Orientadores: Marta Helena Krieger, Regina Celia Spadari-Bratfisch / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-08T07:50:11Z (GMT). No. of bitstreams: 1 Garcia_JoseAntonioDias_D.pdf: 1987747 bytes, checksum: 1ec8d731703522ccf42a42dafd7b13a7 (MD5) Previous issue date: 2006 / Resumo: Recentemente demonstrou-se que S-nitroso-N-acetilcisteína (SNAC) atenua o desenvolvimento da placa de aterosclerose na aorta em cerca de 55% de camundongos deficientes do receptor de lipoproteína de baixa densidade (LDLr-/-). O presente estudo teve como objetivo: i) verificar se a deleção do gene do receptor de LDL pode alterar o perfil hemodinâmico e a resposta inotrópica do coração a agentes adrenérgicos; ii) determinar a capacidade do SNAC na prevenção das alterações estruturais e funcionais do miocárdio e; iii) verificar o efeito do SNAC na pressão arterial de camundongos hipercolesterolêmicos. Camundongos machos C57BL6 (Wild Type = WT) e camundongos LDLr-/- (S) foram alimentados com dieta comercial por 15 dias. Em relação aos camundongos WT, os camundongos S apresentaram aumento de 11% na pressão arterial, diminuição de 62% na contratilidade do átrio esquerdo, e aumento na expressão do CD40L e redução na expressão de NOSe no tecido ventricular esquerdo. Camundongos LDLr-/- alimentados com dieta enriquecida em 1,25% de colesterol, 20% de gordura e 0,5% de ácido cólico por 15dias (Chol) apresentaram hipertrofia ventricular esquerda (HVE) comparados aos camundongos S, a qual foi caracterizada por: a) aumento de 1,25 vezes na razão entre o peso ventricular esquerdo (mg) e o peso corporal (g) (4,17±0,09 vs 3,34±0,07 mg/g, respectivamente; p< 0,05); b) aumento do diâmetro dos cardiomiócitos (25±0,6 vs 19±0,7 µm, p<0.05); c) aumento na expressão das isoformas das NOS (óxido nítrico sintases) e hiperexpressão do CD40L; d) aumento do depósito de colágeno; e) sem alterações na performance contrátil do átrio esquerdo e na responsividade à noradrenalina. A administração do SNAC aos camundongos Chol (chol+SNAC) (0,51 µmol/kg/dia, i.p.), preveniu o aumento na razão do peso ventricular esquerdo (mg) e o peso corporal (g) (3.38±0.23 mg/g), no diâmetro dos cardiomiócitos (20±0.7 µm), no deposito de colágeno, na expressão das isoformas da NOS e a hiperexpressão do CD40L. O SNAC não apresentou efeito no aumento da pressão arterial e nem sobre a hipocontratilidade, mas recuperou a responsividade para noradrenalina. Em conclusão, o presente estudo demonstrou que camundongos com deleção do gene do receptor LDL apresentaram hipertensão e marcada redução contrátil. Essas características podem estar relacionados com o estresse oxidativo resultante do processo inflamatório e da hipoexpressão da NOSe. A dieta hiperlipídica promoveu hipertrofia ventricular esquerda (HVE), devida ao aumento nos processos inflamatório e oxidativo. O SNAC impediu o desenvolvimento da HVE por mecanismos que envolveram efeito antiinflamatório (detectado pela diminuição na expressão do CD40L), a hiperexpressão das NOS, a redução das alterações estruturais ventriculares induzidas pela hipercolesterolemia de maneira independente da hipertensão. No presente estudo, a necessidade de quantificar as análises histológicas exigiu a validação de um software interativo para analisar imagens de amostras teciduais. O software foi projetado para permitir que o usuário altere os tipos de coloração vermelha, verde e azul (RGB) para uma cor padrão que possa ser usada para segmentar a imagem e calcular a fração da área de interesse. Os resultados obtidos com a contagem manual e com a contagem feita com o uso do software foram similares, indicando que são métodos alternativos confiáveis para análises quantitativas de cortes histológicos. Entretanto, o software permite processar as imagens de maneira eficiente e confiável, além de reproduzir o corte tecidual em um menor tempo, e pode ser executado com o microscópio e o computador padrão / Abstract: Recently, it has been that S-nitroso-N-acetylcysteine (SNAC) attenuate in 55% the plaque development in low-density lipoprotein-receptor-deficient (LDLr-/-) mice fed a hypercholesterolemic diet for 15 days. The present study was designed to verify whether deletion of the low-density lipoprotein (LDL) receptor gene may affect the hemodynamic profile and adrenergic inotropic cardiac responses and, particularly, to identify the ability of SNAC to prevent the myocardial alterations and hypertension in hypercholesterolemic mice. C57BL6 wild-type (WT) and LDLr-/- male mice (S) were fed a commercial diet for 15 days. Control mice (S) showed 11 % blood pressure increase, 62% left atrial contractility decrease, CD40L overexpression and eNOS underexpression in comparison to WT. LDLr-/- mice which were fed for 15 days with 1,25% cholesterol, 20% of fat and 0.5% of colic acid enriched diet (Chol), showed significant left ventricular hypertrophy (LVH) versus S, which was characterized by: a) 1.25-fold increase in the LV weight (mg)/body weight (g) ratio (4.17±0.09 vs. 3.34±0.07 mg/g, respectively; p<0.05); b) increased cardiomyocyte diameter (25±0.6 vs. 19±0.7 µm, p<0.05); c) enhanced expression of the constitutive and inducible NOS isoforms and CD40L;d) increased collagen deposit; e) no alteration in the atrial contractile performance or responsiveness to norepinephrine. Administration of SNAC to Chol mice ( Chol +SNAC) (0.51 µmol/kg/day, for 15day, i.p.) prevented increases in the left ventricular weight/body weight ratio (3.38±0.23 mg/g), cardiomyocyte diameter (20±0.7 µm), collagen deposit, NOS isoforms and CD40L overexpression, but had no effect on increased blood pressure or atrial basal hypocontractility, although it recovered responsiveness to norepinephrine. In conclusion, the present study demonstrated that the deletion of the LDL receptor gene in mice determined hypertension and a marked left atrial contractile deficit. These findings may be related to oxidative stress, resulting from inflammation and eNOS underexpression. High-cholesterol diet promoted LVH in LDLr-/- mice associated with enhanced inflammatory and oxidant processes. SNAC prevented LVH by processes that involved decreased CD40L expression and NOS overexpression effects attenuating the ventricular structural alterations induced by hypercholesterolemia independent of hypertension. Histological quantization demanded the development of interactive software for image analysis of tissue samples. The software was designed to allow a user-oriented change of a chosen red, green and blue (RGB) staining in a standardized color that can be used to segment the image and calculate the fractional area of interest. Thus the method allows efficient, reliable and reproducible processing of tissue sections that is less time-consuming than conventional methods and can be performed with standard microscope and computer / Doutorado / Fisiologia / Doutor em Biologia Funcional e Molecular

La rupture du ligament croisé antérieure (LCA) conduit à une instabilité et à des modifications biomécaniques du genou. Actuellement, les patients présentant une lésion du LCA bénéficient généralement une reconstruction du LCA. L'analyse cinématique quantitative est un outil important pour acquérir une compréhension approfondie de la fonction articulaire du genou normal et pathologique au cours de la locomotion humaine. Ce travail est axé sur l'évaluation in vivo de la cinématique du genou chez les patients avec rupture du LCA, avant et après la reconstruction du LCA au cours de toutes les phases de la marche, en utilisant un nouvel outil d'évaluation 3D, quasi-rigide, en temps réel (KneeKGTM). Dans la première étude, des modifications cinématiques significatives ont été identifiées dans les genoux avec une rupture du LCA. Ces changements cinématiques pourraient mener à des contacts fémoro-tibiaux anormaux du cartilage pendant les activités quotidiennes, représentant potentiellement un mécanisme biomécanique de l'usure des articulations après une lésion du LCA. Dans la seconde étude, l'analyse 3D in vivo du mouvement a révélé que les genoux avec LCA reconstruits améliorent significativement leur extension par rapport aux genoux avec LCA déficient, mais il y avait encore une différence par rapport au groupe témoin sain. Dans le plan axial, le tibia reste en position interne significative par rapport à un groupe témoin, tandis qu'il n'y avait pas de différences significatives en translation antéro-postérieur et dans le plan coronal. Ces changements cinématiques pourraient conduire à une charge anormale dans l'articulation du genou et initier le processus d'atteinte dégénérative cartilagineuse à venir. Toutefois, les données cinématiques post-opératoires ont été collectées 10 mois après la chirurgie, ainsi un suivi plus long serait nécessaire pour évaluer si ces changements cinématiques persistent dans le temps, et leurs effets sur l'articulation / Because of the role of the ACL in knee joint’s biomechanics, it is essential to quantify the kinematics of ACL deficient and ACL reconstructed knee. In-vitro models bare the handicap of limited muscle simulation, while static, one-dimensional testing cannot predict the behavior of these groups of patients under realistic loading conditions. Currently, the most widely accepted method for assessing joint movement patterns is gait analysis. Respectively, 3D motion analysis is necessary to provide high reliability movement analysis. The purpose of the study was in-vivo evaluation of the behavior of the anterior cruciate ligament deficient (ACLD) and anterior cruciate ligament reconstructed (ACLR) knees during walking, using 3D, real-time assessment tool. Significant alterations of joint kinematics in the ACLD knee were revealed in this study by manifesting a higher flexion gait strategy and excessive internal tibial rotation during walking that could result in a more rapid cartilage thinning throughout the knee. In the other hand, even though ACLR knees showed some improvements in sagittal plane compared to ACLD knees, in axial plane there still exists difference compared to healthy control knees. These kinematic changes could lead to abnormal loading in knee joint and initiate the process for future chondral degeneration. However, the post-operative kinematic data were collected 10 months after surgery, so a longer follow-up is needed to evaluate if these kinematic changes persist in time, and their effects in joint degeneration

Enzymatic and lipid transfer reactions involved in reverse cholesterol transport were studied in HDL deficient plasma systems. Fasting plasma samples were obtained from control and cholesterol fed guinea pigs as well as from a fish eye disease patient and were used to localize the enzyme LCAT among plasma lipoproteins (VLDL, LDL, and HDL). In both guinea pig and fish eye disease patient plasma, the LCAT activity was found in association with the HDL type particles. Cholesterol feeding in guinea pigs altered the properties of lipoprotein substrates for LCAT resulting in some changes, specifically: 1) decreased fractional rate of plasma cholesterol esterification and, 2) lower transfer of free cholesterol (FC) and esterified cholesterol (CE) within the lipoprotein fractions.

No description available.

Orientador: Ligia Maria Presumido Bracialli / Banca: Mário Antonio Baraúna / Banca: Rita de Cássia Tibério Araújo / Resumo: Alguns indivíduos com encefalopatias podem apresentar movimentos involuntários associados ao quadro de seqüelas. Esses movimentos podem provocar limitações funcionais e vir a prejudicar o desempenho nas atividades escolares e da vida diária. O uso de pulseira estabilizadora com pesos é um recurso que pode minimizar os movimentos involuntários e auxiliar no desempenho desses indivíduos. O presente trabalho teve como objetivo geral analisar o comportamento motor de membro superior, no ato de apontar um alvo, por meio do uso de peso, em indivíduos com seqüelas de encefalopatias, os quais apresentavam movimentos involuntários nos membros superiores. Participaram da pesquisa 8 indivíduos, com seqüelas de encefalopatias, que apresentavam movimentos involuntários em membros superiores, do gênero masculino e feminino. A pesquisa foi realizada no laboratório de análise de movimento do Centro de Estudos da Educação e da Saúde, Unesp, Campus de Marília (CEES). A coleta de dados foi feita por meio de estudo cinemático e eletromiográfico da performance do movimento de membros superiores, no ato de apontar um alvo, com e sem o uso de peso. Foi realizada uma análise quantitativa dos dados obtidos nos registros cinemáticos, em relação ao deslocamento escalar, velocidade média escalar e nos registros eletromiográficos, em relação à intensidade de contração muscular, nos momentos sem o uso de peso contra a ação da gravidade e com o uso de peso. Para cada movimento, foram comparados: a velocidade escalar, o deslocamento escalar, o momento de contração muscular, por meio de análise de dispersão de dados pareados. Os resultados demonstraram respostas mais adequadas de intensidades de contrações musculares e de menor deslocamento escalar, com uso de pulseira de pesos. Pode-se concluir que o uso de pulseira com peso favoreceu a realização... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Some individual with encephalopathy can present involuntary movements associated with features of sequel. These movements can arose functional restrictions and come to prejudice the performance at school and daily activities. The use of stabilized bangle with weight is a resource that can minimize the involuntary movements and relieve this individual performance. The present work had as general objective to analyze the upper limbs motor behavior during the act of aim a target through the use of weight on individual with encephalopathy sequel that presented involuntary movements on upper limbs. Eight individual with encephalopathy sequel participated in this research which presented involuntary movements in upper limbs of male and female gender. The research was accomplished in the laboratory of analyzes of movements at Centro de Estudos da Educação e da Saúde, Unesp, Campus from the city of Marília ( CEES ).The collection of data was accomplished thought the kinematics and electromyographic studies performs of upper limbs in the act of aim a target with and without the use of weight. It was accomplished a quantitative analyses of data obtained to kinematics register related to the dislocation scale, medium scalar velocity in the register of electromyographic related to intensity of muscular contraction during the movements without the use of weight against the gravicity action and also with the use of weight. For each movement was compared: the scalar velocity, scalar dislocation, the movement of muscular contraction through the analyze of electromyographic. The results demonstrated adequate answers to the intensity of muscular contractions and less scalar dislocation with the use of weight. Concluding that the use of bangle with weight exalted the accomplishment of functional and adequate movement during the act of aim a target. / Mestre

Heterotrimeric Gi proteins play a role in signalling activated by lipopolysaccharide (LPS), Staphylococcus aureus (SA) and group B streptococci (GBS), leading to production of inflammatory mediators. We hypothesized that genetic deletion of Gi proteins would alter cytokine and chemokine production induced by LPS, SA and GBS stimulation. LPS-induced, heat-killed SA-induced and heat-killed GBS-induced cytokine and chemokine production in peritoneal macrophages from wild-type (WT), Gαi2-/- or Gαi1/3-/- mice were investigated. LPS induced production of tumour necrosis factor-α (TNF-α), interleukin-6 (IL-6), IL-10 and interferon-γ-inducible protein-10 (IP-10); SA induced TNF-α, and IL-1β production; and GBS induced TNF-α, IL-6, IL-1β, macrophage inflammatory protein-1α (MIP-1α) and keratinocyte chemoattract (KC) production were all decreased (P < 0.05) in Gαi2-/- or Gαi1/3-/- mice compared with WT mice. In contrast to the role of Gi proteins as a positive regulator of mediators, LPS-induced production of MIP-1α and granulocyte-macrophage colony-stimulating factor (GM-CSF) were increased in macrophages from Gαi1/3-/- mice, and SA-induced MIP-1α production was increased in both groups of Gαi protein-depleted mice. LPS-induced production of KC and IL-1β, SA-induced production of GM-CSF, KC and IP-10, and GBS-induced production of IL-10, GM-CSF and IP-10 were unchanged in macrophages from Gαi2-/- or Gαi1/3-/- mice compared with WT mice. These data suggest that Gi2 and Gi1/3 proteins are both involved and differentially regulate murine inflammatory cytokine and chemokine production in response to both LPS and Gram-positive microbial stimuli.

京都大学 / 0048 / 新制・課程博士 / 博士(医科学) / 甲第21030号 / 医科博第91号 / 新制||医科||6(附属図書館) / 京都大学大学院医学研究科医科学専攻 / (主査)教授 生田 宏一, 教授 清水 章, 教授 竹内 理 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

Mesoporous γ-alumina is the most extensively used catalysts support in a wide range of catalytic processes. The usefulness of γ-alumina relies on its favorable combination of physical, textural, thermal, and chemical properties. Pore structure properties are among the most important properties, since high surface area and large pore volume enable higher loading of active catalytic phases, while design of pore size and pore size distribution is critical to optimize pore diffusional transport and product selectivity. In addition, accurate determination of surface area (SA), pore volume (PV) and pore size distribution (PSD) of porous supports, catalysts, and nanomaterials is vital to successful design and optimization of these materials and to the development of robust models of pore diffusional resistance and catalyst deactivation.In this dissertation, we report a simple, one-pot, solvent-deficient process to synthesize mesoporous γ-alumina without using external templates or surfactants. XRD, TEM, TGA and N2 adsorption techniques are used to characterize the morphologies and structures of the prepared alumina nanomaterials. By varying the aluminum salts or the water to aluminum molar ratio in the hydrolysis of aluminum alkoxides, γ-alumina with different morphologies and pore structures are synthesized. The obtained alumina nanomaterials have surface areas ranging from 210 m2/g to 340 m2/g, pore volumes ranging from 0.4 cm3/g to 1.7 cm3/g, and average pore widths from 4 to 18 nm. By varying the alcohols used in the rinsing and gelation of boehmite/bayerite precursors derived from a controlled hydrolysis of aluminum alkoxides, the average pore width of the γ-aluminas can be tuned from 7 to 37 nm. We also report improved calculations of PSD based on the Kelvin equation and a proposed Slit Pore Geometry model for slit-shaped mesopores of relatively large pore size (>10 nm). Two structural factors, α and β, are introduced to correct for non-ideal pore geometries. The volume density function for a log normal distribution is used to calculate the geometric mean pore diameter and standard deviation of the PSD. The Comparative Adsorption (αs) Method is also employed to independently assess mesopore surface area and volume.

Nanotechnology is blossoming into one of the premiere technologies of this century, but the key to its progress lies in developing more efficient nanosynthesis methods. Variations in synthetic technique, however, can cause variations in size, structure, and surface characteristics, thereby altering the physical properties and functionality of the particles. Careful structural characterizations are thus essential for understanding the properties and appropriate applications for particles produced by new synthetic techniques.In this work, a new ‘solvent-deficient’ method is presented for the synthesis of an unprecedentedly wide range of metal oxide nanomaterials including at least one metal oxide from each group in Groups 3-4, 6-15, and the Lanthanides. XRD, BET, and TEM structural characterizations as well as chemical purity analyses of the products are given. The intermediates associated with the method are also investigated, allowing the reaction parameters to be rationalized and culminating in a proposed mechanism for the reaction. Several of the reaction intermediates are themselves useful products, expanding the range of this already versatile method. Optimized synthesis parameters as well as structural characterizations are presented for one such intermediate product, the iron oxyhydroxide called ferrihydrite.The Al2O3 nanoparticles produced by the new method show promise in catalyst support applications, and the synthesis and structural analysis (XRD, X-ray PDF, 27Al NMR, TG/DTA-MS) of these nanoparticles is provided. The XRD, PDF, and NMR analyses reveal that the initial boehmite-like phase transforms to the catalytically useful gamma-Al2O3 phase at unusually low temperatures (300-400°C), but boehmite-like local structure defects remain which heal slowly with increasing temperature up to 800°C. The ‘pure’ gamma-Al2O3 may still contain randomized, non-cubic, local structure distortions, and it transforms directly to alpha-Al2O3 at ~1050°C. To rationalize the local structure and the absence of the delta- and theta-Al2O3 phases during the alpha-phase transition, relationships between the many Al2O3 phases are presented via innovative symmetry-mode analyses, revealing a potential quazi-topotactic mechanism for the gamma-to-alpha transition.To stabilize the gamma-Al2O3 phase to higher temperatures for catalyst applications, 3 wt% of a lanthanum dopant was added via a new, 1-pot process based on the new solvent-deficient method. This process is described and X-ray PDF, TEM, 27Al NMR, and EXAFS analyses of the La-doped gamma-Al2O3 nanoparticles reveal that the dopant resides as isolated, adsorbed atoms on the gamma-Al2O3 surface. The first coordination shell of the isolated La is increasingly La2O3-like as calcination temperature increases but changes drastically to be more LaAlO3-like after the alpha-phase transition, which is delayed ~100°C by the La dopant. Combining the EXAFS, PDF, NMR, and symmetry-mode analyses, we provide new insight into the mechanism of stabilization provided by the La dopant.

No description available.