostrado tener con numerosas variables relevantes, tanto desde el punto de vista científico, como social, e incluso legal. El estudio del déficit en insight en las psicosis, se remonta al siglo XVII, aunque el estudio científico se inicia en el siglo XX, e implica un cambio en la conceptualización del mismo. De modelos categoriales y unitarios, se ha evolucionado a modelos dimensionales y multifacéticos, siendo el modelo de Amador y David uno de los más aceptados en la actualidad. Desde este modelo, se entiende por Insight la conciencia de sufrir una enfermedad, la capacidad para entender las experiencias psicóticas como anormales, los procesos atribucionales o explicaciones del paciente sobre la causa última de estos signos o síntomas, la adherencia al tratamiento y la conciencia sobre las consecuencias sociales de la enfermedad. Por otra parte, se han considerado en los últimos años diversos tipos de insight, además del clínico (conciencia de enfermedad). Así, se considera también el insight cognitivo o metacognitivo, que hace referencia a la conciencia de los pacientes sobre su propio déficit o deterioro cognitivo. En cuanto a la etiología del déficit en insight en la esquizofrenia y otros trastornos psicóticos, se han postulado diversas hipótesis, como son la psicológica, la clínica (o psicopatológica) y la neurocognitiva. Desde la primera se postula que el déficit en insight se relacionaría con un mecanismo de defensa o estilo de afrontamiento frente a los efectos depresógenos del hecho de padecer una enfermedad de este tipo. Desde la segunda, la explicación clínica, se concibe el déficit en insight como un síntoma más del trastorno psicótico, en la línea de las tesis clásicas que lo consideraban en la génesis y desarrollo de la enfermedad. Por último, desde las teorías neurocognitivas, se relaciona el déficit en insight como una consecuencia del déficit cognitivo y/o las alteraciones neuropsicológicas presentes en este tipo de trastornos, que a su vez se relacionan con alteraciones en el neurodesarrollo. El objetivo del presente estudio es aportar información relevante para la validación de las últimas hipótesis etiológicas mencionadas, de modo que se obtengan datos que permitan valorar la relación existente entre el nivel de insight y la sintomatología (positiva, negativa, general y depresiva), así como al relación entre el nivel de insight y el rendimiento en las distintas funciones cognitivas. Para ello se estudiaron un grupo de 70 pacientes ingresados con un primer episodio psicótico. Se realizó una evaluación clínica mediante la administración de la entrevista clínica estructurada para los trastornos del DSM (SCID-I) y el ajuste premórbido mediante la escala de Ajuste premórbido (PAS). Además, se evaluó el funcionamiento cognitivo de los sujetos mediante una batería de instrumentos neuropsicológicos estandarizados, el tipo y gravedad de la sintomatología mediante la administración de la Escala para el síndrome positivo y negativo de la esquizofrenia (PANSS) y se evaluó la sintomatología depresiva mediante la administración de la escala Calgary de depresión en la esquizofrenia. Por otra parte, la evaluación del Insight clínico se llevó a cabo mediante la administración de la escala SUMD de Amador y el Insight metacognitivo se valoró mediante la GEOPTE (escala para la percepción subjetiva del déficit cognitivo, que engloba tanto la percepción subjetiva del déficit en cognición básica como en cognición social). Los resultados pusieron de manifiesto al ausencia de relaciones significativas entre el insight y al sintomatología psicótica y la presencia de algunas asociaciones significativas entre los dos tipos de insight estudiados y algunas variables neurocognitivas. / Insight, or awareness of illness, in psychotic disorders has recently aroused a great interest in Psychiatry and Clinical Psychology because its relationship with numerous relevant variables, both scientifically, socially, and even legal. The study of deficits in insight in psychosis dates from the seventeenth century, but scientific study began in the twentieth century, and involves a change in the conceptualization the concept. Categorical and unitary models, the study has evolved to multifaceted and dimensional models, and Amador & David model’s is one of the most widely accepted today. From the point of view of this model, Insight is conceptualized as the disease awareness, the ability to understand psychotic experiences as abnormal, the attributional processes and patient explanations of the ultimate causes of these signs or symptoms, the treatment adherence and the awareness about social consequences of the disease. On the other hand, have been considered in recent years various types of insight, as well as clinical (awareness of illness). Thus, it is also considered cognitive or metacognitive insight, which refers to the awareness of patients about their own cognitive impairment. As for the etiology of insight deficits in schizophrenia and other psychotic disorders, various hypotheses have been postulated, such as psychological, clinical (or psychopathology) and neurocognitive. Since the first one postulates that the insight deficit would be related to a defense mechanism or coping style against the depressogenic effects of the awareness of illness. The second hypotheses, the clinical explanation, is conceived as a symptom of psychotic disorder, in line with the classical thesis that considered this phenomena in the genesis and development of the disease. Finally, from neurocognitive theories is related insight deficits as a consequence of cognitive deficits and/or neuropsychological abnormalities present in these disorders, which in turn is associated with impaired neurodevelopment. The aim of this study is to provide relevant information for the validation of the last mentioned etiological hypotheses, so as to obtain data to assess the relationship between the level of insight and symptomatology (positive, negative, general and depressive) and the relationship between the level of insight and performance in several cognitive functions. To do this we studied a group of 70 patients admitted with a first psychotic episode. Clinical evaluation was performed by administering the Structured Clinical Interview for DSM Disorders (SCID-I) and premorbid adjustment by the Premorbid Adjustment Scale (PAS). In addition, we evaluated the subject’s cognitive functioning with a battery of standardized neuropsychological instruments, the type and severity of symptoms by the administration of the Scale for Positive and Negative Syndrome of Schizophrenia (PANSS) and depressive symptoms was assessed by the administration of the Calgary Depression Scale in Schizophrenia. Moreover, clinical Insight assessment was conducted by administering the Amador’s SUMD Scale and metacognitive Insight was assessed with the GEOPTE (the Scale for the subjective perception of cognitive deficit), which includes both the subjective perception of the deficit in the basic cognitive functioning and in the subjective perception of deficit in the social cognition. Results showed the absence of significant relationships between insight and psychotic symptoms and the presence of some significant associations between the two types of insight (clinical and metacognitive) with some of the neurocognitive variables.

La presente Tesis desarrolla un estudio acerca de las modalidades discursivas mediante las que tres de los principales periódicos argentinos de capitales privados, Clarín, La Nación y Página/12, se consolidaron como actores políticos durante el primer kirchnerismo. Revalorizando la importancia del periodo presidencial 2003-2007 en la injerencia en la conflictividad entre prensa y kirchnerismo, se constató que los periódicos construyeron sus propios posicionamientos en una tensión permanente entre las respuestas a los acontecimientos políticos que marcaron el perfil del gobierno de Néstor Kirchner y sus propios marcos ideológicos e interpretativos institucionales. Se abordó un área en tensión por su hibridez entre la discursividad periodística y la política, que configuró el vínculo, el solapamiento, la disputa y/o la complementariedad entre los posicionamientos construidos por los periódicos en sus secciones de opinión cuando intervinieron políticamente en los debates y definiciones en torno al poder político, en un contexto en el que fueron ellos mismos cuestionados públicamente respecto de la validez y la transparencia de las prácticas productivas del periodismo.

O objetivo da presente tese foi desenvolver um adesivo com nanocápsulas (NCs), contendo indometacina, e um sistema adesivo com nanocápsulas (NC), contendo indometacina e triclosan, e avaliar suas propriedades. As NCs foram produzidas por meio do método de deposição de polímero, secas e caracterizadas quanto ao tamanho de partículas, à forma, quantidade de fármaco encapsulado e citotoxicidade. Uma resina adesiva foi formulada. Foram adicionadas ao adesivo 1%, 2%, 5% e 10% de NCs em massa, e um grupo permaneceu sem NC. As NCs, contendo indometacina e triclosan, foram também incorporadas no primer comercial a 2% em peso e um grupo permaneceu sem NCs. Os adesivos foram avaliados quanto ao GC imediato e tardio, à degradação em solvente, liberação dos fármacos, difusão de indometacina pela dentina e resistência de união. O adesivo com 10% de NCs, contendo indometacina, foi também avaliado quanto à ação anti-inflamatória em modelo animal. O primer e o adesivo com as diferentes concentrações de NCs, contendo indometacina e triclosan, foram avaliados quanto à liberação dos fármacos, difusão de indometacina pela dentina, ao efeito antimicrobiano, grau de conversão (GC) in situ, ângulo de contato e à resistência de união à microtração. Os dados foram analisados por ANOVA, Tukey e teste t. As NCs apresentaram forma esférica e viabilidade celular acima de 80%. As NCs, contendo indometacina, apresentaram diâmetro médio de 165 nm e as NCs, contendo indometacina e triclosan, 159 nm. O adesivo, contendo 10% de NCs com indometacina, apresentou efeito anti-inflamatório. A incorporação de NCs não alterou o GC, que variou de 63,63 ± 1,01% a 70,50 ± 2,08%. A degradação em solvente não foi alterada com 2% de NCs. Tanto os adesivos quanto o primer apresentaram liberação controlada. A indometacina permeou através da dentina. O adesivo e primer também apresentaram efeito antimicrobiano. A incorporação de NCs no primer e no adesivo não influenciou o GC in situ nem a resistência de união imediata, em comparação aos materiais sem NCs. O uso concomitante do primer e adesivo com NCs aumentou o ângulo de contato e diminuiu a resistência de união longitudinal. Conclui-se que o uso do adesivo com a incorporação de NCs tem potencial para proporcionar ações terapêuticas à adesão dentinária. / The aim of the present thesis was to develop an adhesive with nanocapsules (NCs) containing indomethacin and an adhesive system with nanocapsules containing indomethacin and triclosan and to evaluate their properties. NCs were prepared by the interfacial deposition of preformed polymer technique, dried and characterized regarding particle size, encapsulated drug content and cytotoxicity. Adhesive resin was produced. Concentrations of 1%, 2%, 5% and 10% of NCs were added in the adhesive and a group was maintained with no NCs. Indomethacin and triclosan-loaded NCs were also incorporated into a commercial primer in a concentration of 2% and a group was maintained with no NCs. Adhesives were evaluated regarding immediate and late degree of conversion (DC), degradation in solvent, drug release, indomethacin diffusion through dentin and bond strength. The adhesive with 10% of NCs containing indomethacin was also evaluated regarding the anti-inflammatory effect in an animal model. Primer and adhesive with different concentrations of NCs containing indomethacin and triclosan were evaluated regarding drug release, indomethacin diffusion through dentin, antimicrobial effect, in situ degree of conversion, contact angle and bond strength. Data were analyzed through ANOVA, Tukey post-hoc and t-test.NCs presented a spherical shape and cell viability higher than 80%. NCs containing indomethacin presented an averaged size of 165 nm and NCs containing indomethacin and triclosan, 159 nm. The adhesive with 10% of NCs containing indomethacin presented anti-inflammatory effect. The incorporation of NCs presented no alteration of DC, varying from 63.63 ± 1.01% a 70.50 ± 2.08%. Degradation in solvent suffers no influence of NCs with 2% of NCs. Adhesives and primer presented controlled drug release. Indomethacin diffused through dentin. Adhesive and primer also presented an antimicrobial effect. The incorporation of NCs in adhesive and primer showed no influence on in situ DC and immediate bond strength compared to materials with no NCs. The use in combination of primer and adhesive with NCs resulted in higher contact angle and lower longitudinal bond strength. It is possible to conclude that the use adhesive with incorporation of NCs has potential to provide therapeutic effects on dentin adhesion.

The subject of this thesis is the design of specific LAMP primers and LAMP analysis and its optimization. The purpose of this analysis is to specifically distinguish Xanthomonas gardneri from other pathogenic xanthomonads. Two sets of LAMP primers from the DNA sequences of hrpB and atpD genes were designed. These LAMP primer sets are highly sensitive and the detection limit of LAMP assay was found to be 0.01 l / mg of DNA. The reaction temperature of the LAMP was optimized to 64 °C to reach the maximum of amplification. LAMP can be applied to detection and identification on pathogens in plant tissues. LAMP takes less time than conventional PCR to detect bacteria. This LAMP assay has a great potintial to be applied to detection and identification of Xanthomonas genus bacteria pathogenic for tomato and pepper.

This thesis is based on the use of ISSR method for studying genetic diversity of oilseed rape. Altogether, 180 genotypes were evaluated. Frozen leaves of oilseed rape from three breeding station called: 1/ breeding station Selgen a.s., 2/ OSEVA PRO s.r.o. and 3/ breeding station Slapy u Tábora were used as imput material. DNA isolation was performed by Williams, who modified CTAB method for oilseed rape. We selected for ISSR 4 UBC which showed high variability and stability. The analysis was performed by gel electrophoresis. This thesis is also focused on comparison of gel electrophoresis and chip electrophoresis. In this thesis We show a matrix of simmilarity between varieties. The results were also provided back to breeders as a basis for further breeding.

O objetivo da presente tese foi desenvolver um adesivo com nanocápsulas (NCs), contendo indometacina, e um sistema adesivo com nanocápsulas (NC), contendo indometacina e triclosan, e avaliar suas propriedades. As NCs foram produzidas por meio do método de deposição de polímero, secas e caracterizadas quanto ao tamanho de partículas, à forma, quantidade de fármaco encapsulado e citotoxicidade. Uma resina adesiva foi formulada. Foram adicionadas ao adesivo 1%, 2%, 5% e 10% de NCs em massa, e um grupo permaneceu sem NC. As NCs, contendo indometacina e triclosan, foram também incorporadas no primer comercial a 2% em peso e um grupo permaneceu sem NCs. Os adesivos foram avaliados quanto ao GC imediato e tardio, à degradação em solvente, liberação dos fármacos, difusão de indometacina pela dentina e resistência de união. O adesivo com 10% de NCs, contendo indometacina, foi também avaliado quanto à ação anti-inflamatória em modelo animal. O primer e o adesivo com as diferentes concentrações de NCs, contendo indometacina e triclosan, foram avaliados quanto à liberação dos fármacos, difusão de indometacina pela dentina, ao efeito antimicrobiano, grau de conversão (GC) in situ, ângulo de contato e à resistência de união à microtração. Os dados foram analisados por ANOVA, Tukey e teste t. As NCs apresentaram forma esférica e viabilidade celular acima de 80%. As NCs, contendo indometacina, apresentaram diâmetro médio de 165 nm e as NCs, contendo indometacina e triclosan, 159 nm. O adesivo, contendo 10% de NCs com indometacina, apresentou efeito anti-inflamatório. A incorporação de NCs não alterou o GC, que variou de 63,63 ± 1,01% a 70,50 ± 2,08%. A degradação em solvente não foi alterada com 2% de NCs. Tanto os adesivos quanto o primer apresentaram liberação controlada. A indometacina permeou através da dentina. O adesivo e primer também apresentaram efeito antimicrobiano. A incorporação de NCs no primer e no adesivo não influenciou o GC in situ nem a resistência de união imediata, em comparação aos materiais sem NCs. O uso concomitante do primer e adesivo com NCs aumentou o ângulo de contato e diminuiu a resistência de união longitudinal. Conclui-se que o uso do adesivo com a incorporação de NCs tem potencial para proporcionar ações terapêuticas à adesão dentinária. / The aim of the present thesis was to develop an adhesive with nanocapsules (NCs) containing indomethacin and an adhesive system with nanocapsules containing indomethacin and triclosan and to evaluate their properties. NCs were prepared by the interfacial deposition of preformed polymer technique, dried and characterized regarding particle size, encapsulated drug content and cytotoxicity. Adhesive resin was produced. Concentrations of 1%, 2%, 5% and 10% of NCs were added in the adhesive and a group was maintained with no NCs. Indomethacin and triclosan-loaded NCs were also incorporated into a commercial primer in a concentration of 2% and a group was maintained with no NCs. Adhesives were evaluated regarding immediate and late degree of conversion (DC), degradation in solvent, drug release, indomethacin diffusion through dentin and bond strength. The adhesive with 10% of NCs containing indomethacin was also evaluated regarding the anti-inflammatory effect in an animal model. Primer and adhesive with different concentrations of NCs containing indomethacin and triclosan were evaluated regarding drug release, indomethacin diffusion through dentin, antimicrobial effect, in situ degree of conversion, contact angle and bond strength. Data were analyzed through ANOVA, Tukey post-hoc and t-test.NCs presented a spherical shape and cell viability higher than 80%. NCs containing indomethacin presented an averaged size of 165 nm and NCs containing indomethacin and triclosan, 159 nm. The adhesive with 10% of NCs containing indomethacin presented anti-inflammatory effect. The incorporation of NCs presented no alteration of DC, varying from 63.63 ± 1.01% a 70.50 ± 2.08%. Degradation in solvent suffers no influence of NCs with 2% of NCs. Adhesives and primer presented controlled drug release. Indomethacin diffused through dentin. Adhesive and primer also presented an antimicrobial effect. The incorporation of NCs in adhesive and primer showed no influence on in situ DC and immediate bond strength compared to materials with no NCs. The use in combination of primer and adhesive with NCs resulted in higher contact angle and lower longitudinal bond strength. It is possible to conclude that the use adhesive with incorporation of NCs has potential to provide therapeutic effects on dentin adhesion.

This Masterâs thesis deals with evaluation of quality of Brno Combo combination rifle shotgun rifled barrelâs firing mechanism. It contains a proposal of criteria for evaluation of quality of firearm firing mechanisms. It further focuses on using measurement of primer deformation as a means of assessing the quality of a firing mechanism. Based on measurements made using a Focus Variation microscope, various geometrical parameters of fired and failed primers were analysed. The measurement results were processed using ANOVA and regression analysis. Based on output of these analyses, practical recommendations and further research proposals were formulated.

During the purification of the enzyme glycogen synthase from the muscle of the nematode Ascaris suum, approximately 70% of the glycogen synthase activity can be separated from the bulk of cellular glycogen by centrifugation for 60 min at 105,000 x . The glycogen synthase in the supernatant fraction has an Mr of 1.2 x 106 as determined by Sepharose 4B gel filtration chromatography. The glycogen synthase in this high molecular weight complex (glycogen primer complex) can be further purified by ConA-Sepharose affinity chromatography; the enzyme activity was eluted with 100 .mM a-methylmannoside. The glycogen synthase in glycogen'primer complex is predominately in the glucose 6-phosphatedependent form. The glycogen primer complex can catalyze the transfer of glucosyl units from UDP-glucose to an endogenous acceptor in the absence of exogenous glycogen. Analysis by SDS-PAGE showed three proteins (Mr 140,000, 78,000 and 34,000) and a carbohydrate polymer. The carbohydrate polymer can be partially digested with a-amylase. The glycogen primer complex was further digested by acid hydrolysis, and upon descending paper chromatography analysis, eight different carbohydrates were isolated, two of which were tentatively identified as glucose and sialic acid. The [14 C]-autoradiograph showed that in vitro synthesis of a glycogen-like polysaccharide occurred on this carbohydrate polymer. Polyclonal antibodies have been made to the glycogen primer complex, and Western Blot analysis indicated that all three proteins of the glycogen primer complex were antigenic. Collectively, the data indicate that a glycogen-like polysaccharide is synthesized from a carbohydrate-associated protein primer in the muscle of this worm.

Yes / Magnetic nanoparticles and surface-enhanced Raman scattering (SERS) active nanoparticles were coated with short chain DNA tags. These were then used to identify a target bacterial DNA sequence. The tags function as primers in a standard PCR with the reverse primers and forward primers on the SERS nanoparticles and magnetic nanoparticles, respectively. During the PCR cycles, a composite nanostructure is formed that is both magnetically responsive and SERS active. After magnetic trapping, the intensity of the SERS signal can be related back to the concentration of the target DNA. A test assay was performed that showed a detection limit (based on the signal to noise ratio) of less than 3 zeptomole (41 pg/L). For comparison, a PCR assay based on the standard SYBR Green method was performed. This used the same primers and target DNA and had a detection limit of 10 attomoles (138 ng/L), 3,000 times less sensitive. The work documents the proof of principle study and shows for the first time the use of SERS-NP labels in the quantification of nucleic acid amplification tests and PCR.