Signal transduction pathways involving the hypertension-related WNK1 and WNK4 protein kinases

Dissertação apresentada para obtenção do Grau de Doutor em Biologia, na especialidade de Genética Molecular, pela Universidade Nova de Lisboa, Faculdade de Ciências e Tecnologia

Sintomas somatoformes em Medicina Geral e Familiar

RESUMO: Os doentes que vão à consulta com sintomas físicos para os quais o médico não encontra uma causa orgânica, são frequentes na Medicina Geral e Familiar, embora não sejam específicos, e são o objecto de estudo do presente trabalho. Não deixar uma doença por diagnosticar (erro de tipo II) sem contudo rotular pessoas saudáveis como doentes (erro de tipo I) é um dos mais difíceis problemas da prática clínica diária e para o qual não existe uma orientação infalível e não é previsível que alguma vez venha a existir. Mas se o diagnóstico de doença ou não-doença é difícil, o tratamento dos que não tem doença, embora com sofrimento, também não é mais fácil, sobretudo, se estivermos conscientes do sofrimento que determina a medicalização e a iatrogenia. O presente trabalho está estruturado em 3 partes. Na primeira parte descrevemos a nossa visão integrada do que apreendemos da leitura da literatura publicada e à qual tivemos acesso. À semelhança do que se verifica na maioria das áreas da Medicina esta é também uma em que o conhecimento cresce a ritmo exponencial. No entanto, à falta de conceitos precisos e de definições consensuais sucede um conhecimento, por vezes, pouco consistente, tanto mais que estamos na fronteira entre a cultura leiga e a cultura erudita médica em que os significados devem, a todo o momento, ser validados. Fizemos uma revisão sobre as definições do que está em questão, sobre o que se sabe sobre a frequência dos sintomas físicos na população, quantos recorrem aos serviços de saúde e o que lhes é feito. Passámos por uma revisão da fisiologia destes sintomas e algumas explicações fisiopatológicas para terminarmos sobre o que os doentes pensam sobre os seus sintomas e os cuidados que recebem e o que os profissionais pensam sobre estes doentes. Esta parte termina com uma revisão das propostas de abordagem para este tipo de doentes. Na segunda parte, descrevemos os estudos empíricos focados no problema dos pacientes com sintomas físicos mas sem evidência de doença orgânica. Começa por uma apresentação dos aspectos processuais e metodológicos dos estudos realizados, mais especificamente, de dois estudos quantitativos e um qualitativo. No primeiro estudo pretendeu-se avaliar quais são os sintomas físicos e a sua frequência na população em geral e a frequência de pacientes que procuram (ou não) os serviços de saúde tendo como motivo este tipo de sintomas. O objectivo deste estudo é contribuir para a demonstração que este tipo de sintomas faz parte da vida do dia-a-dia e que, na maioria das vezes, só por si não significa doença, sem contudo negar que representa sofrimento, por vezes até maior do que quando há patologia orgânica. Se no primeiro estudo era demonstrar que os sintomas físicos são frequentes na população, no segundo estudo o objectivo é demonstrar que pacientes com este tipo de sintomas são igualmente frequentes e que o tipo de sintomas apresentados na consulta não difere dos referidos pela população em geral. Pretendia-se ainda saber o que é feito ou proposto pelo médico a estes doentes e se estes doentes traziam ou não, junto com os sintomas, ideias explicativas para os mesmos. Finalmente e não menos importante, é avaliar o grau de fidedignidade do diagnóstico de sintoma somatoforme, chamando assim ao sintoma físico que foi “levado” à consulta e que o médico diagnosticou como não tendo causa orgânica. O terceiro estudo parte do conhecimento adquirido que a Medicina tem muitas respostas para este problema, mas poucas que se possam considerar satisfatórias se usadas isoladamente. Que a maioria das soluções é procurada entre a cultura médica e num paradigma reducionista de separação mente-corpo. Contudo, se o sintoma é “construído” pelo doente, se o principal problema não está no sintoma mas na forma como o paciente o vê, então pareceu-nos lógico que a solução também tem que passar por integrarmos no plano de abordagem o que o doente entende ser melhor para si. Nesta sequência, entrevistaram-se alguns doentes cujo diagnóstico de sintomas somatoformes estava demonstrado pelo teste do tempo. Por isso, entrevistaram-se doentes que já tinham ido à consulta de MGF há mais de 6 meses por sintomas somatoformes e, na data da entrevista, o diagnóstico se mantinha inalterado, independentemente da sua evolução. As entrevistas visaram conhecer as ideias dos doentes sobre o que as motivou a procurarem a consulta, o que pensavam da forma como foram cuidados e que ideias tinham sobre o que os profissionais de saúde devem fazer para os ajudar a restabelecer o equilíbrio com o seu ambiente evitando a medicalização, a iatrogenia e a evolução para a cronicidade. Na terceira parte, discutem-se e integram-se os resultados encontrados no conhecimento previamente existente. Tenta-se teorizar, fazer doutrina sobre o tema e contribuir para abordagens terapêuticas mais personalizadas, abrangentes, variadas e multimodais, baseadas sempre no método clínico centrado no paciente, ou de modo menos correcto mas enfático, baseadas no método centrado na relação. Apresentam-se algumas hipóteses de trabalhos futuros sobre o tema e, sobretudo, esperamos ter contribuído para o reconhecimento da necessidade de a comunicação médico-doente ser uma aprendizagem transversal a todos os profissionais de saúde e ao longo da vida, com a ideia que é sempre possível fazer melhor, caso contrário tenderemos, inexoravelmente, a fazer cada vez pior.-----------ABSTRACT: Patients who go to consultation with physical symptoms, for which the doctor does not find an organic cause, are the subject of the present study. They are common in family medicine, although not specific. Do not let an undiagnosed disease (type II error), but without labeling healthy people as patients with disease (type I error) is one of the most difficult problems in clinical practice and for which doesn’t exist an infallible guide and it is unlikely that any since coming into existence. But, if the diagnosis of disease or non-disease is difficult, the treatment of those who do not have the disease, though suffering, it is not easy, especially if we are aware of the suffering that medicalization and iatrogenic determines. This work is structured in three parts. In the first part we describe our integrated view of what we grasp from reading the published literature and to which we had access. Similar to that found in most areas of medicine, this is also one in which knowledge grows exponentially. However, the absence of precise concepts and consensual definitions determines an inconsistent knowledge, especially because we're on the border between secular culture and medical culture where, at all times, the meaning must be validated. We did a review on the definitions of what is at issue, what is known about the frequency of physical symptoms in the population, how many use the services of health and what they receive as care. We went through a review of the physiology of these symptoms and some pathophysiological explanations, to finish on what patients think about their symptoms and how they perceived the care they received and, finally, what professionals think about these patients. This part ends with a review of the approaches proposed for such patients. In the second part, we describe the empirical studies focused on the problem of patients with physical symptoms but no evidence of organic disease. Begins with a presentation of the procedural and methodological aspects of studies, more specifically, two quantitative and one qualitative. The first study sought to assess which are the physical symptoms, their incidence in the general population and the frequency they seek (or not) health services on behalf of those symptoms. The aim behind this study was to contribute to the demonstration that this type of symptoms is part of life's day-to-day and that, in most cases, does not represent disease by itself, without denying that they represent suffering, sometimes even greater than when there are organic disease. The first study endeavor to demonstrate that the physical symptoms are common in the population. The second study aspires to demonstrate that patients with such symptoms are also common and that the type of symptoms presented in the consultation does not differ from those in the general population. The aim was also to know what is done or proposed by the physician for these patients and if these patients brought or not, along with the symptoms, explanatory ideas for them. Finally and not least, it would try to assess the degree of reliability of diagnosis of somatoform symptoms, thus drawing the physical symptom that patient presents in the consultation and that the doctor diagnosed as having no organic cause. The third study starts from the acquired knowledge that medicine has many answers to this problem, but few can be considered satisfactory if used in isolation. The most solutions are sought in the medical culture and based on a reductionist paradigm of mind-body. However, if the symptom is "built" by the patient, if the main problem is not the symptom but the way the patient sees it, then it seemed logical to us that the solution must integrate the approaches that patients believes are best for them. Subsequently, a few patients, whose diagnosis of somatoform symptoms was demonstrated by the test of time, were interviewed. Therefore, patients who were interviewed had gone to the consultation of family medicine more than 6 months before for somatoform symptoms and. at the moment of the interview, the diagnosis remained unchanged, regardless of their evolution. The interviews aimed to ascertain the patients' ideas about what motivated them to seek consultation, what they thought about the care they got and which ideas they have about what health professionals should do to help these patients to re-establish equilibrium with its environment avoiding medicalization, iatrogenic effects and the evolution to chronicity. In the third section, we discuss and integrate the results found in previously existing knowledge. Attempts to theorize on the subject and contribute to more personalized treatment, comprehensive, varied and multi-modal approaches, always based on patient-centered clinical method, with emphasis on the relationship. We presents some hypotheses for future work on the subject and,above all, defend the recognition of the importance of lifelong learning communication skills for all health professionals, with the idea that we can always do better, otherwise we tend inexorably to do worse.

Biodemes and zymodemes of Trypanosoma cruzi strains: correlations with clinical data and experimental pathology

With the objective of establishing biological and biochemical characteristics of a significant number of Trypanosoma cruzi strains from different geographical areas, 138 strains isolated from naturally infected humans, triatomine or vertebrate hosts were studied; 120 were isolated from different areas of Brazil and 18 from other South and Central American countries. Inocula from triatomine or culture forms were injected into suckling Swiss mice, followed by passages into mice 10 to 12 g. Biological characters and histopathological study permitted the inclusion of the strains into three Types or biodemes: I, II, III. Isoenzymic analysis confirmed a correspondence between the biodemes and zymodemes : Type I and Z2b, Type II and Z2, Type III and Z1. Results showed the ubiquitary distribution of the several types of strains. The predominance of the same Type and zymodeme in one geographical area was confirmed : Type II strains among the human cases from eastern Bahia and east of Goiás; Type III strains from humans of north Brazil and Central America and from silvatic vectors or vertebrates from other geographical areas. The biological types of strains correlate with different histopathological lesions considering cardiac involvement and neuronal lesions. These findings suggest that the biological behavior together with isoenzymes patterns and pathological pictures in the vertebrate host can be an important tool for establishing correlations between strains behavior and clinico-pathological manifestations of Chagas' disease in different geographical areas.

Detection of HTLV-IIa in blood donors in an urban area of the Amazon Region of Brazil (Belém, PA)

The human lymphotropic viruses type I (HTLV-I) and type II (HTLV-II) are members of a group of mammalian retroviruses with similar biological properties, and blood transfusion is an important route of transmission. HTLV-I is endemic in a number of different geographical areas and is associated with several clinical disorders. HTLV-II is endemic in several Indian groups of the Americas and intravenous drug abusers in North and South America, Europe and Southeast Asia. During the year of 1995, all blood donors tested positive to HTLV-I/II in the State Blood Bank (HEMOPA), were directed to a physician and to the Virus Laboratory at the Universidade Federal do Pará for counselling and laboratory diagnosis confirmation. Thirty-five sera were tested by an enzyme immune assay, and a Western blot that discriminates HTLV-I and HTLV-II infection. Two HTLV-II positive samples were submitted to PCR analysis of pX and env genomic region, and confirmed to be of subtype IIa. This is the first detection in Belém of the presence of HTLV-IIa infection among blood donors. This result emphasizes that HTLV-II is also present in urban areas of the Amazon region of Brazil and highlights the need to include screening tests that are capable to detect antibodies for both types of HTLV.

Nevirapine in an animal model of pre-diabetes: study of drug pharmacokinetic and its effects on fasting glycemia and insulin resistance

Dissertação para obtenção do Grau de Mestre em Biotecnologia

Energy-efficient diversity combining for different access schemes in a multi-path dispersive channel

Dissertação para obtenção do Grau de Doutor em Engenharia Electrotécnica e Computadores

Reinfections with strains of Trypanosoma cruzi, of different biodemes as a factor of aggravation of myocarditis and myositis in mice

Reinfections with Trypanosoma cruzi in patients from endemic areas have been claimed to be an aggravation factor of cardiac manifestations in Chagas' disease. In the present study, the influence of triple infections with strains of different biodemes, on cardiac and skeletal muscle lesions was experimentally tested. Fifty eight mice chronically infected with the Colombian strain (Biodeme Type III) were successively reinfected as follows: 1st group - reinfected with 21 SF strain (Type II) followed by Y strain (Type I ); 2nd - group reinfections with Y strain followed by 21SF strain. Isoenzyme analysis of parasites from hemocultures obtained from triple infected mice, revealed the patterns of three distinct zymodemes in the same animal. Each Trypanosoma cruzi strain was reisolated after four passages in mice on either the 7th, 14th or 30th day after inoculation with the blood of triple infected mice. Histopathology results demonstrated a significant exacerbation of cardiac and skeletal muscle inflammatory lesions, confirmed by morphometric evaluation, in mice with triple infection. No aggravation of parasitism was detected. The possibility of an enhancement of cellular response in the triple infected mice is suggested.

Key performance indicators and the balanced scorecard at the Luxury Brand Hotels of Pestana Group

Traditional approaches to evaluate performance in hotels, have mainly used financial measures. Building on Speckbacher et al. (2003), this Work Project aims to design and propose a Balanced Scorecard Type II as a performance measurement/management system for the hospitality industry based on data collected at the Luxury Brand Hotels of Pestana Group. The main contribution is to better align the vision, strategy and financial and non-financial performance measures in this category of hotels, in particular those of Pestana Group, and by doing so, lead their managers to focus on what is really critical and, consequently improve the overall performance.

In vitro chondrogenic commitment of human Wharton's jelly stem cells by co-culture with human articular chondrocytes.

Wharton's jelly stem cells (WJSCs) are a potential source of transplantable stem cells in cartilage-regenerative strategies, due to their highly proliferative and multilineage differentiation capacity. We hypothesized that a non-direct co-culture system with human articular chondrocytes (hACs) could enhance the potential chondrogenic phenotype of hWJSCs during the expansion phase compared to those expanded in monoculture conditions. Primary hWJSCs were cultured in the bottom of a multiwell plate separated by a porous transwell membrane insert seeded with hACs. No statistically significant differences in hWJSCs duplication number were observed under either of the culture conditions during the expansion phase. hWJSCs under co-culture conditions show upregulations of collagen type I and II, COMP, TGFβ1 and aggrecan, as well as of the main cartilage transcription factor, SOX9, when compared to those cultured in the absence of chondrocytes. Chondrogenic differentiation of hWJSCs, previously expanded in co-culture and monoculture conditions, was evaluated for each cellular passage using the micromass culture model. Cells expanded in co-culture showed higher accumulation of glycosaminoglycans (GAGs) compared to cells in monoculture, and immunohistochemistry for localization of collagen type I revealed a strong detection signal when hWJSCs were expanded under monoculture conditions. In contrast, type II collagen was detected when cells were expanded under co-culture conditions, where numerous round-shaped cell clusters were observed. Using a micromass differentiation model, hWJSCs, previously exposed to soluble factors secreted by hACs, were able to express higher levels of chondrogenic genes with deposition of cartilage extracellular matrix components, suggesting their use as an alternative cell source for treating degenerated cartilage.

Aging and Alzheimer's disease: Searching for novel molecular cues

Tese de Doutoramento em Ciências da Saúde

Mechanical fatigue performance of PCL-chondroprogenitor constructs after cell culture under bioreactor mechanical stimulus

In tissue engineering of cartilage, polymeric scaffolds are implanted in the damaged tissue and subjected to repeated compression loading cycles. The possibility of failure due to mechanical fatigue has not been properly addressed in these scaffolds. Nevertheless, the macroporous scaffold is susceptible to failure after repeated loading-unloading cycles. This is related to inherent discontinuities in the material due to the micropore structure of the macro-pore walls that act as stress concentration points. In this work, chondrogenic precursor cells have been seeded in Poly-ε-caprolactone (PCL) scaffolds with fibrin and some were submitted to free swelling culture and others to cyclic loading in a bioreactor. After cell culture, all the samples were analyzed for fatigue behavior under repeated loading-unloading cycles. Moreover, some components of the extracellular matrix (ECM) were identified. No differences were observed between samples undergoing free swelling or bioreactor loading conditions, neither respect to matrix components nor to mechanical performance to fatigue. The ECM did not achieve the desired preponderance of collagen type II over collagen type I which is considered the main characteristic of hyaline cartilage ECM. However, prediction in PCL with ECM constructs was possible up to 600 cycles, an enhanced performance when compared to previous works. PCL after cell culture presents an improved fatigue resistance, despite the fact that the measured elastic modulus at the first cycle was similar to PCL with poly(vinyl alcohol) samples. This finding suggests that fatigue analysis in tissue engineering constructs can provide additional information missed with traditional mechanical measurements.

Relatório de atividade profissional

Relatório de atividade profissional de mestrado em Ciências - Formação Contínua de Professores (área de especialização em Biologia e Geologia)

Autonomic Modulation of the Heart in Systemic Arterial Hypertension

OBJECTIVE: To analyze the heart rate variability in patients with mild to moderate systemic arterial hypertension. METHODS: Thirty-two healthy (group I) and 70 systemic arterial hypertensive (group II) individuals, divided according to age (40 to 59 and 60 to 80 years old, respectively) and with a similar distribution by sex were studied. Thirty-one had left ventricular hypertrophy (LVH), 22 were overweight, and 16 had Type II diabetes mellitus. Smoking, alcohol ingestion, and sedentary habits were the same between groups. Variability in heart rate was analyzed in the time domain, using standard deviations of normal RR intervals (SDNN) and the differences between maximal brady- and tachycardia (D-BTmax) during sustained inspiration. Analysis of the frequency band of the power spectrum between 0.05 and 0.40 Hz at rest and during controlled respiration was chosen for analysis of the frequency domain. RESULTS: In both time and frequency domains, variables were lower in group II than in group I. Within groups, statistically significant variables were only found for individuals in the 40 to 59 year old group. The presence of LVH, overweight, or diabetes mellitus did not influence the variability in heart rate to a significant extent. CONCLUSION: Variability in heart rate was a valuable instrument for analyzing autonomic modulation of the heart in arterial hypertension. The autonomic system undergoes significant losses in cardio-modulatory capacity, more evident in subjects between 40 and 59 years old. In those over 60 years old, reduced variability in heart rate imposed by aging was not significantly influenced by the presence of systemic arterial hypertension.

Modulación de la activación de células dendríticas por antígenos de Fasciola hepatica y ligandos para receptores Toll: Potencial terapéutico en respuestas anti-inflamatorias. Fasciola hepatica antigens and TLR ligands modulate dendritic cells activation: therapeutic potential in anti-inflammatory responses.

Antecedentes: En nuestro laboratorio hemos demostrado que antígenos (Ags) de Fasciola hepatica inducen en células dendríticas murinas (CD), diferentes propiedades tolerogénicas como la incapacidad por si mismos de inducir la maduración de las células, la resistencia a la maduración por ligandos de TLR, el incremento en la producción de IDO y también la capacidad de esta estas células de dirigir la respuesta inmune hacia un perfil Th2 y T reg. Por otra parte ha sido bien documentado que CD con características tolerogénicas, ya sea inmaduras o semimaduras, son útiles para reducir respuestas inflamatorias excesivas tales como las que ocurren en enfermedades autoinmunes. Además hemos demostrado que CD tratadas con Ags del parásito en conjunto con un ligando Toll (CpG-ODN) producen altos niveles de citoquinas anti-inflamatorias (IL-10 y TGF-) bajos de citoquinas proinflamatorias (TNF, IL-6, IL-12). Hipótesis: El fenotipo semimaduro alcanzado en las CDpodría ser utilizado para reducir la inflamación en un modelo de enfermedad autoinmune en donde existe una exacerbada respuesta Th1 y Th17, ya que la producción elevada de IL-10 y TGF- podría inhibir o controlar estas respuestas de manera directa o a través de la inducción de células T regulatorias. Objetivos: En este proyecto nosotros proponemos la inmunización de animales susceptibles (ratones DBA1/j), al desarrollo de artritis inducida por colágeno (AIC) con CD tratadas con Ags de F. hepatica en conjunto con CpG-ODN para reducir los síntomas clínicos de la enfermedad. Materiales a utilizar: En nuestro laboratorio hemos desarrollado un modelo de artritis inducida por colágeno (AIC) mediante dos inmunizaciones de ratones DBA1/j con colágeno tipo II bovino y adyuvante de Freund. El modelo permitió establecer un índice clínico mediante la hinchazón en las patas de los animales. Doce días posteriores a la primera inmunización los animales serán inyectados con CD tratadas con: 1. PBS, 2.Extracto total de F.hepatica (TE) + CII, 3. CpG + CII, 4. TE+CpG+CII Se realizará la observación macroscópica diaria, a partir de los 7 días de la 2a inmunización Luego del sacrificio las articulaciones de las patas se prepararán para realizar un análisis histológico. Se detectará en suero los niveles de anticuerpos IgG1 (perfil Th2) y de IgG2a (perfil Th1) mediante la técnica de ELISA. Se detectará también el perfil de citoquinas en los nódulos drenantes por la técnica de ELISA y adicionalmente la poblaciónes celulares de células T regulatorias (Treg) CD4+CD25+Foxp3 o células Tr1. Resultados esperados: Pensamos que el tratamiento de los animales que desarrollan AIC con CD semimaduras (por el tratamiento con TE y CpG), serán capaces de migrar a los órganos linfaticos y secretar TGF-be(inductora de células T reg), IL-10 (inductoras de células Tr1), IDO inhibitoria de la respuesta de Li T y promotor de células T reg, también podría generarse una respuesta Th2 (por la presencia de antígenos del parásito), y estas respuestas aisladas o en forma sinérgica podrían inhibir las respuestas de tipo Th17 y Th1 asociadas a la patología en esta enfermedad. Importancia del proyecto: En el desarrollo de la artritis existe un aumento de la inmunidad mediada por células, asi como de la respuesta inmune humoral hacia componentes de la matriz del cartílago. El tratamiento convencional de la artritis recae en general en el uso de inmunosupresores no-específicos, los cuales poseen una variedad de efectos adversos y la inhibición de la respuesta inflamatoria no es específica. En este proyecto proponemos el uso de CD tratadas con antígenos del helminto F. hepatica y CpG ligando Tol que capacita a estas células para generar una respuesta adaptativa de tipo regulatoria, útil en la inhibición de las respuestas inflamatorias como la que ocurre durante la progresión de artritis reumatoidea en un modelo experimental en ratones. We have shown that F. hepatica Ags-treated dendritic cells (DC) together with a TLRl ligand (CpG-ODN) produce high levels of anti-inflammatory cytokines (IL-10 and TGF-Beta) and low of proinflammatory cytokines (TNF, IL-6, IL -12). Hypothesis: The semimature phenotype achieved by DC, could be used to reduce inflammation in a model of autoimmune disease. The high production of IL-10 and TGF-Beta by these cells could directly or through the induction of T reg cells inhibit the inflammatory response. Objective: In this project we propose the immunization of DBA1 / j mice, susceptible to the development of collagen-induced arthritis (CIA) with F. hepatica-treated DC in conjunction with CpG-ODN to reduce clinical signs of disease. Materials: In our laboratory, we developed the CIA model by two immunizations of DBA1 / j mice with bovine type II collagen and Freund's adjuvant. The model allowed to stablish a clinical index by swelling in the legs of animals. Twelve days after the first immunization the animals are injected with DC treated with: 1. PBS 2. F.hepatica Extract (TE) + CII, 3. CpG + CII, 4. TE + CpG + CII Macroscopic observation will take place daily from 7 days of the 2nd immunization. After sacrifice the joints of the legs will be prepared for histological analysis. Serum levels of IgG1 antibodies (Th2 profile) and IgG2a (Th1 profile) will be detected by ELISA. It will also detected the cytokine profile in draining lymph nodes by ELISA and additionally the cell populations of regulatory T cells (Treg) CD4 + CD25 + Foxp3 or Tr1 cells. Expected results: We believe that the treatment of animals that had developed CIA with DC will be able to migrate to lymphatic organs and secrete TGF-B (T reg cell-inducing), IL-10 (inducing Tr1 cells), IDO (inhibitory of T cells and inducing of T reg cells) could alone or in synergy inhibit Th17-type responses and Th1 associated with the pathology in this disease.

Association of Peripheral Arterial and Cardiovascular Diseases in Familial Hypercholesterolemia

Background: Familial hypercholesterolemia (FH) is an autosomal dominant genetic disease characterized by an elevation in the serum levels of total cholesterol and of low-density lipoproteins (LDL- c). Known to be closely related to the atherosclerotic process, FH can determine the development of early obstructive lesions in different arterial beds. In this context, FH has also been proposed to be a risk factor for peripheral arterial disease (PAD). Objective: This observational cross-sectional study assessed the association of PAD with other manifestations of cardiovascular disease (CVD), such as coronary artery and cerebrovascular disease, in patients with heterozygous FH. Methods: The diagnosis of PAD was established by ankle-brachial index (ABI) values ≤ 0.90. This study assessed 202 patients (35% of men) with heterozygous FH (90.6% with LDL receptor mutations), mean age of 51 ± 14 years and total cholesterol levels of 342 ± 86 mg /dL. Results: The prevalences of PAD and previous CVD were 17% and 28.2 %, respectively. On multivariate analysis, an independent association between CVD and the diagnosis of PAD was observed (OR = 2.50; 95% CI: 1.004 - 6.230; p = 0.049). Conclusion: Systematic screening for PAD by use of ABI is feasible to assess patients with FH, and it might indicate an increased risk for CVD. However, further studies are required to determine the role of ABI as a tool to assess the cardiovascular risk of those patients.