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Biomedical subjects

F Gentile

Publications and source records attributed to F Gentile.

At least 73 records · Page 4Linked to original sources
PubMed1990

Invasive adenylyl cyclase of Bordetella pertussis. Physical, catalytic, and toxic properties.

A rapid two-step purification to homogeneity of the calmodulin-activated adenylyl cyclase from urea extracts of Bordetella pertussis organisms (strain 114) is described. Catalytic and invasive activities are purified 30- and 177-fold, respectively, and virtually no degraded forms are found. Specific activities are 0.4 mmol/min/mg and 0.5 mumol/mg of enzyme protein/mg of cell protein/min for catalytic and invasive activities, respectively. The 15 amino-terminal amino acids agree with those deduced from the DNA sequence, as does the molecular mass of 175 kDa (guanidine) or 177 kDa (urea) obtained by equilibrium sedimentation. The larger apparent molecular mass seen in sodium dodecyl sulfate-polyacrylamide gel electrophoresis can be ascribed to anomalous migration. Half-maximal cyclase activation occurs at 3-4 X 10(-10) M calmodulin in the presence of Ca2+ and at 2 X 10(-8) M calmodulin in its absence. Ca2+ activation is maximal at 60-100 microM free CaCl2 (at low calmodulin concentrations), and free Ca2+ concentrations above approximately 125 microM are inhibitory at any calmodulin concentration. Extracellular Ca2+ is essential for intoxication. In Chinese hamster ovary cells, exogenous calmodulin does not inhibit penetration of the cyclase.

1-Methyl-3-isobutylxanthine↗
PubMed1990

The earliest site of iodination in thyroglobulin is residue number 5.

In most highly structured native proteins, as well as in thyroglobulin, the reactivity in vitro of the various tyrosyl residues toward iodine is widely different. The present work demonstrates that of nearly 70 tyrosyl residues present in rat thyroglobulin, there is one, residue number 5 from the NH2-terminal end, which has in vivo the highest affinity toward iodine, being the first one to be iodinated. In fact, when 6-(n-propyl)-2-thiouracil (PTU)-treated, iodine-deficient animals were injected with 125I and killed shortly after, we isolated from thyroid glands poorly iodinated thyroglobulin (about 1 iodine atom/thyroglobulin molecule), nearly 90% of the radioactivity of which was found as monoiodotyrosine. Although CNBr cleavage of this protein gave several fragments after gel electrophoresis only one of these, with apparent mass 27,000 Da, contained 125I. This fragment was isolated and fully characterized. Twelve cycles of automated Edman degradation were performed; the sequence found, i.e. N-I-F-E-X-Q-V-X-A-Q-X-L, indicated that the 27,000-Da fragment is the NH2 terminus of thyroglobulin. This portion of the polypeptide chain contains several tyrosyl residues which may well all be potentially involved in the early iodination of the protein. The observation that the removal of seven amino acids from the NH2 terminus is accompanied (at the fifth step) by the total disappearance of radioactivity in the resulting shortened peptide suggested that the fifth residue was the only one iodinated under these conditions. A second, more quantitative experiment was performed on thyroglobulin obtained from 6-(n-propyl)-2-thiouracil-treated animals whose death was postponed 24 h after the injection of 125I. In this case the radioactivity was found not only in a single CNBr fragment (27,000 Da) but also in other discrete species of lower molecular mass. The mixture of these peptides was subjected to seven steps of manual Edman degradation. Fragments before and after partial degradation were run in parallel on a polyacrylamide gel and the distribution of 125I compared. Besides some change in the background, the two profiles were identical except for the absence of the 27,000-Da species. This proves that all the 125I present in the 27,000-Da species was localized at the fifth residue, the same site at which the hormone molecule is preferentially synthesized under normal conditions. This result is not unexpected and is in accord with the known properties of thyroglobulin which has a polypeptide chain designed for efficient synthesis of the hormone even at low levels of iodination.

Amino Acid Sequence↗
PubMed1990

Autonomic neuropathy and secondary hyperparathyroidism in uremia.

To evaluate the relationship between autonomic neuropathy, and biochemical and radiological parameters of secondary hyperparathyroidism, we examined 19 predialysis and 24 hemodialysis non-diabetic uremic patients. Autonomic neuropathy was assessed using four cardiovascular tests. Ten predialysis and 15 hemodialysis patients showed abnormalities in one or more autonomic tests. Ca, Ca2+, P, alkaline phosphatase, immunoreactive parathyroid hormone and osteocalcin did not significantly differ in uremic patients with and without abnormal autonomic test results. Radiological markers of hyperparathyroidism such as acro-osteolysis (score A) and subperiosteal resorption (score B) were more common in patients with abnormalities in autonomic tests than in patients without. When predialysis and hemodialysis patients were considered separately, the correlation between score A, score B and autonomic neuropathy was confirmed only in hemodialysis patients. In conclusion, autonomic neuropathy is not related to biochemical parameters of hyperparathyroidism, while it appears correlated with radiological signs of osteodystrophy, suggesting a possible pathogenetic link between autonomic neuropathy and secondary hyperparathyroidism.

Adult↗
PubMed1990

[A comparison between mitral and aortic outputs evaluated by Doppler echocardiography in a group of healthy subjects: false regurgitation].

Doppler echocardiography is a potentially useful tool for the non invasive evaluation of cardiac output and, therefore, for the quantitative assessment of valvular regurgitation. The aim of our study was to establish the presence of possible pitfalls in the evaluation of mitral and aortic regurgitant fraction obtained by Doppler echocardiography comparing the cardiac output measured at the level of the mitral and aortic valve. For this purpose 19 healthy volunteers, aged between 14-68 years, were studied. Stroke volume and cardiac output were calculated at the level of the mitral and aortic valve. The methods we used for the measurement of both the mitral and aortic cardiac output had already been validated and presumes that the shape of the valve annulus, is circular. No statistically significant differences were found between the parameters obtained at the two different valvular levels. Furthermore, cardiac output values correlated fairly well (r = 0.83, ESS = 0.78 l/min). In 9 subjects the aortic cardiac output was greater than the mitral one, while in the others mitral cardiac output was greater. The average of the differences between the two cardiac outputs was 0.58 +/- 0.48 l/min with a regurgitation fraction of 9.5 +/- 7.9%. Our results show that the mitral and aortic stroke volume and cardiac output, as measured by Doppler echocardiography (considering a circular shaped valve annulus, are not statistically different and correlate fairly well in our normal subjects. Nevertheless, we observed a certain degree of variability between the mitral and the aortic cardiac output.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMed1989

Prevalence of valvular regurgitation by Doppler echocardiography in patients with structurally normal hearts by two-dimensional echocardiography.

The prevalence of valvular regurgitation in a large population with structurally normal hearts remains unknown. From the computer database of the echocardiography laboratory of the hospital, 7209 records containing results of both two-dimensional and Doppler echocardiographic examinations were identified, from which 867 (12%) with no structural abnormality on two-dimensional echocardiograms were obtained for analysis. Of these 867 records, 291 (34%) had evidence of regurgitation by Doppler technique in at least one cardiac valve. Mitral regurgitation was found in 167 (19%), tricuspid regurgitation in 151 (17%), pulmonic regurgitation in 45 (5%), and aortic regurgitation in 29 records (3%). Regurgitation of just one valve was the most common and occurred in 207 records (24%). This was followed by regurgitation of two valves (69 records, 8%), three valves (13 records, 2%), and four valves (two records, 0.2%). The prevalence of mitral, tricuspid, and aortic regurgitation was found to increase significantly with increasing age, as was the prevalence of regurgitation involving multiple valves. In 98% and 95% of mitral and tricuspid regurgitations, respectively, the jets were confined to the proximal one fourth of the atria, suggesting only trivial or mild regurgitation. Thus valvular regurgitation occurs not uncommonly in patients with structurally normal hearts referred for echocardiographic evaluation. These findings caution against the inappropriate diagnosis of clinical disease in the many patients who fall into this category. The increasing prevalence of valvular regurgitation with increasing age suggests that a wear-and-tear phenomenon rather than a congenital cause is involved in most instances.

Adolescent↗
PubMed1989

Modulation of invasiveness and catalytic activity of Bordetella pertussis adenylate cyclase by polycations.

Penetration of Bordetella pertussis adenylate cyclase into CHO cells was monotonically inhibited by polylysines, with a minimum degree of polymerization of greater than 6 and less than or equal to 9 to 10. Above this level, inhibitory potency per lysyl residue was independent of polymer length; 50% inhibition was obtained with 60 microM lysine monomer. Other polycations were also potent inhibitors. The adenylate cyclase itself showed a biphasic (stimulation-inhibition) response, with a similar independence of polymer length above a certain minimum. Half-maximal inhibitory concentrations for cyclic AMP accumulation corresponded to half-maximal stimulatory concentrations of poly-L-lysine for the cyclase. The inhibitory effect of polylysines on cyclic AMP accumulation was not reversed by washing or enzymatic removal of neuraminic acid. We conclude that charge-charge interactions play an important role in the penetration of B. pertussis adenylate cyclase into host cells.

Adenylyl Cyclases↗
PubMed1989

Characteristics and mechanism of action of a heat-stable enterotoxin produced by Klebsiella pneumoniae from infants with secretory diarrhea.

Escherichia coli heat-stable enterotoxins (ST) are classified into STa and STb according to their physicochemical and biologic characteristics. STa induces diarrhea, activating the guanylate cyclase-cGMP system. ST-like enterotoxins can be produced by bacteria other than E. coli, including Klebsiella pneumoniae. A Klebsiella ST has previously been shown to share some chemical and immunologic characteristics with E. coli ST. Aiming to define better the nature of Klebsiella ST, we have screened 237 children with diarrhea and 179 controls for ST-producing Klebsiella, using the SMA. We detected 26 Klebsiella strains from patients, two of which were positive in the SMA, and 36 from controls, all negative for ST. A partial purification was performed using an acetone precipitation followed by ultrafiltration and gel filtration techniques. Klebsiella toxin was heat-stable, methanol-soluble, sensitive to mercaptoethanol, active at acid pH values, but not at pH greater than 8. The time course of Klebsiella toxin in the SMA resembled that of E. coli STa. Klebsiella ST caused reduced Na absorption and net Cl secretion in rabbit ileal mucosa mounted in Ussing chambers. It was found to increase the cGMP but not the cAMP concentration. Finally, Klebsiella ST did not react with anti-E. coli STa MAb in a competitive ELISA. We conclude that K. pneumoniae may induce diarrhea through the production of an STa similar but not identical to E. coli STa.

Bacterial Toxins↗
PubMed1989

[Changes in the brain content of the gonadotropin releasing hormone (GnRH) in caribe colorado, Pygocentrus notatus, during the reproductive cycle].

Immunoreactive gonadotrophic hormone-releasing hormone (ir-GnRH) was detected in extracts from optic tectum, cerebellum and brain stem of the Venezuelan freshwater fish "caribe colorado", Pygocentrus notatus by means of radioimmunoassay. Like most fishes from the area P. notatus, has a reproductive cycle closely related to environmental conditions changes. Our results show a correlation between gonadal maturation, express as Gonosomatic Index (GI) and the beginning of the rainy season (April-May) in males and females of this species. Fluctuations in the ir-GnRH levels that depend of the reproductive state and environmental conditions occurred in the optic tectum and cerebellum extract obtained from female fish. On the other hand, in males no striking variation were found in any of the structures mentioned, except for the optic tectum, were a significant increase in the ir-GnRH content was observed in May.

Animals↗
PubMed1988

Extracellular cAMP formation from host cell ATP by Bordetella pertussis adenylate cyclase.

The effect of exogenously added adenylate cyclase from Bordetella pertussis (strain 114) has been investigated in Y-1 mouse adrenal tumor, chinese hamster ovary (CHO) and several other cells. A partially purified adenylate cyclase was found not to enter cells but, nevertheless, produced large amounts of cAMP in the medium. We could show that this resulted from release of ATP (and not larger molecules). The ATP released by the cells could be (1) directly measured and was replenished after each change of medium; (2) was reciprocally related to the cAMP produced; and (3) was competed for by ATPases present in added serum or by hexokinase and, less effectively, by exoenzymes on the cell surface. The extent of ATP leakage varied widely between different cell lines, being marked in CHO and Y-1 adrenal cells but negligible in transformed lymphocyte lines. The uncertainty of the origin of cAMP found in media of cultured cells requires separate analysis of cell and medium cAMP and an assessment of ATP leakage.

Adenosine Triphosphate↗
PubMed1988

Bordetella pertussis adenylate cyclase. Penetration into host cells.

Exposure of Chinese hamster ovary, mouse adrenal cortex tumor (Y-1), THP-1 and U-937 cells and human erythrocytes to adenylate-cyclase-containing urea extracts of Bordetella pertussis (strain 114) organisms promotes the formation of large concentrations of intracellular cAMP. Accumulation is dependent on dose and temperature, with significant accumulation occurring at 4 degrees C, and is virtually instantaneous, with a doubling at 1 min. There is an absolute Ca2+ requirement but external calmodulin (the activator of cyclase activity) has no effect except in erythrocytes and U-937 cells, where it reduces cAMP accumulation. However, calmodulin antagonists inhibit cAMP accumulation. In Y-1 adrenal cells the urea-extract adenylate cyclase stimulates steroidogenesis. Anti-(B. pertussis) antibodies inhibit cyclase activity and prevent further cAMP accumulation after 10 min in cells previously exposed to urea extract. The same effect is obtained by washing. This suggests that a portion of the cyclase is associated with cells in a form not accessible to antibody or washing but accessible to substrate, which we interpret as internalized enzyme with a short lifetime. Continuing cAMP accumulation thus appears to need a continuing source of external cyclase. Inhibitors of the effect of diphtheria toxin, such as NH4Cl, methylamine, chloroquine or monensin, have no inhibitory effect on the accumulation of intracellular cAMP promoted by the internalized adenylate cyclase of urea extracts of B. pertussis organisms. We conclude that entry of the cyclase into cells is not by receptor-mediated endocytosis.

Adenylate Cyclase Toxin↗
PubMed1988

Sch 37137, a novel antifungal compound produced by a Micromonospora sp. Taxonomy, fermentation, isolation, structure and biological properties.

A new antifungal compound, Sch 37137, was isolated from the cultured broth of a Micromonospora sp., SCC 1792. Sch 37137 is structurally related to A 19009, a dipeptide previously discovered from a Streptomyces sp. The compound was weakly active against species of Candida and dermatophytes (mean MICs greater than or equal to 128 micrograms/ml) in Sabouraud dextrose, yeast-nitrogen and modified Eagles minimum essential media; however activity against Candida sp. (mean MICs greater than or equal to 12 micrograms/ml) and dermatophytes (mean MICs greater than or equal to 0.8 microgram/ml) significantly improved in MA medium.

Amino Acids, Diamino↗
PubMed1988

[Radiological study of the duodenum in celiac disease in adults].

Celiac disease (CD) is the most common form of malabsorption in childhood when it presents with diarrhea and growth failure, a jejunal biopsy is considered the first diagnostic investigation by some authors. In adulthood, clinical symptoms of CD may mimic several different disease, such as peptic ulcer and IBS, and the first diagnostic investigation is an upper GI series. Radiological features of duodenum and small bowel were evaluated in twenty patients with adult onset celiac disease. Sign of duodenitis such as thickened folds, mucosal nodules, dilatation of duodenum and erosions were observed in 19 out of our 20 patients (95%); particularly, thickened folds in 17 (85%), nodularity in 16 (80%), duodenal dilatation in 12 (60%) and erosions in 4 (20%). In celiac disease the lesions are more severe in the upper part of small bowel, and duodenitis may be the unique sign of CD. Duodenitis may be part of a disease involving the entire small bowel; so, a duodenitis observed in the upper gastrointestinal tract requires the study of the entire small bowel--which seems to be very important in the case of celiac disease.

Adolescent↗
PubMed1987

A novel tetracycline from Actinomadura brunnea. Fermentation, isolation and structure elucidation.

A novel tetracycline antibiotic, Sch 33256, was isolated from a culture broth of a new species of Actinomadura. The antibiotic was isolated by solvent extraction, Sephadex G-25 column chromatography and crystallization. The structure was determined by comparison of the spectra with that of chlortetracycline. Spectroscopic analysis of the compound yielded 2'-N-methyl-8-methoxychlortetracycline as the proposed structure.

Bacteria↗
PubMed1987

[Echocardiographic aspects of false intraventricular tendons].

A series of 4260 consecutive echocardiographic examinations, performed in 5 Cardiological Centers was examined, identifying 125 intraventricular false tendons (FTs) in 100 cases (55 normals and 45 with heart disease) of whom 31 were female and 69 male, aged 3 to 82 years. An anatomo-morphological study was possible in 9 cases, dead for stroke or heart failure, on autopsy, and in 7, on heart surgery. A phonocardiogram was performed in all normals and in 20 patients. On echocardiography, FTs appeared as an echo-producing string-like structure, straight between the septum and the ventricular free wall, mobile during the cardiac cycle, without systolic thickening and any relation with the atrioventricular valvular apparatus. The prevalence of FTs was 2.3%; it was 3.2% to 5.3% in younger people. FTs were located in the right ventricle (4 cases), left ventricle (95 cases) or in both (1 case). Their site was left apical (45 FTs), right apical (2 FTs), right (3 FTs) and left (20 FTs), upper septum-to-free wall (55 cases). In 1 case hypertrophy of trabeculae of the left ventricle was detected. FTs were single (79 cases), double (19 cases), multiple (2 cases), short (42 cases), long (58 cases), thick (45 cases) and thin (55 cases). They showed a membrane-like motion (thick FTs-45 cases) and a valve-like motion (thin FTs-55 cases). Innocent murmur was detected in 50 of 55 normals and related to thin FTs. Of 16 cases examined anatomically and histologically, FTs were fibrous in their distal portion and fibro-muscular in the proximal one in 12 cases, whereas they were entirely fibrous-muscular in 4 cases. The site and location of FTs detected by echo were confirmed by anatomy in all cases. In 4 cases other 9 FTs, not detected on echocardiography, were found. These data suggest that echocardiography is a useful tool to detect intraventricular FTs and differentiate them from other echo-producing structures. Although a relationship between FTs and heart disease has not been found, their presence could be responsible of innocent murmur in many normal subjects.

Adolescent↗
PubMed1987

[2-dimensional echocardiography in infarct of the right ventricle: correlation with clinical, hemodynamic and scintigraphic findings].

Right ventricular wall motion was studied in 18 patients hospitalized for acute myocardial infarction by means of cross-sectional echocardiography. Diagnosis of right ventricle myocardial infarction was made on the basis of clinical, electrocardiographic, hemodynamic criteria, respectively present in 66%, 80% and 66% of our patients. Echocardiographic criteria of right ventricle infarction were: 1) right ventricle dilatation; 2) segmental or global right ventricle wall motion abnormalities; 3) paradoxical motion of interventricular septum; 4) tricuspid insufficiency; 5) inferior cava vein dilatation with paradoxical respiratory pattern. Seven patients were studied by means of radionuclide techniques, that confirmed right ventricle infarction in 6 patients. We conclude that: 1) the most accurate echocardiographic views for right ventricle wall motion study are subcostal views and, in particular, the subcostal short-axis (OAS 60 degrees); 2) those patients (12/18) with a specific echocardiographic pattern for right ventricle infarction presented a specific clinical and hemodynamic picture; 3) those patients (6/18) with only segmental right ventricular wall motion abnormalities presented an aspecific clinical and hemodynamic picture. Moreover we showed that an improvement of right ventricular wall motion was observed in some cases within a few days of the infarction. Further observations are needed to assess the frequency and late prognosis of acute right ventricle infarction.

Adult↗
PubMed1987

Hypersideremic and hyperbilirubinemic effect of nicotinic acid in patients with Gilbert's syndrome.

Nicotinic acid test (NA) administration is followed by a rise in unconjugated bilirubin and serum total iron in healthy man. A similar concomitant effect has never been investigated in Gilbert's syndrome (GS) patients, who by definition have a hyperbilirubinemia higher than that of controls. The aim of this study was to verify whether, in addition to the hyperbilirubinemic action, NA could induce parallel hypersideremia in GS subjects. The data we obtained confirm in GS patients: 1. the well-known hyperbilirubinemia; 2. a delayed NA plasma concentration curve, and document that in GS the hypersideremic effect is comparable to that of controls. Previous investigators reported that NA produces an equimolar rise of bilirubin and iron levels in healthy man. In our study the extent of the sideremic response to NA occurring in GS patients is comparable to that seen in controls, and appears unrelated to the bilirubinemic values. If the NA effects are based on the documented hemolytic properties of NA, a disturbance in bilirubin handling by the liver of GS subjects appears to be plausible in the presence of almost normal behavior of NA-induced hypersideremia.

Adolescent↗