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1.
Cardiovasc Pathol ; 48: 107224, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32480283

RESUMO

OBJECTIVE: We used automated image analysis software to determine the proportion of collagen, fat, and myocytes across six histological regions of normal ventricular myocardium. METHODS: Twenty-nine non-cardiac death cases referred to our national cardiac pathology center were included in this study. Whole hearts were macroscopically and microscopically normal following expert histopathological evaluation. Tissue sections from the right ventricular outflow tract, right ventricle (RV), anterior interventricular septum (IVS), posterior IVS, anterior left ventricle (LV), and posterior LV were stained with Picrosirius red for collagen and scanned using a high-resolution slide scanner. Quantification of collagen, fat, and myocyte proportions was performed using Visiopharm software after automated exclusion of perivascular collagen. RESULTS: The majority of decedents were male (25/29; 86%) with a mean age at death of 32.1 ± 9.9 (range 18-54) and mean BMI 28.7 ± 7.3. We report predicted values (collagen %, fat %, myocytes %) for cardiac tissue composition within the RV, IVS, and LV (including epicardial and endocardial layers). The proportion of collagen and fat were higher in the RV compared with the LV (ratios 1.61 [1.45-1.78]; 2.63 [1.99-3.48], respectively) and RV compared with the IVS (ratios 1.77 [1.60-1.97]; 8.41[6.35-11.13], respectively). The ratio of epicardial versus endocardial fat was increased in both ventricles (RV: ratio 4.49 [3.67-5.49]; LV: ratio 3.46 [2.49-4.81]). In multivariable analysis, there was no significant association between collagen or fat proportion and sex (p=0.12; p=0.08, respectively), age at death (p=0.36; p=0.23, respectively), or BMI (p=0.45; p=0.43, respectively). CONCLUSIONS: Our findings provide location and sex-specific proportions of myocardial histological tissue composition that may aid quantitative evaluation of pathology in future studies.


Assuntos
Tecido Adiposo/patologia , Colágeno/análise , Ventrículos do Coração/química , Ventrículos do Coração/patologia , Miócitos Cardíacos/química , Miócitos Cardíacos/patologia , Adolescente , Adulto , Autopsia , Feminino , Fibrose , Humanos , Masculino , Pessoa de Meia-Idade , Fatores Sexuais , Adulto Jovem
2.
BMC Med Genomics ; 12(1): 141, 2019 10 22.
Artigo em Inglês | MEDLINE | ID: mdl-31640709

RESUMO

BACKGROUND: Epigenetic age acceleration, a measure of biological aging based on DNA methylation, is associated with cardiovascular mortality. However, little is known about its relationship with hypertensive target organ damage to the heart, kidneys, brain, and peripheral arteries. METHODS: We investigated associations between intrinsic (IEAA) or extrinsic (EEAA) epigenetic age acceleration, blood pressure, and six types of organ damage in a primarily hypertensive cohort of 1390 African Americans from the Genetic Epidemiology Network of Arteriopathy (GENOA) study. DNA methylation from peripheral blood leukocytes was collected at baseline (1996-2000), and measures of target organ damage were assessed in a follow-up visit (2000-2004). Linear regression with generalized estimating equations was used to test for associations between epigenetic age acceleration and target organ damage, as well as effect modification of epigenetic age by blood pressure or sex. Sequential Oligogenic Linkage Analysis Routines (SOLAR) was used to test for evidence of shared genetic and/or environmental effects between epigenetic age acceleration and organ damage pairs that were significantly associated. RESULTS: After adjustment for sex, chronological age, and time between methylation and organ damage measures, higher IEAA was associated with higher urine albumin to creatinine ratio (UACR, p = 0.004), relative wall thickness (RWT, p = 0.022), and left ventricular mass index (LVMI, p = 0.007), and with lower ankle-brachial index (ABI, p = 0.014). EEAA was associated with higher LVMI (p = 0.005). Target organ damage associations for all but IEAA with LVMI remained significant after further adjustment for blood pressure and antihypertensive use (p < 0.05). Further adjustment for diabetes attenuated the IEAA associations with UACR and RWT, and adjustment for smoking attenuated the IEAA association with ABI. No effect modification by age or sex was observed. CONCLUSIONS: Measures of epigenetic age acceleration may help to better characterize the functional mechanisms underlying organ damage from cellular aging and/or hypertension. These measures may act as subclinical biomarkers for damage to the kidney, heart, and peripheral vasculature; however more research is needed to determine whether these relationships remain independent of lifestyle factors and comorbidities.


Assuntos
Afro-Americanos/genética , Epigênese Genética , Hipertensão/patologia , Fatores Etários , Idoso , Índice Tornozelo-Braço , Anti-Hipertensivos/uso terapêutico , Pressão Sanguínea , Creatinina/urina , Metilação de DNA , Feminino , Taxa de Filtração Glomerular , Ventrículos do Coração/química , Humanos , Hipertensão/tratamento farmacológico , Leucócitos Mononucleares/metabolismo , Modelos Lineares , Masculino , Pessoa de Meia-Idade , Fatores de Risco , Albumina Sérica Humana/urina
3.
Int J Mol Sci ; 20(21)2019 Oct 24.
Artigo em Inglês | MEDLINE | ID: mdl-31653115

RESUMO

Cardiovascular disease (CVD) is common in chronic kidney disease (CKD), while major CV events are rare in young CKD patients. In addition to nitric oxide (NO)-related biomarkers, several surrogate markers have been assessed to stratify CV risk in youth with CKD, including 24-h ambulatory blood pressure monitoring (ABPM), carotid artery intima-media thickness (cIMT), pulse wave velocity (PWV), ABPM-derived arterial stiffness index (AASI), flow-mediated dilatation (FMD), and left ventricular mass index (LVMI). The aim of this study was to identify subclinical CVD through the analysis of indices of CV risk in children and adolescents with CKD. Between 2016 and 2018, the prospective observational study enrolled 125 patients aged 3 to 18 years with G1-G4 CKD stages. Close to two-thirds of young patients with CKD exhibited blood pressure (BP) abnormalities on ABPM. CKD children with abnormal office BP showed lower plasma arginine levels and arginine-to-asymmetric dimethylarginine (ADMA) ratio, but higher ratios of ADMA-to-symmetric dimethylarginine (SDMA) and citrulline-to-arginine. High PWV and AASI, indices of arterial stiffness, both strongly correlated with high BP load. Additionally, LV mass and LVMI exhibited strong correlations with high BP load. Using an adjusted regression model, we observed the citrulline-to-arginine ratio was associated with 24-h systolic and diastolic BP, systolic blood pressure (SBP) load, and diastolic blood pressure (DBP) load. Early assessments of NO-related parameters, BP load abnormalities, arterial stiffness indices, and LV mass will aid in early preventative care toward decreasing CV risk later in life for children and adolescents with CKD.


Assuntos
Pressão Sanguínea , Doenças Cardiovasculares/diagnóstico , Óxido Nítrico/metabolismo , Insuficiência Renal Crônica/patologia , Adolescente , Arginina/análogos & derivados , Arginina/sangue , Biomarcadores/metabolismo , Doenças Cardiovasculares/etiologia , Criança , Pré-Escolar , Feminino , Ventrículos do Coração/química , Ventrículos do Coração/metabolismo , Humanos , Masculino , Estudos Prospectivos , Análise de Onda de Pulso , Insuficiência Renal Crônica/complicações , Fatores de Risco , Índice de Gravidade de Doença , Rigidez Vascular
4.
PLoS One ; 14(8): e0221401, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31461472

RESUMO

Chaotic spiral or scroll wave dynamics can be found in diverse systems. In cardiac dynamics, spiral or scroll waves of electrical excitation determine the dynamics during life-threatening arrhythmias like ventricular fibrillation. In numerical studies it was found that chaotic episodes of spiral and scroll waves can be transient, thus they terminate spontaneously. We show in this study that this behavior can also be observed using models which describe the ion channel dynamics of human cardiomyocytes (Bueno-Orovio-Cherry-Fenton model and the Ten Tusscher-Noble-Noble-Panfilov model). For both models we find that the average lifetime of the chaotic transients grows exponentially with the system size. With this behavior, we classify the systems into the group of type-II supertransients. We observe a significant difference of the breakup behavior between the models, which results in a distinct dynamics during the final phase just before the termination. The observation of a (temporally) stable single-spiral state affects the prevailing description of the dynamics of type-II supertransients as being "quasi-stationary" and also the feasibility of predicting the spontaneous termination of the spiral wave dynamics. In the long term, the relation between the breakup behavior of spiral waves and properties of chaotic transients like predictability or average transient lifetime may contribute to an improved understanding and classification of cardiac arrhythmias.


Assuntos
Arritmias Cardíacas/fisiopatologia , Fenômenos Eletromagnéticos , Canais Iônicos/química , Modelos Cardiovasculares , Potenciais de Ação/fisiologia , Simulação por Computador , Ventrículos do Coração/química , Humanos , Canais Iônicos/fisiologia , Cadeias de Markov , Miócitos Cardíacos/química , Miócitos Cardíacos/fisiologia , Função Ventricular
5.
Int J Legal Med ; 133(6): 1809-1818, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30734118

RESUMO

An increasing number of suicidal asphyxiation with a plastic bag with inert gases, and in particular helium (He), have been reported from numerous countries over the last decade. These cases are differently managed and lead to different and variable interpretations. Based on the 12 last cases analysed in the laboratory and on the review of the most recent literature about this topic, updated autopsy guidelines for sampling have been proposed regarding to the samples choice and analytical challenges required by the gaseous state of this substance. Biological samples from airways (lungs lobe) followed by brain and cardiac blood are the best matrices to take during the autopsy to diagnose He exposure. Gaseous samples from trachea, pulmonary bronchi, gastric and cardiac areas are also recommended as alternative samples. The anatomical site of sampling must be carefully detailed, and to this end, forensic imaging constitutes a beneficial tool. Even if He detection is sufficient to conclude to He exposure, He concentrations in samples may be related to He exposure conditions (duration, breathing rate, etc.). A quantification in biological samples could be helpful to document more precisely the case. He concentrations in gaseous samples are reported up to 6.0 µmol/mL (tracheal gas), 2.4 µmol/mL (pulmonary gas), 0.64 µmol/mL (cardiac gas) and 12 µmol/mL (gastric gas). He concentrations in solid/liquid samples are reported up to 28 µmol/g (lungs) and 0.03 µmol/g (cardiac blood). The other matrices usually sampled during autopsy such as urine, peripheral blood, liver, fat matter and kidney appear as not relevant.


Assuntos
Toxicologia Forense/métodos , Hélio/análise , Asfixia , Química Encefálica , Brônquios/química , Cromatografia Gasosa-Espectrometria de Massas , Ventrículos do Coração/química , Hélio/envenenamento , Humanos , Abuso de Inalantes , Pulmão/química , Envenenamento/diagnóstico , Manejo de Espécimes , Estômago/química , Suicídio , Traqueia/química
6.
J Proteomics ; 191: 107-113, 2019 01 16.
Artigo em Inglês | MEDLINE | ID: mdl-29572163

RESUMO

Right ventricle (RV) remodelling occurs in neonatal patients born with ventricular septal defect (VSD). The presence of a defect between the two ventricles allows for shunting of blood from the left to right side. The resulting RV hypertrophy leads to molecular remodelling which has thus far been largely investigated using right atrial (RA) tissue. In this study we used proteomic and phosphoproteomic analysis in order to determine any difference between the proteomes for RA and RV. Samples were therefore taken from the RA and RV of five infants (0.34 ±â€¯0.05 years, mean ±â€¯SEM) with VSD who were undergoing cardiac surgery to repair the defect. Significant differences in protein expression between RV and RA were seen. 150 protein accession numbers were identified which were significantly lower in the atria, whereas none were significantly higher in the atria compared to the ventricle. 19 phosphorylation sites (representing 19 phosphoproteins) were also lower in RA. This work has identified differences in the proteome between RA and RV which reflect differences in contractile activity and metabolism. As such, caution should be used when drawing conclusions based on analysis of the RA and extrapolating to the hypertrophied RV. SIGNIFICANCE: RV hypertrophy occurs in neonatal patients born with VSD. Very little is known about how the atria responds to RV hypertrophy, especially at the protein level. Access to tissue from age-matched groups of patients is very rare, and we are in the unique position of being able to get tissue from both the atria and ventricle during reparative surgery of these infants. Our findings will be beneficial to future research into heart chamber malformations in congenital heart defects.


Assuntos
Comunicação Interventricular/metabolismo , Miocárdio/química , Proteoma/análise , Átrios do Coração/química , Comunicação Interventricular/patologia , Ventrículos do Coração/química , Ventrículos do Coração/patologia , Humanos , Hipertrofia , Lactente , Fosfoproteínas/análise , Proteômica/métodos
7.
Heart Vessels ; 34(3): 538-544, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30159657

RESUMO

Gold nano-rods, rod-shaped gold nanoparticles, act as contrast agents for in vivo bioimaging, drug delivery vehicles and thermal converters for photothermal therapy. Pro-inflammatory cytokines play critical roles in the development of heart failure. We examined the delivery of GNRs into the failing heart of a transgenic (TG) mouse model of inflammatory cardiomyopathy with the cardiac-specific overexpression of TNF-α. We modified GNRs with polyethylene glycol (PEG) to avoid cytotoxicity and reduce the rapid clearance of nanoparticles from blood. PEG-modified GNRs (4.5 mM as gold atoms, 200 µL) were administered intravenously to TG (n = 7) and wild-type (WT) mice (n = 5). These were killed 24 h later, and the heart, lung, liver, kidney and spleen were excised. A quantitative analysis of gold was performed using inductively coupled plasma mass or optical emission spectrometry. The amount of gold (ng) in the TG heart (3.24 ± 1.56 ng/mg heart weight) was significantly greater than that in the WT heart (1.01 ± 0.19; p < 0.05). No significant differences were observed among the other organs of TG and WT mice. The amount of gold in the TG heart was significantly and positively correlated with the ratio of the ventricular weight to body weight, which is known to be an index of ventricular hypertrophy. In conclusion, PEG-modified GNRs accumulated in the inflammatory TG heart in proportion with the severity of ventricular hypertrophy.


Assuntos
DNA/genética , Regulação da Expressão Gênica , Ouro/análise , Insuficiência Cardíaca/metabolismo , Nanopartículas Metálicas/análise , Miocárdio/química , Fator de Necrose Tumoral alfa/genética , Animais , Modelos Animais de Doenças , Insuficiência Cardíaca/genética , Insuficiência Cardíaca/patologia , Ventrículos do Coração/química , Ventrículos do Coração/patologia , Masculino , Camundongos , Camundongos Transgênicos , Miocárdio/metabolismo , Miocárdio/patologia , Reação em Cadeia da Polimerase , Fator de Necrose Tumoral alfa/biossíntese
8.
J Nutr Sci Vitaminol (Tokyo) ; 65(6): 483-490, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31902861

RESUMO

Calcium supplements were necessary for those people with low calcium intake and high risk of osteoporosis. Recent cohort studies have shown that long-term calcium supplements may raise the risk of cardiovascular disease, but its mechanism is still unclear. In this study, metabonomics were employed to evaluate the changes of metabolism in rats with long-term calcium supplementation and further seek the potential markers of cardiovascular risk. SD rats were divided into two groups including normal control group (calcium intake, 0.50 g/kg bw) and high calcium supplement group (calcium intake, 2.50 g/kg bw). After 6 mo, the cardiovascular system and bone mineral density were observed. UPLC-MS was used to analyze serum metabonomics in rats. The results showed that the contents of total cholesterol and low-density lipoprotein cholesterol in the high calcium group were significantly higher than those in normal control group (p<0.05). The interventricular septum thickness (IVS), left ventricular mass (LVM), left ventricular posterior wall thickness (LVPW) in the high calcium group were higher than those in normal control group (p<0.05). Serum metabonomics analysis showed that there were persistent changes in many metabolites such as sphingosine and its derivatives (p<0.01) in the comparison between the high calcium group and the normal group. These results indicated that long term calcium supplementation can lead to dyslipidemia in rats, such as the rise of cholesterol and low-density lipoprotein, which might induce myocardial hypertrophy. Long-term calcium supplementation can cause the changes of the amount of sphingosine and its derivatives in the body, which many have potential risk to cardiovascular diseases such as myocardial hypertrophy and atherosclerosis.


Assuntos
Densidade Óssea/efeitos dos fármacos , Cálcio na Dieta , Ventrículos do Coração/efeitos dos fármacos , Metaboloma/efeitos dos fármacos , Animais , Biomarcadores/sangue , Cálcio na Dieta/administração & dosagem , Cálcio na Dieta/análise , Cálcio na Dieta/farmacologia , Cardiomiopatia Hipertrófica/induzido quimicamente , Fêmur/química , Fêmur/efeitos dos fármacos , Ventrículos do Coração/química , Masculino , Metabolômica , Osteoporose/metabolismo , Ratos , Ratos Sprague-Dawley
9.
PLoS One ; 13(12): e0208987, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30589871

RESUMO

BACKGROUND: The Nile rat (Arvicanthis niloticus) is an emerging laboratory model of type 2 diabetes. When fed standard rodent chow, the majority of males progress from hyperinsulinemia by 2 months to hyperglycemia by 6 months, while most females remain at the hyperinsulinemia-only stage (prediabetic) from 2 months onward. Since diabetic cardiomyopathy is the major cause of type-2 diabetes mellitus (T2DM)-related mortality, we examined whether sexual dimorphism might entail cardiac functional changes. Our ultimate goal was to isolate the effect of diet as a modifiable lifestyle factor. MATERIALS AND METHODS: Nile rats were fed either standard rodent chow (Chow group) or a high-fiber diet previously established to prevent type 2 diabetes (Fiber group). Cardiac function was determined with echocardiography at 12 months of age. To isolate the effect of diet alone, only the small subset of animals resistant to both hyperinsulinemia and hyperglycemia were included in this study. RESULTS: In males, Chow (compared to Fiber) was associated with elevated heart rate and mitral E/A velocity ratio, and with lower e'-wave velocity, isovolumetric relaxation time, and ejection time. Of note, these clinically atypical types of diastolic dysfunction occurred independently of body weight. In contrast, females did not exhibit changes in cardiovascular function between diets. CONCLUSIONS: The higher prevalence of T2DM in males correlates with their susceptibility to develop subtle diastolic cardiac dysfunction when fed a Western style diet (throughout most of their lifespan) despite no systemic evidence of metabolic syndrome, let alone T2DM.


Assuntos
Diabetes Mellitus Tipo 2/patologia , Dieta , Animais , Glicemia/análise , Peso Corporal , Fibras na Dieta , Modelos Animais de Doenças , Feminino , Frequência Cardíaca , Ventrículos do Coração/química , Ventrículos do Coração/patologia , Masculino , Ratos , Caracteres Sexuais , Função Ventricular Esquerda/fisiologia
10.
Cell Tissue Res ; 373(2): 367-377, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-29691652

RESUMO

Atrial and B-type natriuretic peptides (ANP and BNP) are cardiac hormones important for cardiovascular and body fluid regulation. In some teleost species, an additional member of the natriuretic peptide family, ventricular NP (VNP), has been identified. In this study, we examine tissue distribution of these three NPs in the eel heart. Quantitative real-time PCR showed that anp is almost exclusively expressed in atria, bnp equally in atria and ventricles and vnp three-fold more in ventricles than in atria. The amount of bnp transcript overall in the heart was 1/10 those of anp and vnp. There was no difference in transcript levels between freshwater and seawater-acclimated fishes. Immunohistochemistry using specific antisera and in situ hybridization using gene-specific probes showed that NP signals were detected in most atrial and ventricular myocytes with some regional differences in density. Because of high sequence similarity of the three NPs, each of the three NP antisera individually was pre-incubated with 10-8 M of the other two non-targeted cardiac NPs to increase the specificity. A few atrial myocytes contained all three NPs in the same cell. Immuno-electron microscopy identified many dense-core vesicles containing ANP in atria and VNP in ventricles and some vesicles contained both ANP and VNP as demonstrated using pre-absorbed antisera. Based on these data and those of previous studies, we suggest that in eels ANP is secreted from atria in a regulatory pathway and VNP from ventricles in a constitutive pathway. In addition, VNP, not BNP, is the principal ventricular hormone in eels.


Assuntos
Fator Natriurético Atrial/metabolismo , Enguias/metabolismo , Átrios do Coração/metabolismo , Ventrículos do Coração/metabolismo , Peptídeo Natriurético Encefálico/metabolismo , Sequência de Aminoácidos , Animais , Fator Natriurético Atrial/química , Fator Natriurético Atrial/genética , Enguias/genética , Átrios do Coração/química , Ventrículos do Coração/química , Miócitos Cardíacos/química , Miócitos Cardíacos/metabolismo , Peptídeo Natriurético Encefálico/química , Peptídeo Natriurético Encefálico/genética , RNA Mensageiro/genética , Vesículas Secretórias/química , Vesículas Secretórias/metabolismo , Homologia de Sequência de Aminoácidos
11.
Cell Physiol Biochem ; 45(1): 67-77, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29316552

RESUMO

BACKGROUND/AIMS: Sphingosine-1-phosphate (S1P) is a bioactive lysosphingolipid that is found in high concentration in plasma. The majority of plasma S1P is transported bound to HDL and albumin. Although the major sources of circulating S1P have been identified, it remains obscure what is the contribution of different organs/tissues to S1P homeostasis in plasma. Answering this question was the major aim of the present study. METHODS: The experiment was performed on male Wistar rats from whom blood samples were taken from either: 1) femoral vein, right ventricle of the heart, and abdominal aorta (n=15) or 2) hepatic vein, portal vein, and abdominal aorta (n=11). Plasma was fractionated by sequential flotation ultracentrifugation and sphingolipids were quantified by a HPLC method. RESULTS: Compared to the mixed venous blood sampled from the right ventricle, total plasma and lipoprotein-depleted plasma (LPDP) concentration of S1P in the arterial blood was lower. On the other hand, the level of S1P increased across the leg both in plasma and LPDP. The concentration of S1P, sphingosine, and sphinganine in the plasma, HDL, and LPDP isolated from the blood taken from the hepatic vein was markedly higher compared to both arterial and portal blood. CONCLUSIONS: We conclude that, in contrast to HDL-bound S1P, albumin-associated S1P is very labile in the circulation. It is degraded in the pulmonary, and to a lesser extent, gastrointestinal circulation, and released across the liver and skeletal muscle. We also conclude that liver is an important source of HDL-bound S1P and circulating free sphingoid bases.


Assuntos
Cromatografia Líquida de Alta Pressão , Lisofosfolipídeos/sangue , Esfingosina/análogos & derivados , Animais , Aorta Abdominal/química , Aorta Abdominal/metabolismo , Veia Femoral/química , Veia Femoral/metabolismo , Ventrículos do Coração/química , Ventrículos do Coração/metabolismo , Veias Hepáticas/química , Veias Hepáticas/metabolismo , Lipoproteínas HDL/química , Lipoproteínas HDL/metabolismo , Lipoproteínas LDL/química , Lipoproteínas LDL/metabolismo , Masculino , Veia Porta/química , Veia Porta/metabolismo , Ligação Proteica , Ratos , Ratos Wistar , Esfingolipídeos/sangue , Esfingolipídeos/química , Esfingolipídeos/metabolismo , Esfingosina/sangue
12.
Cardiovasc Res ; 114(2): 247-258, 2018 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-29036603

RESUMO

Aims: Quantitative real-time RT-PCR (RT-qPCR) has become the method of choice for mRNA quantification, but requires an accurate normalization based on the use of reference genes showing invariant expression across various pathological conditions. Only few data exist on appropriate reference genes for the human heart. The objective of this study was to determine a set of suitable reference genes in human atrial and ventricular tissues, from right and left cavities in control and in cardiac diseases. Methods and results: We assessed the expression of 16 reference genes (ACTB, B2M, GAPDH, GUSB, HMBS, HPRT1, IPO8, PGK1, POLR2A, PPIA, RPLP0, TBP, TFRC, UBC, YWHAZ, 18S) in tissues from: right and left ventricles from healthy controls and heart failure (HF) patients; right-atrial tissue from patients in sinus rhythm with (SRd) or without (SRnd) atrial dilatation, patients with paroxysmal (pAF) or chronic (cAF) atrial fibrillation or with HF; and left-atrial tissue from patients in SR or cAF. Consensual analysis (by geNorm and Normfinder algorithms, BestKeeper software tool and comparative delta-Ct method) of the variability scores obtained for each reference gene expression shows that the most stably expressed genes are: GAPDH, GUSB, IPO8, POLR2A, and YWHAZ when comparing either right and left ventricle or ventricle from healthy controls and HF patients; GAPDH, IPO8, POLR2A, PPIA, and RPLP0 when comparing either right and left atrium or right atria from all pathological groups. ACTB, TBP, TFRC, and 18S genes were identified as the least stable. Conclusions: The overall most stable reference genes across different heart cavities and disease conditions were GAPDH, IPO8, POLR2A and PPIA. YWHAZ or GUSB could be added to this set for some specific experiments. This study should provide useful guidelines for reference gene selection in RT-qPCR studies in human heart.


Assuntos
Fibrilação Atrial/genética , Perfilação da Expressão Gênica/métodos , Genes Essenciais , Átrios do Coração/química , Cardiopatias/genética , Ventrículos do Coração/química , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real , Transcriptoma , Proteínas 14-3-3/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Estudos de Casos e Controles , RNA Polimerases Dirigidas por DNA/genética , Europa (Continente) , Feminino , Perfilação da Expressão Gênica/normas , Marcadores Genéticos , Glucuronidase/genética , Gliceraldeído-3-Fosfato Desidrogenases/genética , Humanos , Masculino , Pessoa de Meia-Idade , Peptidilprolil Isomerase/genética , Reação em Cadeia da Polimerase em Tempo Real/normas , Padrões de Referência , Reprodutibilidade dos Testes , beta Carioferinas/genética
13.
PLoS One ; 12(9): e0185125, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28934329

RESUMO

Extracellular matrix plays a role in differentiation and phenotype development of its resident cells. Although cardiac extracellular matrix from the contractile tissues has been studied and utilized in tissue engineering, extracellular matrix properties of the pacemaking sinoatrial node are largely unknown. In this study, the biomechanical properties and biochemical composition and distribution of extracellular matrix in the sinoatrial node were investigated relative to the left ventricle. Extracellular matrix of the sinoatrial node was found to be overall stiffer than that of the left ventricle and highly heterogeneous with interstitial regions composed of predominantly fibrillar collagens and rich in elastin. The extracellular matrix protein distribution suggests that resident pacemaking cardiomyocytes are enclosed in fibrillar collagens that can withstand greater tensile strength while the surrounding elastin-rich regions may undergo deformation to reduce the mechanical strain in these cells. Moreover, basement membrane-associated adhesion proteins that are ligands for integrins were of low abundance in the sinoatrial node, which may decrease force transduction in the pacemaking cardiomyocytes. In contrast to extracellular matrix of the left ventricle, extracellular matrix of the sinoatrial node may reduce mechanical strain and force transduction in pacemaking cardiomyocytes. These findings provide the criteria for a suitable matrix scaffold for engineering biopacemakers.


Assuntos
Matriz Extracelular/metabolismo , Ventrículos do Coração/metabolismo , Nó Sinoatrial/metabolismo , Animais , Membrana Basal/química , Membrana Basal/metabolismo , Membrana Basal/ultraestrutura , Relógios Biológicos/fisiologia , Fenômenos Biomecânicos , Colágeno/metabolismo , Colágeno/ultraestrutura , Elasticidade , Elastina/metabolismo , Elastina/ultraestrutura , Matriz Extracelular/química , Matriz Extracelular/ultraestrutura , Fibronectinas/metabolismo , Fibronectinas/ultraestrutura , Imunofluorescência , Ventrículos do Coração/química , Ventrículos do Coração/ultraestrutura , Espectrometria de Massas , Microscopia de Força Atômica , Microscopia Eletroquímica de Varredura , Miócitos Cardíacos/química , Miócitos Cardíacos/metabolismo , Proteoma , Proteômica , Nó Sinoatrial/química , Nó Sinoatrial/ultraestrutura , Suínos , Resistência à Tração
14.
Circulation ; 136(16): 1528-1544, 2017 Oct 17.
Artigo em Inglês | MEDLINE | ID: mdl-28838933

RESUMO

BACKGROUND: Biochemical DNA modification resembles a crucial regulatory layer among genetic information, environmental factors, and the transcriptome. To identify epigenetic susceptibility regions and novel biomarkers linked to myocardial dysfunction and heart failure, we performed the first multi-omics study in myocardial tissue and blood of patients with dilated cardiomyopathy and controls. METHODS: Infinium human methylation 450 was used for high-density epigenome-wide mapping of DNA methylation in left-ventricular biopsies and whole peripheral blood of living probands. RNA deep sequencing was performed on the same samples in parallel. Whole-genome sequencing of all patients allowed exclusion of promiscuous genotype-induced methylation calls. RESULTS: In the screening stage, we detected 59 epigenetic loci that are significantly associated with dilated cardiomyopathy (false discovery corrected P≤0.05), with 3 of them reaching epigenome-wide significance at P≤5×10-8. Twenty-seven (46%) of these loci could be replicated in independent cohorts, underlining the role of epigenetic regulation of key cardiac transcription regulators. Using a staged multi-omics study design, we link a subset of 517 epigenetic loci with dilated cardiomyopathy and cardiac gene expression. Furthermore, we identified distinct epigenetic methylation patterns that are conserved across tissues, rendering these CpGs novel epigenetic biomarkers for heart failure. CONCLUSIONS: The present study provides to our knowledge the first epigenome-wide association study in living patients with heart failure using a multi-omics approach.


Assuntos
Cardiomiopatia Dilatada/genética , Metilação de DNA , Epigênese Genética , Epigenômica/métodos , Loci Gênicos , Insuficiência Cardíaca/genética , Ventrículos do Coração/química , Cardiomiopatia Dilatada/sangue , Cardiomiopatia Dilatada/diagnóstico , Estudos de Casos e Controles , Ilhas de CpG , Perfilação da Expressão Gênica , Marcadores Genéticos , Predisposição Genética para Doença , Estudo de Associação Genômica Ampla , Insuficiência Cardíaca/sangue , Insuficiência Cardíaca/diagnóstico , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Fenótipo , RNA Mensageiro/genética , Análise de Sequência de RNA
15.
Anal Chem ; 89(9): 4922-4930, 2017 05 02.
Artigo em Inglês | MEDLINE | ID: mdl-28366003

RESUMO

Myosin heavy chain (MHC), the major component of the myosin motor molecule, plays an essential role in force production during muscle contraction. However, a comprehensive analysis of MHC proteoforms arising from sequence variations and post-translational modifications (PTMs) remains challenging due to the difficulties in purifying MHC (∼223 kDa) and achieving complete sequence coverage. Herein, we have established a strategy to effectively purify and comprehensively characterize MHC from heart tissue by combining size-exclusion chromatography (SEC) and middle-down mass spectrometry (MS). First, we have developed a MS-compatible SEC method for purifying MHC from heart tissue with high efficiency. Next, we have optimized the Glu-C, Asp-N, and trypsin limited digestion conditions for middle-down MS. Subsequently, we have applied this strategy with optimized conditions to comprehensively characterize human MHC and identified ß-MHC as the predominant isoform in human left ventricular tissue. Full sequence coverage based on highly accurate mass measurements has been achieved using middle-down MS combining 1 Glu-C, 1 Asp-N, and 1 trypsin digestion. Three different PTMs: acetylation, methylation, and trimethylation were identified in human ß-MHC and the corresponding sites were localized to the N-terminal Gly, Lys34, and Lys129, respectively, by electron capture dissociation (ECD). Taken together, we have demonstrated this strategy is highly efficient for purification and characterization of MHC, which can be further applied to studies of the role of MHC proteoforms in muscle-related diseases. We also envision that this integrated SEC/middle-down MS strategy can be extended for the characterization of other large proteins over 200 kDa.


Assuntos
Miosinas Cardíacas/química , Cromatografia em Gel/métodos , Cadeias Pesadas de Miosina/química , Espectrometria de Massas em Tandem/métodos , Miosinas Cardíacas/isolamento & purificação , Ventrículos do Coração/química , Humanos , Miocárdio/química , Cadeias Pesadas de Miosina/isolamento & purificação , Isoformas de Proteínas , Processamento de Proteína Pós-Traducional
16.
Eur J Cardiothorac Surg ; 51(6): 1063-1071, 2017 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-28329269

RESUMO

OBJECTIVES: We investigated the impact of mechanical unloading with a left ventricular assist device (LVAD) on the myocardial proteome. METHODS: We collected 11 patient-matched samples of myocardial left ventricular tissue of patients with non-ischaemic dilate cardiomyopathy, harvested at time of LVAD implant ('pre-LVAD') and heart transplant ('post-LVAD'). Samples were studied by quantitative proteomics. Further we performed histological assessment of deposited collagens and immune infiltration in both pre- and post-LVAD samples. RESULTS: A core set of >1700 proteins was identified and quantified at a false discovery rate <1%. The previously established decrease post-LVAD of alpha-1-antichymotrypsin was corroborated. We noted a post-LVAD decrease of matricellular proteins and proteoglycans such as periostin and versican. Also, proteins of the complement system and precursors of cardiac peptide hormones were decreased post-LVAD. An increase post-LVAD was evident for individual proteins linked to the innate immune response, proteins involved in diverse metabolic pathways, and proteins involved in protein synthesis. Histological analysis did not reveal significant alterations post-LVAD of deposited collagens or immune infiltration. The proteomic data further highlighted a pronounced inter-patient heterogeneity with regards to the impact of LVAD therapy on the left ventricular myocardial proteome. Finally, the proteomic data showed differential proteolytic processing in response to LVAD therapy. CONCLUSIONS: Our findings underline a strong impact of LVAD therapy on the left ventricular myocardial proteome. Together with previous studies, protein markers of LVAD therapy such as alpha-1-antichymotrypsin are becoming apparent. Further, matricellular proteins are emerging as important components in response to LVAD therapy.


Assuntos
Matriz Extracelular/metabolismo , Ventrículos do Coração , Coração Auxiliar , Proteínas/análise , Proteômica/métodos , Adulto , Idoso , Análise por Conglomerados , Matriz Extracelular/química , Feminino , Ventrículos do Coração/química , Ventrículos do Coração/metabolismo , Ventrículos do Coração/cirurgia , Humanos , Masculino , Pessoa de Meia-Idade , Modelos Cardiovasculares , Proteínas/química , Proteínas/metabolismo
17.
Zhong Nan Da Xue Xue Bao Yi Xue Ban ; 42(1): 41-48, 2017 Jan 28.
Artigo em Chinês | MEDLINE | ID: mdl-28216496

RESUMO

OBJECTIVE: To investigate the evolution of left ventricular global strain in anterior myocardial infarction patients treated with emergency percutaneous coronary intervention (PCI).
 Methods: A total of 54 patients with PCI were enrolled as a PCI group. Forty healthy subjects were enrolled as a control group. Dynamic cardiac images were collected. All of these images were analyzed off-line by velocity vector imaging (VVI) software. N-terminal pro-B-type natriuretic peptide (NT-proBNP) was measured with an electrochemiluminescence immunoassay through the Elecsys 1010/2010 system. Correlation analysis were undertaken between VVI and NT-proBNP levels in blood.
 Results: In PCI group, only globle longitudinal strain (GLS) was significantly increased 3 day after operation (P<0.05). GLS and globle circumferencial strain (GCS) were markedly increased 6 months after operation (P<0.05). In PCI group, left ventricular GLS 1 d to 6 months after PCI shows positive correlation with lgNT-proBNP levels (r=0.66, P<0.001). GLS value was -12.50% at the 3rd day after operation, indicating the improvment of cardiac function in the first and sixth month after PCI.
 Conclusion: The change of Left ventricular globle longitudinal systolic function after emergency PCI may be more sensitive to the improvement of myocardial stunning after STEMI reperfusion; GLS value (-12.50%) at the 3rd day after operation predict the improvment of cardiac function in the first and sixth months after PCI.


Assuntos
Infarto Miocárdico de Parede Anterior/diagnóstico por imagem , Ventrículos do Coração/química , Ventrículos do Coração/diagnóstico por imagem , Peptídeo Natriurético Encefálico/química , Intervenção Coronária Percutânea/reabilitação , Infarto do Miocárdio com Supradesnível do Segmento ST/diagnóstico por imagem , Idoso , Infarto Miocárdico de Parede Anterior/fisiopatologia , Biomarcadores , Diagnóstico por Computador/métodos , Feminino , Coração , Humanos , Masculino , Pessoa de Meia-Idade , Reperfusão Miocárdica/efeitos adversos , Reperfusão Miocárdica/reabilitação , Miocárdio Atordoado/fisiopatologia , Miocárdio Atordoado/terapia , Fragmentos de Peptídeos , Prognóstico , Infarto do Miocárdio com Supradesnível do Segmento ST/fisiopatologia , Sístole , Função Ventricular Esquerda
18.
RNA Biol ; 14(5): 500-513, 2017 05 04.
Artigo em Inglês | MEDLINE | ID: mdl-27124358

RESUMO

Expression of the miR-34 family (miR-34a, -34b, -34c) is elevated in settings of heart disease, and inhibition with antimiR-34a/antimiR-34 has emerged as a promising therapeutic strategy. Under chronic cardiac disease settings, targeting the entire miR-34 family is more effective than targeting miR-34a alone. The identification of transcription factor (TF)-miRNA regulatory networks has added complexity to understanding the therapeutic potential of miRNA-based therapies. Here, we sought to determine whether antimiR-34 targets secondary miRNAs via TFs which could contribute to antimiR-34-mediated protection. Using miRNA-Seq we identified differentially regulated miRNAs in hearts from mice with cardiac pathology due to transverse aortic constriction (TAC), and focused on miRNAs which were also regulated by antimiR-34. Two clusters of stress-responsive miRNAs were classified as "pathological" and "cardioprotective," respectively. Using ChIPBase we identified 45 TF binding sites on the promoters of "pathological" and "cardioprotective" miRNAs, and 5 represented direct targets of miR-34, with the capacity to regulate other miRNAs. Knockdown studies in a cardiomyoblast cell line demonstrated that expression of 2 "pathological" miRNAs (let-7e, miR-31) was regulated by one of the identified TFs. Furthermore, by qPCR we confirmed that expression of let-7e and miR-31 was lower in hearts from antimiR-34 treated TAC mice; this may explain why targeting the entire miR-34 family is more effective than targeting miR-34a alone. Finally, we showed that Acsl4 (a common target of miR-34, let-7e and miR-31) was increased in hearts from TAC antimiR-34 treated mice. In summary, antimiR-34 regulates the expression of other miRNAs and this has implications for drug development.


Assuntos
Cardiomegalia/terapia , Redes Reguladoras de Genes , Insuficiência Cardíaca/terapia , MicroRNAs/antagonistas & inibidores , MicroRNAs/metabolismo , Adulto , Análise de Variância , Animais , Cardiomegalia/metabolismo , Linhagem Celular , Coenzima A Ligases/genética , Coenzima A Ligases/metabolismo , Modelos Animais de Doenças , Regulação da Expressão Gênica , Insuficiência Cardíaca/metabolismo , Ventrículos do Coração/química , Ventrículos do Coração/metabolismo , Humanos , Masculino , Camundongos , Camundongos Endogâmicos , MicroRNAs/análise , Miócitos Cardíacos/química , Miócitos Cardíacos/metabolismo , Placebos , Análise de Sequência de RNA , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
19.
Oxid Med Cell Longev ; 2017: 8156594, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-29333212

RESUMO

Aronia melanocarpa has attracted scientific interest due to its dense contents of different polyphenols. We aimed to analyse effects of Aronia melanocarpa (AME) extract on blood pressure (BP), lipid peroxidation, cytokine level, total NOS activity in the left ventricle (LV), and aorta of L-NAME-induced hypertensive rats. 12-week-old male WKY rats were assigned to the control group and groups treated with AME extract (57.90 mg/kg/day), L-NAME (40 mg/kg/day), or combination of L-NAME (40 mg/kg/day) and AME (57.90 mg/kg/day) in tap water for 3 weeks. NOS activity, eNOS protein expression, and conjugated diene (CD) concentration were determined in the LV and aorta. After 3 weeks of L-NAME treatment, BP was increased by 28% and concomitant treatment with AME reduced it by 21%. NOS activity of the LV and aorta in the L-NAME group was decreased by about 40%, while AME increased it almost on the control level. AME-induced eNOS upregulation may contribute to increase NOS activity. Moreover, AME decreased CD concentration in the LV and aorta and TNF-α and IL-6 production in the plasma were increased by L-NAME treatment. In conclusion, our results showed that active substances of Aronia melanocarpa may have a positive effect on blood pressure, NOS activity, and proinflammatory processes in L-NAME-induced hypertension.


Assuntos
Aorta/efeitos dos fármacos , Ventrículos do Coração/efeitos dos fármacos , Photinia/química , Extratos Vegetais/farmacologia , Regulação para Cima/efeitos dos fármacos , Alcadienos/análise , Alcadienos/metabolismo , Animais , Aorta/química , Aorta/enzimologia , Pressão Sanguínea/efeitos dos fármacos , Ventrículos do Coração/química , Ventrículos do Coração/enzimologia , Hipertensão/etiologia , Hipertensão/prevenção & controle , Hipertensão/veterinária , Interleucina-6/sangue , Masculino , NG-Nitroarginina Metil Éster/toxicidade , Óxido Nítrico Sintase Tipo II/genética , Óxido Nítrico Sintase Tipo II/metabolismo , Óxido Nítrico Sintase Tipo III/genética , Óxido Nítrico Sintase Tipo III/metabolismo , Photinia/metabolismo , Extratos Vegetais/química , Extratos Vegetais/uso terapêutico , Polifenóis/química , Polifenóis/farmacologia , Polifenóis/uso terapêutico , Ratos , Ratos Endogâmicos WKY , Fator de Necrose Tumoral alfa/metabolismo
20.
J Periodontal Res ; 52(3): 603-608, 2017 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-27859254

RESUMO

BACKGROUND AND OBJECTIVE: The role of oxidative stress in the process of cardiac remodeling, hypertrophy and heart failure is a current topic. The purpose of this experimental study was to investigate the influences of periodontitis on levels of cardiac oxidative stress. MATERIAL AND METHODS: Twenty rats were separated into two groups: control and experimental periodontitis (EP). Periodontitis was induced by placing a 3.0 silk suture in the cervix of the left and right mandibular first molar teeth for 5 wk. At the end of the experiment, the animals were killed and blood samples and mandibular and ventricular cardiac tissue samples were collected. Levels of alveolar bone loss were determined using measurements performed on histological slices and radiographies. Left ventricular tissue 8-hydroxy-2'-deoxyguanosine, malonylaldehyde, glutathione peroxidase, total oxidant status, total antioxidant status (TAS) levels and serum paraoxonase-1 activity were evaluated biochemically. RESULTS: Measurements performed on the histological slices and radiographies demonstrated that applying the ligature caused obvious alveolar bone loss. Oxidative damage markers (malonylaldehyde, 8-hydroxy-2'-deoxyguanosine, oxidative stress index: total oxidant status/TAS) were significantly higher, and antioxidant markers (glutathione peroxidase, TAS) were statistically insignificantly higher, in the hearts of rats with EP when compared to the controls. In addition, reduced serum paraoxonase-1 activity was also detected in the EP group. CONCLUSION: The pronounced increase in cardiac oxidative stress caused by periodontitis was due to an excessive increase in the production of reactive oxygen species, rather than due to decreased antioxidant capacity. The results indicate that periodontitis-related cardiac oxidative stress might be one of the mechanisms that contribute to the pathological process that leads to heart failure.


Assuntos
Miocárdio/metabolismo , Estresse Oxidativo , Periodontite/metabolismo , 8-Hidroxi-2'-Desoxiguanosina , Animais , Desoxiguanosina/análogos & derivados , Desoxiguanosina/análise , Glutationa Peroxidase/metabolismo , Ventrículos do Coração/química , Ventrículos do Coração/metabolismo , Ventrículos do Coração/patologia , Masculino , Malondialdeído/análise , Miocárdio/química , Miocárdio/patologia , Periodontite/patologia , Ratos , Ratos Sprague-Dawley
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