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Biomedical subjects

E Giovannini

Publications and source records attributed to E Giovannini.

At least 37 records · Page 2Linked to original sources
PubMed1995

Solubilization, molecular forms, purification and substrate specificity of two acetylcholinesterases in the medicinal leech (Hirudo medicinalis).

Two acetylcholinesterases (AChE) differing in substrate and inhibitor specificities have been characterized in the medical leech (Hirudo medicinalis). A 'spontaneously-soluble' portion of AChE activity (SS-AChE) was recovered from haemolymph and from tissues dilacerated in low-salt buffer. A second portion of AChE activity was obtained after extraction of tissues in low-salt buffer alone or containing 1% Triton X-100 [detergent-soluble (DS-) AChE). Both enzymes were purified to homogeneity by affinity chromatography on edrophonium- and concanavalin A-Sepharose columns. Denaturing SDS/PAGE under reducing conditions gave one band at 30 kDa for purified SS-AChE and 66 kDa for DS-AChE. Sephadex G-200 chromatography indicated a molecular mass of 66 kDa for native SS-AChE and of 130 kDa for DS-AChE. SS-AChE showed a single peak sedimenting at 5.0 S in sucrose gradients with or without Triton X-100, suggesting that it was a hydrophylic monomer (G1). DS-AChE sedimented as a single 6.1-6.5 S peak in the presence of Triton X-100 and aggregated in the absence of detergent. A treatment with phosphatidylinositol-specific phospholipase C suppressed aggregation and gave a 7 S peak. DS-AChE was thus an amphiphilic glycolipid-anchored dimer. Substrate specificities were studied using p-nitrophenyl esters (acetate, propionate and butyrate) and corresponding thiocholine esters as substrates. SS-AChE displayed only limited variations in Km values with charged and uncharged substrates, suggesting a reduced influence of electrostatic interactions in the enzyme substrate affinity. By contrast, DS-AChE displayed higher Km values with uncharged than with charged substrates. SS-AChE was more sensitive to eserine and di-isopropyl fluorophosphate (IC50 5 x 10(-8) and 10(-8) M respectively) than DS-AChE (5 x 10(-7) and 5 x 10(-5) M.

Acetylcholinesterase↗
PubMed1995

Acetylcholinesterase in tentacles of Octopus vulgaris (Cephalopoda). Histochemical localization and characterization of a specific high salt-soluble and heparin-soluble fraction of globular forms.

Transverse sections of Octopus tentacles were stained for acetylcholinesterase (AChE) activity. An intense staining, that was suppressed by preincubation in 10(-5) M eserine, was detected in a number of neuronal cells, nerve fibres and neuromuscular junctions of intrinsic muscles of the arm. Octopus acetylcholinesterase was found as two molecular forms: an amphiphilic dimeric form (G2) sensitive to phosphatidylinositol phospholipase C and a hydrophilic tetrameric (G4) form. Sequential solubilization revealed that a significant portion of both G2 and G4 forms was recovered only in a high salt-soluble fraction (1 M NaCl, no detergent), Heparin (2 mg/ml) was able to solubilize G2 and G4 forms with the same efficiency than 1 M NaCl. The solubilizing effect of heparin was concentration-dependent and was reduced by protamine (2 mg/ml). This suggests that heparin operates through the dissociation of ionic interactions existing in situ between globular forms of AChE and cellular or extracellular polyanionic components. Interaction of AChE molecular forms with heparin has been reported so far in only a few instances and its physiological meaning is uncertain. G2 and G4 forms, interacting or not with heparin, all belong to a single pharmacological class of AChE. This suggests the existence of a single AChE gene. Amphiphilic and hydrophilic subunits thus likely result either from the processing of a single AChE transcript by alternative splicing (as in vertebrate AChE) or from a post-translation modification of a single catalytic peptide.

Acetylcholinesterase↗
PubMed1995

N,S-bis-fluorenylmethoxycarbonylglutathione: a new, very potent inhibitor of mammalian glyoxalase II.

A very potent competitive inhibitor of mammalian glyoxalase II activity, N,S-bis-fluorenylmethoxycarbonylglutathione (DiFMOC-G) has been synthesized and characterized. The Ki value for inhibition of glyoxalase II purified from calf liver is 0.08 microM. The Ki values for glyoxalase I inhibitions range from 285 to 500 fold higher than the values obtained for glyoxalase II inhibitions, depending on the source of the enzyme. Among other enzymes involved in glutathione metabolism, such as glutathione S-transferase, glutathione reductase, and glutathione peroxidase, only glutathione S-transferase is inhibited to a small extent by DiFMOC-G. Diesters of DiFMOC-G were prepared in order to improve transport of DiFMOC-G into mammalian tumor cells (rat adrenal pheochromocytoma, PC-12) in culture. Among the diesters synthesized, diisopropyl DiFMOC-G was found to be the most inhibitory to cell viability, with a [I]0.5 value of 3 microM.

Animals↗
PubMed1993

Dimeric forms of cholinesterase in Sipunculus nudus.

In developing a research on the cholinesterase (ChE) evolution in Invertebrata, this enzyme was studied in the unsegmented marine worm Sipunculus nudus. ChE activity was solubilized through three successive steps of extraction. These fractions are noted as low-salt (LSS), detergent (DS) and high-salt soluble (HSS) and represent 27%, 68% and 5% of total activity, respectively. LSS and DS ChE were purified to homogeneity by affinity chromatography on edrophonium-Sepharose gel. Purification factors of 1700 (LSS) and 1090 (DS) were obtained. The small amount of HSS ChE prevented a similar purification and an extensive characterization. Based on SDS/PAGE and density-gradient centrifugation, both LSS and DS enzymes show a M(r) value of about 130,000 and are likely G2 globular dimers of a 67,000 subunit. Moreover, LSS ChE seems to be an amphiphilic form including a hydrophobic domain, while DS ChE is probably linked to the cell membrane by a phosphatidylinositol anchor. Both LSS and DS enzymes hydrolyze at the highest rate propionylthiocholine. However, they also show a fairly high catalytic efficiency with other thiocholine esters as substrates, thus suggesting a wide and little-specialized conformation of the active site. Based on immunological cross-reactivity trials, LSS and DS ChE from S. nudus show a reduced structural affinity with a molluscan (Murex brandaris) enzyme. HSS ChE, an acetylcholinesterase, is also solubilized by heparin, like typical vertebrate HSS asymmetric enzymes. However, it lacks fast-sedimenting forms and an enzyme-anchoring collagenous structure.

Animals↗
PubMed1993

Dipyridamole echocardiography evaluation of acute inferior myocardial infarction with concomitant anterior ST segment depression.

The significance of anterior ST segment depression in inferior acute myocardial infarction (AMI) remains controversial. The aim of this study was to relate precordial ST segment depression to the topography of residual myocardial ischaemia, with myocardial mapping of the asynergic area and coronary anatomy. Twenty-five patients with first inferior AMI (15 patients with anterior ST segment depression: group A and 10 patients without anterior ST segment shift: group B), all underwent: (1) electrocardiographic evaluation on admission to the Coronary Care Unit and at 24 h intervals thereafter; (2) 2D-echocardiographic study within 3 h of CCU admission; (3) dipyridamole echocardiographic test (DET) (doses of dipyridamole up to 0.84 mg.kg-1 i.v. over 10 min) 4 days after AMI; (4) coronary arteriography within 14 days from AMI. To assess regional left ventricular wall motion, a 16 segment model was used and a wall motion score index (WMSI) was derived. The results of DET were correlated to the anatomy of the infarct-related vessel. Compared to group B, group A patients showed a significantly greater maximal ST segment elevation in inferior limb leads (lead III: 3.9 +/- 1.9 mm vs 2.2 +/- 1.1 mm, P < 0.05; aVF: 3.5 +/- 1.3 mm vs 1.7 +/- 0.8 mm, P < 0.001). Group A patients showed greater WMSI (1.35 +/- 0.22 vs 1.17 +/- 0.12, P < 0.05), with more frequent postero-lateral wall involvement (72% vs 20%, P < 0.05). No patient of either group showed asynergy of the anterior, anterolateral or anteroseptal segments. No differences in the distribution of coronary artery disease were observed.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1992

[Follow-up of patients undergoing surgery for aortic dissection: evaluation with transesophageal echocardiography].

BACKGROUND: Transesophageal echocardiography (TEE) is a useful means in the diagnosis of acute aortic dissection (AD), owing to its very high sensibility and specificity. In this study, TEE was performed to assess post-surgical evolution. PATIENTS: Between 1982 and 1991, 119 pts. were operated on in our institution for AD (De Bakey I and II type): 87 pts. underwent replacement of the ascending aorta with a composite tubular graft bearing a mechanical valve; 26 had a simple tubular graft and 6 had aortic reconstruction. Sixty-eight of 72 discharged pts. were followed for up to 9.5 years (mean 4.5 +/- 2.6). Nine years after surgery actuarial survival of discharged pts. was 75%. Seven pts. died after a mean period of 3.4 years from surgery: only one died from postoperative complication (dehiscence of proximal anastomosis), none for aortic rupture distal to the graft. TEE was performed in 32 of these pts. and in other two operated on elsewhere, after 4.4 +/- 2.7 years from surgery; before the operation, type I AD was diagnosed in 23 pts. and type II in 11 pts. RESULTS: In 10/11 pts. with type II AD the aortic arch and the descending aorta looked normal; in one patient a localized intimal flap was found up to the arch. The descending aorta diameter was somewhat higher than in normal subjects (25.2 +/- 2.8 vs 21.9 +/- 3.7 mm), but in only one case was it beyond 2DS (32 mm). In all type I pts. an intimal flap persisted distal to the graft, along the whole thoracic aorta. Within the false lumen a flow was detected by color-Doppler in 14/23 pts. (61%), and spontaneous echo-contrast was noted in 14 pts. (61%). A thrombus was observed in 7 pts. (30%) and it was generally localized; in only one case it was extensive with total obliteration of the false lumen. In 16 pts. (70%) communications between the two lumina were found. The descending aorta diameter ranged from 25 to 53 mm, and mean value was higher than in normal subjects (34.2 +/- 6.2 vs 21.9 +/- 3.7 mm). CONCLUSIONS: In most pts. with type II AD, surgery can be a definitive treatment, as the remaining aorta keeps to normal size and appearance. In type I AD, operation is only palliative, as the dissection persists: the false lumen is often perfused through one or more communications with the true lumen and seldom its obliteration is noted. The persistence of dissection does not necessarily seem to be an ominous finding, as the survival of the study population was high and no patient died from aortic rupture. Nevertheless, long-term prognosis can be affected by aorta dilation that often (but not always) follows the persistence of wall dissection. For its high reliability, easy feasibility and low cost TEE is a very useful method for following up patients operated on for AD and for detecting those who are at higher risk of aortic rupture because of lumen dilation.

Adult↗
PubMed1991

[The incidence and clinical implications of left ventricular thrombosis in 769 patients with acute myocardial infarct treated with antithrombotics and fibrinolytics].

Seven hundred sixty-nine patients (pts) admitted to the Coronary Care Unit (CCU) between January 1987 and January 1990 suffering from first acute myocardial infarction (AMI) were studied. The presence of left ventricular thrombosis (LVT) was evaluated by two-dimensional echocardiography (2D-echo). The relation of LVT to site, size and intra-CCU clinical outcome of AMI, in terms of systemic embolic events, Killip class and mortality, was also assessed. AMI was transmural in 707 pts (92%), anterior in 446 pts (58%) and inferior in 261 pts (34%), non-Q in 62 pts (8%). Two hundred sixty-one pts (34%) were treated with IV thrombolytic therapy followed by IV heparin 1000 IU/h over 12 hrs and then calcium heparin (CH) 12500 IU s.c. bid; 508 pts (66%) were given only antithrombotic therapy (CH 12500 IU s.c. bid). 2D-Echo was performed within 48 hours and on day 5-7 from the onset of AMI. In 41 pts (5.3%) LVT was observed: 39 had anterior AMI (8.7% of all anterior AMI pts), one had inferior AMI (0.4% of all inferior AMI pts), and one had non-Q AMI (1.6% of all non-Q AMI pts) [p less than 0.001 for anterior AMI vs inferior and non-Q AMI]. Pts with LVT had a greater infarct size (number of akinetic plus dyskinetic segments/total number of segments x 100) compared to pts without LVT (32.3 +/- 12.6% vs 16.4 +/- 5.7%, p less than 0.001). In pts treated with thrombolytic therapy, LVT incidence was not significantly different from that of pts treated with antithrombotic therapy (4.2% vs 5.9%) alone.(ABSTRACT TRUNCATED AT 250 WORDS)

Chi-Square Distribution↗
PubMed1991

[Echocardiographic evaluation of HIV-positive subjects].

The purpose of the study was to assess the prevalence and the type of cardiac abnormalities in patients with HIV infection. Echocardiographic examination (M-mode, two-dimensional and Doppler) was performed in 51 patients (40 male, 11 female), whose mean age was 29 +/- 10 years; 48 of them (94%) were intravenous drug addicts, 3 (6%) homosexuals. Diagnosis was AIDS in 19 (37%) patients, AIDS related complex in 19 (37%) and asymptomatic infection in 13 (26%). Echocardiography was normal in 13 subjects. Pericardial effusion was found in 19 patients (in 8 of them, this was the only cardiac abnormality). Valve vegetations were found in 16 patients (3 of them had pericardial effusion, 5 had ventricular dilatation or wall motion abnormalities, 1 had both pericardial and myocardial impairment). Myocardial dysfunction was found in 18 patients: 11 had left ventricular dilatation (5 with wall hypokinesia), 1 had right ventricular enlargement, 1 had biventricular dilatation and 5 had only wall motion abnormalities (diffuse or localized). During the follow-up 9 patients died: 8 had AIDS, 1 was asymptomatic. Eight subjects died during hospitalization (none because of cardiac causes) and one at home for sudden unexplained death. Echocardiography had displayed myocardial dysfunction in 6 of them, thickened pericardium in 1 and was normal in 2. Pathologic examination (performed in 8 subjects) showed cardiac enlargement in 3 subjects, thickened pericardium in 2 and valve vegetation in 1. One subject had histopathologic diagnosis of myocarditis and 7 had non specific histologic abnormalities. The study shows a cardiac involvement in 75% of HIV infected patients: 35% had myocardial dysfunction, 37% pericardial disease, 31% infective endocarditis.(ABSTRACT TRUNCATED AT 250 WORDS)

AIDS-Related Complex↗
PubMed1990

Presence of glyoxalase II in mitochondria from spinach leaves: comparison with the enzyme from cytosol.

Glyoxalase II has been purified from cytosol and mitochondria of spinach leaves. Electrophoresis and isoelectric focussing have resolved cytosolic and mitochondrial glyoxalase II in multiple forms: pl 5.3, 5.8 and 6.2 (cytosol) and pl 4.8 (mitochondria). The enzyme of both localizations is a monomer showing a relative molecular mass of about 26 kDa. The values of kinetic constants using several glutathione thiolesters as substrates, are similar for the enzymes from cytosol and mitochondria. These results extend also to plant the presence in mitochondria of peculiar forms of glyoxalase II, likewise recently demonstrated in mammalians.

Cytosol↗
PubMed1989

Isolation of glyoxalase II from two different compartments of rat liver mitochondria. Kinetic and immunochemical characterization of the enzymes.

Two separate pools of glyoxalase II were demonstrated in rat liver mitochondria, one in the intermembrane space and the other in the matrix. The enzyme was purified from both sources by affinity chromatography on S-(carbobenzoxy)glutathione-Affi-Gel 40. From both crude and purified preparations polyacrylamide gel-electrophoresis resolved multiple forms of glyoxalase II, two from the intermembrane space and five from the matrix. Among the thioesters of glutathione tested as substrates, S-D-lactoylglutathione was hydrolyzed most efficiently by the enzymes from both sources. Significant differences were observed in the specificities between the intermembrane space and matrix enzymes with S-acetoacetylglutathione, S-acetylglutathione, S-propionylglutathione and S-succinylglutathione as substrates. Pure glyoxalase II from rat liver cytosol was chemically polymerized and used as antigen. Antibodies were raised in rabbits and the antiserum was used for comparison of the two purified mitochondrial enzymes with cytosolic glyoxalase II by immunoblotting. The enzyme purified from the intermembrane space cross-reacted with the antiserum, but the matrix glyoxalase II did not. The results give evidence for the presence in rat liver mitochondria of two species of glyoxalase II with differing characteristics. Only the enzyme from the intermembrane space appears to resemble the cytosolic glyoxalase II forms.

Animals↗
PubMed1989

Doppler study of precordial musical murmurs.

Using phonocardiography, continuous- and pulsed-wave Doppler, 51 patients with precordial "musical" murmurs (49 with cardiac abnormalities) and 21 patients with noisy murmurs were examined. With M-mode echocardiography, fine fluttering of the structure generating the murmur was evident in 23 patients with musical murmurs and in 5 with noisy murmurs. A continuous-wave Doppler spectral signal characterized by parallel harmonics (Doppler musical signal) was evident in all patients with musical murmurs and in none with a noisy murmur. With pulsed-wave Doppler, the musical signal had less defined spectral features because of range ambiguity. Such a signal was experimentally reproduced by activating a diapason bathed in saline solution. The source of the musical murmur was established in all 51 patients by Doppler. The musical signal was associated with a valvular regurgitation signal in 36 patients and with a ventricular septal defect in 1 patient. The musical signal always disappeared when the pulsed-wave Doppler sample volume was placed 2 cm away from the generating structure. In 11 patients with musical murmur examined by color Doppler, no abnormal bidirectional flow signal was observed in the structures generating the signal. In 6 of the patients without valvular regurgitation, no flow disturbance was found. In conclusion, Doppler is valuable in determining the source of musical murmurs, and musical murmurs are caused by a vibrating structure even in the absence of flow turbulence.

Aortic Valve Insufficiency↗
PubMed1989

Use of defibrotide in the treatment of acute myocardial infarction.

The results of this preliminary clinical trial confirm the thrombolytic effect of defibrotide demonstrated in preclinical models; demonstrate the positive influence of the product on the natural history of early AMI; suggest that the optimal dosage range should include not less than 1.6 gm of defibrotide during the first hour of treatment; and justify further commitment in the study of defibrotide also beyond the scope of treating AMI.

Dose-Response Relationship, Drug↗
PubMed1989

Effects of some S-blocked glutathione derivatives on the prevalent glyoxalase II (a form) of rat liver.

The prevalent glyoxalase II (S-2-hydroxyacylglutathione hydrolase, EC 3.1.2.6, a form) of rat liver cytosol has been studied with a series of seven S-blocked glutathione derivatives. At pH 7.4 and 20 degrees C, only p-nitrobenzyl-S-glutathione was found completely inactive. All the other derivatives are linear competitive inhibitors of the enzyme. Ki values using S-D-lactoylglutathione as substrate are reported. Alkyl-S-glutathiones are weak inhibitors and their inhibition increases with the decrease of the length of the alkyl chain. The best inhibitors are those glutathione derivatives which contain a thioester bond (carbobenzoxy- and p-nitrocarbobenzoxy-S-glutathione) or a carbonyl group (p-chlorophenacyl-S-glutathione). Inhibition by carbobenzoxy-S-glutathione seems to be more complex since the double reciprocal plot shows deviation from linearity at low substrate concentration.

Animals↗
PubMed1988

Demonstration of glyoxalase II in rat liver mitochondria. Partial purification and occurrence in multiple forms.

Glyoxalase II (S-(2-hydroxyacyl)glutathione hydrolase, EC 3.1.2.6), which has been regarded as a cytosolic enzyme, was also found in rat liver mitochondria. The mitochondrial fraction contained about 10-15% of the total glyoxalase II activity in liver. The actual existence of the specific mitochondrial glyoxalase II was verified by showing that all of the activity of the crude mitochondrial pellet was still present in purified mitochondria prepared in a Ficoll gradient. Subfractionation of the mitochondria by digitonin treatment showed that 56% of the activity resided in the mitochondrial matrix and 19% in the intermembrane space. Partial purification of the enzyme (420-fold) was also achieved. Statistically significant differences were found in the substrate specificities of the mitochondrial and the cytosolic glyoxalase II. Electrophoresis and isoelectric focusing of either the crude mitochondrial extract or of the purified mitochondrial glyoxalase II resolved the enzyme activity into five forms with the respective pI values of 8.1, 7.5, 7.0, 6.85 and 6.6. Three of these forms (pI values 7.0-6.6) were exclusively mitochondrial, with no counterpart in the cytosol. The relative molecular mass of the partially purified enzyme, as estimated by Superose 12 gel chromatography, was 21,000. These results give evidence for the presence of mitochondrial glyoxalase II which is different from the cytosolic enzymes in several characteristics.

Animals↗
PubMed1988

[Echo-Doppler study of musical heart murmurs].

The origin of systolic or diastolic musical murmurs was investigated by means of echo-doppler examination in 51 patients with various cardiac diseases. In all cases a typical doppler spectrum was identified, showing bi-directional clusters of frequencies which were concentric in systole and parallel in diastole. The doppler audio signal was musical. A similar echo-doppler signal was obtained by a diapason vibrating in isotonic solution. These data allowed us to identify the site of the vibrating cardiac structure causing the typical echo-doppler spectrum and characteristic audio signal.

Echocardiography↗