Atrial natriuretic factor (ANF) is a cardiac hormone that helps maintain cardiovascular homeostasis. ANF secretion is linked to the constitutive, regulated and constitutive-like pathways. Presence of a monensin-sensitive pool that may follow constitutive-like secretion has previously been identified in an isolated atrial perfusion study. The intracellular ANF storage pools linked to each secretory pathway have not been identified. In this study, ANF storage and secretion was characterized in HL-1 atrial cardiomyocytes through the use of pharmacological agents, density gradient and RP- HPLC analysis. Treatment of HL-1 cells with monensin followed by cell fractionation was unsuccessful in identifying the monensin-sensitive pool. RP-HPLC analysis identified presence of low molecular weight ANF in low density gradient fractions that were defined by the presence of organelle markers of Golgi, early endosome, clathrin and corin. Since the monensin-sensitive pool was thought to be of a constitutive-like nature, targeting this pathway with pharmacological inhibitors of clathrin coat vesicle (CCV) formation and endosomal trafficking failed to prevent stimuli-independent secretion. Based on an inability to prevent ANF secretion by targeting the constitutive-like pathway and the presence of low molecular weight ANF in low density gradient fractions, stimuli- independent ANF secretion seems to be through a constitutive pathway.

Atrial natriuretic factor (ANF) is a cardiac hormone that helps maintain cardiovascular homeostasis. ANF secretion is linked to the constitutive, regulated and constitutive-like pathways. Presence of a monensin-sensitive pool that may follow constitutive-like secretion has previously been identified in an isolated atrial perfusion study. The intracellular ANF storage pools linked to each secretory pathway have not been identified. In this study, ANF storage and secretion was characterized in HL-1 atrial cardiomyocytes through the use of pharmacological agents, density gradient and RP- HPLC analysis. Treatment of HL-1 cells with monensin followed by cell fractionation was unsuccessful in identifying the monensin-sensitive pool. RP-HPLC analysis identified presence of low molecular weight ANF in low density gradient fractions that were defined by the presence of organelle markers of Golgi, early endosome, clathrin and corin. Since the monensin-sensitive pool was thought to be of a constitutive-like nature, targeting this pathway with pharmacological inhibitors of clathrin coat vesicle (CCV) formation and endosomal trafficking failed to prevent stimuli-independent secretion. Based on an inability to prevent ANF secretion by targeting the constitutive-like pathway and the presence of low molecular weight ANF in low density gradient fractions, stimuli- independent ANF secretion seems to be through a constitutive pathway.

A infestação por carrapatos é uma das principais parasitoses que afetam a bovinocultura prejudicando a saúde do animal por meio de hábito hematófago, além de gerar perdas econômicas para o seu controle, sendo o uso de carrapaticidas o principal método utilizado. O uso negligenciado destes produtos induz a geração de resistência tornando necessário o desenvolvimento de métodos alternativos, como a imunização por vacinas e o uso de probióticos. Probióticos são alimentos funcionais, compostos de micro-organismos vivos, que quando ingeridos em doses adequadas e com frequência, conferem diversos efeitos benéficos à saúde do hospedeiro por meio de diferentes mecanismos de ação. Desta forma, o presente trabalho teve como objetivo avaliar o efeito imunomodulador e carrapaticida de micro-organismos probióticos em bovinos da raça Girolando. Para tanto, durante 154 dias, foram estudados 30 animais distribuídos em 06 grupos e submetidos a diferentes tratamentos, a saber: Grupo A (controle) os animais não receberam tratamento específico; Grupo B (efeito vacina) nos dias inicial, 14 e 28 os animais foram imunizados com a proteína rGST-Hl; Grupo C (efeito probiótico EEL) os animais receberam, durante 70 dias, uma dose diária de 10 mL, da preparação probiótica EEL; Grupo D (efeito vacina + probiótico EEL) nos dias inicial, 14 e 28 os animais foram vacinados e receberam durante 70 dias, 10 mL da preparação probiótica EEL; Grupo E (efeito probiótico PT) os animais receberam, durante 70 dias, uma dose de 10 mL da preparação probiótica ProbioTick e Grupo F (efeito vacina + probiótico PT) nos dias inicial, 14 e 28 os animais foram vacinados e receberam diariamente, durante 70 dias, 10 mL da preparação probiótica ProbioTick. A cada 14 dias foram realizadas coletas de sangue para avaliação da hemostasia, do estado de anemia e o desenvolvimento da resposta imune humoral. Os resultados demonstraram que os tratamentos não afetaram significativamente a hemostasia, bem como os parâmetros eritrocíticos dos animais. No tocante ao sistema imunológico, verificou-se que os animais tratados com as preparações probióticas apresentaram maior número de linfócitos quando comparado ao grupo controle diferindo estatisticamente no 70º dia, sendo que os percentuais encontrados para os grupos C, D, E e F foram respectivamente de 55,8%; 63,5%; 60,8% e 59,4%. Os animais que foram vacinados (grupo B) e tratados com as preparações probióticas (Grupos D e F), apresentaram níveis superiores de anticorpos totais IgG (72,769; 93,735 e 98,825 mg/dL) e quanto aos anticorpos IgG anti rGST-Hl, os animais dos Grupos D e F tiveram seus valores aumentados quando comparados aos do Grupo B por um período mais prolongado, onde no 98º dia, as densidades ópticas medidas foram de 0,868 e 0,915. Desta forma, conclui-se que as preparações probióticas foram capazes de modular a resposta imunológica dos animais. / Tick infestation is the main parasitic diseases that affect the cattle farming. In addition to impairing the health of the animal by means of a hematophagous habit, it also generates economic losses regarding its control, being the use of acaricides the main method used. Its neglected use has led to cases of resistance, which turn necessary the development of alternative methods such as the immunization with vaccines and the use of probiotics. Probiotics are functional food that contains live microorganisms that when ingested in appropriate amount bring benefits to the hosts through different mode of action. Thus, the present work aimed to evaluate the immunomodulatory and acaricide effect of probiotic microorganisms in Girolando cattle. Therefore, during 154 days, 30 animals were distributed in 6 groups, and submitted to different treatments: Group A (control), the animals did not received any treatment. Group B (vaccine effect), on the initial, 14 and 28 days, the animals were immunized with rGST-H1. Group C (probiotic effect EEL, the animals received, for 70 days, a daily dose of 10 mL of the probiotic EEL preparation. Group D (vaccine + probiotic EEL effect), on the initial, 14 and 28 days, the animals were vaccinated and received for 70 days, 10 mL probiotic preparation EEL. Group E (probiotic effect PT), the animals received, for 70 days, a dose of 10 mL of probiotic preparation ProbioTick and Group F (vaccine + PT probiotic effect), on the initial, 14 and 28 days, the animals were vaccinated and received daily for 70 days, 10 mL ProbioTick. In each 14 days blood samples were collected and exams were performed to evaluate hemostasis, anemia status and development of the immune response. The results showed that the different treatments did not affect hemostasis and the erythrocytic parameters of the animals. Regarding the immune system, the results showed that the animals treated with probiotic preparations presented a higher number of lymphocytes when compared to the control group, differing statistically on the 70th day, being that the percentages found for groups C, D, E and F were respectively 55.8%; 63.5%; 60.8% and 59.4%. In addition, animals treated with the probiotic (Groups D and F) and vaccinated (B) showed higher levels of total IgG antibodies (72.769, 93.735 and 98.825 mg/dL) and as for the IgG anti rGST-H1 antibodies, only the animals in Groups D and F had their values increased for a longer period, where on the 98th day, The optical densities measured were 0.868 and 0.915. Therefore, the probiotic preparations evaluated were able to modulate the imune response of animals.

Atrial natriuretic factor (ANF) is a cardiac hormone that helps maintain cardiovascular homeostasis. ANF secretion is linked to the constitutive, regulated and constitutive-like pathways. Presence of a monensin-sensitive pool that may follow constitutive-like secretion has previously been identified in an isolated atrial perfusion study. The intracellular ANF storage pools linked to each secretory pathway have not been identified. In this study, ANF storage and secretion was characterized in HL-1 atrial cardiomyocytes through the use of pharmacological agents, density gradient and RP- HPLC analysis. Treatment of HL-1 cells with monensin followed by cell fractionation was unsuccessful in identifying the monensin-sensitive pool. RP-HPLC analysis identified presence of low molecular weight ANF in low density gradient fractions that were defined by the presence of organelle markers of Golgi, early endosome, clathrin and corin. Since the monensin-sensitive pool was thought to be of a constitutive-like nature, targeting this pathway with pharmacological inhibitors of clathrin coat vesicle (CCV) formation and endosomal trafficking failed to prevent stimuli-independent secretion. Based on an inability to prevent ANF secretion by targeting the constitutive-like pathway and the presence of low molecular weight ANF in low density gradient fractions, stimuli- independent ANF secretion seems to be through a constitutive pathway.

Financial analysis of Dopravní podnik hl. m. Prahy, a.s.

The diploma thesis sets out to deal with the role of local governments for the protection of cultural values. The theoretical part discusses what cultural values are, what are their contributions and why it is necessary to protect them. This is followed by the analysis of instruments and institutions regarding the protection of culture values in the Czech Republic with the focus on Prague, being a significant cultural centre with many cultural values within its territory. It can be seen that local governments have many options on how to protect their cultural values, for which their level of initiative is crucial. The Prague government has many institutions and instruments aiming to protect the cultural values of the area, however there are some shortcomings. The most current one is a missing updated cultural strategy in Prague. From a long-term point of view, it can also be seen that there is a decline in spending on culture and lack of education and awareness in the cultural sphere.

As part of the Large Hadron Collider high luminosity upgrade it is proposed to include crab cavities and large aperture niobium tin final focussing magnets in the lattice in order to enhance the luminosity. In this thesis the dynamics of a proposed cavity design were considered in terms of their impact upon the dynamic aperture of the machine. Taylor maps for the cavity were created and used to perform this analysis with a full assessment of their validity. A set of symplectic thin cavity models were also developed and cross checked with the Taylor maps. Finally, dynamic aperture studies were performed using these models in order to determine which components of the crab cavity dynamics are important when considering the long term stability of the beam in the LHC upgrade. It is shown that crab cavities exhibit little impact on the LHC beam stability. For the final focussing magnets a preliminary study was conducted into the importance of including their fringe fields in a model of the LHC upgrade. A technical study was carried out into developing a symplectic model which was compatible with the current magnet models use for dynamic aperture studies. A preliminary dynamic aperture study was performed with the inclusion of fringe fields for the final focussing magnets from which the fringe fields are shown to have a negative impact on the long term beam stability.

This diploma thesis deals with processing of high strength aluminum alloy EN AW 7075 by Selective Laser Melting and verify the influence of process parameters on relative density and mechanical properties. The theoretical part contains an introduction to additive manufacturing of aluminum alloys, the influence of process parameters and description of processes occuring during SLM production. Based on the theoretical part were prepared experiments and method of evaluation. Samples were produced by melting metallurgical powder using ytterbium laser with a maximum output power of 400 W. This diploma thesis also describes the formation and growth of cracks inside the material, which so far in the literature for alloy EN AW 7075 were not described. As the result, the process parameters dependence on the relative density and an overview of this aluminum alloy processing by SLM technology is determined.

This diploma thesis deals with finding and verification of appropriate technological parameters of SLM technology for the processing of aluminum alloy 2618. In the theoretical part, an introduction to additive manufacturing of aluminum alloys and general description of processes occurring during SLM production is given. Based on general knowledge were designed different types of testing samples produced by sintering the metallurgical powder using 400 W ytterbium fiber laser, which so far in the literature for aluminum alloy 2618 were not described. As the result, the technological parameters dependence on relative density and the detailed overview of the 2618 alloy processing by SLM technology is determined.

This diploma thesis deals with finding copper alloy suitable for processing SLM technology and determining the process parameters leading to a relative density close to the full material. The theoretical part provides an insight into additive technology and the processing of new alloys in SLM. Work also contains a search report of processed copper alloys used in SLM. Based on the theoretical part were designed test samples and method of evaluation. Samples were produced by melting metallurgical powder using ytterbium laser with an output power 400 W. The testing is divided into three stages; Determination of the parameters of the SLM process, Debug strategies for larger parts, Geometric precision and mechanical testing. Based on the results was determined dependence of relative density on the input parameters. For the best parameters were determined geometric precision correction and mechanical properties.