Introdução: Pacientes com esquizofrenia geralmente apresentam obesidade e diversos distúrbios metabólicos, como estresse oxidativo e inflamação. Esses pacientes tem expectativa de vida mais baixa e a principal causa de mortalidade são as doenças cardiovasculares Além disso, a função cognitiva é um dos determinantes mais críticos da qualidade de vida nesta patologia. Neste contexto, o resveratrol é um composto polifenólico natural que tem sido associado à diminuição do estresse oxidativo e da inflamação, bem como à ação neuro e cardioprotetora. No entanto, não existem estudos sobre o resveratrol em esquizofrenia. Por isso, o objetivo deste estudo foi determinar a eficácia da suplementação com resveratrol em fatores de risco cardiovasculares, estresse oxidativo e cognição em indivíduos com esquizofrenia. Métodos: Este estudo é um ensaio clínico randomizado que incluiu 19 pacientes com diagnóstico de esquizofrenia (DSM-IV e CID-10) selecionados por conveniência, de acordo com os critérios de inclusão. Os pacientes foram randomizados para receber resveratrol (200mg/dia) ou placebo (200mg/dia) e orientados a utilizar a suplementação durante um mês. Todas as aferições foram realizadas no início e ao final do estudo. Os participantes tiveram suas medidas antropométricas aferidas (peso, altura, circunferência abdominal, percentual de gordura) e responderam verbalmente a um questionário de anamnese clínica e alimentar. Foram coletadas amostras de sangue para avaliação dos parâmetros de marcadores cardiovasculares (citocinas, proteína C-reativa - PRC), de estresse oxidativo (glutationa peroxidase - GPx, thiobarbituric acid reactive species - TBARS) e exames sanguíneos de rotina (perfil lipídico, glicose de jejum). Os pacientes participaram de uma entrevista psicológica para avaliação da cognição (testes Hopkins, Wais-R, Stroop) e de sintomas da doença (teste BPRS). Todos assinaram o Termo de Consentimento Livre e Esclarecido e estavam em tratamento crônico com antipsicótico atípico (apenas clozapina). Resultados: Os participantes apresentaram características antropométricas e bioquímicas homogêneas no início do estudo. Após um mês de suplementação, observamos redução dos triglicerídeos no grupo resveratrol e piora significativa do perfil lipídico no grupo placebo. Em relação aos marcadores de estresse oxidativo, houve redução do TBARS e da PCR no grupo resveratrol e aumento do TBARS e da PCR no grupo placebo. Houve aumento da GPx no grupo resveratrol e redução no grupo placebo. Não houve melhora na performance cognitiva após suplementação com resveratrol. Conclusão: Este estudo traz evidências clínicas de grande importância na relação terapêutico-nutricional do paciente com esquizofrenia. Após um mês de suplementação com resveratrol (200 mg/dia), as variáveis e os marcadores analisados não melhoraram significativamente porém, pudemos observar diversas alterações relevantes na prática clínica. Dosagens do perfil lipídico pioraram no grupo placebo, mas não no grupo resveratrol. Embora não foram encontradas diferenças significativas, podemos assumir que o resveratrol pode prevenir danos no perfil lipídico. Além disso, a suplementação com resveratrol demonstrou uma tendência à diminuição da peroxidação lipídica, estresse oxidativo e inflamação. Não houve melhora na performance cognitiva dos participantes. Estes resultados indicam que o resveratrol merece atenção adicional para cuidados clínicos com esquizofrenia devido ao seu papel na prevenção de comorbidades, risco cardiovascular e estresse oxidativo. Há a necessidade de mais estudos para determinação de dose-efeito, duração do efeito, efeitos a curto e longo prazo. / Background: Patients with schizophrenia are generally obese and have several metabolic disorders, such as increased oxidative stress and inflammation. Additionally, such patients have a lower life expectancy and the main cause of their increased mortality is cardiovascular disease. Cognitive function is one of the most critical determinants of quality of life in this pathology. Resveratrol is a natural polyphenolic compound has been reported to decrease oxidative stress, attenuate inflammation, neuroprotective and cardioprotective action but there are no studies regarding resveratrol on schizophrenia. The objective of this study was to determine the efficacy of resveratrol supplementation on cardiovascular risk factors, oxidative stress and cognition in individuals with schizophrenia. Methods: This is a 1-month randomized, double-blind controlled trial in which 19 men with a diagnosis of schizophrenia were assigned to either a resveratrol supplement group (200 mg/day) or a placebo group (200 mg/day). All measurements were taken on the first and last day (day 1 and day 30) of the 1-month follow-up. Nutritional evaluation was conducted by anthropometric data (weight, height, waist and hip circumference, body fat percentage) and food intake was estimated through a 24-h Recall Survey. Clinical evaluation included thiobarbituric acid-reactive substances (TBARS), glutathione peroxidase (GPx), and Creactive protein (CRP), lipid profile and glucose levels. Trained psychologists with expertise in psychiatric disorders assessed psychopathology severity (BPRS) and neuropsychology performance (Hopkins, Wais-R, Stroop). All participants had been on a stable dose of clozapine (an atypical antipsychotic) for at least 6 months and provided signed informed consent. Results: The resveratrol and placebo groups were similar in baseline characteristics. Triglycerides decreased in the resveratrol group after 1 month of supplementation. In the placebo group, we found a significant worsened in lipid profile. TBARS and CRP show a clear increasing trend in the placebo group and a decreasing trend in the resveratrol group whereas GPx increased in the resveratrol group and decreased in the placebo group. There were no significant improvements in neuropsychology performance and psychopathology severity. Conclusion: We did not observe significant changes after 1- month of resveratrol supplementation (200 mg daily). It was possible to note that the lipid profile in the placebo group worsened and, although no significant differences were found, we can assume that resveratrol might prevent lipid profile damage. Resveratrol supplementation demonstrated a tendency to reduce lipid peroxidation, oxidative stress and inflammation. Resveratrol did not improve neuropsychology performance and psychopathology severity. These findings indicate that resveratrol deserves additional attention for the clinical care of schizophrenia due to its role in comorbidity and cardiovascular disease prevention. Further studies are needed to determine dose-effect, effect duration, short and long term effects.

Introdução: Poucos estudos investigaram o papel da periodontite apical (PA) como fator de risco para doenças cardiovasculares (DCV) e mortalidade associada. Esta tese teve o objetivo de avaliar a associação entre variáveis endodônticas, com ênfase na PA, e DCV, estando dividida em quatro diferentes artigos: 1- uma revisão sistemática e meta-análise investigando se indivíduos com PA apresentam alteração nos níveis séricos de marcadores inflamatórios (MI); 2- um coorte retrospectivo avaliando a associação entre variáveis endodônticas e eventos cardiovasculares (ECV) incidentes, incluindo morte por ECV, em participantes do Estudo Longitudinal de Envelhecimento de Baltimore (BLSA); 3- um estudo transversal verificando se perda dentária e a condição de saúde bucal autorrelatada (SROH) estão associados com a carga aterosclerótica (CAB) em indivíduos do sul do Brasil; 4- um estudo de validação do auto-relato de tratamento endodôntico (SRHET) como método de identificar a presença de tratamento endodôntico (TE) e a presença de PA detectável radiograficamente, em participantes do BLSA. Métodos: No Artigo 1, as bases de dados MEDLINE, Embase, Cochrane e Pubmed foram pesquisadas, sem restrição de idioma. A partir de critérios de inclusão e exclusão, dois revisores avaliaram a qualidade dos estudos pela Escala de Newcastle-Ottawa. A principal variável de desfecho da meta-análise foi o nível sérico de MI em indivíduos com PA versus controles saudáveis ou em indivíduos com PA antes versus após o tratamento da PA. No Artigo 2, 278 indivíduos com exames médicos e odontológicos foram avaliados. Doença periodontal (DP) e perda dentária foram aferidas. O número total de dentes com PA e TE foi apurado através de radiografias panorâmicas. A carga endodôntica (EB) foi calculada a partir da soma de PA e TE. O desfecho ECV incidentes incluiu angina, infarto agudo do miocárdio e morte por ECV. Os participantes foram monitorados por até 44 anos (média=17,4±11,1 anos). Riscos relativos (RR) foram calculados por modelos de regressão de Poisson, estimando a relação entre PA, TE, EB e ECV incidentes. No Artigo 3, 382 indivíduos submetidos a coronarioangiografia foram incluídos. Dados sociodemográficos, fatores de risco cardiovascular e condição de saúde bucal foram coletados por questionário, incluindo dados acerca do SROH. O número de dentes a as medidas antropométricas foram coletadas através de exames clínicos. CAB foi quantificada através do escore de Friesinger (FS). Razões de prevalência (PR) foram calculadas por regressão de Poisson. No Artigo 4, o SRHET foi coletado através de questionário em 247 participantes do BLSA. TE e PA foram determinados através de radiografias panorâmicas. O número total de TE, PA e dentes perdidos foi registrado para cada indivíduo. A validação de SRHET foi determinada com base em ET e PA, separadamente. Acurácia, eficiência, sensibilidade, especificidade, valores preditivos positivos e negativos (+PV, -PV) e razões de verossimilhança positivos e 13 negativos (+LR, -LR) foram calculados por métodos padrão. Resultados: No Artigo 1, dos 531 estudos inicialmente identificados, 20 integraram a análise final. Trinta e um MI foram analisados, sendo IgA, IgM, IgG e CRP os mais comumente investigados. CRP, IL-1, IL-2, IL-6, ADMA, IgA, IgG, e IgM demonstraram níveis elevados em pacientes com PA em comparação com os controles na maioria dos estudos. A meta-análise revelou que: a) os níveis séricos de IgA (P=0,001), IgG (P=0,04), e IgM (P<0,00001) apresentaram-se aumentados em humanos com PA comparados com indivíduos saudáveis; b) os níveis séricos de CRP, IgA, IgE, IgG e IgM não foram estatisticamente diferentes entre pacientes com PA antes e após o tratamento (P>0.05). No Artigo 2, a idade média foi de 55,0±16,8 anos, sendo 51,4% homens. Ao total, 62 participantes (22,3%) desenvolveram ECV. Modelos multivariados, ajustados para idade, gênero, IMC, fumo, hipertensão, dislipidemia, diabetes, DP e perda dentária demonstraram que EB (P=0,035) esteve associada de forma independente com ECV incidentes. No Artigo 3, a idade média foi de 60,3±10,8 anos, com 63,2% de homens. Modelos multivariados, ajustados para idade, sexo, fumo, hipertensão, diabetes e dislipidemia mostraram que a condição ruim de SROH (P=0,03) e perda dentária (P=0,02) foram associados de modo independente com a CAB. No Artigo 4, 229 participantes compuseram as análises de TE e 129 as análise de PA. Os valores de validação do SRHET foram: sensibilidade (TE=0,91; PA=0,78), especificidade (TE=0,89; PA=0,69), +PV (TE=0,82; PA=0,35), -PV (TE=0,95; PA=0,94), +LR (TE=8,39; PA=2,51) e -LR (TE=0,09; PA=0,32). Conclusões: O Artigo 1 mostrou que a evidência disponível é limitada mas consistente, sugerindo que a PA está associada a um aumento nos níveis séricos de CRP, IL-1, IL-2, IL-6, ADMA, IgA, IgG, e IgM em humanos. Estes achados indicam que a PA pode contribuir para uma resposta imune em nível sistêmico, com possível aumento no risco vascular global. Os resultados do Artigo 2 demonstraram que embora as variáveis PA e ET não estiveram associadas com o desenvolvimento de ECV, a variável EB foi um preditor independente da incidência de ECV entre participantes do BLSA. No Artigo 3, perda dentária e uma condição ruim de SROH foram associados de modo independente com a CAB, sugerindo que a associação entre doenças bucais e aterosclerose está presente entre indivíduos do sul do Brasil. O Artigo 4 demonstrou que o SRHET foi um método acurado para identificar indivíduos com TE, mas um preditor fraco da presença de PA entre indivíduos do BLSA. O uso do SRHET como método de predizer a presença de PA em estudos populacionais deve ser vista com cautela. Esta tese identifica a necessidade do desenvolvimento de estudos prospectivos controlados de larga escala que avaliem a possível redução no risco de DCV frente ao tratamento da PA. / Introduction: Few studies have investigated the role of apical periodontitis (AP) as a risk factor for cardiovascular diseases (CVD) and related mortality. This thesis aimed to evaluate the association between apical periodontitis (AP) and cardiovascular diseases (CVD) and was divided in four different articles: 1- a systematic review and meta-analysis investigating evidence to support whether AP can modify systemic levels of inflammatory markers (IM) in humans; 2- a retrospective cohort evaluating whether the presence of AP and endodontic treatment (ET) was associated with incident cardiovascular events (CVE) on participants in the Baltimore Longitudinal Study of Aging (BLSA); 3- a cross-sectional study testing the hypothesis that poor self-reported oral health (SROH) and tooth loss are positively associated with coronary atherosclerotic burden (CAB) in a group of southern Brazilian patients; 4- a study quantifying the validity of self-reported history of endodontic treatment (SRHET), as reported in the BLSA, as a method to identify individuals who experienced ET and individuals who present with AP. Methods: In Article 1, the MEDLINE, Embase, Cochrane and Pubmed databases were searched, with no language restriction. Based on inclusion and exclusion criteria, two reviewers rated the quality of each study based on the Newcastle-Ottawa Scale. The primary outcome variable for meta-analysis was determined by the serum levels of IM in AP subjects versus healthy controls or in AP subjects before versus after treatment intervention. In Article 2, 278 subjects who received complete medical and dental examinations were evaluated. Periodontal disease (PD) and missing teeth were recorded. Total number of AP and ET were determined from panoramic radiographs. Endodontic burden (EB) was calculated as the sum of AP and ET. Main outcome incident CVE included angina, myocardial infarction and cardiovascular-related death. Participants were monitored for up to 44 years (mean=17.4±11.1 years) following the dental examination. Relative Risks (RR) were calculated through Poisson regression models, estimating the relationship between AP, ET, EB and incident CVE. In Article 3, 382 subjects undergoing coronary angiography were included. Socio-demographic characteristics, cardiovascular risk factors and oral health status were collected using a standardized questionnaire, including data on SROH. Number of teeth and anthropometric measures were collected through clinical examinations. CAB at coronary angiography was quantified using the Friesinger score (FS). Prevalence ratios (PR) were calculated with Poisson regression analyses. In Article 4, SRHET was collected through the BLSA questionnaire in 247 participants. Data on ET and AP were determined from panoramic radiographs. The total number of ET, AP and missing teeth were recorded for each individual. Validity of SRHET was determined based on ET and AP, separately. Accuracy, efficiency, sensitivity, specificity, positive and negative predictive values (+PV, -PV) and positive and negative likelihood ratios (+LR, -LR) were 15 calculated according to standard methods. Results: In Article 1, among the 531 initially identified papers, 20 comprised the final analysis. Thirty-one IM were analyzed, with IgA, IgM, IgG and CRP being most commonly investigated. CRP, IL-1, IL-2, IL-6, ADMA, IgA, IgG, and IgM were shown to be increased in patients with AP compared to controls in most studies. Meta-analyses showed that: a) serum levels of IgA (P=0.001), IgG (P=0.04), and IgM (P<0.00001) were increased in humans with AP compared to healthy controls and b) serum levels of CRP, IgA, IgE, IgG and IgM were not significantly different between patients with AP before and after treatment (P>0.05). In Article 2, mean age at dental examination was 55.0±16.8, with 51.4% males. A total of 62 participants (22.3%) developed CVE. Multivariate models, adjusted for covariates age, sex, BMI, smoke, hypertension, dyslipidemia, diabetes, PD and tooth loss showed that EB (P=0.035) was independently associated with incident CVE. In Article 3, mean age was 60.3±10.8 years, with 63.2% males. Multivariate models, adjusted for age, gender, smoking, hypertension, diabetes and dyslipidemia showed that poor SROH (P=0.03) and tooth loss (P=0.02) were independently associated with CAB. In Article 4, 229 participants were available for ET analysis and 129 for AP analysis. The SRHET validity values were: sensitivity (ET=0.915; AP=0.782), specificity (ET=0.891; AP=0.689), +PV (ET=0.824; AP=0.353), -PV (ET=0.949; AP=0.936), +LR (ET=8.394; AP=2.514) and -LR (ET=0.095; AP=0.316). Conclusions: The systematic review showed that available evidence is limited but consistent, suggesting that AP is associated with increased levels of CRP, IL-1, IL-2, IL-6, ADMA, IgA, IgG, and IgM in humans. These findings indicate that AP may contribute to a systemic immune response not confined to the localized lesion, with a possible role in patient's global vascular risk. Results from Article 2 demonstrated that although AP and ET were not individually associated with the development of CVE, EB in mid-life was an independent predictor of CVE among community-resident participants in the BLSA. In Article 3, poor SROH and tooth loss were independently associated with CAB, suggesting that the association between oral diseases and atherosclerosis is also present among southern Brazilian individuals. In Article 4, SRHET was found to be a highly accurate method to predict ET but a weak predictor of the presence of AP among participants in the BLSA. Thus, the use of SRHET as a predictor of AP in large populational studies must be considered with careful. Finally, future large-scale prospective controlled studies are required to evaluate CVD risk reduction with the treatment of AP.

De nombreux facteurs de risque cardiovasculaire comme l’hypercholestérolémie, l’hypertension, et le diabète sont comptés parmi les facteurs de risque modifiables les plus importants pour le déclin cognitif et la démence. L’exposition à ces facteurs de risque au cours de la vie en particulier avant l’âge de 65 ans ainsi que leur agrégation contribuent de manière plus importante au déclin cognitif. Peu d’études se sont intéressées aux mesures composites de risque cardiovasculaire par rapport à la fonction cognitive chez les sujets de moins de 65 ans. L’objectif de cette thèse était d’étudier l’association entre les mesures composites de risque et le déclin cognitif au cours de la phase précocedu vieillissement. Les données de la cohorte Whitehall II dans laquelle les fonctions cognitives ont été mesurées à trois reprises ont été utilisées pour étudier l’association entre le syndrome métabolique et deux scores de risque de Framingham (de maladie cardiovasculaire globale et d’AVC), et la fonction cognitive et le déclin cognitif sur 10 ans. Les scores de risque cardiovasculaire de Framingham ont aussi été comparés avec un score de risque de démence. De toutes les mesures composites de risque étudiées, les scores de risque de Framingham montraient la plus forteassociation avec le déclin cognitif. Un risque cardiovasculaire plus élevé était associé à un déclin plus rapide dans de multiples tests cognitifs dont la fluence verbale, le vocabulaire et la cognition globale. Ces résultats suggèrent d’une part qu’un risque cardiovasculaire plus élevé contribue au déclin cognitif dès la phase précoce de vieillissement et d’autre part que l’estimation du risque cardiovasculaire et son effet sur la fonction cognitive nécessite une approche multifactorielle.L’identification et la réduction de facteurs de risque cardiovasculaire peuvent avoir un impact important sur la réduction du déclin cognitif et de la démence. / Several cardiovascular disease risk factors including, dyslipidemia, high blood pressure, and diabetes have been proposed as important modifiable risk factors for cognitive decline and dementia. These risk factors often co-occur and their aggregation is associated with increased risk of cardiovascular disease and dementia. However, studies of composite measures of cardiovascular disease risk in relation to cognitive outcomes in non-elderly populations are scarce. The aim of this thesis was to examine composite measures of risk in relation to cognition and longitudinal cognitive change amongmiddle-aged adults. Data from the Whitehall II study were used to study the associations between the metabolic syndrome, two Framingham risk scores; the Framingham stroke and general cardiovascular disease risk scores, and cognition, based on three cognitive assessments over 10 years. In addition, these two (cardio)vascular risk scores were compared with the CAIDE dementia risk score. Of all composite measures of risk examined, the two Framingham risk scores were the best predictors of 10-year cognitive decline. Higher cardiovascular risk was associated with faster 10-year decline inmultiple cognitive tests including verbal fluency, vocabulary and global cognition. These results suggest that multiple cardiovascular disease risk factors contribute to cognitive decline starting in midlife and that multi-risk factor models such as cardiovascular risk scores may be better suited to assessing risk of cognitive decline. Early identification and treatment of cardiovascular disease risk factors may offer the possibility of markedly delaying or preventing cognitive decline.

Hjärt-kärlsjukdom är idag en av de största folksjukdomarna i Sverige med högst mortalitet till följd. Flera studier visar att uppföljning av patienter, informationsbehov och stöd till patienter efter utskrivning från sjukhusen till primärvården är otillfredsställande. Det finns endast ett fåtal studier som visar på patienternas egna upplevelser av att leva med hjärtinfarkt. För att kunna förbättra vården till dessa patienter är det viktigt att ta del av patienternas egna upplevelser av att leva med en hjärtsjukdom.Syftet med studien är att beskriva patienters upplevelser av att leva med hjärtsjukdom ur ett patient- och livsvärldsperspektiv. Metoden i studien är kvalitativ intervju. Studien har genomförts med fenomenologisk ansats och utifrån ett livsvärldsperspektiv. Eftersom det är ett sätt att förstå den omgivande världen och ger möjlighet att beskriva informantens levda erfarenhet. För att få tillgång till patienternas livsvärld intervjuades åtta patienter. Resultat tydliggörs med tre huvudkategorier; Förändrad livssituation, Behov av kunskap och utbildning vid hjärtsjukdom och Tankar på återinsjuknande och med fem tillhörande sub-kategorier; Hälsosammare vardagsliv, Att leva som vanligt, Hjärtskola ger tilltro till sin egen förmåga. Min tillgängliga sjuksköterska, Symtomfrihet minskar oro och tankar på döden. / <p>Program: Fristående kurs</p><p>Uppsatsnivå: D</p>

Proprotein convertase subtilisin / kexin type 9 (PCSK9) is a negative regulator of the low-density lipoprotein receptor, and PCSK9 inhibition has become an important cholesterol-lowering therapeutic strategy. PCSK9 also associates with LDL particles, and evidence suggests that the activity of PCSK9 may be regulated by LDL binding. We have investigated the biochemistry of the interaction between PCSK9 and lipoproteins. Through mutagenesis and in-vitro binding assays, we found conserved motifs in the PCSK9 N-terminus that play a role in LDL binding. Through secondary structure studies using circular dichroism and computational modelling, we determined that the N-terminal region of the PCSK9 prodomain undergoes an environment-dependent structural shift that affects the ability of PCSK9 to bind LDL. We also found that the commonly found loss-of-function polymorphism R46L is capable of modulating this structural shift. Importantly, we found a surface-exposed region of the PCSK9 prodomain that maps a cluster of gain-of-function mutations (L108R, S127R, and D129G) that severely disrupt LDL binding. Through gel shift assays and density gradient centrifugation, we observed that PCSK9 shows remodeling-dependent ability to bind different classes of lipoprotein particles in vitro, binding strongly to LDL and IDL but showing barely detectable association to VLDL. Further, in human plasma, we found that lipoprotein-bound populations of PCSK9 shifted in response to differences in lipoprotein profiles between normolipidemic and hypercholesterolemic or hypertriglyceridemic subjects. Overall, elucidation of how lipoproteins regulate PCSK9 activity will reveal new targets for designing cholesterol-lowering therapeutics.

Chronic kidney disease (CKD) is growing global public health problem affecting 1 in 10 adults in developed countries and recognised as an important risk factor for cardiovascular disease (CVD) development. CVD is the main cause of death among CKD patients. Endothelial injury and dysfunction are critical steps in atherosclerosis, a major CVD. Oxidative stress (increased level of reactive oxygen species, ROS) has been associated with CVD development. Intravenous (IV) iron preparations are widely used in the management of CKD mediated anaemia, and have been associated with increased oxidative stress and cellular dysfunction. This study examined the effect of pharmacologically-relevant concentrations of IV Venofer (iron sucrose) or IV Ferinject (Ferric carboxymaltose, FCM) on primary human umbilical vein endothelial cell (HUVEC) activation/damage and on intracellular ROS generation as well as studying the potential mechanisms responsible. Data from TUNEL assay and Annexin V-FITC/PI staining showed that, IV FCM had no effect, but IV iron sucrose increased HUVEC apoptosis at 24hr. IV iron sucrose inhibited cell proliferation and reduced cell viability. Both compounds induced EC activation through sustained activation of p38 MAPK and up-regulation of ICAM-1 and VCAM-1. Additionally, the compounds induced significant increase in total ROS and superoxide anion production, which was attenuated by the anti-oxidant N-acetylcysteine (NAC). P38 MAPK showed up-regulation of pro-apoptotic protein Bax and down-regulation of antiapoptotic Bcl-2 protein in HUVEC treated with IV iron sucrose and p38 inhibition reversed these effects. In summary, these results suggest that IV iron sucrose causes more severe EC injury than IV FCM. However, both IV iron preparations induced intracellular ROS and superoxide anion generation in HUVEC leading to EC activation/dysfunction, providing a potential explanation for vascular damage in CKD patients.

Small heat-shock proteins (sHsps) are part of a broad cellular sys- tem that functions to maintain a stable proteome under stress. They also perform a variety of regulatory roles at physiological conditions. Despite the multitude of sHsp targets, their interactions with partners are not well understood due to highly dynamical structures. In this thesis, I apply a variety of biophysical and structural approaches to examine distinct interactions made by the abundant human sHsps αβ-crystallin and Hsp27. First, I find that αβ-crystallin binds a cardiac-specific domain of the muscle sarcomere protein titin. A cardiomyopathy-causative variant of αβ-crystallin is shown to disrupt this interaction, with demonstrated implications for tissue biomechanics. Next, I investigate the conformation and unfolding behaviour of another sarcomere-associated protein, filamin C, finding support for the hypothesis that it is mechanosensitive. This leads into an interrogation of the interaction between filamin C and Hsp27, which we find is modulated by phosphorylation of Hsp27. This modulation only manifests during filamin C unfolding, pointing toward a protective chaperoning mode against over-extension during mechanical stress. This finding is bolstered by up-regulation and interaction of both proteins in a mouse model of heart failure. I establish a system for similar studies of a third sHsp, cvHsp, which is muscle-specific and implicated in various myopathies but scantly understood at the molecular level compared to αβ-crystallin and Hsp27. Finally, I probe the stoichiometries and kinetics of complexes formed between αβ-crystallin and Hsp27 themselves, which co-assemble into a highly polydisperse ensemble. This involved the development of a high-resolution native mass spectrometry method for disentangling heterogeneous systems. Together these findings add to our understanding of the roles and mechanisms of ATP-independent molecular chaperones.

Approximately 30% of people admitted to ICU in the UK have co-existing cardiovascular disease (CVD), and this may rise as life-expectancy increases. Patients with CVD have impaired compensatory mechanisms to enable maximum oxygen delivery to the tissues in the event of critical illness, which itself increases global oxygen demand, further stressing the heart. This is exacerbated by tachycardia and hypotension, which may relatively reduce blood flow to the coronary arteries, and catecholamines which increase myocardial oxygen demand. The myocardium extracts 75% of the oxygen supplied by the coronary arteries at rest, and atheroma-related flow limitation further compromises myocardial oxygen delivery. However, the diagnosis of acute coronary syndrome in critical illness is not straightforward, due to patient inability to communicate symptoms, non-specific ECG changes, and poorly understood cardiac biomarker troponin elevation. My overall hypothesis is that patients with CVD benefit from increased oxygen delivery to the myocardium during critical illness. A focus is the importance of anaemia. The aims of the studies presented in this thesis are (i) to systematically review the literature regarding blood transfusion thresholds specifically in patients with CVD; (ii) to explore the association between Troponin I (TnI) within 24 hours of ICU admission and hospital mortality (iii) to describe and quantify the dynamics of TnI in patients with CVD during the first ten days after ICU admission; and (iv) to define myocardial infarction in the context of critical illness. I have performed a systematic review and meta-analysis of randomised controlled trials comparing a restrictive with liberal transfusion threshold and that included patients with CVD. In total, 11 trials enrolling patients with CVD (n=3033) were included for meta-analysis (restrictive n=1514, liberal=1519). The pooled risk ratio for the association between a restrictive transfusion threshold and 30 day mortality was 1.15 (95% CI 0.88 to 1.50, p=0.50, I2=14%). The risk of acute coronary syndrome in patients managed with restrictive compared with liberal transfusion was increased (nine trials, risk ratio 1.78, 95% CI 1.18 to 2.70, p=0.0, I2=0%). In contrast to broader literature supporting restrictive thresholds, our systematic review shows that a restrictive transfusion threshold of less than 80g/l may not be safe in patients with co-existing CVD, and highlights the variability in diagnostic definitions of ACS and the potential for ascertainment bias in transfusion trials. I undertook a retrospective cohort study in two independently collected cohorts of general ICU patients who had TnI measured within 24 hours of ICU admission. Importantly, the majority of TnI samples were collected routinely rather than for clinical indications. We used the Abbott ARCHITECT Stat assay (limit of detection 0.01mcg/l. We performed multivariable regression, adjusting for components of the APACHE II model. We derived the risk prediction score from the multivariable model with TnI. TnI was associated with all cause hospital mortality (OR per doubling TnI 1.16, 95% CI 1.13 to 1.20, p < 0.001) which persisted after adjustment for APACHE II model components (OR TnI 1.05, 95% CI 1.01 to 1.09, p=0.003). TnI correlated highly with the Acute Physiological Score component of APACHE II (r=0.39), suggesting that TnI release may be largely explained by acute physiological stress. Addition of TnI to the APACHE II model did not improve the performance of the risk prediction model and we would not advocate the adoption of a routine single troponin sample at admission. I designed, set up, and recruited 279 patients to a prospective cohort study TROPonin I in Cardiovascular patients in CriticAL care (TROPICCAL, UKCRN 19253) in 11 UK centres. The aims were to (i) determine the incidence of Myocardial Injury and Infarction, defined by the Third Universal Definition of Myocardial Infarction; (ii) explore factors associated with Injury and Infarction from multivariable analyses; and (iii) explore the relationship between Injury/Infarction and outcome in unadjusted and adjusted analyses. We recorded baseline characteristics, and took daily hs-TnI for ten days after ICU admission, severity of illness measures and ECGs for 5 days. There was a wide range of peak TnI (med 114ng/l (min 3, Q1 27, Q3 412, max 58820ng/l)) and a high prevalence of myocardial injury on systematic screening: 71% of patients had peak TnI greater than the sex-specific diagnostic threshold ('Injury'), and 24% had peak TnI greater than the sex-specific diagnostic threshold and dynamic changes on ECG consistent with ischaemia ('Infarction'). TnI consistently showed a rise-and-fall pattern consistent with an acute myocardial 'hit' rather than persisting injury, which peaked early during ICU stay. Importantly, only 12 (4.4%) patients were diagnosed with MI by the clinicians looking after the patients. Independent predictors of peak TnI in the preceding 24 hours were SOFA score, dynamic ECG ischaemia, lactate, haemoglobin, and age. The lack of association with CRP (representing systemic inflammation), with stronger association with lactate (representing inadequate perfusion/oxygen supply), Hb and ECG ischaemia support the conjecture that injury results in part from an acute ischaemic hit in this population. Patients with Infarction had similar baseline demographics to patients with Injury, but had higher peak TnI concentrations, and higher hospital and six month mortality (Figure 2). This supports the importance of including systematic assessment of dynamic ECG changes in the myocardial injury 'construct' in ICU. My work has shown an increased risk of ACS in patients with CVD randomised to restrictive transfusion thresholds. TnI elevation is prevalent in general ICU patients, and is independently associated with hospital mortality. A systematic approach to the detection of myocardial injury in critically ill patients with co-existing CVD who are unable to communicate symptoms, can identify a high risk population who have poorer survival than patients with no injury. Markers of ischaemia are more associated with TnI rise than markers of inflammation, supporting the hypothesis that myocardial injury in this population is at least in part due to oxygen supply-demand imbalance 'myocardial infarction'. From this work, I would recommend (i) a more liberal transfusion threshold of at least 80g/l in patients with coexisting CVD; (ii) systematic use of sequential ECGs in ICU to screen for myocardial injury in 'at risk' patients; and (iii) manipulation of physiological parameters such as anaemia, hypotension and tachycardia should be considered for patients with dynamic ECG changes plus troponin increase consistent with Infarction. Future research should include 'precision medicine' trials in the substantial cohort of ICU patients with co-existing CVD to explore whether interventions that increase myocardial oxygen supply and/or treat infarction alter outcomes.

Trata-se de estudo descritivo com abordagem de análise quantitativa que tem por objetivo identificar o perfil de clientes portadores de angina pectoris instável atendidos numa instituição privada, de um município do interior paulista. O referencial teórico utilizado foi o modelo de “Campo de Saúde” de Lalonde que analisa elementos como a biologia humana, meio ambiente, estilo de vida e organização dos serviços de saúde diante da ocorrência de patologias. A amostra constituiu-se de 58 clientes, com idade entre 34 e 88 anos, de ambos os sexos, a maioria aposentados, que reinternaram na Unidade Coronariana da referida instituição, no período de 01 de setembro de 2002 a 31 de março de 2003. A coleta de dados se deu no domicílio dos clientes, através de entrevista semi-estruturada; aplicada pela pesquisadora. Quanto à biologia humana 32,7% clientes apresentaram obesidade; os pais destes tiveram como causa mortis o infarto agudo do miocárdio e angina pectoris com índices de 24,1% e 20,7%, respectivamente; os antecedentes familiares mais freqüentes quanto às patologias foram a ocorrência de infarto agudo do miocárdio em 22,4% dos pais e hipertensão arterial sistêmica em 41,3% dos parentes próximos; 84,4% clientes possuíam hipertensão arterial sistêmica; 46,5% eram hipercolesterolêmicos; 27,5% tiveram doença vascular periférica e 17,2% apresentaram quadro de acidente vascular cerebral; valores de PAS&#8805;140 mmHg e PAD&#8805;90 mmHg, identificados em 71,1% e 55,7% dos clientes, respectivamente; além de valores de glicemia de jejum >110 mg/dl apresentados por 34,5% da amostra. Com relação ao meio ambiente, 55,1% dos clientes possuíam 1º grau incompleto e 8,6% eram analfabetos; 82,7% dos clientes eram casados, 65,5% não exerciam atividade remunerada e tinham renda familiar entre 03 e 06 salários mínimos. No que concerne ao estilo de vida, 100% dos clientes relacionaram a doença com fatores de risco como a hipertensão arterial sistêmica, o estresse, história familiar, dieta inadequada, tabagismo e falta de atividade física; 24,1% referiram o consumo de bebida alcoólica; 55,2% eram ex-fumantes; 37,9% dos clientes realizavam atividade física; 48,3% referiram fatos ocorridos antes da dor anginosa; 55,2% dos clientes relataram alterações de sono; inatividade sexual em 43,1% dos clientes sendo que 88,0% das clientes já estavam na menopausa e apenas 13,6% faziam terapia de reposição hormonal. No tocante ao atendimento de saúde, todos os clientes eram conveniados mas apenas 13,8% dos clientes utilizavam o serviço de medicina preventiva disponibilizado pelo convênio para prevenir doenças; o serviço de saúde pública é referência para a amostra no tocante à aquisição de medicamentos. Os achados confirmaram a interferência dos elementos referenciados pelo modelo de “Campo de Saúde” na ocorrência das patologias e identificaram a necessidade de trabalhar a mudança no estilo de vida dos clientes, através da prevenção dos fatores de risco para as doenças cardiovasculares e promoção da saúde, em geral. / Descriptive study which was carried out in order to identify customer’s profiles with unstable pectoris angina attended in a Sao Paulo’s state’s town. The theoretical model used was the “Lalonde’s Health Field Model”, which analyses elements like human biology, environment, lifestyle, and health services’ organization ahead diseases occurrences. The sample was composed by 58 customers, aged between 34 and 88 year old, masculine and feminine, most of them retired, which were readmitted in a the mentioned Coronary Unity’s institution. The data were collected at the customer’s residences and a semi- structured interview was used. Concerning about human biology, 32.7% male customers presented obesity; the parents died under acute infarct and pectoris angina and the percentual was 24,1% and 20,7%, respectively. The most common relatives’ diseases were related to the parent’s acute infarct (22,4%) and systemic arterial hypertension for the near relatives; 84.4% of the costumers suffered by systemic arterial hypertension; 46.5% had high cholesterol; 27.5% had peripheral vascular diseases and 17.2% demonstrated cerebral vascular accidents. Values like PAS&#8805;140mmHg and PAD&#8805;90 mmHg, identified in 71,1% and 55,7% among the customers, respectively; beyond glycemia’s values over 110mg/d, presented by 34,4% among the costumers. According to the environment, 55,1% customers didn’t study high school and 8,6% were illiterate; 82,7% were married and among them 65.5% didn’t have remunerate activity and their familiar remuneration was between 03 and 06 Brazil’s minimum salary. Related to the life style, 100% customers related the disease with risk factors’diseases as systemic arterial hypertension, stress, family history, inadequate diet, smoking and physical activity’s lack; 24,1% referred alcoholics drinks consume; 55,2% were ex-smokers; 37,9% practiced physical activity; 55.2% related sleeping problems; 43,1% related sexual inactivity and among them 88,0% female costumers already had menopause and only 13.6% were doing hormonal replacement therapy. About health services, all the costumers were had health policies but only 13.8% customers had used preventive medicine. The public health service is a reference in medicines’ acquisition. The data confirm the mentioned health field model elements in the happening of diseases and identify the need of working on changes in the costumers’ life style, through risk factors prevention for the cardiovascular diseases and health promotion.

Estudos recentes sugeriram associação entre doença periodontal e doenças cardiovasculares e respiratórias. O objetivo deste estudo foi investigar a presença de bactérias periodontopatogênicas em placas ateromatosas humanas. Foram incluídos no estudo 24 pacientes de ambos os sexos, 40-60 anos de idade, internados na UTI do Hospital Nossa Senhora da Conceição em Tubarão-SC com indicação para endarterectomia. Após a coleta das placas ateromatosas, procedeu-se a identificação de sete diferentes espécies bacterianas periodontopatógenos por meio da reação em cadeia da polimerase em tempo real (RT-PCR). Para realização da análise estatistica foi verificada a associação entre as duas variáveis qualitativas nominais (presença ou ausência de bácterias X tipo de bactérias) por meio do teste Qui-quadrado, com nível designificância de 5 %. Os resultados desse trabalho mostraram haver associação entre presença ou ausência de bactérias e dos diferentes tipos de bactérias nos ateromas analisados (p = 0001). Dos ateromas avaliados 75% apresentaram Porphyromonas gingivalis., 58,3% Treponema denticola., 37,5% apresentaram Fusobacteriumnucleatum., 33,3% Prevotella intermedia., 20,8% Agreggatibacter actinomycetencomitanse. 0,4% de Prevotella nigrescens. Esses achados permitem concluir que bactérias periodontopatogênicas apresentam a capacidade de invadir a parede das artérias, podendo ter papel significativo no desenvolvimento ou ruptura das placas ateroescleróticas. / Recent studies suggested an association between periodontal disease and cardiovascular and respiratory diseases. The aim of this study was to investigate the presence of periodontopathogens in human atherosclerotic plaque. It were included in this study 24 non-smoking patients, both genders, 40-60 years, admitted at intensive unit care with indication of endarterectomy. After collection of carotid samples containing the lesions, the identification of seven different bacterial species was performed by real time polymerase chain reaction (RT-PCR). For statistical analysis, the association between two nominal qualitative variables (presence or absence of bactérias vs. bacterial specieis) was verified by Chi-squared test, with a 5% significance level. The results obtained showed an association between the presence or absence and bacterial species found in atheroma (p= .0001). Among the atheroma examined, 75% showed the presence of Porphyromonas gingivalis, 58.3% showed Treponema denticola, 37.5% showed Fusobacteriumnucleatum, 33.3% showed Prevotella intermedia, 20.8% Agreggatibacter actinomycetencomitans and 0.4% Prevotella nigrescens. These findings suggest that periodontopatoghens are capable of invading artery wall and could play a significant role in the development or rupture of atherosclerotic lesions.