Background: N-terminal pro-brain natriuretic peptide (NT-proBNP) is secreted by cardiac ventricular myocytes upon pressure and volume overload and is a prognostic marker to monitor the severity of precapillary pulmonary hypertension and the extent of right heart failure. Objectives: The impact of physical exercise on NT-proBNP levels in patients with left heart disease was demonstrated previously. No data regarding patients with isolated right heart failure and the influence of acute exercise on NT-proBNP serum levels exist. Methods: Twenty patients with precapillary pulmonary hypertension were examined. Hemodynamic parameters were measured during right heart catheterization. Serum NT-proBNP of patients was measured at rest, after a 6-min walking test, during ergospirometry and during recovery, all within 7 h. Significant differences in sequential NT-proBNP values, relative changes compared to values at rest and the correlation between NT-proBNP and obtained parameters were assessed. Results: At rest, the mean serum level of NT-proBNP was 1,278 ± 998 pg/ml. The mean level of NT-proBNP at maximal exercise was increased (1,592 ± 1,219 pg/ml), whereas serum levels decreased slightly during recovery (1,518 ± 1,170 pg/ml). The relative increase of serum NT-proBNP during exercise correlated with pulmonary vascular resistance (r = 0.45; p = 0.026) and cardiac output (r = –0.5; p = 0.015). Conclusions: In this study, we demonstrated acute changes in NT-proBNP levels due to physical exercise in a small group of patients with precapillary pulmonary hypertension. Our results also confirm the predominant usefulness of NT-proBNP as an intraindividual parameter of right heart load. / Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG-geförderten) Allianz- bzw. Nationallizenz frei zugänglich.

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

Nos últimos anos, o sistema do peptídeo natriurético do tipo C (CNP) e seu receptor (NPR-B) foi apontado como um importante regulador do processo de ossificação endocondral. Vários estudos em animais evidenciam o seu papel de estímulo à proliferação e diferenciação de condrócitos e secreção de matriz extracelular. Mutações bialélicas com perda de função do gene do NPR-B (NPR2) levam a uma doença denominada displasia acromesomélica do tipo Maroteaux (AMDM), uma displasia esquelética caracterizada por baixa estatura extrema. Observa-se que familiares de pacientes com AMDM carreadores de mutação no NPR2 têm estatura abaixo da média da população a qual pertencem, sugerindo um papel de mutações em heterozigose do NPR2 como causadoras de baixa estatura idiopática (BEI). Os objetivos deste estudo foram avaliar a presença de mutações no gene NPR2 em um grupo de pacientes com BEI e correlacionar os achados moleculares com o fenótipo dos pacientes e familiares. A região codificadora do gene NPR2 foi sequenciada pelo método de Sanger em 60 pacientes com diagnóstico de BEI. Foram identificadas cinco diferentes variantes alélicas missense em heterozigose no NPR2, cada uma em um único paciente. Essas variantes foram submetidas à análise funcional in vitro para avaliação da atividade da guanililciclase e microscopia confocal para localização intracelular dos receptores NPR-B. As variantes c.226T > C / p.Ser76Pro, c.788G > C / p.Arg263Pro e c.2455C > T / p.Arg819Cys segregam com o fenótipo de baixa estatura dentro das famílias e determinam um prejuízo funcional ao NPR-B. As três variantes geram proteínas que exercem efeito dominante negativo e os receptores NPR-B com as mutações p.Ser76Pro e p.Arg263Pro não se localizam na membrana celular. As variantes c.491C > G / p.Ala164Gly e c.1636A > T / p.Asn546Tyr não segregam com o fenótipo de baixa estatura nas famílias e não se evidenciou um efeito dominante negativo. O escore-Z da altura dos indivíduos carreadores das variantes funcionalmente deletérias variou de -4,5 a -1,7. Um dos pacientes e dois familiares apresentam desproporção corporal e um paciente tem metacarpos curtos. Como conclusão, mutações em heterozigose no gene NPR2 são causa de baixa estatura em 3 de 60 pacientes com diagnóstico inicial de BEI (5% da nossa casuística). Os indivíduos afetados têm graus variados de baixa estatura, sem um fenótipo característico / Over the past several years, C-type natriuretic peptide (CNP) and its receptor (NPR-B) system has emerged as an important regulator of endochondral bone growth. Animal models showed a CNP/NPR-B role in promoting chondrocyte proliferation and differentiation and matrix synthesis. Biallelic loss-of-function mutations in NPR-B gene (NPR2) cause acromesomelic dysplasia type Maroteux (AMDM), a skeletal dysplasia with extreme short stature. Relatives of patients with AMDM, heterozygous for NPR2 mutations, were noted to be shorter than expected for their population of origin, suggesting that heterozygous mutations in NPR2 could be a cause of idiopathic short stature (ISS). The objective of this study was to investigate the presence of NPR2 mutations in a group of patients with ISS and to correlate molecular findings with phenotype. The NPR2 coding region was sequenced by Sanger\'s method in 60 patients with ISS. Five different heterozygous missense variants in NPR2 were identified in five patients. The functional consequences of those variants were established using in vitro cell-based assay to determine guanylate cyclase activity and confocal microscopy to determine intracellular localization of NPR-B. The variants c.226T > C / p.Ser76Pro, c.788G > C / p.Arg263Pro and c.2455C > T / p.Arg819Cys segregated with short stature phenotype and were functionally deleterious. NPR-B receptors with these three variants have a dominantnegative effect and p.Ser76Pro and p.Arg263Pro NPR-B were not localized in the cell membrane. Cosegregation analysis of the variants c.491C > G / p.Ala164Gly and c.1636A > T / p.Asn546Tyr was inconclusive and they did not have a dominant negative effect. Carriers of functionally deleterious variants have a height SD score that ranged from -4.5 to -1.7. One of these patients and two relatives have disproportionate short stature and one has shortened metacarpal. In conclusion, heterozygous mutations in NPR2 gene are cause of short stature in 3 of 60 patients initially classified as ISS (5% of our cohort). Affected individuals have variable degrees of short stature without a distinct phenotype

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

Nos últimos anos, o sistema do peptídeo natriurético do tipo C (CNP) e seu receptor (NPR-B) foi apontado como um importante regulador do processo de ossificação endocondral. Vários estudos em animais evidenciam o seu papel de estímulo à proliferação e diferenciação de condrócitos e secreção de matriz extracelular. Mutações bialélicas com perda de função do gene do NPR-B (NPR2) levam a uma doença denominada displasia acromesomélica do tipo Maroteaux (AMDM), uma displasia esquelética caracterizada por baixa estatura extrema. Observa-se que familiares de pacientes com AMDM carreadores de mutação no NPR2 têm estatura abaixo da média da população a qual pertencem, sugerindo um papel de mutações em heterozigose do NPR2 como causadoras de baixa estatura idiopática (BEI). Os objetivos deste estudo foram avaliar a presença de mutações no gene NPR2 em um grupo de pacientes com BEI e correlacionar os achados moleculares com o fenótipo dos pacientes e familiares. A região codificadora do gene NPR2 foi sequenciada pelo método de Sanger em 60 pacientes com diagnóstico de BEI. Foram identificadas cinco diferentes variantes alélicas missense em heterozigose no NPR2, cada uma em um único paciente. Essas variantes foram submetidas à análise funcional in vitro para avaliação da atividade da guanililciclase e microscopia confocal para localização intracelular dos receptores NPR-B. As variantes c.226T > C / p.Ser76Pro, c.788G > C / p.Arg263Pro e c.2455C > T / p.Arg819Cys segregam com o fenótipo de baixa estatura dentro das famílias e determinam um prejuízo funcional ao NPR-B. As três variantes geram proteínas que exercem efeito dominante negativo e os receptores NPR-B com as mutações p.Ser76Pro e p.Arg263Pro não se localizam na membrana celular. As variantes c.491C > G / p.Ala164Gly e c.1636A > T / p.Asn546Tyr não segregam com o fenótipo de baixa estatura nas famílias e não se evidenciou um efeito dominante negativo. O escore-Z da altura dos indivíduos carreadores das variantes funcionalmente deletérias variou de -4,5 a -1,7. Um dos pacientes e dois familiares apresentam desproporção corporal e um paciente tem metacarpos curtos. Como conclusão, mutações em heterozigose no gene NPR2 são causa de baixa estatura em 3 de 60 pacientes com diagnóstico inicial de BEI (5% da nossa casuística). Os indivíduos afetados têm graus variados de baixa estatura, sem um fenótipo característico / Over the past several years, C-type natriuretic peptide (CNP) and its receptor (NPR-B) system has emerged as an important regulator of endochondral bone growth. Animal models showed a CNP/NPR-B role in promoting chondrocyte proliferation and differentiation and matrix synthesis. Biallelic loss-of-function mutations in NPR-B gene (NPR2) cause acromesomelic dysplasia type Maroteux (AMDM), a skeletal dysplasia with extreme short stature. Relatives of patients with AMDM, heterozygous for NPR2 mutations, were noted to be shorter than expected for their population of origin, suggesting that heterozygous mutations in NPR2 could be a cause of idiopathic short stature (ISS). The objective of this study was to investigate the presence of NPR2 mutations in a group of patients with ISS and to correlate molecular findings with phenotype. The NPR2 coding region was sequenced by Sanger\'s method in 60 patients with ISS. Five different heterozygous missense variants in NPR2 were identified in five patients. The functional consequences of those variants were established using in vitro cell-based assay to determine guanylate cyclase activity and confocal microscopy to determine intracellular localization of NPR-B. The variants c.226T > C / p.Ser76Pro, c.788G > C / p.Arg263Pro and c.2455C > T / p.Arg819Cys segregated with short stature phenotype and were functionally deleterious. NPR-B receptors with these three variants have a dominantnegative effect and p.Ser76Pro and p.Arg263Pro NPR-B were not localized in the cell membrane. Cosegregation analysis of the variants c.491C > G / p.Ala164Gly and c.1636A > T / p.Asn546Tyr was inconclusive and they did not have a dominant negative effect. Carriers of functionally deleterious variants have a height SD score that ranged from -4.5 to -1.7. One of these patients and two relatives have disproportionate short stature and one has shortened metacarpal. In conclusion, heterozygous mutations in NPR2 gene are cause of short stature in 3 of 60 patients initially classified as ISS (5% of our cohort). Affected individuals have variable degrees of short stature without a distinct phenotype

L’hypertension artérielle et l’obésité sont deux composantes conjointement reliées du syndrome métabolique. Les récepteurs de l’ANP (GCA) et de l’oxyde nitrique (GCs) ont des propriétés diurétiques, natriurétiques, vasodilatatrices et sont liés au contrôle de la pression. Des études récentes ont démontré leur implication dans l’obésité. Hypothèse : Une différence génétique au niveau du gène GCA pourrait contribuer à des différences physiologiques. La composante lipidique et/ou sodique de la diète pourrait influencer la fonction rénale, cardiaque et les valeurs anthropométriques différemment chez les souches congéniques. Objectifs : (1) Déterminer l’effet de la composante lipidique et sodique des diètes; (2) Évaluer l’influence de GCA sur la réponse physiologique des souches congéniques; (3) Expliquer les mécanismes physiologiques procurant une réduction de la pression artérielle chez la souche SM9. Méthodologie : Des modèles congéniques du rat Dahl (DSS) hypertendu, nourri avec une diète riche en gras (HF) ou normale (NF), ont été utilisés pour démontrer l’impact d’un segment chromosomique d’origine normotendue. Résultats : La souche SM9 a une prise de poids plus importante que SM12 et DSS sur diète HF malgré un apport alimentaire équivalent. La souche SM9 présente également un ratio masse adipeuse/masse maigre plus élevé que SM12 et DSS. Nous n’avons observé aucune augmentation de la pression artérielle en réponse à la diète HF pour les 3 souches malgré une augmentation du dommage rénal pour les 3 souches. Le dommage rénal est plus important chez DSS que pour les 2 congéniques. La réponse diurétique à l’ANP est plus élevée chez SM9 et est influencée par le contenu en sel dand la diète. La perte glomérulaire plus importante chez le rat DSS semble compensée par une augmentation de la réponse à l’ANP par les glomérules résiduels. Il y a une corrélation entre l’activité de GCA en réponse à l’ANP, les niveaux d’ARNm et le nombre de répétition du dinucléotide TA dans son promoteur. Le rat DSS présente une hypertrophie cardiaque plus importante que les deux souches congénique et ceci n’est pas modifié par la diète HF. Conclusion : Nos études ont permis de mettre en évidence un effet génétique impliquant le segment chromosomique normotendu contenant GCA dans la réponse à une diète HF chez le rat DSS. / Hypertension and obesity are two related components of the metabolic syndrome. The ANP receptor (GCA) and nitric oxide receptor (sGC) have diuretic, natriuretic, vasodilatory properties, and are linked to blood pressure control. Furthermore a recent study has demonstrated the implication of GCA and sGC in the development of obesity. Hypothesis: A genetic difference in GCA gene could contribute to physiological differences. The differencial lipid and/or sodium composition of the diet could influence the renal, cardiac and anthropometric values. Objectives: (1) To determine the effect of fat and sodium on the physiological parameters; (2) To evaluate the influence of GCA on the physiological response of the congenic rat; (3) To explain the mechanisms of the blood pressure reduction in SM9 rats. Methodology: Congenic model of DSS rat, fed with either high fat (HF) or normal (NF) diet, were used to demonstrate the impact of a chromosome segment from normotensive origin on physiological functions. C2SM9 contains GCA and sGC from normotensive origin while C2SM12 harbours only sGC from normotensive origin. Results: HF diet had negative feature on body composition, renal damage, creatinine clearance and inhibited the diuretic/natriuretic effect of ANP. The normotensive segment including GCA and sGC has reduced the blood pressure, improve the renal damage and increased the diuretic/natriuretic capacity of SM9 in response to ANP injection when compared to SM12 and DSS. GCA mRNA and the clearance receptor ratio were reduced in SM9 in the renal cortex and retroperitoneal fat. SM12 and SM9, containing the chromosomal segment that includes sGC, improve their lipid profile compared with DSS. Conclusion: Our results suggested a compensatory increase in the GCA levels for SM12 and DSS that is insufficient to improve their pathophysiologic status as observed in SM9. HF diet increases the metabolic syndrome in those rats.

En dépit de nombreux essais thérapeutiques, les syndromes septiques sont la première cause de mortalité en service de soins intensifs. La population septique étant très hétérogène, une meilleure caractérisation des patients serait essentielle afin de mieux individualiser et cibler les thérapeutiques potentiellement bénéfiques. Une approche multiparamétrique de l’utilisation des biomarqueurs est une alternative qui viserait à appréhender la situation de manière plus globale. Notre travail s’inscrit dans ce contexte au travers de l’étude plus spécifique de la défaillance des systèmes cardio-vasculaire et immunitaire. Au-delà de la confirmation de l’intérêt des biomarqueurs présentement étudiés (prohormones cardio-vasculaires et PD-1) dans la prédiction de la mortalité et du risque d’infections nosocomiales, nos résultats apportent des éléments nouveaux. Nous avons montré que (1) la sur-expression des molécules PD-1 est associée à l’énergie leucocytaire, (2) un même biomarqueur peut apporter une information différente au cours du temps, (3) l’information apportée par l’analyse simultanée de deux biomarqueurs est supérieure à celle de la somme de leurs valeurs individuelles, et (4) l’expression dynamique d’un biomarqueur est meilleure que son expression à un temps donné. Au total, notre travail illustre l’intérêt potentiel d’un panel de biomarqueurs pour mieux appréhender la complexité des états septiques et leur rapide évolution. Il reste néanmoins à développer des outils biostatistiques capables de donner au clinicien une vision globale en temps réel des processus en cours. Cela constituera une étape clé pour mieux stratifier et cibler les prochains essais cliniques dans le domaine. / Septic syndromes remain the leading cause of death in the intensive care units despite numerous clinical trials. Septic patients constitute a very heterogeneous population. Therefore improved characterisation of patients in order to better target and personalize potential new therapeutics is highly desirable. A multiparametric biomarker-based approach could be an attractive alternative to obtain a global view of the pathophysiologic situation. In this context, we worked specifically on cardio-vascular and immune dysfunctions. We first confirmed the predictive value of biomarkers for mortality or nosocomial infections, and showed new elements. We observed that (1) PD-1 overexpression is associated with leukocyte anergy, (2) one biomarker could give different information over time, (3) information provided by the association of two biomarkers is more interesting than the addition of their individual values, and (4) dynamic expression of one biomarker is more informative than its expression at a given time point. Finally, our results illustrate the potential interest of biomarker panels to improve our understanding of the septic syndrome complexity and to reflect their rapid evolution. Consequently, next step will depend on our capacity to develop biostatistic tools that enable clinicians to get, in real time, a global view of the process over time. This key step is likely necessary to decrease the heterogeneity of septic patient population in order to better stratify and target next clinical trials in the field.

<p>Uvod: Pacijenti na hemodijalizi (HD) imaju visoku stopu ukupnog i kardiovaskularnog morbiditeta i mortaliteta. Preko 80% bolesnika na HD ima neki tip kardiovaskularne bolesti. Hipervolemija značajno doprinosi njihovom nastanku, dovodeći do hipertenzije, hipertrofije miokarda leve komore, srčane insuficijencije i nastanka plućnog edema. Procena stanja volemije kod pacijenata na HD najče&scaron;će se vr&scaron;i kliničkim pregledom, uprkos nezadovoljavajućoj specifičnosti i senzitivnosti. Hipervolemija je prisutna kod određenog broja pacijenata, uprkos normotenziji, odsustvu edema i urednom auskultatornom nalazu na plućima. Različite metode se koriste za procenu stanja volemije, svaka sa određenim manama. Upotreba analize bioelektrične impedanse zahteva skupu opremu i potro&scaron;ni materijal, vrednosti B-tipa natriuretskog peptida (BNP) i njegovog N terminalnog propeptida (NT-proBNP) zavise i od stanja volemije i od srčane funkcije, kao i od tipa dijalizne membrane. Ehokardiografija (EHO) i ultrazvuk donje &scaron;uplje vene (UZ ICV) sa određivanjem dijametara u inspirijumu i ekspirijumu (IVCDi i IVCDe) zahteva posebno obučen kadar. Ultrazvuk pluća (UZ pluća) je jednostavna, brza i jeftina metoda za detekciju ekstravaskularne plućne tečnosti (EVLW). EVLW predstavlja onu količinu tečnosti koja se nalazi u plućnom intersticijumu. UZ pluća detektuje EVLW kao UZ artefakte zvane &bdquo;B linije&ldquo;. Količina EVLW zavisna je od pritiska punjenja leve komore i povećava se u stanjima hipervolemije, &scaron;to se na UZ pluća manifestuje kao veći broj detektovanih &bdquo;B linija&ldquo;. Zbir svih &bdquo;B linija&ldquo; detektovanih na definisanim mestima na grudnom ko&scaron;u naziva se &bdquo;skor B linija&ldquo; (BLS) i koristi se za kvantifikaciju EVLW pomoću UZ pluća. Cilj: Uporediti adekvatnost i efikasnost UZ pluća u proceni stanja volemije kod pacijenata na HD u odnosu na standardne tehnike UZ donje &scaron;uplje vene, EHO i BNP-a. Proceniti mogućnost pojednostavljenja protokola UZ pregleda pluća redukcijom broja analiziranih plućnih polja. Materijal i metode: Istraživanje je sprovedeno kao studija preseka od aprila 2016. do juna 2017. godine na 83 pacijenta koji su se nalazili na hroničnom programu HD u Službi za HD Odeljenja za internu medicinu Op&scaron;te bolnice Kikinda. Ispitanicima je prvog dana HD u nedelji neposredno pre HD urađen UZ pluća, UZ IVC, EHO, i uzorkovanje krvi za određivnje vrednosti BNP-a. Potom su pacijenti dijalizirani prema svojim utvrđenim HD protokolima. Neposredno nakon HD ponovljeni su UZ pluća, UZ IVC, EHO, a uzorkovanje krvi za BNP je ponovljeno pre započinjanja sledeće HD u nedelji, da bi se izbegao neposredni postdijalizni skok BNP-a. Za poređenje varijabli kori&scaron;ćeni su T test parova odnosno Vilkoksonov test, a za ispitivanje korelacije Pirsonov odnosno Spirmanov test, u zavisnosti od distribucije varijabli. Razlike između grupa ispitanika ispitivane su pomoću jednofaktorske analize varijanse (ANOVA) za kontinuirane varijable, a za kategorijske je kori&scaron;ćena analiza kontingencijskih tabela. Analiza glavnih komponenata (PCA) je kori&scaron;ćena za procenu mogućnosti redukcije broja ispitivanih plućnih polja. Rezultati: Utvrđena je signifikantna razlika između predijaliznih (pre HD) srednjih vrednosti BLS-a (18,85) i postdijaliznih (post HD) srednjih vrednosti BLS-a (7,30); između srednjih vrednosti BNP-a pre HD (894,89 pg/ml) i post HD (487,74 pg/ml); između srednjih vrednosti IVCDe pre HD (10,45 mm) i post HD (7,85 mm); između srednjih vrednosti IVCDi pre HD (7,20 mm) i post HD (4,41 mm); između srednjih vrednosti indeksa kolapsibilnosti IVC pre HD (32%) i post HD (45%). Utvrđene su i signifikantne razlike između srednjih vrednosti sledećih EHO parametara: dijametar leve pretkomore pre HD (3,78 cm) i post HD (3,53 cm), dijametra leve komore u dijastoli pre HD (5,21 cm) i post HD (4,96 cm), dijametra leve komore u sistoli pre HD (3,69 cm) i post HD (3,43 cm) i zapremine leve pretkomore u sistoli pre HD (60,54 ml) i post HD (52,36 ml). Sve razlike su bile signifikantne na nivou p&lt;0,0001. Dokazana je signifikantna pozitivna korelacija između BLS-a pre HD i BNP-a pre HD (&rho;=0,49, p&lt;0,01) i BNP-a post HD (0,43, p&lt;0,01); BLS-a pre HD i IVCDe pre HD (&rho;=0,29, p&lt;0,01), IVCDi pre HD (&rho;=0,30, p&lt;0,05) i IVCDi post HD (&rho;=0,23, p&lt;0,05) kao i između BLS-a post HD i BNP-a pre HD (&rho;=0,44, p&lt;0,01) i BNP-a post HD (&rho;=0,42, p&lt;0,01), između BLS-a post HD i IVCDe pre HD (&rho;=0,29, p&lt;0,05) IVCDi pre HD (&rho;=0,33, p&lt;0,05) i IVCDi post HD (&rho;=0,23, p&lt;0,05). Utvrđeno je da su bolesnici sa vi&scaron;im BLS-om imali niže vrednosti hemoglobina (p=0,006) i vi&scaron;e vrednosti visoko senzitivnog troponina T (p=0,02), kao i veće dijametre leve komore u sistoli (p=0,04). Pomoću PCA utvrđeno je da je moguća redukcija broja ispitivanih plućnih polja na 4 do 12 plućnih polja, koja bi bila odgovorna za 75,38% odnosno 84,51% varijabilnosti BLS-a. Zaključak: UZ pluća može adekvatno i efikasno da proceni stanje volemije i može se koristiti za ovu svrhu kod pacijenata na hroničnom programu HD. UZ pluća je brz, jednostavan i jeftin pregled koji se može izvoditi u bolesničkoj postelji i koji daje pouzdan podatak o bolesnikovom statusu volemije u realnom vremenu. UZ pluća bez većih te&scaron;koća mogao uključiti u kliničke protokole u svim centrima sa dostupnom opremom. Postoji mogućnost redukcije broja ispitivanih plućnih polja i time pojednostavljenja samog UZ pregleda pluća. UZ pluća može koristiti u proceni srčane funkcije kod pacijenata na HD. Pacijenti koji su procenjeni kao hipervolemični pomoću UZ pluća imaju povećan kardiovaskularni rizik, kao i pacijenti procenjeni kao hipervolemični pomoću vrednosti BNP-a i dijametra D&Scaron;V.</p> / <p>Introduction: Patients on hemodialysis (HD) have a high general morbidity and all-cause mortality, as well as high cardiovascular morbidity and mortality. More than 80% of patients on HD have some cardiovascular disease. Hypervolemia plays a significant role here, contributing to hypertension, left ventricular hypertrophy, heart failure and pulmonary edema. Fluid status assessment in HD is still mostly clinical, despite having low specificity and sensitivity. A number of patients remain hypervolume, despite being normotensive, without edema or bibasilar crackles on lung auscultation. Different methods are used for volume status assessment in HD setting, no method without its flaws. Bioelectric impedance analysis requires expensive equipment and supplies. B type natriuretic peptide (BNP) values, and those of its terminal propeptide (NT-proBNP) depend on volume status, cardiac function as well as type of dialysis membrane used. Echocardiography (ECHO) and ultrasonography of inferior vena cava (IVC US) with measurements of its diameters in inspirum and expirium (IVCDi and IVCDe) require trained medical personnel. Lung ultrasound (LUS) is a simple, fast and inexpensive method for detection of extravascular lung water (EVLW), which is the water contained in the lung interstitium. LUS detects EVLW as ultrasonographic artefacts called &bdquo;B lines&ldquo;. EVLW is dependent on left ventricular filling pressures and is increased in volume overload, manifesting as more &bdquo;B lines&ldquo; on LUS. The sum of all &bdquo;B lines&ldquo; detected on predetermined places on the chest is called &bdquo;B line score&ldquo; (BLS) and is used to quantify EVLW using LUS. Goal: Compare the adequacy and efficacy of LUS in assessment of volume status in patients on HD to other methods (IVC US, ECHO, BNP). Assess the possibility of simplifying LUS by reducing the number of examined lung fields. Materials and methods: A cross-section study was performed from April 2016 to June 2017. on 83 dialysis patients in Dialysis unit of Internal medicine department of General hospital Kikinda. LUS, ECHO, IVC US and blood sampling for BNP were performed on the first dialysis day of the week, just prior to HD. Patients were then dialyzed according to their dialysis protocols. After HD all tests were repeated, except blood sampling for BNP, which was sampled just prior to the next HD session in order to avoid elevated BNP values after HD. Variables were compared using double sample T test or Wilcoxon test. Correlation was assessed using Pearson&rsquo;s or Spearman&rsquo;s test, depending on variable distribution. Differences between groups were tested using one-way analysis of variance for continuous variables and contingency tables for categorical variables. Principal component analysis (PCA) was used to assess the possibility of lung field reduction. Results: There was a significant difference between BLS predialysis (pre HD) (mean 18,85) and BLS postdialysis (post HD) (mean 7,30); between IVCDe pre HD (mean 10,45 mm) and IVCDe post HD (mean 7,85 mm); between IVCDi pre HD (mean 7,20 mm) and IVCDi post HD (mean 4,41 mm); between CCI pre HD (mean 32%) and CCI post HD (mean 45%), between BNP pre HD (mean 894,89 pg/ml) and BNP post HD (mean 487,74 pg/ml). There was also a significant difference between the following ECHO parameters: left atrial diameter pre HD (mean 3,78 cm) and post HD (mean 3,53 cm), left ventricular internal diameter in diastole pre HD (mean 5,21 cm) and post HD (mean 4,96 cm) and left ventricular internal diameter in sistole pre HD (mean 3,69 cm) and post HD (mean 3,43 cm), left atrial volume in sistole pre HD (mean 60,54 ml) and post HD (mean 52,36 ml). All differences were significant at a level of p&lt;0,0001. There was a significant positive correlation between BLS pre HD and BNP pre HD (&rho;=0,49, p&lt;0,01) and BNP post HD (&rho;=0,43, p&lt;0,01); BLS pre HD and IVCDe pre HD (&rho;=0,29, p&lt;0,01) IVCDi pre HD (&rho;=0,30, p&lt;0,05) and IVCDi post HD (&rho;=0,23, p&lt;0,05); between BLS post HD and BNP pre HD (&rho;=0,44, p&lt;0,01) and BNP post HD (&rho;=0,42, p&lt;0,01); between BLS post HD and IVCDe pre HD (&rho;=0,29, p&lt;0,05), IVCDi pre HD (&rho;=0,33, p&lt;0,05) and IVCDi post HD (&rho;=0,23, p&lt;0,05). Subjects with higher BLS had lower hemoglobin levels (p=0,006), higher troponin T levels (p=0,02) and greater left ventricular internal dimensions in sistole (p=0,04). PCA showed that there is a possibility of lung field reduction to 12 lung fields and even down to 4 lung fields, which would account for 84,51% or 75,38% of BLS variability. Conclusion: LUS can be used to adequately and effectively assess volume status in patients on HD. LUS is simple, fast and inexpensive exam with bedside capability, which gives accurate volume status data in real time. The exam can be implemented into dialysis unit protocols without difficulty. There is a possibility of simplifying LUS by reducing the number of examined lung fields. LUS can be used in assessment of cardiac function in patients on HD. Patients rated as hypervolemic by LUS have increased cardiovascular risk, as well as patients rated as hypervolemic by BNP levels or IVC diameters.</p>

HINTERGRUND: Im Zustand der Herzinsuffizienz sind die Plasmaspiegel des atrialen natriuretischen Peptids (ANP) erhöht. ANP wird durch Herzmuskelzellen bei atrialer (und dadurch kardiomyozytärer) Dehnung freigesetzt. Eine Reihe von Plasmafaktoren, welche bei der Herzinsuffizienz aktiviert sind, bewirken ebenfalls eine ANP-Freisetzung durch die Interaktion mit G(alpha-q)-gekoppelten Rezeptoren und der daraus resultierenden Aktivierung der Proteinkinase C (PKC). Ziel der vorliegenden Studie war es, im Vergleich von normalen und insuffizienten Rattenherzen die dehnungsinduzierte ANP-Sekretion sowie die Rolle der PKC bei der basalen ANP-Freisetzung zu analysieren. METHODIK: Durch Anlegen eines infrarenalen aortokavalen Shunts (4 Wochen) wurde eine volumeninduzierte Herzinsuffizienz in Wistar Ratten erzeugt. Die ANP-Freisetzung wurde am isoliert, retrograd perfundierten Herzmodell analysiert. Durch Umstellen auf anterograde Perfusion (Perfusionsdruck: 10mmHg) wurde eine atriale De! hnung induziert. Die Aktivierung der PKC fand durch Zugabe von PMA (Phorbol-12-Myristat-13-Azetat) zum Perfusat im Vergleich zu Vehikel statt. Alle Versuchsreihen wurden mit Herzen von shunt- sowie scheinoperierten Tieren durchgeführt. ERGEBNISSE UND DISKUSSION: Nach Shuntoperation war die ANP-Basalfreisetzung deutlich gesteigert und könnte die erhöhten ANP-Plasmaspiegel bei Patienten mit Herzinsuffizienz erklären. Dagegen wurde durch atriale Dehnung (als adäquaten Freisetzungsreiz am gesunden Herzen) keine weitere Steigerung der ANP-Freisetzung in der Shuntgruppe bewirkt. Die ANP-Freisetzung am insuffizienten Herzen ist im Vergleich zur normalen Herzfunktion somit verändert. Unter PKC-Stimulation sank die Freisetzung von ANP in der Shuntgruppe erheblich ab, wogegen es in der Kontrollgruppe zu keiner signifikanten Änderung der ANP-Sekretion kam. Damit wurde der Verdacht erhärtet, dass die Proteinkinase C in die abweichende Regulation der ANP-Freisetzung bei Herzinsuffizie! nz involviert ist. / BJECTIVE: Plasma levels of atrial natriuretic peptide (ANP) are markedly increased in congestive heart failure. ANP has been shown to be released by cardiomyocytes during atrial (and thereby cardiomyocytical) stretch. Several factors that are activated in heart failure as well enhance ANP release through the interaction with G(alpha-q)-coupled receptors and the consequent activation of proteinkinase C (PKC). The goal of this study was to analyse stretch induced ANP secretion and to investigate the involvement of PKC in the regulation of ANP release in heart failure compared to normal hearts. METHODS: Volume overload induced heart failure was produced by an infrarenal aortocaval shunt (4 weeks) in Wistar rats. ANP release was analysed in an isolated retrogradly perfused heart preparation. Atrial stretch was induced by switching to anterograd perfusion (perfusion pressure: 10mmHg). For PKC activation PMA (phorbol 12 myristate 13 acetate) was added to the perfusate and compar! ed to vehicle treatment. All experiments were performed with shunt and sham operated rats. RESULTS AND CONCLUSIONS: After shunt operation ANP baseline release was considerably augmented and could thus explain elevated ANP plasma levels in heart failure patients. In contrast, atrial stretch as an adequate secretion stimulus in control hearts did not further enhance ANP release in hearts of shunt operated animals. The ANP secretion in heart failure therefore seems to differ from the ANP secretion in the healthy state. Stimulation of PKC markedly decreased ANP release in the shunt group, whereas it did not have any significant effect on ANP release in the control group. In conclusion, the role of the proteinkinase C in ANP release differs between normal and failing hearts and seems to be involved in the deviating regulation of ANP release in states of heart failure.

Introdução: A Organização Mundial da Saúde estimou em 2015 que 23 milhões de pessoas em todo o mundo sofrem de insuficiência cardíaca (IC), com taxas de mortalidade equivalentes às do câncer. Níveis mais elevados de HDL-colesterol têm sido associados com maior sobrevivência na IC. É consensual que as várias funções protetoras da HDL devem ser exploradas além da concentração de HDL-colesterol. Transferência de lípides para HDL, mediada por proteínas de transferência CETP e PLTP, é uma etapa importante no transporte reverso de colesterol e metabolismo de HDL.,Desenvolvemos um ensaio in vitro para avaliar as transferências de lípides para a HDL, mostrando que esse fenômeno é alterado em várias condições, como na doença arterial coronária, no diabetes mellitus e pelo estilo de vida sedentário. Recentemente, tem sido descrito que a HDL transporta pequenos RNAs não codificadores de proteína, os chamados microRNAs (miRNAs). Alguns miRNAs foram descritos como reguladores críticos do metabolismo das lipoproteínas. O objetivo deste estudo foi comparar lípides plasmáticos, transferência lipídica para HDL, perfil inflamatório, miRNAs relacionados ao metabolismo de lipoproteínas obtidos de pacientes com IC e de pacientes sem IC (sem-IC). Métodos: Quarenta e oito pacientes com IC foram avaliados, 25 em classe funcional NYHA I e II (IC-I/II) e 23 em NYHA III e IV (IC-III/IV), bem como 50 pacientes sem-IC pareados por gênero e idade. Todos os pacientes com IC apresentavam uma fração de ejeção <=40%. Foram determinadas as concentrações plasmáticas de CETP, LCAT, LDL oxidada (LDLox) e atividade de paraoxonase 1 (PON-1). Transferências de lípides para a HDL foi avaliada a partir da incubação de uma nanopartícula artificial com plasma total. A expressão de miRNAs circulantes envolvidos no metabolismo das lipoproteínas também foi analisada. Resultados: Os níveis de colesterol total, LDL e HDL e triglicérides não diferiram entre os três grupos. A concentração da apolipoproteína A-I foi menor no grupo IC-I/II em comparação ao grupo sem-IC (125±23 versus 142±19; p < 0,05), enquanto que a concentração da apolipoproteína B foi menor em ICIII/ IV comparado ao sem-IC (81±35 versus 114±40; p < 0,001). A transferência de colesterol esterificado (5,44±1,76 versus 6,26±0,85), fosfolípides (19,05±2,5 versus 20,21±1,45) e de triglicérides (6,29±2,05 versus 7,40±1,47) foi menor no grupo IC-III/IV do que no grupo sem-IC (p < 0,05). No entanto, não houve diferença nas transferências entre IC-I/II e sem-IC. A concentração de LDLox foi menor em ambos os grupos com IC comparados ao sem-IC (p < 0,0001). A massa de CETP foi menor em IC-III/IV do que em IC-I/II (2,77±1,3 versus 3,78±1,3; p=0,021). A concentração de LCAT e a atividade de PON-1 não foram diferentes entre os grupos. A análise da expressão dos miRNAs circulantes miR-33a, miR-144, miR-185, miR-125, miR-758, miR-26a, miR- 106b, miR-122 e miR-30c, mostrou-se significantemente aumentada nos indivíduos com IC em comparação aos indivíduos sem-IC, ao passo que o miR- 10b foi o único encontrado diminuído na IC comparado com indivíduos sem-IC (p=0,007). Conclusão: Em pacientes com IC mais severa e sintomática da IC, o processo de transferência de lípides para a HDL está deficiente, bem como alguns dos mecanismos que o regulam, e possivelmente estas alterações influenciem no transporte reverso do colesterol e nas funções protetoras da HDL desses pacientes / Background: World Health Organization estimated that there were twentythree million subjects worldwide suffering from heart failure (HF) in 2015, with mortality rates equivalent to those of cancer. Higher HDL-cholesterol levels have been associated with longer survival in HF. It is now consensual that the various protective functions of HDL should be explored beyond HDLcholesterol. Transfer of lipids to HDL, mediated by transfer proteins CETP and PLTP, is an important step in reverse cholesterol transport and HDL metabolism. Previously, we developed an in vitro assay to test those lipid transfers and showed that transfer of cholesterol to HDL is altered in several conditions, such as coronary artery disease (CAD), diabetes and sedentary lifestyle. Recently, HDL transports small non-coding RNA molecule, called micro RNAs (miRNAs). Some miRNA are critical regulators of lipoprotein metabolism. The aim of this study was compare plasma lipids, lipid transfers to HDL, inflammatory profile, miRNAs related to plasma lipids from patients with HF with those from patients with without HF (non-HF). Methods: Forty-eight HF patients were studied, 25 with functional class NYHA I and II (HF I/II) and 23 with NYHA III and IV (HF III/IV), as well as 50 non-HF patients matched for gender, age and BMI. All HF had ejection fraction <= 40%. CETP, LCAT, oxidized LDL (oxLDL) and paraoxonase 1 (PON-1) activity were determined. Transfers of lipids from a donor artificial nanoparticle to HDL was determined by an in vitro assay in which the emulsion was incubated with whole plasma. Expression of circulating miRNAs involved in cholesterol metabolism was also analyzed. Results: Total, LDL and HDL cholesterol and triglycerides did not differ among the 3 groups. Apolipoprotein A-I was lower in NYHA I/II group compared to non- HF (125±23 versus 142±19; p < 0.05) and apo B was lower in NYHA III/IV group compared to non-HF (81±35 versus 114±40, p < 0.001). The transfer of esterified cholesterol (5.44±1.76 versus 6.26±0.85), phospholipids (19.05±2.5 versus 20.21±1.45) and of triglycerides (6.29±2.05 versus 7.40±1.47) to HDL was lower in HF-III/IV than in non-HF (p < 0.05), but lipid transfers were not different between HF-I/II and non-HF. oxLDL was lower in both HF groups compared to non-HF (p < 0.0001). CETP mass was lower in HF-III/IV than in HF-I/II (2.77±1.3 versus 3.78±1.3; p=0.021). LCAT and PON-1 activity was not different among the groups. Regarding to miRNA, miR-33a, miR-144, miR-185, miR-125, miR- 758, miR-26a, miR-106b, miR-122 e miR-30c were significantly increased in HF compared to non-HF subjects, whereas miR-10b was the only one found to be decreased in HF compared to non-HF subjects (p=0.007). Conclusion: In patients with the more severe and symptomatic HF, the lipid transfer to HDL is deficient, as well as some mechanisms that regulate it, and possibly these changes influence reverse cholesterol transport and the protective functions of HDL in these patients