Evaluation of the Chagas Disease Control Program in Açucena Municipality, Rio Doce Valley, State of Minas Gerais, Brazil

Introduction Açucena Municipality, Rio Doce Valley, State of Minas Gerais, Brazil temporarily (2001-2005) interrupted epidemiological surveillance for Chagas disease. The objective of this work was to evaluate the Chagas Disease Control Program (CDCP) in Açucena and to offer suggestions for improving local epidemiological surveillance. Methods This study was conducted in three phases: I) a serological investigation of schoolchildren aged 5 to 15 years using an enzyme-linked immunosorbent assay (ELISA) test performed on blood collected on filter paper followed by ELISA, indirect immunofluorescence (IIF) and indirect hemaglutination (IHA) on venous blood for borderline cases and those in the gray zone of reactivity; II) vector evaluation using the data obtained by local health agents during 2006-2010; and III) examination by ELISA, IIF and IHA of serum samples from the inhabitants of houses where infected Triatoma vitticeps was found and evaluation of their knowledge about Chagas disease. Results Five individuals had inconclusive results in the ELISA screening but were seronegative for Chagas disease. The triatomine evaluation revealed the presence of three species: Triatoma vitticeps, Panstrongylus megistus and Panstrongylus diasi. Triatoma vitticeps was the most prevalent and widespread, with a higher (67%) index of Trypanosoma cruzi flagellates and evidence of colonization. Most of the inhabitants of the infested houses recognized triatomines and had basic knowledge about Chagas disease. Conclusions Although T. vitticeps is not clearly associated with Chagas disease transmission, these results highlight the importance of maintaining CDCP in endemic areas and the need for greater emphasis on epidemiological surveillance, especially in areas with important vectorial changes or that have been modified by human intervention.

Risk factors for hepatitis C virus transmission in the municipality of Catanduva, State of São Paulo: a case-control study

Introduction Hepatitis C virus (HCV) is primarily transmitted via contact with the blood of infected patients, although the form of contact has not been identified for a significant percentage of carriers. The present study evaluated possible risk factors for HCV transmission in a medium-sized town located in the northwest region of the State of São Paulo. Methods This was a case-control study, with the case group consisting of 190 chronic HCV carriers older than 18 years residing in the municipality of Catanduva. The control group also consisted of 190 individuals with HCV-negative serology. The groups were paired (1:1) for gender, age range (± five years), and place of residence. The same structured questionnaire was applied to all subjects, who gave written informed consent to participate in the study. The data were statistically analyzed using crude and adjusted logistic regression, and the results were expressed as odds ratios with a 95% confidence interval. Results The demographic profiles of the groups indicated a predominance of males (68.9%) and mean ages of 47.1 years (case group) and 47.3 years (control group). After adjusting for conditional regression, the following factors were found to represent risks for HCV: history of sexually transmitted disease (STD) and blood transfusion; accidents with syringes and/or needles; tattoos; and the use of non-injectable drugs and injectable medications. Conclusions The transmission of HCV via the blood route has been well characterized. Other forms of contact with human blood and/or secretions are likely to transmit the virus, although with a lower frequency of occurrence.

Evaluation of preventative and control measures for congenital syphilis in State of Mato Grosso

Introduction Congenital syphilis is an important health problem in Brazil. This study assessed measures aimed at the prevention and control of syphilis in the State of Mato Grosso and its capital, Cuiabá. Methods A descriptive study cross-sectional and of time trends assessing the congenital syphilis was performed in Cuiabá and Mato Grosso between 2001 and 2011. We compared maternal sociodemographic characteristics and health care utilization related to cases of congenital syphilis during the periods from 2001 to 2006 and from 2007 to 2011. We assessed the temporal trends in this disease's incidence using a simple linear regression. Results Between 2001 and 2006 in Mato Grosso, 86.8% of the mothers who had live births with congenital syphilis received prenatal care, 90.6% presented with a nontreponemal test reagent at delivery, 96.2% had no information regarding a treponemal confirmatory test at delivery, and 77.6% received inadequate treatment for syphilis; additionally, 75.8% of their partners were not treated. There was a statistically significant reduction in prenatal visits (p = 0.004) and an increase in the proportion of mothers reactive to nontreponemal tests at delivery (p = 0.031) between the two periods. No other variables were found to differ significantly between the periods. In Cuiabá, we observed a similar distribution of variables. In the state and in the capital, the increasing trend of congenital syphilis was not statistically significant. Conclusions The high incidence of congenital syphilis in Mato Grosso and the low levels of health care indicators for pregnant women with syphilis suggest the need to improve the coverage and quality of prenatal care.

Impact of insecticide resistance on the field control of Aedes aegypti in the State of São Paulo

IntroductionThe need to control dengue transmission by means of insecticides has led to the development of resistance to most of the products used worldwide against mosquitoes. In the State of São Paulo, the Superintendência de Controle de Endemias(SUCEN) has annually monitored the susceptibility of Aedes aegypti to insecticides since 1996; since 1999, surveys were conducted in collaboration with the National Network of Laboratories (MoReNAa Network) and were coordinated by the Ministry of Health. In this study, in addition to the biological characterization of insecticide resistance in the laboratory, the impact of resistance on field control was evaluated for vector populations that showed resistance in laboratory assays.MethodsField efficacy tests with larvicides and adulticides were performed over a 13-year period, using World Health Organization protocols.ResultsData from the field tests showed a reduction in the residual effect of temephos on populations with a resistance ratio of 3. For adults, field control was less effective in populations characterized as resistant in laboratory qualitative assays, and this was confirmed using qualitative assays and field evaluation.ConclusionsOur results indicated that management of resistance development needs to be adopted when insect populations show reduced susceptibility. The use of insecticides is a self-limiting tool that needs to be applied cautiously, and dengue control requires more sustainable strategies.

Potential for entomopathogenic fungi to control Triatoma dimidiata (Hemiptera: Reduviidae), a vector of Chagas disease in Mexico

Introduction The use of entomopathogenic fungi to control disease vectors has become relevant because traditional chemical control methods have caused damage to the environment and led to the development of resistance among vectors. Thus, this study assessed the pathogenicity of entomopathogenic fungi in Triatoma dimidiata. Methods Preparations of 108 conidia/ml of Gliocladium virens, Talaromyces flavus, Beauveria bassiana and Metarhizium anisopliae were applied topically on T. dimidiata nymphs and adults. Controls were treated with the 0.0001% Tween-80 vehicle. Mortality was evaluated and recorded daily for 30 days. The concentration required to kill 50% of T. dimidiata (LC50) was then calculated for the most pathogenic isolate. Results Pathogenicity in adults was similar among B. bassiana, G. virens and T. flavus (p>0.05) and differed from that in triatomine nymphs (p=0.009). The most entomopathogenic strains in adult triatomines were B. bassiana and G. virens, which both caused 100% mortality. In nymphs, the most entomopathogenic strain was B. bassiana, followed by G. virens. The native strain with the highest pathogenicity was G. virens, for which the LC50 for T. dimidiata nymphs was 1.98 x108 conidia/ml at 13 days after inoculation. Conclusions Beauveria bassiana and G. virens showed entomopathogenic potential in T. dimidiata nymphs and adults. However, the native G. virens strain presents a higher probability of success in the field, and G. virens should thus be considered a potential candidate for the biological control of triatomine Chagas disease vectors.

Compaction quality control on site of earthworks - A comparative study

The capacity to use geologic materials (soil and rock) that are available in the surrounding environment is inherent to the human civilization and has contributed to the evolution of societies throughout the course of history. The use of these materials in the construction of structures such as houses, roads, railways or dams, stirred the improvement of socioeconomic and environmental conditions. Several reports of structural problems on embankments can be found throughout history. A considerable number of those registers can be linked to inadequate compaction, demonstrating the importance of guaranteeing a suitable quality of soil compaction. Various methodologies and specifications of compaction quality control on site of earthworks, based on the fill moisture content and dry unit weight, were developed during the 20th century. Two widely known methodologies are the conventional and nuclear techniques. The conventional methods are based on the use of the field sand cone test (or similar) and sampling of material for laboratory-based testing to evaluate the fill dry unit weight and water content. The nuclear techniques measure both parameters in the field using a nuclear density gauge. A topic under discussion in the geotechnical community, namely in Portugal, is the comparison between the accuracy of the nuclear gauge and sand cone test results for assessing the compaction and density ratio of earth fills, particularly for dams. The main purpose of this dissertation is to compare both of them. The data used were acquired during the compaction quality control operations at the Coutada/Tamujais dam trial embankment and core construction. This is a 25 m high earth dam located in Vila Velha de Rodão, Portugal. To analyse the spatial distribution of the compaction parameters (water content and compaction ratio), a 3D model was also developed. The main results achieved are discussed and finally some considerations are put forward on the suitability of both techniques to ensure fill compaction quality and on additional research to complement the conclusions obtained.

Estudo do controlo motor via Sincronias entre sinais de EEG e de EMG / Motor control study through sincrony between EEG and EMG signals

Um dos maiores desafios da neurofisiologia é o de compreender a forma como a informação é transmitida através do sistema nervoso. O estudo do sistema nervoso tem várias aplicações, tanto na neurologia, permitindo avanços ao nível clínico, como noutras áreas, e.g., nos sistemas de processamento de informação baseados em redes neuronais. A transmissão de informação entre neurónios é feita por via de sinais elétricos. A compreensão deste fenómeno é ainda incompleta e há projectos a nível europeu e mundial com o objetivo de modular o sistema nervoso no seu todo de forma a melhor o compreender. Uma das teses que se desenvolve hoje em dia é a de que a transmissão de sinais elétricos no sistema nervoso é influenciada por fenómenos de sincronia. O objetivo desta dissertação é o de otimizar um protocolo de aquisição e análise de dados reais de eletroencefalograma e eletromiograma com o propósito de observar fenómenos de sincronia, baseando-se num algoritmo (análise por referência de fase, ou RPA, do inglês reference phase analysis) que deteta sincronias de fase entre os sinais de eletroencefalograma (EEG) e um sinal de referência, que é, no caso presente, o eletromiograma (EMG). A otimização deste protocolo e sua validação indicaram a existência de fenómenos significativos de sincronia no sinal elétrico, transmitido entre os músculos da mão e o córtex motor, no decorrer da ação motora.

Theranostic applications of phage display to control leishmaniasis: selection of biomarkers for serodiagnostics, vaccination, and immunotherapy

AbstractPhage display is a high-throughput subtractive proteomic technology used for the generation and screening of large peptide and antibody libraries. It is based on the selection of phage-fused surface-exposed peptides that recognize specific ligands and demonstrate desired functionality for diagnostic and therapeutic purposes. Phage display has provided unmatched tools for controlling viral, bacterial, fungal, and parasitic infections, and allowed identification of new therapeutic targets to treat cancer, metabolic diseases, and other chronic conditions. This review presents recent advancements in serodiagnostics and prevention of leishmaniasis -an important tropical parasitic disease- achieved using phage display for the identification of novel antigens with improved sensitivity and specificity. Our focus is on theranostics of visceral leishmaniasis with the aim to develop biomarker candidates exhibiting both diagnostic and therapeutic potential to fight this important, yet neglected, tropical disease.

Control of a white organic light emitting diode’s emission parameters using a single doped RGB active layer

White Color tuning is an attractive feature that Organic Light Emitting Diodes (OLEDs) offer. Up until now, there hasn’t been any report that mix both color tuning abilities with device stability. In this work, White OLEDs (W-OLEDs) based on a single RGB blend composed of a blue emitting N,N′-Di(1-naphthyl)-N,N′-diphenyl-(1,1′-biphenyl)-4,4′-diamine (NPB) doped with a green emitting Coumarin-153 and a red emitting 4-(Dicyanomethylene)-2-methyl-6-(4-dimethylaminostyryl)-4H-pyran (DCM1) dyes were produced. The final device structure was ITO/Blend/Bathocuproine (BCP)/ Tris(8-hydroxyquinolinato)aluminium (Alq3)/Al with an emission area of 0.25 cm2. The effects of the changing in DCM1’s concentration (from 0.5% to 1% wt.) allowed a tuning in the final white color resulting in devices capable of emitting a wide range of tunes – from cool to warm – while also keeping a low device complexity and a high stabilitty. Moreover, an explanation on the optoelectrical behavior of the device is presented. The best electroluminescense (EL) points toward 160 cd/m2 of brightness and 1.1 cd/A of efficiency, both prompted to being enhanced. An Impedance Spectroscopy (IS) analysis allowed to study both the effects of BCP as a Hole Blocking Layer and as an aging probe of the device. Finally, as a proof of concept, the emission was increased 9 and 64 times proving this structure can be effectively applied for general lighting.

Universal banks and corporate control: Evidence from the global syndicated loan market

We investigate the effects of bank control over borrower firms whether by representation on boards of directors or by the holding of shares through bank asset management divisions. Using a large sample of syndicated loans, we find that banks are more likely to act as lead arrangers in loans when they exert some control over the borrower firm. Bank-firm governance links are associated with higher loan spreads during the 2003-2006 credit boom, but lower spreads during the 2007-2008 financial crisis. Additionally, these links mitigate credit rationing effects during the crisis. The results are robust to several methods to correct for the endogeneity of the bank- firm governance link. Our evidence, consistent with intertemporal smoothing of loan rates, suggests there are costs and benefits from banks’ involvement in firm governance.

Development of process control strategies exploiting knowledge from systems biology: application to MDCK suspension cells

Madine Darby Canine Kidney (MDCK) cell lines have been extensively evaluated for their potential as host cells for influenza vaccine production. Recent studies allowed the cultivation of these cells in a fully defined medium and in suspension. However, reaching high cell densities in animal cell cultures still remains a challenge. To address this shortcoming, a combined methodology allied with knowledge from systems biology was reported to study the impact of the cell environment on the flux distribution. An optimization of the medium composition was proposed for both a batch and a continuous system in order to reach higher cell densities. To obtain insight into the metabolic activity of these cells, a detailed metabolic model previously developed by Wahl A. et. al was used. The experimental data of four cultivations of MDCK suspension cells, grown under different conditions and used in this work came from the Max Planck Institute, Magdeburg, Germany. Classical metabolic flux analysis (MFA) was used to estimate the intracellular flux distribution of each cultivation and then combined with partial least squares (PLS) method to establish a link between the estimated metabolic state and the cell environment. The validation of the MFA model was made and its consistency checked. The resulted PLS model explained almost 70% of the variance present in the flux distribution. The medium optimization for the continuous system and for the batch system resulted in higher biomass growth rates than the ones obtained experimentally, 0.034 h-1 and 0.030 h-1, respectively, thus reducing in almost 10 hours the duplication time. Additionally, the optimal medium obtained for the continuous system almost did not consider pyruvate. Overall the proposed methodology seems to be effective and both proposed medium optimizations seem to be promising to reach high cell densities.

A historical overview of leprosy epidemiology and control activities in Amazonas, Brazil

Leprosy is an ancient infectious disease caused by Mycobacterium leprae. According to comparative genomics studies, this disease originated in Eastern Africa or the Near East and spread with successive human migrations. The Europeans and North Africans introduced leprosy into West Africa and the Americas within the past 500 years. In Brazil, this disease arrived with the colonizers who disembarked at the first colonies, Rio de Janeiro, Salvador and Recife, at the end of the sixteenth century, after which it was spread to the other states. In 1854, the first leprosy cases were identified in State of Amazonas in the north of Brazil. The increasing number of leprosy cases and the need for treatment and disease control led to the creation of places to isolate patients, known as leprosaria. One of them, Colonia Antônio Aleixo was built in Amazonas in 1956 according to the most advanced recommendations for isolation at that time and was deactivated in 1979. The history of the Alfredo da Matta Center (AMC), which was the first leprosy dispensary created in 1955, parallels the history of leprosy in the state. Over the years, the AMC has become one of the best training centers for leprosy, general dermatology and sexually transmitted diseases in Brazil. In addition to being responsible for leprosy control programs in the state, the AMC has carried out training programs on leprosy diagnosis and treatment for health professionals in Manaus and other municipalities of the state, aiming to increase the coverage of leprosy control activities. This paper provides a historical overview of leprosy in State of Amazonas, which is an endemic state in Brazil.

Control in distribution networks with demand side management

The way in which electricity networks operate is going through a period of significant change. Renewable generation technologies are having a growing presence and increasing penetrations of generation that are being connected at distribution level. Unfortunately, a renewable energy source is most of the time intermittent and needs to be forecasted. Current trends in Smart grids foresee the accommodation of a variety of distributed generation sources including intermittent renewable sources. It is also expected that smart grids will include demand management resources, widespread communications and control technologies required to use demand response are needed to help the maintenance in supply-demand balance in electricity systems. Consequently, smart household appliances with controllable loads will be likely a common presence in our homes. Thus, new control techniques are requested to manage the loads and achieve all the potential energy present in intermittent energy sources. This thesis is focused on the development of a demand side management control method in a distributed network, aiming the creation of greater flexibility in demand and better ease the integration of renewable technologies. In particular, this work presents a novel multi-agent model-based predictive control method to manage distributed energy systems from the demand side, in presence of limited energy sources with fluctuating output and with energy storage in house-hold or car batteries. Specifically, here is presented a solution for thermal comfort which manages a limited shared energy resource via a demand side management perspective, using an integrated approach which also involves a power price auction and an appliance loads allocation scheme. The control is applied individually to a set of Thermal Control Areas, demand units, where the objective is to minimize the energy usage and not exceed the limited and shared energy resource, while simultaneously indoor temperatures are maintained within a comfort frame. Thermal Control Areas are overall thermodynamically connected in the distributed environment and also coupled by energy related constraints. The energy split is performed based on a fixed sequential order established from a previous completed auction wherein the bids are made by each Thermal Control Area, acting as demand side management agents, based on the daily energy price. The developed solutions are explained with algorithms and are applied to different scenarios, being the results explanatory of the benefits of the proposed approaches.

Medium access control design for distributed opportunistic radio networks

Existing wireless networks are characterized by a fixed spectrum assignment policy. However, the scarcity of available spectrum and its inefficient usage demands for a new communication paradigm to exploit the existing spectrum opportunistically. Future Cognitive Radio (CR) devices should be able to sense unoccupied spectrum and will allow the deployment of real opportunistic networks. Still, traditional Physical (PHY) and Medium Access Control (MAC) protocols are not suitable for this new type of networks because they are optimized to operate over fixed assigned frequency bands. Therefore, novel PHY-MAC cross-layer protocols should be developed to cope with the specific features of opportunistic networks. This thesis is mainly focused on the design and evaluation of MAC protocols for Decentralized Cognitive Radio Networks (DCRNs). It starts with a characterization of the spectrum sensing framework based on the Energy-Based Sensing (EBS) technique considering multiple scenarios. Then, guided by the sensing results obtained by the aforementioned technique, we present two novel decentralized CR MAC schemes: the first one designed to operate in single-channel scenarios and the second one to be used in multichannel scenarios. Analytical models for the network goodput, packet service time and individual transmission probability are derived and used to compute the performance of both protocols. Simulation results assess the accuracy of the analytical models as well as the benefits of the proposed CR MAC schemes.

The role of s-nitrosothiols and rho-kinase 1 in glucose and lipid metabolism

RESUMO: Na sociedade contemporânea a diabetes tipo 2 e a obesidade estão a aumentar exponencialmente, representando um grave problema de saúde pública. De acordo com a IDF “A diabetes e a obesidade são o principal problema de saúde pública do século XXI’. Para além destas duas patologias, a prevalência de esteatose hepática não-alcoólica (NAFLD), entre a população obesa e diabética, é de cerca de 90%. O aumento da obesidade, diabetes e NAFLD tem uma forte correlação com o aumento do consumo de gorduras e açúcares, acompanhado de um decréscimo acentuado da actividade física. A obesidade, diabetes e NAFLD tem sido escrupolosamente investigada mas as terapêuticas disponíveis continuam a ser muito limitadas. Tendo em conta o número crescente e alarmante de obesos e diabéticos o conhecimento detalhado da patofisiologia da obesidade, diabetes e NAFLD, tendo em vista a necessidade extrema de desenvolvimento de novas estratégias terapêuticas, é da mais elevada urgência. O fígado é reconhecido como um orgão primordial no controlo da homeostase. No estado pós-prandial, o fígado converte a glucose em glicogénio e lípidos. Em contraste, no estado de jejum, o fígado promove a produção de glucose. Sistemas neuronais e hormonais, bem como o estado metabólico do fígado, controlam de forma muito precisa a alternância entre os diferentes substratos metabólicos, dependente do estado prandial. A insulina tem um papel central no controlo do metabolismo energético no fígado; se, por um lado, inibe a produção hepática de glucose e corpos cetónicos, por outro, promove a glicólise e a lipogénese. O metabolismo energético no fígado é também regulado por vários factores de transcrição e co-reguladores que, por sua vez, são regulados pela insulina, glucagina e outras hormonas metabólicas. Em conjunto, todos estes factores e reguladores vão controlar de forma muito estreita a gluconeogénese, a β-oxidação e a lipogénese, no fígado. Para além dos já conhecidos reguladores do metabolismo hepático, novas moléculas têm sido estudadas como tendo um papel fundamental na regulação do metabolismo energético no fígado. Qualquer desequilíbrio no metabolismo hepático vai contribuir para a insulino-resistência, NAFLD e diabetes tipo 2. O principal objectivo do trabalho de investigação aqui apresentado é o contributo para o estudo detalhado da patogénese da diabetes e obesidade, num contexto de dietas ricas em açúcares e gorduras, e com a perspectiva de explorar novas estratégias terapêuticas. Os objectivos específicos deste trabalho eram: primeiro, determinar se o tratamento com glutationo (GSH) e óxido nítrico (NO) era suficiente para melhorar a insulino-resistência associada ao elevado consumo de sacarose; segundo, determinar o papel da Rho-kinase 1 (ROCK1) na regulação do metabolismo hepático da glucose e dos lípidos; e terceiro, estudar o efeito do metilsulfonilmetano (MSM) em doenças metabólicas associadas à obesidade. Na primeira parte deste trabalho de investigação foram utilizados ratos Wistar machos sujeitos a uma dieta rica em sacarose (HS). Tal como esperado, estes animais apresentavam insulino-resistência e hiperinsulinémia. A dieta HS levou ao aumento dos níveis hepáticos de NO e ao decréscimo dos níveis de GSH no fígado. Em jejum, a administração intraportal de GSH e NO, a animais saudáveis promoveu um aumento significativo da sensibilidade à insulina. Também nestes animais, a administração intravenosa de S-nitrosotióis, compostos orgânicos que contém um grupo nitroso acoplado a um átomo de enxofre de um tiol, promoveu o aumento significativo da sensibilidade à insulina. Pelo contrário, em animais sujeitos à dieta HS, as doses padrão de GSH + NO e de S-nitrosotióis não conseguiram promover o aumento da sensibilidade à insulina. No entanto, ao aumentar a dose de S-nitrosotióis administrados por via intravenosa, foi possível observar o aumento da sensibilidade à insulina dependente da dose, indicando um possível papel dos S-nitrosotióis como sensibilizadores de insulina. O estudo detalhado do papel dos S-nitrosotióis na via de sinalização da insulina revelou que há um aumento da fosforilação do receptor da insulina (IR) e da proteína cinase B (Akt), sugerindo um efeito dos S-nitrosotióis nesta via de sinalização. Os resultados apresentados nesta primeira parte sugerem que os S-nitrosotióis promovem a correcta acção da insulina, podendo vir a ser importantes alvos terapêuticos. Na segunda parte deste trabalho de investigação utilizámos murganhos, com uma delecção específica da ROCK1 no fígado, e sujeitos a uma dieta rica em lípidos (HFD). Foi possível concluir que a ausência da ROCK1 no fígado previne a obesidade, melhora a sensibilidade à insulina e protege contra a esteatose hepática. A ausência de ROCK1 no fígado levou a um decréscimo significativo da expressão génica de genes associados à lipogénese, com uma diminuição acentuada do fluxo metabólico associado a esta via. Pelo contrário, a sobreexpressão de ROCK1, exclusivamente no fígado, promove a insulino-resistência e a esteatose hepática no contexto de obesidade induzida pela dieta. Para além disto, a delecção da ROCK1 no fígado de animais obesos e diabéticos, os murganhos deficientes em leptina, corroborou os dados obtidos no primeiro modelo animal, com a franca melhoria da hiperglicémia, hiperinsulinémia e esteatose hepática. Os dados que compõem esta parte do trabalho de investigação sugerem que a ROCK1 tem um papel crucial na regulação do metabolismo lipídico. Na terceira e última parte deste trabalho de investigação foi investigado o efeito do composto metilsulfunilmetano (MSM), um composto organosulfúrico naturalmente presente em plantas e utilizado também como suplemento dietético, em murganhos obesos e insulino-resistentes, por exposição a uma dieta rica em lípidos (DIO). O tratamento com MSM melhorou a insulino-resistência e protegeu contra a esteatose hepática. O conteúdo hepático em triglicéridos e colesterol também diminuíu de forma significativa nos animais DIO sujeitos ao tratamento com MSM, bem como a expressão génica associada à lipogénese. Para além disto, o tratamento com MSM levou a uma diminuição da expressão génica associada à inflamação. De realçar que o tratamento com MSM levou a uma melhoria do perfil hematopoiético destes animais, tanto na medula óssea como no sangue. Para comprovar o efeito benéfico do MSM na obesidade e insulino-resistência utilizámos murganhos deficientes no receptor da leptina, e por isso obesos e diabéticos, tendo observado um perfil semelhante ao obtido para murganhos sujeitos a uma dieta rica em lípidos e tratados com MSM. Concluímos, através dos dados recolhidos, que o MSM como suplemento pode ter efeitos benéficos na hiperinsulinémia, insulino-resistência e inflamação que caracterizam a diabetes tipo 2. Em resumo, os dados obtidos neste trabalho de investigação mostram que os S-nitrosotióis podem ter um papel importante como sensibilizadores da insulina, promovendo um aumento da sensibilidade à insulina num contexto de dietas ricas em sacarose. Para além disto, estudos in vitro, sugerem que os S-nitrosotióis regulam, especificamente, a via de sinalização da insulina. Este trabalho teve também como objectivo o estudo da ROCK1 como regulador do metabolismo da glucose e dos lípidos no fígado. Através do estudo de animais com uma delecção ou uma sobreexpressão da ROCK1 no fígado mostrou-se que esta tem um papel crucial na patogénese da obesidade e diabetes tipo 2, especificamente através do controlo da lipogénese de novo. Finalmente, foi também objectivo deste trabalho, explorar o efeito do MSM em animais DIO e deficientes em leptina. O tratamento com MSM protege de forma evidente contra a obesidade e insulino-resistência, com especial enfâse para a capacidade que esta molécula demonstrou ter na protecção contra a inflamação. Em conjunto os vários estudos aqui apresentados mostram que tanto os S-nitrosotióis como a ROCK1 têm um papel na patogénese da obesidade e diabetes tipo 2 e que a utilização de MSM como suplemento às terapêuticas convencionais pode ter um papel no tratamentos de doenças metabólicas.-------------------------------ABSTRACT: In modern western societies type 2 diabetes and obesity are increasing exponentially, representing a somber public concern. According to the International Diabetes Federation (IDF) ‘Diabetes and Obesity are the biggest public health challenges of the 21st century’. Aside from these the prevalence of nonalcoholic fatty liver disease (NAFLD), among the diabetic and obese population, is as high as 90%. It is now well established that the increase in obesity, diabetes and NAFLD strongly correlates with an increase in fat and sugar intake in our diet, alongside physical inactivity. The pathogenesis of obesity, diabetes and NAFLD has been thoroughly studied but the treatment options available are still narrow. Considering the alarming number in the obese and diabetic population the complete understanding of the pathogenesis, keeping in mind that new therapeutic strategies need to be attained, is of the highest urgency. The liver has been well established as a fundamental organ in regulating whole-body homeostasis. In the fed state the liver converts the glucose into glycogen and lipids. Conversely, in the fasted state, glucose will be produced in the liver. Neuronal and hormonal systems, as well as the hepatic metabolic states, tightly control the fast to fed switch in metabolic fuels. Insulin has a central role in controlling hepatic energy metabolism, by suppressing glucose production and ketogenesis, while stimulating glycolysis and lipogenesis. Liver energy metabolism is also regulated by various transcription factors and coregulators that are, in turn, regulated by insulin, glucagon and other metabolic hormones. Together, these regulators will act to control gluconeogenesis, β-oxidation and lipogenesis in the liver. Aside from the well-established regulators of liver energy metabolism new molecules are being studied has having a role in regulating hepatic metabolism. Any imbalance in the liver energy metabolism is a major contributor to insulin resistance, NAFLD and type 2 diabetes. The overall goal of this research work was to contribute to the understanding of the pathogenesis of diabetes and obesity, on a setting of high-sucrose and high-fat diets, and to explore potential therapeutic options. The specific aims were: first, to determine if treatment with glutathione (GSH) and nitric oxide (NO) was sufficient to ameliorate insulin resistance induced by high-sucrose feeding; second, to determine the physiological role of rho-kinase 1 (ROCK1) in regulating hepatic and lipid metabolism; and third, to study the effect of methylsulfonylmethane (MSM) on obesity-linked metabolic disorders. In the first part of this research work we used male Wistar rats fed a high-sucrose (HS) diet. As expected, rats fed a HS diet were insulin resistant and hyperinsulinemic. HS feeding increased hepatic levels of NO, while decreasing GSH. In fasted healthy animals administration of both GSH and NO, to the liver, was able to increase insulin sensitivity. Intravenous administration of S-nitrosothiols, organic compounds containing a nitroso group attached to the sulfur atom of a thiol, in fasted control animals also increased insulin sensitivity. Under HS feeding the standard doses of GSH + NO and S-nitrosothiols were unable to promote an increase in insulin sensitivity. However, the intravenous administration of increasing concentrations of S-nitrosothiols was able to restore insulin sensitivity, suggesting that S-nitrosothiols have an insulin sensitizing effect. Investigation of the effect of S-nitrosothiols on the insulin signaling pathway showed increased phosphorylation of the insulin receptor (IR) and protein kinase B (Akt), suggesting that S-nitrosothiols may have an effect on the insulin signaling pathway. Together, these data showed that S-nitrosothiols promote normal insulin action, suggesting that they may act as potential pharmacological tools. In the second part of this research work we used liver-specific ROCK1 knockout mice fed a high-fat (HF) diet. Liver-specific deletion of ROCK1 prevented obesity, improved insulin sensitivity and protected against hepatic steatosis. Deficiency of ROCK1 in the liver caused a significant decrease in the gene expression of lipogenesis associated gene, ultimately leading to decreased lipogenesis. Contrariwise, ROCK1 overexpression in the liver promoted insulin resistance and hepatic steatosis in diet-induced obesity. Furthermore, liver-specific deletion of ROCK1 in obese and diabetic mice, the leptin-deficient mice, improved the typical hyperglycemia, hyperinsulinemia and liver steatosis. Together, these data identify ROCK1 as a crucial regulator of lipid metabolism. In the third and final part of this research work we investigated the effect of MSM, an organosulfur compound naturally found in plants and used as a dietary supplement, on diet-induced obese (DIO) and insulin resistant mice. MSM treatment ameliorated insulin resistance and protected against hepatosteatosis. Hepatic content in triglycerides and cholesterol was significantly decreased by MSM treatment, as well as lipogenesis associated gene expression. Furthermore, MSM treated mice had decreased inflammation associated gene expression in the liver. Importantly, FACS analysis showed that MSM treatment rescued the inflammatory hematopoietic phenotype of DIO mice in the bone marrow and the peripheral blood. Moreover, MSM treatment of the obese and diabetic mice, the leptin-deficient mice, resulted in similar effects as the ones observed for DIO mice. Collectively, these data suggest that MSM supplementation has a beneficial effect on hyperinsulinemia, insulin resistance and inflammation, which are often found in type 2 diabetes. In conclusion, this research work showed that S-nitrosothiols may play a role as insulin sensitizers, restoring insulin sensitivity in a setting of high-sucrose induced insulin resistance. Furthermore, in vitro studies suggest that S-nitrosothiols specifically regulate the insulin signaling pathway. This research work also investigated the role of hepatic ROCK1 in regulation of glucose and lipid metabolism. Using liver-specific ROCK 1 knockout and ROCK1 overexpressing mice it was shown that ROCK1 plays a role in the pathogenesis of obesity and type 2 diabetes, specifically through regulation of the de novo lipogenesis pathway. Finally, this research work aimed to explore the effect of MSM in DIO and leptin receptor-deficient mice. MSM strongly protects against obesity and insulin resistance, moreover showed a robust ability to decrease inflammation. Together, the individual studies that compose this dissertation showed that S-nitrosothiols and ROCK1 play a role in the pathogenesis of obesity and type 2 diabetes and that MSM supplementation may have a role in the treatment of metabolic disorders.