Akademiks avhandling--Uppsala. / Bibliography: pref. p. 4.

Akademisk avhandling--Uppsala. / Extra t.p., with thesis statement in full, inserted. Bibliography: p. [xi]-xx.

Peroxisome Proliferator-Activated Receptors (PPARs) belong to the nuclear receptor superfamily of transcription factors. PPARs, like some other nuclear receptors, bind their cognate DNA binding sites as heterodimers with Retinoid X Receptor (RXR). Like most members of this superfamily, PPARs regulate transcription of target genes in response to specific lypophilic ligands. Given the fact, however, that many members of the nuclear receptor superfamily possess a ligand independent activation function (AF -1) in their amino-terminal A/B domains, we identified and characterized the AF-1 activity in the A/B domains of mPPARα and mPPARγ2. When fused to the GAL4 DNA Binding Domain (DBD), the A/B domains of both mPPARα and mPPARγ2 subtypes are transcriptionally active both in mammalian and yeast cells in the absence of cognate PPAR activators, indicating that both PPAR subtypes do in fact contain AF -1 activities in their respective A/B domains. Our deletion studies localizing the region in the A/B domains of the PPAR subtypes indicate that the amino-terminal region of A/B domains of both PP AR subtypes is necessary, but not sufficient, for the AF -1 activity. After having identified and characterized the AF-1 activities of both mPPARα and mPPARγ subtypes, we proceeded to investigate how this activity is controlled. Generally, nuclear receptors, including PPARs, do not activate transcription when ligand is absent, suggesting that the ligand-independent AF -1 activity is somehow masked. We used the GAL4 DBD to investigate this phenomenon. When fused to GAL4 DBD, the full-length mPPARα receptor is not active in mammalian cells in absence of ligand despite the presence of the AF -1 activity in the A/B domain. Only when the PPAR activator Wy-14 is added, does the receptor become active. Carboxy-terminal deletion of the fusion at the junction of the DBD (C domain) and the Hinge region (D domain) results in partially active protein whose activity does not depend on the presence of ligand, suggesting a role of the hinge region in the masking of the AF-1 activity. The transcriptional activity of PPARs is ligand independent when PPARs are expressed as GAL4 DBD fusions or allowed to bind reporter genes as dimers with RXR. Yeast do not possess nuclear receptors or any homologues of known accessory proteins which interact with nuclear receptors. Given our observations and those of others made in yeast and mammalian cells, we suggest that the masking of the AF -1 activity occurs in the inactive state of the receptor, and that mammalian cells contain unidentified factor(s) which is able to maintain PPARs in the inactive state, thus enabling the masking of the AF-1 activity. Nuclear receptors including PPARs are known to interact with a host of accessory proteins which modulate their activity and are thought to act as a bridge between the receptors and the basal transcription machinery. One of these proteins is RIP140 whose function has not been determined. Our studies suggest that RIP 140 is a co-activator for the PPAR/RXR heterodimer, as well as for the AF-1 activity of mPPARα and mPP ARγ2. / Thesis / Master of Science (MSc)

Orientador: Reinaldo Fernandes Cooke / Resumo: Vacas leiteiras passam por muitas mudanças fisiológicas e metabólicas durante o período de transição (Gummer, 1995; Drackely, 1999). Essas prejudicam o sistema imune, expondo os animais a distúrbios metabólicos e doenças infecciosas (Mallard et al., 1998). Omingen-AF® (OMN) é um produto comercial que demonstrou potencial de modular a resposta imune de animais de produção (Wang et al., 2009). Com o objetivo de avaliar o potencial de OMN como suplemento nutricional para vacas leiteiras durante o período de transição, este estudo comparou parâmetros produtivos, fisiológicos e de saúde em vacas leiteiras suplementadas ou não com OMN durante o período de transição. Trinta e oito vacas mestiças Holandês-Gir prenhas, multíparas e não-lactantes foram classificadas por peso corporal (PC) e escore de condição corporal (ECC) e designadas a receber (OMN; n=19) ou não (CON; n=19) 56 g/vaca/dia de OMN iniciando 35 dias antes da data estimada para o parto. Omnigen foi suplementado individualmente às vacas sobre o concentrado oferecido durante o período da manhã. Antes do parto, o PC e ECC das vacas foi aferido semanalmente e amostras de sangue foram coletadas a cada 5 dias, iniciando no d-35 em relação à data prevista para a parição. Após o parto e até 46 dias em leite (DEL), o PC e ECC foram aferidos também semanalmente. O registro da produção de leite individual e a coleta de amostras de leite para análise de sólidos totais e Contagem de Células Somáticas (CCS) e de amostras de sangue de ... (Resumo completo, clicar acesso eletrônico abaixo) / Mestre

Submitted by Alice Poggi Brandão null (alicepbrandao@gmail.com) on 2016-12-30T00:28:42Z No. of bitstreams: 1 Dissertação Mestrado Alice.pdf: 1977086 bytes, checksum: 6fcceacacc9b5a4bac4bdb99a6c2b5d5 (MD5) / Rejected by LUIZA DE MENEZES ROMANETTO (luizamenezes@reitoria.unesp.br), reason: Solicitamos que realize uma nova submissão seguindo a orientação abaixo: O arquivo submetido está sem a ficha catalográfica. A versão submetida por você é considerada a versão final da dissertação/tese, portanto não poderá ocorrer qualquer alteração em seu conteúdo após a aprovação. Orientamos que indique o número de processo do financiamento FAPESP nos agradecimentos. Corrija estas informações e realize uma nova submissão com o arquivo correto. Agradecemos a compreensão. on 2017-01-03T16:31:54Z (GMT) / Submitted by Alice Poggi Brandão (alicepbrandao@gmail.com) on 2017-01-04T15:44:55Z No. of bitstreams: 1 Dissertação Mestrado Completa.pdf: 2053776 bytes, checksum: 0d72a35c8158fb28fb236bdfa3476e1c (MD5) / Approved for entry into archive by Juliano Benedito Ferreira (julianoferreira@reitoria.unesp.br) on 2017-01-06T12:39:56Z (GMT) No. of bitstreams: 1 brandao_ap_me_bot.pdf: 2053776 bytes, checksum: 0d72a35c8158fb28fb236bdfa3476e1c (MD5) / Made available in DSpace on 2017-01-06T12:39:56Z (GMT). No. of bitstreams: 1 brandao_ap_me_bot.pdf: 2053776 bytes, checksum: 0d72a35c8158fb28fb236bdfa3476e1c (MD5) Previous issue date: 2016-12-19 / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Vacas leiteiras passam por muitas mudanças fisiológicas e metabólicas durante o período de transição (Gummer, 1995; Drackely, 1999). Essas prejudicam o sistema imune, expondo os animais a distúrbios metabólicos e doenças infecciosas (Mallard et al., 1998). Omingen-AF® (OMN) é um produto comercial que demonstrou potencial de modular a resposta imune de animais de produção (Wang et al., 2009). Com o objetivo de avaliar o potencial de OMN como suplemento nutricional para vacas leiteiras durante o período de transição, este estudo comparou parâmetros produtivos, fisiológicos e de saúde em vacas leiteiras suplementadas ou não com OMN durante o período de transição. Trinta e oito vacas mestiças Holandês-Gir prenhas, multíparas e não-lactantes foram classificadas por peso corporal (PC) e escore de condição corporal (ECC) e designadas a receber (OMN; n=19) ou não (CON; n=19) 56 g/vaca/dia de OMN iniciando 35 dias antes da data estimada para o parto. Omnigen foi suplementado individualmente às vacas sobre o concentrado oferecido durante o período da manhã. Antes do parto, o PC e ECC das vacas foi aferido semanalmente e amostras de sangue foram coletadas a cada 5 dias, iniciando no d-35 em relação à data prevista para a parição. Após o parto e até 46 dias em leite (DEL), o PC e ECC foram aferidos também semanalmente. O registro da produção de leite individual e a coleta de amostras de leite para análise de sólidos totais e Contagem de Células Somáticas (CCS) e de amostras de sangue de 0 a 7 DEL foram realizados diariamente. De 9 a 21 DEL, amostras foram coletadas em dias alternados e de 26 a 46 DEL, a cada 5 dias. Todas as amostras de sangue, do pré e pós-parto, foram avaliadas para cortisol, ácidos graxos não-esterificados (AGNE), insulina, glicose, haptoglobina, cortisol e IGF-1. Nos dias 30 e 46 após o parto vacas foram avaliadas para endometrite através da técnica do cytobrush, baseada na porcentagem de células polimorfonucledas (PMN) no total de uma contagem de 100 células (PMN + células endometriais). No dia 48,7 ± 1,6 após o parto, 9 vacas de cada tratamento receberam uma injeção de lipopolissacarídeo (LPS) (0,25μg/kg de PC) e amostras de sangue foram coletadas a cada hora de -2h até 8h após a injeção às 0h; a cada 12h das 12 às 72h e a cada 24h das 96 às 120h. Amostras de sangue coletadas durante o desafio de LPS foram analisadas para haptoglobina e de fator de necrose tumoral alfa (TNF-α). Não foi observado efeito de tratamento sobre o PC (P ≥ 0,15), ECC (P ≥ 0,55), concentrações séricas de cortisol, AGNE, insulina, glicose, haptoglobina, cortisol e IGF-1. Vacas recebendo OMN apresentaram maior (P = 0,02) produção de leite (30,3kg/d vs. 27,1kg/d) e maior (P = 0,04) porcentagem de células PMN na população celular endometrial (12,2% vs. 3,9%) quando comparadas com vacas recebendo CON. Após a administração do LPS, vacas do grupo OMN apresentaram concentração sérica média superior (P = 0,04) de haptoglobina (212 μg/mL vs. 94 μg/mLpara vacas dos grupos OMN e CON, respectivamente) assim como maior concentração plasmática de TNF-α (P ≤ 0,04) nos momentos 1 (1,34 vs. 2,35 ng/mL para OMN e CON respectivamente), 2 (0,79 vs. 2,01 ng/mL, para OMN e CON respectivamente) e 3h (0,41 vs. 1,34 ng/mL, para OMN e CON respectivamente) em relação à injeção de LPS. Em conclusão, a suplementação de Omnigen-AF® durante o período de transição elevou a concentração plasmática de marcadores da resposta imune inata e aumentou a produção de leite em vacas leiteiras. / Dairy cows undergo several physiological and metabolic changes during the transition period. (Grummer, 1995; Drackley, 1999). These changes impair the immune function and cows become more susceptible to infectious and metabolic diseases (Mallard et al., 1998). Omnigen-AF® (OMN) is a commercial feed ingredient that has been shown to modulate the immune response in livestock animals (Wang et al., 2009). Aiming to evaluate the potential of OMN as a feed additive for transition dairy cows, this study compared physiological, health, and productive parameters in dairy cows supplemented or not with OMN during the transition period. Thirty-eight non-lactating, multiparous, pregnant Holstein × Gir cows were ranked by body weight (BW) and body condition score (BCS), and assigned to receive (OMN; n = 19) or not (CON; n = 19) 56 g/cow daily (as-fed basis) of OMN beginning 35 d prior to expected calving date. Omnigen was individually offered to cows as top-dressing into the morning concentrate feeding. Before calving, cow BW and BCS were recorded weekly and blood samples collected every 5 d beginning on d -35 relative to expected calving date. After calving and until 46 DIM, BW and BCS were recorded weekly. Individual milk production was recorded and milk samples were collected daily for total solids and SCC analyses. Blood was sampled daily from 0 to 7 DIM, every other day from 9 to 21 DIM, and every 5 d from 26 to 46 DIM. All blood samples were evaluated for cortisol, non-esterified fatty acids (NEFA), insulin, glucose, haptoglobin and IGF-1. On 30 and 46 DIM, cows were evaluated for endometritis via cytobrush technique, based on the % of polymorphonuclear (PMN) cells within 100 total cell count (PMN + endometrial cells). On 48.7 ± 1.6 DIM, 9 cows/treatment received a lipopolysaccharide (LPS) injection (0.25 μg/kg of BW), and blood was sampled hourly from -2 to 8 h, at 12-h intervals from 12 to 72 h, and at 24-h intervals form 96 to 120 h relative to LPS administration at hour 0. Blood samples collected during the LPS challenge were analyzed for haptoglobin and tumor necrosis factor alpha (TNF-α). No treatment differences were detected on BW (P ≥ 0.15), BCS (P ≥ 0.55), serum concentrations of cortisol, NEFA, insulin, glucose, haptoglobin, cortisol, and IGF-I. Cows receiving the OMN diet had greater milk yield (30.3 vs. 27.1 kg/d, P = 0.02) and percentage of PMN cells in endometrial cell population (12.2 vs. 3.9%, P = 0.04) compared with CON cows. After LPS administration, OMN cows had greater (P = 0.04) mean serum haptoglobin (212 vs. 94 µg/mL, for OMN and CON, respectively), as well as greater (P ≤ 0.04) plasma concentrations of TNF-α at 1, (1.34 vs. 2.35 ng/mL for OMN e CON respectively), 2 (0.79 vs. 2.01 ng/mL, for OMN e CON respectively) and 3h (0.41 vs. 1.34 ng/mL, for OMN e CON respectively) relative to LPS injection. In conclusion, supplementing Omnigen-AF® during the transition period increased plasma concentrations of innate immunity markers and increased milk yield of transition dairy cows. / FAPESP: 2015/05536-1

Made available in DSpace on 2014-06-12T18:08:13Z (GMT). No. of bitstreams: 2 arquivo8044_1.pdf: 436930 bytes, checksum: 4f958f44689dc03881cbb9851fa8e45e (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) Previous issue date: 2002 / Nesta dissertação são investigadas as propriedades magnéticas da interface constituída por um material ferromagnético (FM) e um material antiferromagnético (AF). As amostras investigadas foram preparadas por sputtering dc e foram caracterizadas por ressonância ferromagnética (FMR), magnetometria por efeito Kerr magneto-óptico (MOKE) e susceptibilidade ac (cAC). Os resultados experimentais foram interpretados a partir de um modelo fenomenológico no qual são consideradas a energia livre da camada FM (Zeeman, anisotropia e desmagnetização); a energia devido ao acoplamento na interface (exchange bias) e a energia da parede de domínio que se forma na camada AF. Parâmetros tais como: campo de anisotropia, campo de acoplamento na interface, campo da parede de domínio no AF, etc. são extraídos a partir dos resultados experimentais. É mostrado que os parâmetros extraídos a partir dos resultados de MOKE e cAC são iguais (dentro do erro experimental), por outro lado esses mesmos parâmetros são diferentes daqueles obtidos por FMR. O modelo foi testado em três sistemas com composições diferentes, produzindo resultados consistentes para todas as amostras. Com o objetivo de investigar o deslocamento que ocorre no campo de ressonância ferromagnética em bicamadas FM/AF, investigamos tricamadas nas quais um espaçador não-magnético é depositado entre a camada FM e a camada AF. Os espectros de absorção apresentaram dois modos de ressonância. Um deles apresenta uma variação angular com simetria C2 correspondendo ao modo uniforme do filme FM sem acoplamento. O outro modo ocorre em valores de campo correspondentes ao modo uniforme do filme FM acoplado. Porém a variação angular do campo de ressonância deste segundo modo apresenta nenhuma simetria. Também foram investigados, por susceptibilidade ac, filmes simples policristalinos de permalloy. Os resultados foram interpretados utilizando-se teorias fenomenológicas. Foi mostrado que a introdução termos que levam em conta os grãos policristalinos reproduzem os resultados experimentais

Class of 2015 Abstract / Objectives: Warfarin has historically been the anticoagulant used for the primary prevention of stroke in atrial fibrillation (AF), however three target specific oral anticoagulants, dabigatran, rivaroxaban, and apixaban, have recently been approved for use in this setting. Current literature lacks a comparison of these four drugs in relation to post-stroke outcomes, and this study aims to compare their performance in a natural setting. Methods: This retrospective cohort study identified stroke patients admitted to an academic medical center between January 2013 and December 2014 using the Quintiles, Inc.-American Heart Association Get With The Guidelines-Stroke database; pertinent data was collected from the database and patient electronic medical records. Primary endpoints measured were length of stay, 30-day readmission, and discharge disposition; secondary endpoints included rates of admission to the intensive care unit (ICU) and complications. Results: Of 940 stroke admissions, 53 ischemic stroke patients were identified as receiving an oral anticoagulant for stroke prevention in AF. The warfarin (n=40) and non-warfarin (dabigatran, rivaroxaban, and apixaban; n=13) groups were well matched regarding admission demographics, however patients taking warfarin were more likely to have an elevated INR at hospital admission (P=0.0053) and receive tPA (P=0.047). Patients in the warfarin group were also statistically significantly more likely to receive warfarin on discharge (P=0.004). No endpoints achieved statistical significance. Conclusions: No differences in post-stroke outcomes between warfarin and non-warfarin oral anticoagulants used for stroke prevention in AF were found.

Background: Atrial fibrillation (AF) is common in patients presenting with myocardial infarction (MI). Percutaneous coronary intervention (PCI) has been shown to improve cardiovascular outcomes in MI. However, outcomes of PCI in AF patients presenting with MI remains largely unknown. Methods: We analyzed the Nationwide Inpatient Sample (NIS) database to calculate the age adjusted mortality rate for PCI in AF patients presenting with MI between 2002 and 2011, in adults over 40 years of age. This was then compared to the mortality rate for PCI in non-AF patients with MI. Specific ICD-9-CM codes were used to identify patients and outcomes. Results: Of 3,226,405 PCIs done during the study period, 472,609 (14.6%) PCIs were done on AF patients of which 137,870 PCIs were for MI. About 60% of these patients were male. Patients with AF were older (71.3 ± 10.6 years). Overall the number of PCIs shows a declining trend from 2002 to 2011, but for MI patients the number of PCIs appears stable over the years. The age adjusted in-hospital mortality following PCI in MI was significantly higher in AF group compared to the non-AF group (190.24 ± 17.21vs 109.08 ± 5.89 per 100,000; P < 0.01). This trend was seen during the entire study period. Conclusions: AF is prevalent in MI patients undergoing PCI. AF is associated with increased mortality following PCI for acute MI. AF is not a benign arrhythmia in MI patients and close attention is warranted in these patients to improve mortality.

Ferrichrome is a hydroxamate-containing siderophore produced by the pathogenic fungus Aspergillus fumigatus during infection. This siderophore includes N5-hydroxylated L-ornithine in the peptide backbone that serve as iron chelators. Af SidA is the L-ornithine N5-hydroxylase, which performs the first enzymatic step in the biosynthesis of ferrichrome. In this study, Af SidA was recombinantly expressed and purified as a soluble tetramer with a bound FAD cofactor. The enzyme demonstrated typical Michaelis-Menten kinetics in a product formation assay with respect to L-ornithine, but similar experiments as a function NADH and NADPH indicated inhibition at high coenzyme concentrations. Af SidA is highly specific for its substrate; however, it is promiscuous with respect to its coenzyme requirement. A multi-functional role of NADPH is observed since NADP+ is a competitive inhibitor with respect to NADPH and steady-state kinetic experiments indicate that Af SidA forms a ternary complex with NADP+ and L-ornithine for catalysis. Furthermore, in the absence of substrate, Af SidA forms a stable C4a-(hydro)peroxyflavin intermediate that is stable on the second time scale. Af SidA is also inhibited by several halides and the arginine-reactive reagent, phenylglyoxal. Biochemical comparison of Af SidA to other flavin-containing monooxygenases reveal that Af SidA likely proceeds by a sequential-ordered mechanism. / Master of Science in Life Sciences

Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Nela Kuchařová Supervisor: PharmDr. Jana Pourová, Ph.D. Title of diploma thesis: Vasodilatory effects of bisphenol AF ex vivo Bisphenols are organic compounds used in the manufacture of plastics, resins, varnishes and lot of other products. However, their effects on human body are associated with a number of adverse effect that need to be investigated in more detail. The aim of this diploma thesis was to determine whether bisphenol AF has vasodilating effects ex vivo and to verify the mechanism of the relaxing effect. The experiments were performed on an isolated thoracic aorta of a Wistar rat. Bisphenol AF was added cumulatively to the precontracted aorta at increasing concentrations and we monitored whether vasodilation was induced. In testing the mechanism of action, we used inhibitors of the mechanisms tested during the experiment. The obtained results were evaluated using the GraphPad Prism program. The results show that bisphenol AF has a dose-dependent ex vivo vasodilatory effect (EC50 = 57.16 μmol/l). The mechanism of this effect is the blockage of voltagegated calcium channels on the vascular smooth muscle cell. The participation of other tested mechanisms has not been confirmed. Key...