Les maladies cardiovasculaires sont la principale cause de mortalité dans les pays occidentaux et représentent une complication majeure du syndrome métabolique. Il est maintenant largement admis que l’athérosclérose est une maladie inflammatoire chronique et que l’inflammation joue un rôle pathogénique majeur dans l’initiation et la progression de la maladie athéromateuse. Il a été démontré qu’une augmentation des niveaux sériques de la protéine c-réactive (CRP), une protéine de la phase aigüe et un important constituant de la réponse immunitaire de type inné, est associée à un risque cardiovasculaire accru. Ainsi, il a été documenté qu’une augmentation de CRP, tant chez les sujets sains que chez les sujets diabétiques, était associée à une augmentation du risque de morbidité et de mortalité cardiovasculaires. De multiples évidences suggèrent que la CRP puisse non seulement constituer un marqueur de risque des maladies cardiovasculaires mais aussi représenter un facteur pro-athérogénique direct. La dysfonction endothéliale représente un des stades les plus précoces du processus athérosclérotique et un rôle de la CRP dans la pathogenèse de la dysfonction endothéliale est postulé. Outre son origine systémique, la CRP est produite dans la lésion athérosclérotique et par diverses cellules vasculaires, dont les cellules endothéliales. Afin d’élucider le rôle de la CRP vasculaire dans l’altération de la fonction endothéliale associée au syndrome métabolique, nous avons étudié la régulation de l’expression endothéliale de la CRP par les acides gras libres (AGL) et le rôle de la CRP endothéliale dans l’inhibition de la synthèse d’oxyde nitrique (NO) par les AGL. Nos résultats démontrent que :1) l’acide palmitique (PA) induit l’expression génique de CRP au niveau de cellules endothéliales aortiques humaines (HAECs) en culture et, augmente, de manière dose-dépendante, l’expression protéique de la CRP; 2) La pré-incubation des HAECs avec des antioxydants et des inhibiteurs de la i) protéine kinase C (PKC), ii) du facteur nucléaire-kappa B, iii) des Janus kinases et des protéines de transduction et de régulation de la transcription et iv) des protéines kinases activées par les mitogènes prévient l’effet stimulant du PA sur l’expression protéique et génique de la CRP; 3) Le traitement des HAECs par le PA induit une augmentation de la production des espèces réactives oxygénées, un effet prévenu par les inhibiteurs de la PKC et par l’AICAR(5-amino-4-imidazole carboxamide 1-β-D-ribofuranoside), un activateur de la protéine kinase activée par l’AMP; 4) L’incubation des HAECs en présence de PA résulte enfin en une diminution de la production basale endothéliale de NO, un effet abrogé par la préincubation de ces cellules avec un anticorps anti-CRP. Dans l’ensemble, ces données démontrent un effet stimulant du PA sur l’expression de la CRP endothéliale via l’activation de kinases et de facteurs de transcription sensibles au stress oxydatif. Ils suggèrent en outre un rôle de la CRP dans la dysfonction endothéliale induite par les AGL. / Atherosclerotic cardiovascular disease is the leading cause of death in western countries and the major complication of metabolic syndrome. It is now widely accepted that atherosclerosis is a chronic inflammatory disease and that inflammation plays a major pathogenic role in the initiation and progression of atherosclerotic disease. It has been demonstrated that increased serum levels of C-reactive protein (CRP), a protein of the acute phase and a major constituent of the innate immune response, is associated with increased cardiovascular risk and that, in both healthy subjects and diabetic patients, high CRP enhances the risk of cardiovascular morbidity and mortality. Several evidences suggest that CRP may not only be a cardiovascular risk marker but may also represent a direct pro-atherogenic factor. Endothelial dysfunction is a characteristic feature of early-state atherosclerosis and a role of CRP in the pathogenesis of endothelial dysfunction has been proposed. In addition to its systemic origin, CRP is produced in atherosclerotic lesions and by various vascular cells, including endothelial cells. To elucidate the role of CRP in endothelial dysfunction associated with the metabolic syndrome, we studied the regulation of endothelial CRP expression by free fatty acids (FFA) and the role of endothelial CRP as mediator of the inhibitory effect of FFA on nitric oxide (NO) production. Our results demonstrated that: 1) Palmitic acid (PA) induced CRP gene expression in cultured human arterial endothelial cells (HAECs) and increased CRP protein expression in a dose-dependent manner; 2) Pretreatment of HAECs with antioxidants and inhibitors of i) protein kinase C (PKC), ii) nuclear factor-kappa B, iii) Janus kinase and signal transducer and activator of transcription and iv) mitogen-activated protein kinases prevented the stimulatory effect of PA on CRP protein and gene expression; 3) Treatment of HAECs by PA led to an increased production of reactive oxygen species, an effect prevented by PKC inhibitors and by AICAR (5-amino-4-imidazole carboxamide 1-β-D-ribofuranoside), an AMP- activated protein kinase activator; 4) Decreased production of NO was finally observed in PA-treated HAECs, an effect prevented by preincubating endothelial cells with an anti-CRP. Overall, these data indicate a stimulatory effect of PA on endothelial CRP expression through the activation of oxidative stress-sensitive kinases and transcription factors. They further suggest a role of CRP in FFA-induced endothelial dysfunction.

La découverte du système des peptides natriurétiques (NP), au début des années 80, fut une découverte majeure qui révéla le rôle endocrinien du cœur. Les connaissances sur la relaxation vasculaire, la diurèse et la natriurèse provoquées par ce système ont évolué vers un niveau de complexité insoupçonné à cette époque. Nous savons à présent que les NP sont impliqués dans plusieurs autres mécanismes dont la prolifération cellulaire, l’apoptose, l’inhibition du système rénine-angiotensine-aldostérone (RAAS) et le métabolisme des adipocytes. Le métabolisme des lipides est maintenant devenu une cible de choix dans la lutte contre l’obésité. Cette condition aux proportions pandémiques est un facteur de risque majeur dans l’apparition de l’hypertension et du syndrome métabolique (MetS). La compréhension des mécanismes et des défauts de la voie des NP pourrait avoir un impact positif sur le contrôle du MetS et de l’hypertension. L’expression du récepteur des peptides natriuretiques de type 1 (NPR1/GCA) est contrôlée par plusieurs agents incluant son propre ligand, le peptide natriurétique de l’oreillette (ANP). La découverte d’une boucle de retro-inhibition, dans les années 90, a été un événement majeur dans le domaine des NP. En effet, suite à une stimulation à l’ANP, le NPR1/GCA peut inhiber l’activité transcriptionnelle de son propre gène par un mécanisme dépendant du cGMP. Notre groupe a identifié un élément cis-régulateur responsable de cette sensibilité au cGMP et mon projet consistait à identifier la ou les protéine(s) liant cet élément de réponse au cGMP (cGMP-RE). Nous avons identifié un clone liant le cGMP-RE en utilisant la technique du simple hybride chez la levure et une banque d’ADN complémentaire (ADNc) de rein humain. Ce clone provient d’un ADNc de 1083-bp dont le gène est localisé sur le chromosome 1 humain (1p33.36) et codant pour une protéine dont la fonction était inconnue jusqu’ici. Nous avons nommé cette nouvelle protéine GREBP en raison de sa fonction de cGMP Response Element Binding Protein. Des essais de liaison à l’ADN ont montré que cette protéine possède une affinité 18 fois plus élevée pour le cGMP-RE que le contrôle, tandis que des expériences de retard sur gel (EMSA) ont confirmé la spécificité des interactions protéine-ADN. De plus, l’immuno-précipitation de la chromatine (ChIP) a prouvé que GREBP lie le cGMP-RE dans des conditions physiologiques. La liaison de GREBP au cGMP-RE inhibe l’expression du gène rapporteur luciférase sous contrôle du promoteur de npr1/gca. L’inhibition de GREBP à l’aide d’ARN interférant active le promoteur de npr1/gca. Dans les cellules NCI-H295R, l’ANP stimule l’expression de grebp de 60% après seulement 3 heures et inhibe l’expression de npr1/gca de 30%. GREBP est une protéine nucléaire surtout exprimée dans le cœur et ayant le facteur eIF3F comme partenaire. Les variations nucléotidiques du gène sont plus fréquentes chez les patients hypertendus que chez des patients normotendus ou hypertendus souffrant de MetS. Nous rapportons ici l’existence d’un gène spécifique à l’humain qui agit comme répresseur transcriptionnel de npr1/gca et potentiellement impliqué dans le développement de l’hypertension. / The natriuretic peptide (NP) system was a milestone discovery that revealed the endocrine role of the heart for the first time in the early 1980s. From its vasodilatory, natriuretic and diuretic actions, knowledge about this system has evolved to a degree of complexity unsuspected at that time. Now, through cGMP generation, NPs are involved in several other mechanisms, such as cell proliferation, apoptosis, renin-angiotensine-aldosterone system (RAAS) inhibition, and fat cell function. The latter point is of growing interest in lipid metabolism and has become an important issue in the fight against obesity. This pandemic condition is one of the main risk factors leading to hypertension development and metabolic syndrome (MetS) progression. Thus, understanding, at least in part, the lipid mobilization pathways controlled by NPs could have a positive impact in MetS management. As with hypertension, identifying defects in signaling pathways will certainly help to identify mechanisms implicated in lost sensitivity of the NP system. Natriuretic peptide receptor 1 (npr1/gca) expression is controlled by several agents including its own ligand, the atrial natriuretic peptide (ANP). A major finding in NPs field occured in the mid-90s when a mechanism involving a retro-inhibition loop was described. Indeed, after ANP stimulation, NPR1/GCA down-regulates the transcriptional activity of its gene via a cGMP-dependent mechanism. Since our group previously identified a cis-acting element responsible for this cGMP sensitivity, I proceeded to explore novel putative protein binding to the cGMP-response element (cGMP-RE). Using the yeast-one-hybrid technique with a human kidney cDNA library, we identified a strongly positive clone able to bind cGMP-RE. The clone was derived from a 1083-bp long cDNA of a gene of yet unknown function localized on human chromosome 1 (1p33.36). We named this new protein GREBP for cGMP-Response Element-Binding Protein. DNA-binding assays showed 18-fold higher cGMP-RE-binding capacity than the controls while electromobility shift assay (EMSA) indicated a specific binding for the cGMP-RE and chromatin immuno-precipitation (ChIP) confirmed the binding of GREBP to the element under physiological conditions. By acting on cGMP-RE, GREBP inhibited the activity of a luciferase-coupled NPR1 promoter construct. In H295R cells, ANP heightened GREBP expression by 60% after just 3 hours of treatment while inhibiting npr1/gca expression by 30%. Silencing GREBP with specific small interfering RNA increased the activity of the luciferase-coupled NPR1/GCA promoter and NPR1/GCA mRNA levels. GREBP is a nuclear protein mainly expressed in the heart and has the eIF3F factor as partner. Its nucleotide variations are more frequent in non-obese hypertensive patients than normotensive subjects or hypertensive patients suffering from MetS. We report here the existence of a human specific gene acting as a transcriptional repressor of npr1/gca gene that could be implicated in hypertension development.

Dietary fat quality influences the risk of type 2 diabetes and cardiovascular disease. A low-grade inflammation is suggested to contribute to the disease development, often accompanied by obesity. Whereas n-3 polyunsaturated fatty acids (PUFA) have been considered anti-inflammatory, n-6 PUFA have been proposed to act pro-inflammatory. Saturated fatty acids (SFA) act pro-inflammatory in vitro. This thesis aimed to investigate effects of different fatty acids on low-grade inflammation in observational and interventional studies. In Paper I and II, fatty acid composition in serum cholesterol esters was used as objective marker of dietary fat quality and related to serum C-reactive protein (CRP) and other circulating inflammatory markers in two population-based cohorts, conducted in middle-aged men and elderly men and women, respectively. In Paper III and IV, the impact of diets differing in fat quality on inflammation and oxidative stress was investigated in randomised controlled studies, in subjects with metabolic syndrome and abdominal obesity. In Paper I and II, a low proportion of linoleic acid (18:2 n-6) in serum was associated with higher CRP concentrations, indicating that a low intake of vegetable fats may be related to low-grade inflammation. High CRP concentrations were also associated with high proportions of palmitoleic (16:1) and oleic (18:1) acids and high stearoyl coenzymeA desaturase index, possibly reflecting altered fat metabolism and/or high SFA intake in this population. When comparing two high-fat diets rich in either saturated or monounsaturated fat, and two low-fat diets with or without long-chain n-3 PUFA supplementation during 12 weeks (Paper III), no differences in inflammation or oxidative stress markers were observed. Moreover, a 10-week intervention (Paper IV) with high linoleic acid intake showed no adverse effects on inflammation or oxidative stress. Instead, interleukin-1 receptor antagonist and tumor necrosis factor receptor-2 decreased after linoleic acid intake compared with a diet high in SFA. The results in this thesis indicate that dietary n-6 PUFA found in vegetable fats is associated with lower inflammation marker levels, and to some extent reduces systemic inflammation when compared with SFA. Supplementation of n-3 PUFA did not exert any systemic anti-inflammatory effects, maybe due to a relatively low dose.

Increased possibilities during the last decades for early detection of prostate cancer have sparked research on preventable or treatable risk factors and on improvements in therapy. Treatments of the disease still entail significant side effects potentially affecting men during the rest of their lives. The studies of the present thesis concern different aspects of prostate cancer from etiological risk factors and factors influencing treatment to an improved methodology for the detection of treatment side effects. Papers I, II, both based in the population based cohort ULSAM (Uppsala Longitudinal Study of Adult Men), investigate possible risk factors of prostate cancer with options for intervention: selenium levels and the metabolic syndrome. The phenomenon of competing risk of death from other causes than prostate cancer and its impact on and importance for choice of statistical methods is also exemplified and discussed for the first time in prostate cancer research. -Smokers with low selenium status have an increased future risk of later development of prostate cancer. Influence of genetic variability appears plausible. -The metabolic syndrome and especially its increased waist circumference component are associated with later development of prostate cancer – taking competing risks of death from other causes into account. Papers III and IV using pharmacoepidemiological methods investigate aspects of drug utilisation in prostate cancer using nationwide and international databases. In Paper III factors influencing anti-androgen use in prostate cancer are investigated, both from a prescriber- and patient perspective.  The age and disease risk group of the patient, unsupported scientifically, influence both the prescribers’ choice of dose and the patients’ adherence to treatment. -Adherence, not previously investigated in male cancer patients, was considerably higher than reported for adjuvant breast cancer treatment. Subgroups of men suitable for intervention to increase adherence were identified. Paper IV, investigates the feasibility of improving an established method for screening large adverse drug reactions databases, the proportional reporting ratio (PRR), this by using restricted sub-databases according to treatment area (TA), introducing the concept of PRR-TA. -The PRR-TA method increases the signal-noise relationship of analyses; a finding highly relevant for possibly conserving manual resources in Pharmacovigilance work in a drug-authority setting.

Introdução: A Doença Hepática Gordurosa Não Alcoólica (DHGNA) tem como causa principal a obesidade. Atualmente não existe tratamento medicamentoso específico para DHGNA. A cirurgia bariátrica surge como uma alternativa de tratamento em pacientes morbidamente obesos. Objetivos: Analisar, através de biópsia hepática intra-operatória, o grau de comprometimento hepático em obesos submetidos à cirurgia bariátrica, correlacionando os achados histopatológicos com o estado glicêmico dos pacientes. Métodos: Estudo de coorte prospectivo incluindo 521 pacientes submetidos à cirurgia bariátrica de julho de 2001 até dezembro de 2016. Os pacientes foram classificados em três grupos de acordo com o status glicêmico basal: 167(32,05%) diabéticos tipo 2 (G1), 132 (25,33%) pré-diabéticos (G2) e 222 (42,61%) obesos normoglicêmicos (G3). Foram obtidas biópsias hepáticas transoperatórias, as quais foram classificadas conforme os critérios de Brunt e do NASH-CRN. As variáveis clínicas e bioquímicas e histológicas foram comparadas antes da cirurgia e durante o seguimento pós-operatório. Resultados: A prevalência de DHGNA nesta coorte foi de 95%. Não houve diferença quanto ao gênero e IMC entre os grupos. Observaram-se taxas mais altas de fibrose (56,4% G1 vs 29,2% G2 vs 28,6% G3 p<0,001) e Esteatohepatite Não Alcoólica (EHNA) (59,4% G1vs 49,2% G2 vs 36% G3 p <0,001) nos pacientes diabéticos. Apenas 1,5 %, dos diabéticos apresentaram histologia normal (vs 7,76% G2 vs 15,7% G3). / Introduction: Obesity is the main cause of nonalcoholic fatty liver disease (NAFLD), for which there is currently no specific medical treatment. Bariatric surgery is a treatment alternative for morbidly obese patients. Objectives: Use an intraoperative liver biopsy to analyze the degree of liver damage in obese patients submitted to bariatric surgery and correlates the histopathological findings with glucose status. Methods: Prospective cohort study of 521 morbid obese patients undergoing bariatric surgery from July 2001 to December 2016, classified into three groups according to their baseline glucose status: 167 (32.05%) type 2 diabetes (G1), 132 (25.33%) pre-diabetic (G2) and 222 (42.61%) normoglycemic obese (G3). Patients using potentially hepatotoxic medications and a history of ethanol consumption or viral hepatitis were excluded. Intraoperative liver biopsies were obtained and classified in accordance with Brunt and NASH-CRN criteria. Clinical, biochemical and histopathological variables were compared before surgery and during postoperative follow-up. Results: The prevalence NAFLD was 95%. There was no intergroup difference for sex and BMI. Higher rates of fibrosis (56.4% G1 vs. 29.2% G2 vs. 28.6% G3 p<0.001) and nonalcoholic steatohepatitis (NASH) (59.4% G1vs 49.2% G2 vs. 36% G3 p <0.001) were observed in the diabetic patients. Only 1.5 % of diabetics showed normal histology (vs. 7.76% G2 and 15.7% G3).

Introdução: A Doença Hepática Gordurosa Não Alcoólica (DHGNA) tem como causa principal a obesidade. Atualmente não existe tratamento medicamentoso específico para DHGNA. A cirurgia bariátrica surge como uma alternativa de tratamento em pacientes morbidamente obesos. Objetivos: Analisar, através de biópsia hepática intra-operatória, o grau de comprometimento hepático em obesos submetidos à cirurgia bariátrica, correlacionando os achados histopatológicos com o estado glicêmico dos pacientes. Métodos: Estudo de coorte prospectivo incluindo 521 pacientes submetidos à cirurgia bariátrica de julho de 2001 até dezembro de 2016. Os pacientes foram classificados em três grupos de acordo com o status glicêmico basal: 167(32,05%) diabéticos tipo 2 (G1), 132 (25,33%) pré-diabéticos (G2) e 222 (42,61%) obesos normoglicêmicos (G3). Foram obtidas biópsias hepáticas transoperatórias, as quais foram classificadas conforme os critérios de Brunt e do NASH-CRN. As variáveis clínicas e bioquímicas e histológicas foram comparadas antes da cirurgia e durante o seguimento pós-operatório. Resultados: A prevalência de DHGNA nesta coorte foi de 95%. Não houve diferença quanto ao gênero e IMC entre os grupos. Observaram-se taxas mais altas de fibrose (56,4% G1 vs 29,2% G2 vs 28,6% G3 p<0,001) e Esteatohepatite Não Alcoólica (EHNA) (59,4% G1vs 49,2% G2 vs 36% G3 p <0,001) nos pacientes diabéticos. Apenas 1,5 %, dos diabéticos apresentaram histologia normal (vs 7,76% G2 vs 15,7% G3). / Introduction: Obesity is the main cause of nonalcoholic fatty liver disease (NAFLD), for which there is currently no specific medical treatment. Bariatric surgery is a treatment alternative for morbidly obese patients. Objectives: Use an intraoperative liver biopsy to analyze the degree of liver damage in obese patients submitted to bariatric surgery and correlates the histopathological findings with glucose status. Methods: Prospective cohort study of 521 morbid obese patients undergoing bariatric surgery from July 2001 to December 2016, classified into three groups according to their baseline glucose status: 167 (32.05%) type 2 diabetes (G1), 132 (25.33%) pre-diabetic (G2) and 222 (42.61%) normoglycemic obese (G3). Patients using potentially hepatotoxic medications and a history of ethanol consumption or viral hepatitis were excluded. Intraoperative liver biopsies were obtained and classified in accordance with Brunt and NASH-CRN criteria. Clinical, biochemical and histopathological variables were compared before surgery and during postoperative follow-up. Results: The prevalence NAFLD was 95%. There was no intergroup difference for sex and BMI. Higher rates of fibrosis (56.4% G1 vs. 29.2% G2 vs. 28.6% G3 p<0.001) and nonalcoholic steatohepatitis (NASH) (59.4% G1vs 49.2% G2 vs. 36% G3 p <0.001) were observed in the diabetic patients. Only 1.5 % of diabetics showed normal histology (vs. 7.76% G2 and 15.7% G3).

This thesis focuses on the effects of n-3 polyunsaturated fatty acids (n-3 PUFA) on development of non-alcoholic fatty liver disease (NAFLD) in experiment, on prevalence of this condition in patients with type 2 diabetes mellitus and metabolic syndrome and also on non-invasive diagnostics. The aim was to study the effect of n-3 PUFA on NAFLD development in an experimental model and based on analysis of a group of patients with type 2 diabetes and metabolic syndrome to assess the prevalence of this condition. Lastly we aimed to evaluate non-invasive diagnostic methods of liver fibrosis and NASH. We demonstrated beneficial effects of n-3 PUFA administration on NAFLD development in a C57/Bl6 mice high fat methionin-cholin defficient dietary model of NAFLD. n-3 PUFA administration led to biochemical improvement, decrease of lipid accumulation in the liver as well as improvement of histology. These effects are determined by complex modulation of lipid metabolism, mainly due to decrease in availability of fatty acids for triglyceride synthesis in the liver, changes of adipokine levels and amelioration of proinflammatory status in the liver. In a group of type 2 diabetics we found NAFLD prevalence of almost 80%, 14% of these patients had also signs of liver fibrosis or cirrhosis. Non-invasive methods...

INTRODUÇÃO: A obesidade é uma das causas de hipogonadismo (HG) secundário no homem. A terapia de reposição padrão de testosterona (TRT) é associada à melhora dos parâmetros metabólicos, mas pode levar à infertilidade. Apenas recentemente indicou-se que não há novas evidências nível 1 para apoiar uma conexão definitiva entre TRT e eventos cardiovasculares (CV). OBJETIVO: Avaliar os efeitos do Citrato de Clomifeno (CC) em homens jovens com hipogonadismo associado à obesidade diagnosticado por testosterona total (TT) <= 300 ng/dL em duas ocasiões, sintomas positivos no questionário ADAM, hormônio Luteinizante (LH) baixo ou inadequadamente normal (VR: 1,7 - 8,6 UI/L). MÉTODOS: Estudo randomizado, duplo cego, controlado por placebo (PLB), longitudinal em centro único. Setenta e oito pacientes com idade entre 36,5±7,8 anos, índice de massa corporal (IMC) 46,2±8,5 kg/m2 foram randomizados (1:1) para receber CC 50 mg ou PLB durante 12 semanas. Os pacientes foram avaliados através de: 1) Parâmetros clínicos: Questionário ADAM, número de intercursos sexuais, queixa de insatisfação com a vida sexual; 2) Parâmetros hormonais: dosagem sérica de TT, testosterona livre, Estradiol (E2), LH, hormônio folículo estimulante (FSH), SHBG, relação TT:E2; 3) Parâmetros de composição corporal: IMC, circunferência abdominal (CA) e análise de bioimpedanciometria; 4) Parâmetros metabólicos: pressão arterial sistólica e diastólica, glicemia em jejum (GJ), hemoglobina glicada (HbA1c), índice HOMA-IR, colesterol total e frações, triglicérides; 5) Parâmetros de resposta CV: dilatação fluxo mediada artéria braquial (FMDAB), níveis circulantes de sICAM-1, sVCAM-1, Selectina-sE e quantificação de células endoteliais progenitoras (CEPs) por citometria de fluxo; 6) Efeitos adversos: hematócrito, antígeno prostático específico sérico (PSA), questionário internacional de sintomas prostáticos (I-PSS), dosagem sérica de alanina aminotransferase (ALT), aspartato aminotransferase (AST), e efeitos adversos autorreferidos. RESULTADOS: Na randomização os dois grupos foram semelhantes em relação à idade (CC: 35,5±7,8 anos, PLB: 35,6±7,8; p= 0,951), IMC (CC: 45,5±11,3 kg/m2; PLB: 47,2±9,6; p= 0,470), CA (CC: 137,5±17,9 cm; PLB: 140,2±19,6; p= 0,526) e testosterona total (CC: 225,8±70,0 ng/dL; PLB: 216,0±72,1; p= 0,543). Não houve diferenças nos parâmetros de resposta clínica, exceto com relação à queixa de perda de vigor nas ereções (p < 0,001). Observou-se elevação significativa (p= < 0,001) de TT, Testosterona livre, E2, LH, FSH e SHBG no grupo CC em comparação com PLB. Houve um aumento significativo (p < 0,001) na massa magra e na massa muscular; e também na massa livre de gordura (p= 0,004). O CC reduziu HDL em comparação com PLB (p < 0,001) e não mostrou efeito em outros parâmetros metabólicos. Não houve significância estatística nos parâmetros CV, indicando efeito nulo do tratamento. CC reduziu ALT (p < 0,001) e aumentou o PSA (p= 0,023) dentro dos limites da normalidade. CONCLUSÕES: CC foi efetivo para melhorar os parâmetros de resposta hormonal e afetou positivamente um parâmetro de resposta clínica (perda de vigor nas ereções). Apesar das alterações na composição corporal, não se observou melhora do perfil metabólico. No entanto, o CC não ocasionou resposta adversa nos parâmetros CV. O tratamento CC para HG parece ser uma alternativa efetiva em jovens obesos que desejam preservar sua fertilidade, mas ensaios clínicos de seguimento em longo prazo e com maior número de participantes são necessários para melhor análise do perfil metabólico e de sintomas, além de impactos CV / INTRODUCTION: Obesity can cause secondary hypogonadism in man. The standard testosterone replacement therapy (TRT) improves metabolic parameters but can lead to infertility. Only recently TRT was not clearly associated with adverse cardiovascular (CV) events, but its impacts on endothelial function are still controversial. AIM: To evaluate the effects of Clomiphene Citrate (CC) in out clinic young man with obesity related hypogonadism: total testosterone (TT) <= 300 ng/dL on two occasions, positive symptoms in ADAM questionnaire, Luteinizing Hormone (LH) low or inappropriate normal (RV: 1.7-8.6 IU/liter). METHODS: This is a randomized, double blind, placebo-controlled, parallel group, single-center study. Seventy eight patients aged 36.5±7.8 years, Body mass index (BMI) 46.2±8.5 kg/m2 were randomized (1:1) to receive CC 50 mg or Placebo (PLB) during 12 weeks. MAIN OUTCOME MEASURES: 1) Clinical symptomology: ADAM Questionnaire, number of sexual intercourses and satisfaction with sexual life; 2) Hormonal monitoring: serum TT, Free testosterone, Estradiol (E2), LH and Follicle-stimulating hormone (FSH), SHBG, TT/E2 ratio; 3) Body composition and anthropometric measurements: BMI, waist circumference (WC) and Bioelectric Impedance analysis parameters; 4) Metabolic response parameters: systolic and diastolic blood pressure, fasting blood glucose (FBG), glycated hemoglobin (HbA1c), serum cholesterol and fractions, triglycerides; 5) CV assessment by endothelial function parameters: Flowmediated dilatation of the brachial artery (FMDAB), circulating levels of sICAM-1, sVCAM-1, E-selectin and flow cytometry endothelial progenitor cells (EPCs); 6) Adverse outcomes: Hematocrit, serum Prostate-Specific Antigen (PSA), International Prostate Symptom Score (I-PSS), Alanine Aminotransferase (ALT), Aspartate Aminotransferase (AST) and Selfreported Adverse Effects. RESULTS: Two groups were similar with regard to age (CC: 35.5±7.8 years; PLB: 35.6±7.8; P=0.951), BMI (CC: 45.5±11.3 kg/m2; PLB: 47.2±9.6; P=0.470), WC (CC: 137.5±17.9 cm; PLB: 140.2±19.6; P=0.526) and total testosterone (CC: 225.8±70.0 ng/dL; PLB: 216.0±72.1; P=0.543) in baseline data. There was an improvement in one sexual complaint (weaker erections) (P < 0.001) and there were significant improvements (P < 0.001) in TT, Free Testosterone, E2, LH, FSH and SHBG in CC group (vs. PLB). There was a gain in lean mass (P < 0.001), free fat mass (P=0.004) and muscle mass (P < 0.001). CC reduced HDL compared to PLB (P < 0.001) and showed no effect in other metabolic parameters. No statistical significance was seen in CV parameters. CC reduced ALT (P < 0.001) and increased PSA (P=0.023). CONCLUSIONS: CC was effective in increase hormonal response parametersand improved one sexual complaint (weaker erections). Despite body composition changes, CC did not improved metabolic profile and lowered LDL cholesterol. CC showed no adverse response in CV parameters. CC treatment for HG appears to be an effective alternative in young obese men wishing to preserve their fertility but long-term follow-up trials to better analyze the metabolic profile and CV outcomes are needed

<p>Izvod: UVOD: Metabolički zdrave gojazne osobe su okarakterisane odsustvom metaboličkog sindroma i/ili insulinske rezistencije i sistemske inflamacije. Mali je broj podataka o ulozi aktivnosti autonomnog nervnog sistema u razvoju kardiometaboličkih komplikacija kod gojaznih osoba, kao i o njegovoj vezi sa specifičnom distribucijom masnog tkiva. CILJ: Analiza varijabilnost srčane frekvencije (HRV) kod metabolički zdravih (MHO) i gojaznih osoba sa metaboličkim rizikom (MUO), analiza povezanosti HRV sa metaboličkim faktorima i distribucijom masnog tkiva, kao i analiza uzrasne dinamike HRV kod gojaznih osoba različitih kardiometaboličkih profila. MATERIJAL I METODE: Ukupno 125 gojaznih ispitanika oba pola podvrgnuto je antropometrijskim merenjima u cilju procene mase i distribucije masnog tkiva, izvr&scaron;ena je analiza telesne kompozicije, uzeti su uzorci krvi u cilju određivanja lipidskog i lipoproteinskog statusa, stanja glikoregulacije i nivoa inflamatornih markera, meren je krvni pritisak i procenjena je HRV tokom petominutne digitalne elektrokardiografije. Podaci su statistički obrađeni kori&scaron;ćenjem paketa SPSS 11.5. REZULTATI: HRV mere se nisu razlikovale statistički značajno među MHO i MUO mu&scaron;karcima. MHO žene su imale vi&scaron;e vrednosti RRNN, SDNN, RMSSD, pNN50, LF, HF i TP u odnosu na MUO žene, na čega metabolički profil utiče sa 6,6-11,2%(p˂0,01), predstavljeno kroz parcijalnu deljenu varijansu. Nakon antropometrijskih faktora uzetih u obzir, perzistirale su vi&scaron;e vrednosti HF kod MHO žena. Razlika u RRNN, pNN50 i TP između MHO i MUO premenopauzalnih žena (vi&scaron;e vrednosti za MHO, p˂0,05) se izgubila nakon kontrole za krvni pritisak. Insulinemija je uticala na pojave razlika u RRNN između MHO i MUO premenopauzalnih žena, parcijalna deljena varijansa 7,6%. SAD kod žena se negativno povezivao sa LF/HF i LFnorm, a pozitivno sa HFnorm, parcijalne deljene varijanse 8,4-11,9% (p˂0,05). Prednji nabor podlaktice kod žena se pozitivno povezivao sa LF i LF/HF, a negativno sa HFnorm (p˂0,01). Visceralna masna masa je predviđala značajno HRV mere mu&scaron;karaca, parcijalna deljena varijansa 13-34% (p˂0,01). U okviru gornjeg tercila HRV mera RMSSD, pNN50 i LF MUO osoba, HOMA indeks je statistički značajno niži (p˂0,05). Kod MUO osoba SDNN, RMSSD, lnpNN50, lnLF, lnHF i TP značajno su se smanjivali u uzrastu od 19-29 do 40-49 godina. Kod MHO osoba primetna je uzrasna promena HF mere u četvrtoj deceniji života. ZAKLJUČAK: MHO osobe ženskog pola imaju značajno vi&scaron;e vrednosti markera varijabilnosti srčane frekvencije u odnosu na MUO. Razlike u HRV merama su uslovljene kriterijumima metaboličke podele, predominantno insulinemijom, vrednostima krvnog pritiska i centralnom masnom masom. Kod žena centralna distribucija masnog tkiva korelira sa smanjenom srčanom simpatičkom aktivno&scaron;ću dok se periferna distribucija masnog tkiva povezuje obrnuto sa komponentama aktivnosti autonomnog nervnog sistema. Kod mu&scaron;karaca centralna masna masa, ali ne i periferna, je značajno povezana sa HRV. MUO osobe sa nižom HRV imaju veći stepen insulinske rezistencije, dok HRV ne utiče na insulinsku senzitivnost MHO osoba. Značajniji uzrasno zavisni pad HRV mera primetan je kod MUO osoba, pogađajući obe komponente autonomnog nervnog sistema za razliku od MHO osoba.</p> / <p>Abstract: INTRODUCTION: Metabolically healthy obese (MHO) individuals are characterized by absence of metabolic syndrome and/or insulin resistance and inflammation. Little is known about the role of autonomic nervous system in development of cardiometabolic complications in obese people and about its influence on the specific adipose tissue distribution. AIM: Analysis of the hearth rate variability (HRV) in metabolically healthy (MHO) and unhealthy (MUO) obese people, its connection with adipose tissue distribution, and age dependent dynamics of HRV. MATERIAL AND METHODS: A total of 125 obese patients of both sexes underwent anthropometric measurements in order to assess adipose tissue mass and distribution, body composition was assessed, blood samples were taken in order to analyze parameters of lipid and lipoprotein profile, condition of glycoregulation and inflammatory markers, blood pressure was measured and short term HRV was conducted. Data were statisticaly analyzed using SPSS 11.5. RESULTS: HRV measures did not differ significantly between MHO and MUO men. MHO women had higher values of RRNN, SDNN, RMSSD, pNN50, LF, HF and TP compared to the MUO women, influence of metabolic profile was 6,6-11,2% (p˂0,01), presented through partial shared variance. After controlling for anthropometric factors higher HF persisted in MHO women. Differences in RRNN, pNN50 and TP between MHO and MUO premenopausal women (higher values of MHO, p˂0,05) were lost after controlling for blood pressure. Insulinemia influenced the difference in RRNN between MHO and MUO premenopausal women, partial shared variance 7,6%. SAD in women was connected negatively with the LF/HF and LFnorm, and positively with HFnorm, partial shared variance 8,4-11,9% (p˂0,05). Anterior forearm skinfold in women correlated positively with LF and LF/HF, and negatively with HFnorm (p˂0,01). Visceral fat mass predicted significantly HRV in men, partial shared variance 13-34% (p˂0,01). Within the upper tertile of HRV measures RMSSD, pNN50 and LF in MUO people, HOMA was significantly lower (p˂0,05). In MUO SDNN, RMSSD, lnpNN50, lnLF,lnHF and TP significantly decreased in the period from 19-29 to 40-49 years. In MHO people the change in HF was noticeable in the fourth decade of life. CONCLUSION: MHO women have significantly higher levels of HRV markers comparing to the MUO. The differences in HRV measures are influenced by metabolic criteria used, predominantly by insulinemia, blood pressure and central fat mass. In women, central distribution of adipose tissue correlates with reduced cardiac sympathetic activity, while the connection of peripheral fat mass distribution with components of autonomic nervuos system activity is reverse. In men, central fat mass, but not peripheral, is significantly associated with HRV. MUO people with lower HRV have a higher degree of insulin resistance, while the level of HRV measures does not affect insulin sensitivity in MHO individuals. Significant age-dependent decrease in both ANS representatives of HRV measures was noticed in MUO people, unlike MHO individuals.</p>

This thesis investigates how air pollution, both natural and anthropogenic, affects changes in the proximal small intestine and ileum microbiota profile, as well as intestinal barrier integrity, histological changes, and inflammation. APO-E KO mice on a high fat diet were randomly selected to be exposed by whole body inhalation to either wood smoke (WS) or mixed vehicular exhaust (MVE), with filtered air (FA) acting as the control. Intestinal integrity and histology were assessed by observing expression of well- known structural components tight junction proteins (TJPs), matrix metallopeptidase-9 (MMP-9), and gel-forming mucin (MUC2), as well known inflammatory related factors: TNF-α, IL-1β, and toll-like receptor (TLR)-4. Bacterial profiling was done using DNA analysis of microbiota within the ileum, utilizing 16S metagenomics sequencing (Illumina miSeq) technique. Overall results of this experiment suggest that air pollution, both anthropogenic and natural, cause a breach in the intestinal barrier with an increase in inflammatory factors and a decrease in beneficial bacteria. This evidence suggests the possibility of air pollution being a potential causative agent of intestinal disease as well as a possible contributing mechanism for induction of systemic inflammation.