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Biomedical subjects

G Ricci

Publications and source records attributed to G Ricci.

At least 289 records · Page 16Linked to original sources

Evidence for glycoprotein abnormality in platelets from patients with May-Hegglin anomaly.

In the present research protein analysis on SDS-polyacrylamide gel electrophoresis (PAGE) has been used to study the glycoprotein pattern of the blood platelets of four members from a family affected by May-Hegglin anomaly. In order to characterize the glycoprotein fractions, lactoperoxidase-iodination and immunoprecipitation procedures were used. N,N'diallyltartardiamide (DATD) cross-linked gel electrophoresis was shown to improve the glycoprotein pattern resolution, although with the lactoperoxidase-iodination the glycoprotein characterization of May-Hegglin platelets overlapped the normal. On the other hand with the immunoprecipitation the specific antiserum precipitated all the five major fractions of normal membrane glycoproteins, but it was shown to react quite poorly with the component V of the May-Hegglin glycoprotein pattern.

Antigens↗

Gas-chromatographic mass-spectrometric detection of 1,4-hexahydrothiazepine-3,5-dicarboxylic acid (cyclothionine) in bovine brain.

Cystathionine has been reported to undergo enzymatic changes leading to the formation of seven membered cyclic products (Ricci, G., Santoro, L., Achilli, M., Matarese, R. M., Nardini, M., and Cavallini, D. (1983) J. Biol. Chem. 258, 10511-10517; Cavallini, D., Costa, M., Pensa, B., and Coccia, R. (1985) Biochem. Int. 10, 641-646). Gas-chromatographic and mass-spectrometric evidence reported in this paper indicates that the cyclic derivative of cystathionine, 1,4-hexahydrothiazepine-3,5-dicarboxylic acid, here simply named cyclothionine, is a normal component of bovine brain. This finding together with the detection of the same compound in the urine of cystathioninuric patients (Kodama, H., Sasaki, K., Mikasa, H., Cavallini, D., and Ricci, G. (1984) J. Chromatogr. 311, 183-188) supports the conclusion that cystathionine, apart from its role in trans-sulfuration, is converted also into cyclic compounds whose biochemical significance is as yet unknown.

Animals↗

1,4-Thiomorpholine-3,5-dicarboxylic acid, a novel cyclic imino acid detected in bovine brain.

Gas-liquid chromatography of enriched bovine brain extract revealed the occurrence of several sulfur-containing compounds. By co-chromatography with authentic product and by mass-spectrometric analysis, one of these compounds has been identified as 1,4-thiomorpholine-3,5-dicarboxylic acid (TMDA). The possible derivation of TMDA from lanthionine is discussed. This represents the second S-containing cyclic amino acid so far discovered in a mammalian brain whose physiological significance has not yet been explored.

Animals↗

In vitro generation of procoagulant activity by Corynebacterium parvum-stimulated mononuclear leukocytes.

Thromboembolic complications are known in cancer patients after i.v. administration of Corynebacterium parvum. We examined the ability of this organism to induce production of procoagulant activity by human blood mononuclear leukocytes in vitro. After 4 hours incubation Corynebacterium parvum was an effective stimulant for mononuclear leukocytes, behaving in the same way as the typical gram-negative bacterium Escherichia coli, whereas mononuclear cells incubated with Staphylococcus aureus were not affected. Corynebacterium parvum used was found devoid of endotoxin by the Limulus assay and was not affected by Polymyxin B, which, on the contrary, inhibited Escherichia coli-induced production of procoagulant activity. Intact Corynebacterium parvum may be required for the production of procoagulant activity and, although this specific aspect of the research will require further study, from the exposed data it is concluded that such a production could be a factor contributing to the pathogenesis of the coagulopathy following Corynebacterium parvum therapy.

Blood Coagulation Tests↗

[Tissue polypeptide antigen: significance and prospective use in the clinical laboratory].

Sequential determination of tissue polypeptide antigen were carried out in patients with neoplastic diseases. Blood specimens were taken before surgery and one week, one month and every months after the operation. The values obtained were compared with those of a control group of blood donors and those of a control group of healthy persons. The values obtained in control groups suggest that alcohol could cause significant increase of tissue polypeptide antigen values.

Adult↗

Enzymatic production of S-(2-hydroxy-2-carboxyethyl)homocysteine.

The monodeamination product of L-cystathionine, S-(2-oxo-2-carboxyethyl)homocysteine is enzymatically reduced by human and hog L-lactate dehydrogenase to S-(2 hydroxy-2-carboxyhethyl)homocysteine which has been found in the urine of cystathioninuric patients. Km values are 6.6 mM and 13 mM for human and hog enzymes respectively. The reaction is not complete at alkaline pH values because of cyclization of the ketoacid into a 1,4 thiazepine derivative ring. By a coupled enzymatic system of L-amino acid oxidase and L-lactate dehydrogenase S-(2-hydroxy-2-carboxyethyl)homocysteine is also formed from L-cystathionine.

Animals↗

Behçet syndrome: report of two early-onset cases treated with transfer factor.

Report of two cases of Behçet syndrome in children, one with the features of a neuro-Behçet syndrome and the other with chiefly severe ocular changes. Both children were treated with transfer factor when previous treatment had proved ineffective. Some years after the beginning of treatment, which is still continuing, transfer factor may be said to be of value in Behçet syndrome.

Behcet Syndrome↗

Determination of rhodanese activity by tetrazolium reduction.

A colorimetric method for the assay of rhodanese activity based on the continuous determination of the sulfite product is described. 5-Ethylphenazinium ethyl sulfate is used as the intermediate electron carrier between sulfite and nitroblue tetrazolium to produce the colored reduced species. The present method is more sensitive than the usual procedure based on the colorimetric determination of thiocyanate. Furthermore, the color developed by nitroblue tetrazolium reduction affords a straightforward means to locate rhodanese activity in polyacrylamide gels.

Animals↗

Continuous spectrophotometric assay of pantetheinase activity.

A continuous spectrophotometric assay for pantetheinase determination using S-pantetheine-3-pyruvate as substrate is described. The enzymatic hydrolysis of this new substrate leads to the formation of S-cysteamine-3-pyruvate, which cyclizes in a non-rate-limiting step to give 2H-1,4-thiazin-5,6-dihydro-3-carboxylic acid (aminoethylcysteine ketimine), a compound exhibiting a strong absorption at 296 nm. The assay is optimized with respect to pH, buffer, and substrate concentration. Prereduction of the enzyme and some properties of the reaction are also studied. The assay is simple, rapid, very sensitive, and specific.

Amidohydrolases↗

Detection of glutathione transferase activity on polyacrylamide gels.

A simple and sensitive assay for glutathione transferase activity on polyacrylamide gel is described. The method is based on the fast reduction of nitroblue tetrazolium salt by glutathione. Blue insoluble formazan colors the gel except in the glutathione transferase area. The stable and defined colorless zone is still detectable with 0.005 unit enzyme. This technique has been successfully applied with enzyme preparations of human heart and other tissues.

Animals↗

Interaction between 1,4-thiazine derivatives and D-amino-acid oxidase.

Aminoethylcysteine-ketimine (2H-1,4-thiazine-5,6-dihydro-3-carboxylic acid) strongly inhibits D-amino-acid oxidase (D-amino-acid:oxygen oxidoreductase (deaminating), EC 1.4.3.3). The inhibition is purely competitive (Ki = 3.3 X 10(-7) M). Aminoethylcysteine-ketimine modifies the visible spectrum of the enzyme: the absorption maxima of bound FAD shift from 375-455 nm to 385-445 nm with a definite shoulder at 465 nm; the appearance of a large absorption band centered at 750 nm may be due to a charge-transfer complex formation. The dissociation constant for the aminoethylcysteine-ketimine-enzyme complex, calculated by a photometric procedure (4 X 10(-7) M), is in good agreement with kinetic data. The dicarboxylic analogue of this inhibitor (lanthionine-ketimine) is ineffective in D-amino-acid oxidase inhibition and does not produce any spectral modification of the enzyme. These results confirm structural requirements for D-amino-acid oxidase inhibitor reported by other researchers. Ketimine reduced forms (thiomorpholine-2-carboxylic acid and thiomorpholine-2,6-dicarboxylic acid) are chemically synthesized and checked as D-amino-acid oxidase substrates: only thiomorpholine-2-carboxylic acid is oxidized to aminoethylcysteine-ketimine (Km = 2 X 10(-4) M).

Amino Acids, Sulfur↗

Similarity of the oxidation products of L-cystathionine by L-amino acid oxidase to those excreted by cystathioninuric patients.

L-Cystathionine is oxidized by snake venom L-amino acid oxidase at a rate about half that with L-leucine at pH 8.5. The appearance of an absorbance at 296 nm and quantitation of the products of oxidation in the presence of catalase indicate formation in the solutions of a seven-membered ketimine ring produced by cyclization of the monoamino monoketo derivative of cystathionine. A limited double deamination has also been observed. In the absence of catalase, S-(carboxymethyl)homocysteine and S-(beta-carboxyethyl)cysteine have been identified together with ninhydrin-unreactive compounds yielding the above mentioned carboxy compounds upon hydrolysis with HCl. Authentic samples of the monoamino monoketo analogs of cystathionine have been prepared and compared with the enzymatic products. Cyclization of the synthetic products into the ketimine ring is pH-dependent as established by UV spectrum and other assays. Compounds derived from either the oxidation or the reduction of the ketimine have been prepared. It was found that many products of enzymatic and chemical changes of cystathionine and its ketimine described in the present paper are identical with those identified in the urine of cystathioninuric patients. This result indicates the occurrence in humans of secondary metabolic routes of cystathionine centered on the production of cystathionine ketimine, in equilibrium with the open form, which in cystathioninurics is revealed by the lack of cystathionase.

Amino Acid Metabolism, Inborn Errors↗

Plasma and erythrocyte fatty acids: a methodology for evaluation of hypocholesterolemic dietary interventions.

A major source of concern in the management of hypercholesterolemia is the careful evaluation of adherence to dietary prescriptions. Routine analysis of plasma fatty acids to monitor adherence of hyperlipidemic outpatients to diets varied in lipid composition is a major task of our research group. Plasma fatty acid patterns are determined by gas-liquid chromatography 60 days after starting an experimental normocaloric diet with 37% fat calories and a polyunsaturated/saturated (P/S) ratio of 3.4. Where poor adherence is observed, the dietary message is administered more incisively or changed accordingly. It is also possible to identify with certainty diet-resistant hypercholesterolemic individuals, i.e., the true nonresponders, who represent only a minority. The determination of erythrocyte fatty acid patterns represents a useful index of adherence in long-term feeding studies and has been extensively carried out in subsamples of persons examined at the fourth rescreening of a controlled trial of primary prevention of coronary heart disease. Increases in mean erythrocyte linoleic/oleic (L/O) and P/S ratios were observed in persons undergoing collective or individual hypocholesterolemic treatment, mean changes being proportional to the different type of nutritional approach. In our opinion, this methodology is the most reliable technique not only for the identification of the different responses to hypocholesterolemic dietary recommendations but also to monitor fat consumption in population samples.

Cholesterol, Dietary↗

Fractionated total body irradiation in marrow transplantation for leukaemia.

Thirty consecutive patients with leukaemia were prepared for bone marrow transplantation (BMT) with cyclophosphamide (CY) 120 mg/kg followed by total body irradiation (TBI). TBI was delivered in a single dose (sTBI) of 10 Gy, at a dose rate of 0.06-0.08 Gy/min, or in fractionated doses (fTBI) of 3.3 Gy/d, on each of 3 consecutive days, at the same dose rate. Lung shielding was adopted for all patients, in order to obtain a homogeneous dose delivered to the lung and at midline. The first 12 patients were prepared with sTBI and the following 18 with fTBI. Several variables (remission status, mode of irradiation, prevention of graft versus host disease (GvHD) with methotrexate (MTX) or cyclosporin A(CyA) were then analysed for their influence on (a) incidence and mortality of GvHD, (b) incidence and mortality of interstitial pneumonia (IP), (c) 100-d survival, and (d) relapses. Our data suggest that fTBI is the single most important factor associated with a lower incidence of IP (P = 0.002), a lower mortality from GvHD and IP (P = 0.03 and 0.001) and a better 100-d survival (P = 0.03). Remission status had no significant influence on GvHD, IP and acute mortality. When compared to MTX, the use of CyA was associated with less GvHD and IP, but when only patients in remission given fTBI were analysed, this was no longer true. Relapses were mostly influenced by the remission status of the patient at transplant (P = 0.001). Although this is not a randomized study, our data suggest that fTBI can reduce the acute risks of marrow transplantation in leukaemia.

Adolescent↗

Pathogenetic relationships between renal tubular acidosis and sodium metabolism alterations in liver cirrhosis.

5 cirrhotic patients with latent distal renal tubular acidosis (RTA) and 5 cirrhotic patients with normal distal acidification ability were studied. All the patients were maintained on a 80 mEq/day sodium diet for at least 5 days before the study. Only 2 of the 5 patients with RTA showed a reduced daily sodium excretion while the 3 other patients with RTA had a normal natriuresis. Acidification ability was retested in all the patients after increasing sodium distal delivery by intravenous administration of ethacrynic acid. Some hours after the maximal natriuretic effect induced by ethacrynic acid, a normalization of the acidification test was seen in all patients with RTA. These findings support the concept that although sodium metabolism alterations are not the only factor in the pathogenesis of RTA in liver cirrhosis, reduced availability of sodium at the distal tubule may impair tubular acidification, probably by influencing the distal transtubular potential difference.

Acidosis, Renal Tubular↗