RESUMEN
Chromatin condensation paralleled by DNA fragmentation is one of the most important nuclear events occurring during apoptosis. Histone modifications, and in particular phosphorylation, have been suggested to affect chromatin function and structure during both cell cycle and cell death. We report here that phosphate incorporation into all H1 subtypes decreased rapidly after induction of apoptosis, evidently causing a strong reduction in phosphorylated forms of main H1 histone subtypes. H1 dephosphorylation is accompanied by chromatin condensation preceding the onset of typical chromatin oligonucleosomal fragmentation, whereas H2A.X hyperphosphorylation is strongly correlated to apoptotic chromatin fragmentation. Using various kinase inhibitors we were able to exclude some of the possible kinases which can be involved directly or indirectly in phosphorylation of histone H2A.X. Neither DNA-dependent protein kinase, protein kinase A, protein kinase G, nor the kinases driven by the mitogen-activated protein kinase (MAP) pathway appear to be responsible for H2A.X phosphorylation. The protein kinase C activator phorbol 12-myristate 13-acetate (PMA), however, markedly reduced the induction of apoptosis in TNFalpha-treated cells with a simultaneous change in the phosphorylation pattern of histone H2A.X. Hyperphosphorylation of H2A.X in apoptotic cells depends indirectly on activation of caspases and nuclear scaffold proteases as shown in zVAD-(OMe)-fmk- or zAPF-cmk-treated cells, whereas the dephosphorylation of H1 subtypes seems to be influenced solely by caspase inhibitors. Together, these results illustrate that H1 dephosphorylation and H2A.X hyperphosphorylation are necessary steps on the apoptotic pathway.
Asunto(s)
Anticuerpos Monoclonales/farmacología , Apoptosis/fisiología , Histonas/metabolismo , Péptidos y Proteínas de Señalización Intracelular , Factor de Necrosis Tumoral alfa/farmacología , Células 3T3 , Animales , Anticuerpos Monoclonales de Origen Murino , Apoptosis/efectos de los fármacos , Cantaridina/farmacología , Proteínas Portadoras/farmacología , Electroforesis en Gel de Poliacrilamida , Activadores de Enzimas/farmacología , Inhibidores Enzimáticos/farmacología , Histonas/química , Ratones , Fosforilación/efectos de los fármacos , Inhibidores de Proteasas/farmacologíaRESUMEN
Brain natriuretic peptide (BNP), NT-proBNP and NT-pro-atrial natriuretic peptide (NT-proANP) were measured in blood samples from 57 patients using immunoassays and immunoradiometric assays to evaluate the usefulness as diagnostic markers for the detection of heart failure. For the detection of impaired left ventricular ejection fraction (LVEF), receiver operating characteristic curves showed that BNP had the best diagnostic performance with an area under curve (AUC) of 0.75+/-0.06. However, NT-proBNP (AUC: 0.67+/-0.07) and NT-proANP (AUC: 0.69+/-0.08) showed no significant difference to BNP. In a further analysis for the detection of resting LVEF <40%, BNP again was the best marker with an AUC of 0.83+/-0.06. NT-proBNP showed only a slightly smaller AUC (0.79+/-0.07). The AUC for NT-proANP was significantly smaller (0.65+/-0.08) compared to BNP. Additionally, BNP and NT-proBNP correlated negatively with the resting LVEF (BNP: -0.472, p<0.001; NT-proBNP: -0.306, p=0.026), whereas NT-proANP showed no significant correlation. In summary, BNP was the best marker to detect patients with impaired LVEF compared to NT-proBNP and NT-proANP. However, NT-proBNP showed no significant differences to BNP and it is therefore a new promising alternative marker for the detection of left ventricular dysfunction.
Asunto(s)
Factor Natriurético Atrial/sangre , Péptido Natriurético Encefálico/sangre , Proteínas del Tejido Nervioso/sangre , Fragmentos de Péptidos/sangre , Precursores de Proteínas/sangre , Disfunción Ventricular Izquierda/sangre , Disfunción Ventricular Izquierda/diagnóstico , Anciano , Anciano de 80 o más Años , Biomarcadores/sangre , Humanos , Persona de Mediana EdadRESUMEN
The long-predicted endocrine function of the heart has been proven by the discovery of atrial natriuretic peptide (atrial natriuretic factor, A-type natriuretic peptide; ANP) 20 years ago. This subsequently led to the description of a whole family of structurally similar but genetically distinct peptides, the natriuretic peptide family, which contributes to cardiovascular homeostasis. These looped peptides promote natriuresis and diuresis, act as vasodilators, and exert antimitogenic effects on cardiovascular tissues. Two members, ANP and brain natriuretic peptide (B-type natriuretic peptide; BNP) are secreted by the heart mainly in response to myocardial stretch induced by volume load. The natriuretic peptides are synthesized as preprohormones. The C-terminal endocrinological active peptides (ANP, BNP) and their N-terminal prohormone fragments are found in plasma. The natriuretic peptide system is activated to its highest degree in ventricular dysfunction. However, natriuretic peptides are increased in all patients with edematous disorders which lead to an increase in atrial tension or central blood volume, such as renal failure or ascitic liver cirrhosis. It could be demonstrated that in chronic heart failure patients and during the subacute phase of myocardial infarction, of all tested neurohormones, the cardiac natriuretic peptides were best markers to identify heart failure and the most powerful predictors of morbidity and mortality. Natriuretic peptides are independent markers for risk assessment. In comparative studies BNP was superior to ANP and its N-terminal prohormone fragments in myocardial infarction as well as in chronic heart failure patients. Less data on N-terminal proBNP (NT-proBNP) is available, but BNP and NT-proBNP appear to be equivalent markers. For primary care physicians natriuretic peptide measurement is useful to decide which patient with suspected heart failure warrants further investigation, particularly when assessment of left ventricular function is not readily available. Natriuretic peptides have an excellent negative predictive value, particularly in high risk patients. An increase in BNP is serious enough to warrant follow-up examinations. For the cardiologists the natriuretic peptides are helpful for guidance of therapy and monitoring disease course in heart failure patients and for risk stratification in heart failure and myocardial infarction.
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Factor Natriurético Atrial/sangre , Factor Natriurético Atrial/química , Insuficiencia Cardíaca/diagnóstico , Factor Natriurético Atrial/metabolismo , Química Clínica/métodos , Dimerización , Insuficiencia Cardíaca/sangre , Humanos , Modelos Biológicos , Péptido Natriurético Encefálico/sangre , Péptido Natriurético Encefálico/química , Valores de Referencia , Factores de RiesgoRESUMEN
The aim of this study was to examine the relationship between myosin heavy chain (MHC) release as a specific marker of slow-twitch muscle fibre breakdown and magnetic resonance imaging (MRI) of skeletal muscle injury after eccentric exercise. The effects of a single series of 70 high-intensity eccentric contractions of the quadriceps femoris muscle group (single leg) on plasma concentrations of creatine kinase and MHC fragments were assessed in 10 young male sport education trainees before and 1 and 4 days after exercise. To visualize muscle injury, MRI of the loaded thigh was performed before and 4 days after the eccentric exercise. All participants recorded an increase (P < 0.05) in creatine kinase after exercise. In five participants, T2 signal intensity was unchanged post-exercise compared with pre-exercise and MHC plasma concentration was normal; however, they showed an increase (P < 0.05) in creatine kinase after exercise. For the remaining five participants, there was an increase in T2 signal intensity of the loaded vastus intermedius and vastus lateralis. These changes in MRI were accompanied by an increase in MHC plasma concentration (P< 0.01) as well as an increase in creatine kinase (P < 0.01). We suggest that changes in MRI T, signal intensity after muscle damage induced by eccentric exercise are closely related to damage to structurally bound contractile filaments of some muscle fibres. Additionally, MHC plasma release indicates that this damage affects not only fast-twitch fibres but also some slow-twitch fibres.
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Creatina Quinasa/sangre , Ejercicio Físico/fisiología , Imagen por Resonancia Magnética , Músculo Esquelético/metabolismo , Cadenas Pesadas de Miosina/sangre , Adulto , Humanos , Masculino , Contracción Muscular/fisiología , Fibras Musculares de Contracción Lenta/patología , Músculo Esquelético/diagnóstico por imagen , Músculo Esquelético/lesiones , RadiografíaRESUMEN
Natriuretic peptides, atrial natriuretic peptide and brain natriuretic peptide, are key regulators in the homeostasis of salt and water excretion and in the maintenance of blood pressure. During heart failure, these peptides are highly activated because of volume overload and increased myocardial wall tension. Among all natriuretic peptides and neurohormones, brain natriuretic peptide and its N-terminal prohormone fragment have been shown to be the best markers to identify patients with heart failure. They are useful prognostic markers as well. The stability of brain natriuretic peptides and N-terminal prohormones in ethylene-diamine-tetraacetic-acid whole blood is sufficient for routine use. Sensitive and specific assays without the need for plasma extraction are commercially available. The available data indicate that natriuretic peptides are powerful diagnostic and prognostic markers in heart failure patients. First data on treatment guidance are promising.
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Factor Natriurético Atrial/sangre , Biomarcadores/sangre , Gasto Cardíaco Bajo/diagnóstico , Péptido Natriurético Encefálico/sangre , Anciano , Anticoagulantes , Factor Natriurético Atrial/fisiología , Gasto Cardíaco Bajo/etiología , Estabilidad de Medicamentos , Ácido Edético , Femenino , Humanos , Masculino , Péptido Natriurético Encefálico/fisiología , Pronóstico , Sensibilidad y EspecificidadRESUMEN
OBJECTIVE: Heat shock proteins (HSPs) are molecular chaperones which are essential for cell survival. Heat shock and hypoxia markedly increase the expression of several HSPs in various tissues, i.e. heart. In our in vitro study, we investigated whether HSPs are inducible in human vessels which are used as coronary artery bypass grafts. METHODS: We used remnants of the saphenous vein and the internal mammary artery from 34 patients undergoing coronary artery bypass surgery. Each vessel was divided into segments, one for control conditions at 37 degrees C (5% CO(2)-95% air), the remaining ones for thermal (30 min at 42 degrees C) or hypoxic treatment (6 h oxygen deprivation with nitrogen). The expression of Hsp60, Hsp72 and Hsp73 was investigated by immunohistochemistry and Western-blot analysis. RESULTS: Compared to controls, segments of the saphenous vein undergoing heat treatment showed significantly increased expression of Hsp72 in the intima (P=0.035) and Hsp73 in the media (P=0.003). In the internal mammary artery, Hsp72 and Hsp73 were expressed in the intima at significantly higher levels (P=0.042 each). A 6 h oxygen deprivation with nitrogen resulted in elevated levels of Hsp60 (media: P=0.048), of Hsp72 (intima: P<0.001 and media: P=0.004) and of Hsp73 (intima: P=0.029) in the saphenous vein. In the internal mammary artery, Hsp73 expression was significantly enhanced (intima: P=0.048 and media: P=0.017). The results were confirmed by Western-blot analysis in representative veins. CONCLUSIONS: These findings demonstrate the common cellular defense mechanism of HSP expression in response to stress in coronary artery bypass grafts. Hypoxia and heat treatment strongly induce Hsp72 and Hsp73 expression in human coronary artery bypass grafts.
Asunto(s)
Puente de Arteria Coronaria , Proteínas HSP70 de Choque Térmico , Proteínas de Choque Térmico/metabolismo , Hipoxia/metabolismo , Arterias Mamarias/metabolismo , Vena Safena/metabolismo , Anciano , Western Blotting , Proteínas Portadoras/metabolismo , Femenino , Proteínas del Choque Térmico HSC70 , Proteínas del Choque Térmico HSP72 , Calor , Humanos , Masculino , Persona de Mediana Edad , Técnicas de Cultivo de ÓrganosRESUMEN
Cardiac troponin T (cTnT), cardiac troponin I (cTnI), myosin heavy chains (MHC), myoglobin, creatine kinase (CK), and creatine kinase isoenzyme MB (CKMB), were measured in blood samples from 39 polymyositis (PM) or dermatomyositis (DM) patients without clinical evidence for cardiac involvement to evaluate their clinical usefulness in this patient population. MHC, myoglobin, and CKMB were frequently elevated and correlated with each other and with disease severity. Undetectable cTnI in all but one patient indicated that MHC was released from skeletal muscle, thereby providing the first laboratory evidence of frequent slow-twitch muscle fibre-necrosis in patients with PM or DM. CKMB was elevated in 51%, cTnT in 41%, and cTnI in only 2.5% of patients. cTnI did not correlate with other markers or with disease severity scores. The close correlations found between cTnT and skeletal muscle damage markers and the relationship between cTnT with disease severity without clinical evidence for myocardial damage suggest a release of cTnT from skeletal muscle. The relationship of cTnT with disease severity indicates a possible role of the marker for risk stratification. However, the prognostic values of cardiac troponins and other muscle damage markers in PM/DM patients remain to be compared in prospective outcome trials.
Asunto(s)
Dermatomiositis/sangre , Miocardio/metabolismo , Cadenas Pesadas de Miosina/sangre , Polimiositis/sangre , Troponina I/sangre , Troponina T/sangre , Adolescente , Adulto , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Masculino , Persona de Mediana Edad , RadioinmunoensayoRESUMEN
BACKGROUND: Because of controversial earlier studies, the purpose of this study was to provide novel experimental and additional clinical data regarding the possible reexpression of cardiac troponin T (cTnT) in regenerating skeletal muscle in Duchenne muscular dystrophy (DMD). METHODS: Plasma from 14 patients (mean age, 7.5 years; range, 5.7-19.4 years) with DMD was investigated for creatine kinase (CK), the CK MB isoenzyme (CKMB), cTnT and cardiac troponin I (cTnI), and myoglobin. cTnT concentrations were measured by an ELISA (second-generation assay; Roche) using the ES 300 Analyzer. cTnI, myoglobin, and CKMB were measured by an ELISA using the ACCESS System (Beckman Diagnostics). Troponin isoform expression was studied by Western blot analysis in remnants of skeletal muscle biopsies of three patients with DMD and in an animal model of DMD (mdx mice; n = 6). RESULTS: There was no relation of cTnT and cTnI to clinical evidence for cardiac failure. cTnI concentrations remained below the upper reference limit in all patients. cTnT was increased (median, 0.11 microg/L; range, 0.06-0.16 microg/L) in 50% of patients. The only significant correlation was found for CK (median, 3938 U/L; range, 2763-5030 U/L) with age (median, 7.5 years; range, 6.8-10.9 years; r = -0.762; P = 0.042). Western blot analysis of human or mouse homogenized muscle specimens showed no evidence for cardiac TnT and cTnI expression, despite strong signals for skeletal muscle troponin isoforms. CONCLUSIONS: We found no evidence for cTnT reexpression in human early-stage DMD and in mdx mouse skeletal muscle biopsies. Discrepancies of cTnT and cTnI in plasma samples of DMD patients were found, but neither cTnT nor cTnI plasma concentrations were related with other clinical evidence for cardiac involvement.
Asunto(s)
Distrofia Muscular de Duchenne/metabolismo , Miocardio/metabolismo , Troponina I/análisis , Troponina T/análisis , Adolescente , Adulto , Animales , Western Blotting , Niño , Preescolar , Humanos , Inmunoensayo , Masculino , Ratones , Músculo Esquelético/metabolismo , Distrofia Muscular de Duchenne/sangre , RegeneraciónRESUMEN
The release of muscle proteins after downhill running, which mainly includes eccentric muscle action, was compared in females (F; n=9) and males (M; n=9). They performed 20 min of downhill treadmill running with 16% decline with a target heart rate of 70% of the individual VO2peak, which was determined two weeks before. Blood samples were drawn before, 6 and 24 h after exercise to measure plasma levels of skeletal troponin I (sTnI), myosin heavy chain fragments (MHC), creatine kinase (CK), and myoglobin (Mb). Baseline levels before exercise were significantly higher in males compared to females for the cytoplasmic proteins CK and Mb, but the difference for MHC and sTnI was not significant. Both groups displayed marked and significant early (6 h) increases (P<0.05) for sTnI (median: F: 8.2 microg/L; M: 22.0 microg/L), Mb (median: F: 86.8 microg/L; M: 407 microg/L), and CK (median: F: 162 U/L; M: 339 U/L). A significant (P<0.05) but delayed (24 h) increase was found for MHC (median: F: 482 microU/L; M: 651 microU/L). The absolute values for all four parameters were significantly (P<0.05) higher in males compared to females; however, no difference was found for the relative increases and the time course of all parameters between females and males. We conclude 1) that there were no significant differences in the basal concentrations of predominantly bound proteins, and 2) that there were no differences in the relative muscle protein release between females and males before and after one bout of high-intensive eccentric exercise. The higher plasma concentrations of all measured muscle proteins in males are probably caused by the higher muscle mass compared to females.
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Creatina Quinasa/análisis , Músculo Esquelético/fisiología , Mioglobina/análisis , Cadenas Pesadas de Miosina/análisis , Carrera/fisiología , Troponina I/análisis , Adulto , Composición Corporal , Femenino , Humanos , Masculino , Factores SexualesRESUMEN
OBJECTIVE: It has been shown previously that the internal mammary artery releases more cyclic guanosine monophosphate after stimulation with atrial natriuretic peptide than the saphenous vein, and that C-type natriuretic peptide possesses a cyclic guanosine monophosphate stimulating potential on saphenous vein bypass grafts. The present study was undertaken to investigate intracellular content and extracellular release of cyclic guanosine monophosphate, by the internal mammary artery and saphenous vein, after challenge with further members of the natriuretic peptide family. METHODS: Specimens of human internal mammary artery and saphenous vein from 29 patients were cut into segments and stimulated with 10(-6) M concentrations of atrial natriuretic peptide (internal mammary artery n=8, saphenous vein n=10), brain natriuretic peptide (internal mammary artery n=9, saphenous vein n=13), C-type natriuretic peptide (internal mammary artery n=12, saphenous vein n=15), and urodilatin (internal mammary artery n=8, saphenous vein n=12). Intracellular content and extracellular release of cyclic guanosine monophosphate were determined using an (125)I radioimmunoassay. RESULTS: The following median stimulated intracellular cyclic guanosine monophosphate concentrations were measured in the internal mammary artery and saphenous vein: 35358 and 8672 fmol/cm(2) (P<0.001) after atrial natriuretic peptide, 45632 and 7830 fmol/cm(2) (P=0.003) after brain natriuretic peptide, 10144 and 13216 fmol/cm(2) (P=NS for intergraft comparison) after C-type natriuretic peptide, and 20949 and 6690 fmol/cm(2) (P=0.001) after urodilatin. Stimulation with atrial natriuretic peptide, brain natriuretic peptide and urodilation also led to a significant increase of extracellular cyclic guanosine monophosphate release by the internal mammary artery. CONCLUSIONS We conclude that brain natriuretic peptide and urodilatin exhibit a similarly effective cyclic guanosine monophosphate-stimulating potential on the internal mammary artery as atrial natriuretic peptide. In contrast, C-type natriuretic peptide shows comparable effects on the internal mammary artery and saphenous vein.
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Factor Natriurético Atrial/farmacología , GMP Cíclico/metabolismo , Arterias Mamarias/metabolismo , Péptido Natriurético Encefálico/farmacología , Vena Safena/metabolismo , Humanos , Técnicas In Vitro , Péptido Natriurético Tipo-C/farmacología , Fragmentos de Péptidos/farmacologíaRESUMEN
We report here on the HPCE separation of two chicken H5 histones, which do not show the heterogeneity (Gln/Arg) at residue 15 first found by Greenaway and Murray [Greenaway and Murray (1971) Nat. New Biol. 229, 233-238]. The two subfractions obtained were identified using reversed-phase HPLC, hydrophilic interaction HPLC, Edman degradation, and MALDI-MS analysis. We found that the two H5 subcomponents differ only by an acetylated (designated H5a) and an unacetylated N-terminus (H5b). In contrast to the N-terminally acetylated form of rat kidney histone H1(o), which increased by about 40% with aging of the animal, the corresponding form of chicken H5 did not: the ratio N-terminally acetylated: unacetylated remained constant (30:70) when histone H5 was extracted from erythrocytes of newly hatched chickens and from adult chickens, respectively. The HPCE technique used in this investigation represents a quick and convenient method for analyzing N-terminally acetylated proteins in the presence of unacetylated forms.
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Envejecimiento/fisiología , Histonas/química , Histonas/metabolismo , Acetilación , Animales , Pollos , Cromatografía Líquida de Alta Presión , Quimotripsina/metabolismo , Electroforesis Capilar , Eritrocitos/química , Histonas/aislamiento & purificación , Riñón/química , Riñón/citología , Ratas , Espectrometría de Masa por Láser de Matriz Asistida de Ionización DesorciónRESUMEN
Muscular overuse after high force eccentric muscle action is associated with structural damage of the contractile apparatus that can be observed as Z-line steaming and myofibrillar disruption. Mechanical stress is the major contributing factor for inducing muscle injury, which initiates a cascade of processes resulting in skeletal muscle damage. Disturbances in Ca2+ homeostasis with elevated intracellular [Ca2+] activates the nonlysomal cysteine protease, calpain. Calpain is assumed to play an important role in triggering the response of skeletal muscle protein breakdown, of inflammatory changes, and of regeneration processes in response to eccentric muscle action. The inflammatory response is attributed to changes in hormone and cytokine levels in blood and skeletal muscle. To assess the amount of skeletal muscle damage, plasma CK activity and plasma myoglobin levels have been widely used as markers for muscle injury. As the cytosolic proteins do not necessarily reflect the amount of structural damage, structurally bound proteins such as myosin heavy chains and troponin have been investigated. This paper briefly reviews the cascade of events causing muscle cell injury after unaccustomed eccentric muscle action and the potential of muscle proteins as markers of skeletal muscle damage.
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Proteínas Musculares/metabolismo , Músculo Esquelético/lesiones , Proteínas de Neoplasias , Proteínas Supresoras de Tumor , Animales , Biomarcadores , Proteínas Portadoras/metabolismo , Creatina Quinasa/metabolismo , Ejercicio Físico/fisiología , Proteína de Unión a los Ácidos Grasos 7 , Proteínas de Unión a Ácidos Grasos , Humanos , Fibras Musculares Esqueléticas/metabolismo , Músculo Esquelético/metabolismo , Proteína P2 de Mielina/metabolismo , Mioglobina/metabolismo , Cadenas Pesadas de Miosina/metabolismo , Esfuerzo Físico/fisiología , Troponina I/metabolismoRESUMEN
The heart secrets two different natriuretic peptides, atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP), which have potent vasorelaxant, diuretic, and natriuretic actions. They are main tools in the body's defense against volume overload and hypertension. The natriuretic peptides (NP) are synthetized as prohormones. The C-terminal endocrinological active peptides and their N-terminal prohormone fragments are found in plasma. The NP system is maximally activated in ventricular dysfunction. However, NP:s are also increased in patients with renal failure or pulmonary hypertension, and increases may be found in arterial hypertension or liver cirrhosis. Among all NP and prohormone fragments currently BNP is the most promising candidate analyte for routine diagnosis. BNP is also superior to other neurohormones for diagnosis of left-ventricular dysfunction (LVD) or estimating prognosis in LVD or during the subacute phase of myocardial infarction. For primary care physicians BNP measurement is useful to decide which patient with suspected heart failure warrants further investigation, particularly when assessment of left ventricular function is not readily available. BNP has an excellent negative predictive value particularly in high risk patients. For the cardiologists the NP:s are helpful for monitoring therapy and disease course in LVD patients and for estimating prognosis in LVD and myocardial infarction patients. There is now sufficient evidence to encourage physicians to gain experience with NP as a supplement in the diagnosis of patients suspected of having heart failure. An increase in BNP is serious enough to warrant follow-up examinations.
Asunto(s)
Factor Natriurético Atrial , Péptido Natriurético Encefálico , Disfunción Ventricular Izquierda/sangre , Disfunción Ventricular Izquierda/diagnóstico , Biomarcadores , Insuficiencia Cardíaca/sangre , Insuficiencia Cardíaca/diagnóstico , Insuficiencia Cardíaca/fisiopatología , Humanos , Disfunción Ventricular Izquierda/fisiopatologíaRESUMEN
BACKGROUND: Proteolytic enzymes and oxygen free radicals released from activated leucocytes contribute significantly to the organ dysfunction associated with cardiopulmonary bypass. Leucocyte depletion during extracorporeal circulation should reduce the release of these toxic compounds and thereby improve postbypass myocardial and pulmonary function. Recently, a leucocyte-specific arterial line filter to achieve leucocyte depletion during clinical perfusion has become commercially available. The aim of this study, therefore, was to evaluate the influence of the leucocyte depleting arterial line filter on proteolytic enzyme release, oxygen free radical release and postbypass pulmonary and myocardial function in patients undergoing bypass surgery. METHODS: Forty patients undergoing elective aortocoronary bypass surgery were included into this prospective, randomized clinical study, 20 in the leucocyte depletion (LG-6 group, leucocyte-specific arterial line filter) and 20 in the control group (AV-6 group, standard arterial line filter). White cell count, differential white cell count, plasma elastase concentration, plasma malondialdehyde concentration and C-reactive protein were determined before, twice during and immediately after cardiopulmonary bypass, at the end of surgery and 6 and 20 h thereafter. RESULTS: White cell count, differential white cell count, malondialdehyde and C-reactive protein were not significantly different between LG-6 and control patients. Plasma elastase concentrations were significantly (P < or = 0.03) higher during and immediately after extracorporeal circulation in LG-6 group patients. Need for inotropic support, arterial pO2 after extracorporeal circulation and perioperative CK MB mass and troponin I release were not different between the two groups of patients. CONCLUSION: The use of a leucocyte depleting arterial line filter is associated with an increased release of the proteolytic enzyme elastase, but does not reliably and consistently achieve effective leucocyte depletion during clinical perfusion. In contrast to previous studies, we could not demonstrate any significant difference in postbypass pulmonary or myocardial function between patients perfused with the leucocyte-specific arterial line filter and control patients. Our data do not support the routine use of a leucocyte depleting arterial line filter during clinical perfusion in patients undergoing elective aortocoronary bypass surgery.
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Puente de Arteria Coronaria , Leucaféresis/instrumentación , Péptido Hidrolasas/sangre , Especies Reactivas de Oxígeno/metabolismo , Anciano , Proteína C-Reactiva/análisis , Puente Cardiopulmonar , Cardiotónicos/uso terapéutico , Creatina Quinasa/sangre , Procedimientos Quirúrgicos Electivos , Circulación Extracorporea , Femenino , Filtración/instrumentación , Estudios de Seguimiento , Radicales Libres/sangre , Corazón/fisiopatología , Humanos , Isoenzimas , Recuento de Leucocitos , Pulmón/fisiopatología , Masculino , Malondialdehído/sangre , Persona de Mediana Edad , Oxígeno/sangre , Elastasa Pancreática/sangre , Estudios Prospectivos , Troponina I/sangreRESUMEN
Sixteen patients with heart failure who underwent right heart catherization were studied with regard to plasma natriuretic peptide levels and hemodynamic parameters at rest and immediately after symptom-limited ergometry. Atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) were measured and correlated with left ventricular ejection fraction (LVEF), mean pulmonary arterial pressure (MPAP), pulmonary arterial wedge pressure (PAWP) and cardiac index (CI). Compared to normal controls, ANP and BNP were elevated at rest. During exercise ANP and BNP increased. BNP was highly significantly correlated with LVEF (P = 0.005) at rest, whereas ANP did not (P = 0.082). BNP significantly correlated with MPAP and PAWP after exercise and showed a significant inverse correlation with CI. Our data provide evidence that BNP might be a better indicator for LVEF at rest than ANP. In addition, BNP correlates very well with MPAP and PAWP, after exercise. This close correlation is likely to reflect a correlation of BNP with left ventricular enddistolic pressure in heart failure when patients are exposed to physical exercise.
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Factor Natriurético Atrial/sangre , Ejercicio Físico , Insuficiencia Cardíaca/sangre , Hemodinámica , Péptido Natriurético Encefálico/sangre , Ciclismo , Femenino , Insuficiencia Cardíaca/fisiopatología , Humanos , Masculino , Persona de Mediana EdadAsunto(s)
Biomarcadores/sangre , Gasto Cardíaco Bajo/sangre , Infarto del Miocardio/diagnóstico , Animales , Gasto Cardíaco Bajo/diagnóstico , Creatina Quinasa/sangre , Humanos , Isoenzimas , Persona de Mediana Edad , Mioglobina/sangre , Péptido Natriurético Encefálico/sangre , Troponina I/sangre , Troponina T/sangreRESUMEN
The binding of all known linker histones, named H1a through H1e, including H1(0) and H1t, to a model chromatin complex based on a DNA fragment containing the mouse mammary tumor virus long terminal repeat promotor was systematically studied. As for the histone subtype H1b, we found a dissociation constant of 8-16 nM to a single mononucleosome (210 base pairs), whereas the binding constant of all other subtypes varied between 2 and 4 nM. Most of the H1 histones, namely H1a, H1c, H1d/e, and H1(0), completely aggregate polynucleosomes (1.3 kilobase pairs, 6 nucleosomes) at 270-360 nM, corresponding to a molar ratio of six to eight H1 molecules per reconstituted nucleosome. To form aggregates with the histones H1t and H1b, however, greater amounts of protein were required. Furthermore, our results show that specific types of in vivo phosphorylation of the linker histone tails influence both the binding to mononucleosomes and the aggregation of polynucleosomes. S phase-specific phosphorylation with one to three phosphate groups at specific sites in the C terminus influences neither the binding to a mononucleosome nor the aggregation of polynucleosomes. In contrast, highly phosphorylated H1 histones with four to five phosphate groups in the C and N termini reveal a very high binding affinity to a mononucleosome but a low chromatin aggregation capability. These findings suggest that specific S phase or mitotic phosphorylation sites act independently and have distinct functional roles.
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Ciclo Celular/fisiología , Histonas/metabolismo , Virus del Tumor Mamario del Ratón/genética , Nucleosomas/ultraestructura , Nucleosomas/virología , Regiones Promotoras Genéticas , Secuencias Repetitivas de Ácidos Nucleicos , Células 3T3 , Animales , Cromatina/fisiología , Cromatina/ultraestructura , Cromatografía Líquida de Alta Presión , ADN Viral/aislamiento & purificación , ADN Viral/metabolismo , Fase G1 , Histonas/química , Histonas/aislamiento & purificación , Hígado/ultraestructura , Hígado/virología , Masculino , Ratones , Mitosis , Fosforilación , Isoformas de Proteínas/química , Isoformas de Proteínas/aislamiento & purificación , Isoformas de Proteínas/metabolismo , Fase S , Testículo/ultraestructura , Testículo/virologíaRESUMEN
We investigated the net myocardial release of creatine kinase isoenzyme MB (CKMB), myoglobin, cardiac troponin T (cTnT), cardiac troponin I (cTnI), and cardiac beta-type myosin heavy chain (beta-MHC) into the coronary circulation after cardioplegic cardiac arrest in humans. Cardiac markers were measured in paired arterial, central venous, and coronary sinus blood in 19 patients undergoing elective coronary artery bypass grafting (CABG) before aortic cross-clamping and 1, 5, 10, and 20 min after aortic declamping. cTnT and cTnI were released into the coronary sinus in parallel to each other and almost simultaneously to myoglobin and CKMB within 20 min of reperfusion. In contrast, no beta-MHC was released in the same patients during the study period. The average soluble cTnT and cTnI pools in right atrial appendages of 11 patients with right atrial and right ventricular pressures within reference values were comparable and were approximately 8% of total myocardial troponin content. The soluble beta-MHC pool was <0.1% in all patients. Our results demonstrate the impact of the different intracellular compartmention of regulatory and contractile proteins on their early release from damaged myocardium.
Asunto(s)
Compartimento Celular , Daño por Reperfusión Miocárdica/sangre , Miocardio/metabolismo , Cadenas Pesadas de Miosina/sangre , Troponina I/sangre , Troponina/sangre , Adulto , Anciano , Puente de Arteria Coronaria , Circulación Coronaria , Creatina Quinasa/sangre , Procedimientos Quirúrgicos Electivos , Femenino , Paro Cardíaco Inducido , Humanos , Isoenzimas , Masculino , Persona de Mediana Edad , Daño por Reperfusión Miocárdica/fisiopatología , Miocardio/ultraestructura , Mioglobina/sangre , Troponina TRESUMEN
BACKGROUND: Normovolemic hemodilution is a well-accepted method for intraoperative blood salvage. However, some controversy exists concerning the possible risk of myocardial fiber injury as consequence of the reduced oxygen content. Laboratory diagnosis of perioperative myocardial fiber injury is difficult, since biochemical markers are elevated postoperatively due to the surgical trauma. Cardiac troponin I (cTnI) is a new, highly sensitive and specific marker for the detection of myocardial injury. The aim of our study was to investigate whether normovolemic hemodilution in patients with major orthopedic surgery (13 hemodiluted patients, 15 control) induces a release of cTnI. METHODS: cTnI as a highly specific and sensitive cardiac parameter, as well as total creatine kinase (CK), creatine kinase isoenzyme MB mass (CKMB mass) and myoglobin were measured after induction of anesthesia, after normovolemic hemodilution, prior to retransfusion of blood components, 3 h after surgery, and on the first and third postoperative days. RESULTS: Prior to retransfusion of blood components the hematocrit was decreased to 25.4 +/- 1.2% (mean +/- SEM; range: 18%-34%) in the control group and to 20.2 +/- 0.8% (mean +/- SEM; range: 17%-24%) in the hemodilution group. Total CK, CKMB mass as well as myoglobin concentration increased significantly in both groups, reaching their maxima within the first day of surgery. In contrast, cTnI was below the detection limit of assay (< 0.5 micrograms/L) at any time. CONCLUSIONS: We suggest that pre- and intraoperative hemodilution to a hematocrit of approximately 20% by maintaining normovolemia does not induce myocardial fiber injury in patients without preexisting cardiac diseases.
Asunto(s)
Hemodilución/métodos , Miocardio/metabolismo , Procedimientos Ortopédicos , Troponina I/sangre , Adulto , Biomarcadores/sangre , Transfusión de Sangre Autóloga , Volumen Sanguíneo , Creatina Quinasa/sangre , Estudios de Seguimiento , Hematócrito , Hemodilución/efectos adversos , Humanos , Cuidados Intraoperatorios , Complicaciones Intraoperatorias , Isoenzimas , Persona de Mediana Edad , Fibras Musculares Esqueléticas/patología , Isquemia Miocárdica/etiología , Mioglobina/sangre , Factores de Riesgo , Sensibilidad y EspecificidadRESUMEN
An insertion/deletion polymorphism in the gene coding for the angiotensin-converting enzyme (ACE) is strongly associated with ACE activity. This polymorphism may be a marker for an increased risk for cardiovascular events. Our study examined a possible relationship between the D/I polymorphism and myocardial release of atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP). Ninety-six individuals with normal or impaired left ventricular function were included in the study. ANP and BNP plasma levels were measured at rest and after exposure to physical stress. At rest no association of ACE genotypes with ANP and BNP was found. After exercise homozygotes with the genotype DD had significantly higher ANP plasma levels than homozygotes with the genotype II. In contrast to ANP, BNP levels were not significantly different between genotype groups after exercise. Differences in site of production and mode of release between ANP and BNP might explain this difference. We hypothesize that our result might represent a variability gene effect of the ACE gene locus on endocrine processes in the heart during exposure to physical stress.