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Biomedical subjects

M Chiariello

Publications and source records attributed to M Chiariello.

At least 55 records · Page 3Linked to original sources
PubMed1999

Interaction cloning and characterization of the cDNA encoding the human prenylated rab acceptor (PRA1).

Rab proteins are small GTPases involved in the regulation of intracellular membrane traffic in mammalian cells. In order to find Rab-interacting proteins we performed a two-hybrid screening using a human brain cDNA library. Here we report the isolation of a full-length human cDNA clone coding for a protein of 185 amino acids. This protein interacts strongly with the Rab4b, Rab5a, and Rab5c proteins and weakly with Rab4a, Rab6, Rab7, Rab17, and Rab22 in the two-hybrid assay. Comparison with the Data Bank revealed that this clone represents the human homolog of the previously isolated rat Prenylated Rab Acceptor (rPRA1). Analysis of mRNA expression shows a single abundant mRNA of about 0.8 kb ubiquitously expressed. Western blot analysis of the overexpressed protein shows a band of the expected size equally distributed between cytosol and membranes.

Amino Acid Sequence↗
PubMed1999

Influence of left ventricular cavity size on clinical presentation in hypertrophic cardiomyopathy.

The aim of this study was to assess whether left ventricular (LV) cavity size relates to functional impairment and syncope in patients with hypertrophic cardiomyopathy (HC). LV diastolic dysfunction influences functional limitation in HC. A reduced LV end-diastolic dimension may underlie impaired diastolic properties and be implicated in hemodynamic syncope. Eighty-two consecutive patients with HC (off drugs, in sinus rhythm) underwent echocardiography to measure LV end-diastolic dimension in the short-axis view (indexed to the body surface area) and radionuclide angiography (n = 50) to calculate peak filling rate (normalized to stroke counts/s). Patients in New York Heart Association functional classes II to IV had smaller LV end-diastolic dimension (23.2 +/- 2.6 vs 25.5 +/- 2.5 mm/M2, p = 0.0001) and lower peak filling rate (4.3 +/- 1.4 vs 5.1 +/- 1.3 stroke counts/s, p = 0.036) than those in New York Heart Association class I. LV end-diastolic diameter was correlated to peak filling rate (r = 0.37; p = 0.008). The most potent predictors of functional limitation were LV end-diastolic dimension (relative risk [RR] 0.63, confidence interval [CI] 0.45 to 0.88; p = 0.008), age (RR 1.09, CI 1.03 to 1.17; p = 0.003), and LV thickness score (RR 1.08, CI 1.02 to 1.13; p = 0.003). LV cavity size was smaller in patients with functional limitation irrespective of obstruction and hypertrophy. Patients with differed from those without a history of syncope for a smaller LV end-diastolic dimension (23.2 +/- 2.5 vs 25.0 +/- 2.7 mm/M2, p = 0.008), which was the only independent predictor of syncope (RR 0.77, CI 0.63 to 0.95; p = 0.013). Thus, a small LV cavity size is associated with functional limitation and history of syncope in HC.

Adolescent↗
PubMed1999

Age-related decrease in cardiac tolerance to oxidative stress.

Oxygen radical-mediated injury has been implicated in the process of cardiac aging. We investigated the tolerance to oxidative stress of hearts from rats of different age. Langendorff-perfused hearts from 3-12-month-old rats received a 30 min infusion of the reactive oxygen specie hydrogen peroxide (H2O2, 60 microM), followed by a 15 min wash-out. H2O2 infusion resulted in a significantly larger increase in end-diastolic pressure in hearts of 6- and 12-month-old rats than 3 months (P<0.01). In addition, developed pressure and rate-pressure product decreased more evidently in 12-month-old hearts (P<0.01 vs 3 and 6 months). Arrhythmia analysis showed higher score in hearts of 12- and 6-month-old rats with respect to 3-month-old animals (P<0.05). Cardiac release of oxidized glutathione (an index of the ability of the heart to inactivate oxygen metabolites) was significantly lower in hearts from rats of 6 and 12 months than in younger animals (P<0.001). Finally, cardiac concentrations of the scavenger enzymes glutathione peroxidase and Mn-superoxide dismutase also significantly decreased with age. In conclusion, in rat heart metabolic and functional tolerance toward oxidative stress decreases with age. This phenomenon may contribute to the development of cardiovascular alterations with increasing age.

Aging↗
PubMed1999

Determinants of aortic artifacts during transesophageal echocardiography of the ascending aorta.

BACKGROUND: The origin of artifacts of the ascending aorta during transesophageal echocardiography has not been widely studied. This study was undertaken to investigate in vivo whether anatomic features could determine the appearance of artifacts. METHODS AND RESULTS: Transesophageal echocardiograms of 46 patients studied for suspected dissection with proven diagnosis (30 patients with and 16 without ascending aortic dissection) were reviewed. The incidence of artifacts was 46%, and it was similar in patients with and those without dissection (chi-square 0.516; P = not significant). Artifacts were located in the aortic lumen twice as far from the transducer as the atrial-aortic interface. The aortic diameter was larger in patients with than in those without artifacts (6.4 +/- 1.1 vs 4.2 +/- 0.9 cm, P <.001). An aortic diameter >5 cm and an atrial-aortic ratio </=0.6 predicted the artifact appearance with good sensitivity, specificity, positive and negative predictive values, and diagnostic accuracy; these parameters reached a value of 100% by analysis only of patients without dissection. CONCLUSIONS: An ascending aortic diameter >5.0 cm that exceeds the left atrial diameter with an atrial-aortic ratio </=0.6 creates in vivo the conditions for the reverberation of the atrial-aortic interface within the aorta. Therefore, in patients with such anatomic features, artifacts must be suspected in the presence of linear structures within the aorta.

Aortic Dissection↗
PubMed1999

Activated platelets and leucocytes cooperatively stimulate smooth muscle cell proliferation and proto-oncogene expression via release of soluble growth factors.

BACKGROUND: Previous studies indicate that platelets and leucocytes might contribute to the development of neointimal hyperplasia following arterial injury. The present study was aimed at further investigating the role of platelets and leucocytes, alone or in combination, in promoting vascular smooth muscle cell (SMC) proliferation in vitro, focusing on the relative contribution of different soluble growth factors released by these cells, and on the ability to induce proto-oncogene expression, such as c-fos. METHODS: SMCs from rabbit aortas, made quiescent by serum deprivation, were stimulated with either activated platelets, leucocytes, or both, separated from SMCs by a membrane insert. SMC proliferation was evaluated by measuring the incorporation of 3H-thymidine. The relative contribution of different platelet-derived mediators to SMC growth was evaluated by adding either ketanserin, a 5-HT2 receptor antagonist, R68070, a TxA2 receptor antagonist, BN52021, a platelet activating factor (PAF) receptor antagonist, and trapidil, a platelet derived growth factor (PDGF) receptor antagonist. The role of different leucocyte sub-populations (neutrophils and monocytes + lymphocytes) was also determined in additional experiments. RESULTS: SMC proliferation was significantly increased by activated platelets to 360 +/- 9% of control values (P < 0.05). This effect was reduced by ketanserin, R68070, BN 52021 or trapidil. Whole leucocytes, neutrophils or lymphocytes + monocytes also increased SMC proliferation with respect to control experiments. Simultaneous stimulation of SMCs by platelets and whole leucocytes was associated with a significant greater increase in SMC proliferation as compared to SMC stimulated with platelets or leucocytes alone. c-fos expression, almost undetectable in unstimulated SMCs, was markedly increased by activated platelets or leucocytes. CONCLUSIONS: Activated platelets promote SMC proliferation in vitro via release of soluble mediators, including serotonin, thromboxane A2 PAF and PDGF; activated leucocytes also induce a significant SMC proliferation and exert an additive effect when activated together with platelets; SMCs stimulated with activated platelets and leucocytes show an early expression of the proto-oncogene c-fos.

Animals↗
PubMed1999

Determinants of efficacy of atrial pacing in preventing atrial fibrillation recurrences.

INTRODUCTION: Several studies have shown that single or dual site atrial pacing is effective in reducing the frequency of recurrent atrial fibrillation (AF) in selected patients. However, it is still unclear what the best predictors are of long-term efficacy of atrial pacing. METHODS AND RESULTS: Forty-seven patients with paroxysmal AF requiring demand pacing underwent electrophysiologic study and dual chamber pacemaker implant. After 4 months of follow-up, patients were divided into two groups according to the presence (group 1) or absence (group 2) of symptomatic AF recurrences. Atrial pacing markedly reduced AF recurrences in all patients. Twenty-four patients were free of arrhythmia. The basal state conduction times (CTs) and the incremental conduction times (ICTs), during programmed electric stimulation between the high right atrium (HRA) and the coronary sinus ostium (CSos), but not between the HRA and the His-bundle region, were significantly longer in group 1. There was no statistical difference in the effective refractory period (ERP) recorded at the HRA, the low right atrium (LRA), and the CSos between the two groups, whereas the differences between the greatest and least recorded ERPs measured from the HRA, LRA, and CSos (deltaERP) were significantly greater in group 1 patients. Two parameters were selected by discriminant multivariate analysis, namely deltaCTos (ICT-CT between HRA and CSos) and deltaERP. The first had a greater relative importance in predicting AF recurrence (r2 = 0.33 and r2 = 0.1, respectively). CONCLUSION: Single site atrial pacing is effective in reducing AF recurrences, with decreasing efficacy in patients with greater right atrial conduction delay and wider refractoriness dispersion.

Aged↗
PubMed1999

A network of mitogen-activated protein kinases links G protein-coupled receptors to the c-jun promoter: a role for c-Jun NH2-terminal kinase, p38s, and extracellular signal-regulated kinase 5.

The expression of the c-jun proto-oncogene is rapidly induced in response to mitogens acting on a large variety of cell surface receptors. The resulting functional activity of c-Jun proteins appears to be critical for cell proliferation. Recently, we have shown that a large family of G protein-coupled receptors (GPCRs), represented by the m1 muscarinic receptor, can initiate intracellular signaling cascades that result in the activation of mitogen-activated protein kinases (MAPK) and c-Jun NH2-terminal kinases (JNK) and that the activation of JNK but not of MAPK correlated with a remarkable increase in the expression of c-jun mRNA. Subsequently, however, we obtained evidence that GPCRs can potently stimulate the activity of the c-jun promoter through MEF2 transcription factors, which do not act downstream from JNK. In view of these observations, we set out to investigate further the nature of the signaling pathway linking GPCRs to the c-jun promoter. Utilizing NIH 3T3 cells, we found that GPCRs can activate the c-jun promoter in a JNK-independent manner. Additionally, we demonstrated that these GPCRs can elevate the activity of novel members of the MAPK family, including ERK5, p38alpha, p38gamma, and p38delta, and that the activation of certain kinases acting downstream from MEK5 (ERK5) and MKK6 (p38alpha and p38gamma) is necessary to fully activate the c-jun promoter. Moreover, in addition to JNK, ERK5, p38alpha, and p38gamma were found to stimulate the c-jun promoter by acting on distinct responsive elements. Taken together, these results suggest that the pathway linking GPCRs to the c-jun promoter involves the integration of numerous signals transduced by a highly complex network of MAPK, rather than resulting from the stimulation of a single linear protein kinase cascade. Furthermore, our findings suggest that each signaling pathway affects one or more regulatory elements on the c-jun promoter and that the transcriptional response most likely results from the temporal integration of each of these biochemical routes.

3T3 Cells↗
PubMed1999

[Therapy with nitro derivatives and the development of tolerance: a comparative study with stress ECG and dipyridamole ECG].

The ECG stress test represents the most commonly-used technique to evaluate the occurrence of nitroglycerin tolerance. It acts by increasing cardiac O2 demand with resulting insufficient blood flow through a stenotic coronary artery and development of cardiac ischemia. However, other tests are also potentially suitable, such as the ECG-dipyridamole test. The aim of the present study was to evaluate the acute response of ECG-dipyridamole and ECG-stress tests to nitroglycerin. In particular, the development of nitroglycerin tolerance during chronic therapy was evaluated with both tests in patients with stable angina. Eleven patients (8 men and 3 women) with CAD proven by a previous coronarography, a known history of stable angina within at least six months and a positive response to both the tests were studied. At the end of a seven-day wash-out period, all patients were positive to initial ECG-stress and ECG-dipyridamole tests; after 3 days a new evaluation was carried out (Effort 0 and Dip 0) and this confirmed the previous results. We performed a randomized trial in two phases: acute and chronic therapy. In the acute phase, all patients underwent ECG-stress and ECG-dipyridamole tests (Effort 1 and Dip 1) in a randomized fashion one day apart, four hours after administration of a 10 mg/24 h nitroglycerin patch. The chronic phase consisted of 25 days of continuous treatment with a nitroglycerin patch. The two tests (Effort 2 and Dip 2) were always repeated after four hours of the morning therapy. Nitroglycerin does not modify the hemodynamic response to dipirydamole in either acute or chronic treatment. Lastly, our data confirm the efficacy of nitroglycerin on stress and dipyridamole tests after acute administration. Nitroglycerin tolerance is confirmed by both tests although with different patterns. ECG stress test showed nitroglycerin tolerance because time to ischemia and max ST deteriorated during chronic therapy. Moreover, the ECG-dipyridamole test showed nitroglycerin tolerance because five patients with a negative acute test (Dip 1) became positive during chronic therapy (Dip 2).

Administration, Cutaneous↗
PubMed1999

Rest-injected 201thallium in the evaluation of myocardial viability.

In recent years 201thallium scintigraphy at rest has been used for evaluating myocardial viability in patients with chronic ischemic coronary artery disease and left ventricular dysfunction. Based on the assumption that reversible myocardial dysfunction arises from chronic hypoperfusion (hibernation), resting 201thallium scintigraphy is performed by acquiring two sets of images, one early after tracer injection and a second following 3 to 4 hours to allow for the redistribution process to take place. However, redistribution of 201thallium following injection at rest rarely occurs, and in many studies it does not significantly contribute to the identification of reversibly dysfunctional myocardium. In fact, current interpretation of resting 201thallium scintigraphy is based on the measurement of regional tracer uptake on the redistribution images, using a fixed threshold value (most commonly from 50 to 65% of maximal uptake) that arbitrarily identify viable (presumably reversible) and nonviable (presumably irreversible) dysfunctional myocardium. The practical implication of this approach is relevant as it implies that analysis of a single set of images is adequate for viability information. As with other nuclear techniques, sensitivity of 201thallium scintigraphy for predicting functional recovery following revascularization is very high, but specificity is suboptimal, reflecting the identification of substantial residual tracer uptake in territories that will remain dysfunctional at rest following successful revascularization. Inadequate timing of follow-up functional evaluation, ongoing degeneration of hibernating myocytes, admixture of necrotic and normal myocardium in dysfunctional areas are among factors that likely explain this discrepancy. It remains to be evaluated in future studies whether revascularization of these areas that contain viable myocardium but where resting function does not change, may also contribute to the beneficial effects of revascularization in patients with left ventricular dysfunction.

Cardiomyopathies↗
PubMed1999

Emerging antithrombotic treatments for acute coronary syndromes.

In the last few years the hypothesis of coronary thrombosis, frequently triggered by plaque ulceration or fissuration, has gained wide acceptance as one of the key events in the pathophysiology of acute coronary syndromes. Plaque ulceration may activate both platelets and the coagulation cascade via exposure of a variety of substances, such as von Willebrand factor and tissue factor. It has been demonstrated that aspirin reduces mortality and improves the prognosis of patients with such syndromes. More recently, newer drugs have been identified for the treatment of acute coronary syndromes; in particular, platelet glycoprotein IIb/IIIa inhibitors have been found to be more effective than aspirin in a variety of clinical conditions, such as unstable angina, acute myocardial infarction, and coronary angioplasty. Other drugs with different mechanisms of action will be soon available for large scale clinical trials.

Acute Disease↗
PubMed1998

Signalling of the Ret receptor tyrosine kinase through the c-Jun NH2-terminal protein kinases (JNKS): evidence for a divergence of the ERKs and JNKs pathways induced by Ret.

The RET proto-oncogene encodes a functional receptor tyrosine kinase (Ret) for the Glial cell line Derived Neurotrophic Factor (GDNF). RET is involved in several neoplastic and non-neoplastic human diseases. Oncogenic activation of RET is detected in human papillary thyroid tumours and in multiple endocrine neoplasia type 2 syndromes. Inactivating mutations of RET have been associated to the congenital megacolon, i.e. Hirschprung's disease. In order to identify pathways that are relevant for Ret signalling to the nucleus, we have investigated its ability to induce the c-Jun NH2-terminal protein kinases (JNK). Here we show that triggering the endogenous Ret, expressed in PC12 cells, induces JNK activity; moreover, Ret is able to activate JNK either when transiently transfected in COS-1 cells or when stably expressed in NIH3T3 fibroblasts or in PC Cl 3 epithelial thyroid cells. JNK activation is dependent on the Ret kinase function, as a kinase-deficient RET mutant, associated with Hirschsprung's disease, fails to activate JNK. The pathway leading to the activation of JNK by RET is clearly divergent from that leading to the activation of ERK: substitution of the tyrosine 1062 of Ret, the Shc binding site, for phenylalanine abrogates ERK but not JNK activation. Experiments conducted with dominant negative mutants or with negative regulators demonstrate that JNK activation by Ret is mediated by Rho/Rac related small GTPases and, particularly, by Cdc42.

3T3 Cells↗