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Biomedical subjects

E Marinello

Publications and source records attributed to E Marinello.

At least 127 records · Page 7Linked to original sources

Effects of testosterone on the liver of castrated rats: morphologic study.

It is well known that castration in rats causes atrophy of the sexual organs and, to a lesser extent, of other organs. We have observed that castration causes a reduction of the liver weight due to a reduction (-10%) of the numerical density of the hepatocytes after castration (p less than 0.01). Testosterone administration at high concentration (2.5 mg/100 g b. wt.) as well as at low concentration (0.025 mg/100 g) in the castrated rats causes an increase of the liver weight due to an increase (+30%) in the cellular area (p less than 0.001) as well as in the nuclear area (+20%) (p less than 0.01) of the hepatocytes, revealing a powerful hypertrophic effect of the hormone in the liver.

Animals↗

Purine nucleotide content of lymphocytes from healthy subjects and asymptomatic HIV-1 seropositive patients.

The purine nucleotide content of lymphocytes of normal subjects and asymptomatic HIV-1 seropositive patients was analyzed by HPLC. An increase in IMP and a decrease in ADP, GDP, ATP and GPT were observed in the HIV-infected patients with respect to healthy subjects. The changes may depend on different factors and indicate that the virus influences purine nucleotide metabolism of the cell. The finding sheds light on the metabolic situation of the infected cell, and could be applied to monitoring the disease.

Adult↗

An improved method for purification of L-threonine deaminase from rat liver.

L-threonine deaminase was obtained at a high degree of purity from rat liver. Two main steps were added to the previously reported procedure: cryoprecipitation and chromatofocusing (in the presence of a specific KCl concentration). The purification factor was 3,090 and the specific activity 989. The method is very reproducible and convenient. It gives the highest specific activity and the highest degree of purity of the enzyme recorded to date.

Animals↗

Plasma oxypurines in gastric and colorectal cancer.

Reverse phase high performance liquid chromatography has been used for the determination of plasma hypoxanthine, xanthine, and uric acid in normal subjects and in patients with gastric and colorectal cancer. Plasma oxypurines are significantly elevated in either type of cancer, while uric acid concentration is only higher in gastric cancer. The variations are related to the stage of the tumors, and the physiopathology of their occurrence is discussed.

Adenocarcinoma↗

Influence of testosterone on purine nucleotide turnover in rat kidney.

The influence of testosterone on purine nucleotide metabolism in rat kidney has been investigated in adult and in prepubertal castrated rats. Results have been evaluated through biomathematical model. Castration enhanced the turnover of purine nucleotides in adult rats and reduced it in young castrated rats. Treatment with testosterone in the castrated rats further enhanced nucleotide turnover both in the adult rats and also in the second group, with an oscillatory profile. A clear effect on the inosinic branch point was demonstrated, and specifically on GMP formation, which was opposite according to the age of the animal. The different behavior in the two groups after castration was partially ascribed to the action of other hormones in the absence of testosterone. The observed changes show that the action of the hormone is not limited to sexual organs; they might be at the basis of variations in cellular size and number which probably occur in the kidney after orchiectomy and following androgen administration.

Animals↗

In vitro regulation of rat liver L-threonine deaminase by different effectors.

The regulation of liver L-threonine deaminase by different effectors--bile acids, bile pigments and monocarbon molecules--was investigated. Total inhibition of the enzyme was observed with physiological concentrations of bile acids and biliverdin. Purely competitive inhibition of the holoenzyme by several monocarbon molecules was demonstrated; the mechanism was partially competitive for bicarbonate. Inhibition was more pronounced in the case of the dialyzed enzyme. From the higher Km values for pyridoxal-5'-phosphate (PLP), obtained in the presence of the inhibitors, the results are explained on the basis of interference in the association reaction: apoprotein + PLP----holoenzyme. The various effects determined by bicarbonate may play a specific role in vivo since they occur at physiological concentrations of this compound.

Animals↗

[Enzymes of the inosinic crossing point in human lymphocytes].

At the "inosinic branch point", inosinic acid (IMP) can be channelled either to guanylic acid (GMP) or to adenylic acid (AMP). The 4 enzymes involved in these processes are IMP-dehydrogenase (IMP-DH) and GMP synthetase for the formation of GMP and adenylosuccinate (AMP-S) synthetase and lyase for the formation of AMP. The Authors study the behavior of these enzymes in peripheral blood lymphocytes from normal and leukemic patients. The cells were isolated as previously reported. GMP synthetase was assayed with radiochemical method, IMP-DH and AMP-S synthetase with a radiochemical method coupled to HPLC, while AMP-S lyase was determined following the formation of AMP separated by AMP-S by HPLC, without using labelled precursors. Except for GMP synthetase, which was very low, no activity was detectable in normal lymphocytes; while AMP-S was absent also in leukemic cells, the remaining three activities were well evident. The results open the possibility of using the inosinic branch point enzymes as tumor markers.

Adenylosuccinate Lyase↗

[Purine metabolism: determination of adenyl deaminase in human lymphocytes].

Adenylic acid (AMP) deaminase is a "catabolic enzyme" involved in nucleotide degradation, transforming AMP into inosinic acid (IMP). We present a simple method for the determination of the enzyme activity, which combines high sensitivity with requirement of low quantities of lymphocytes. Human lymphocytes were isolated with a Lymphocyte Separation Medium from FLOW and sonicated. After centrifugation at 2,000 rpm x 10 min and treatment with Norit A, the cells were incubated at 37 degrees C with ATP 0.8 mM and 14C-AMP 0.1 mM (specific activity 12 microCi/mumole) in potassium phosphate 100 mM (pH 7.4). 14C-IMP and 14C-AMP were separated through HPLC by an isocratic elution, with 20 mM KH2PO4 (pH 5.5) at a 1.5 ml/min flow rate. Identification of the nucleotides was carried out through retention time, coelution with internal standards: their evaluation by determining the radioactivity of the collected peaks. The enzyme activity is decreased in patients affected by CLL: the decrease is evident only when data are referred to the single cells and not when they are referred to the protein.

AMP Deaminase↗

[Behavior of the enzymes of the salvage pathway of purine bases in leukemia lymphocytes].

The Authors present a procedure for the determination of adenine phosphoribosyltransferase (APRT) and hypoxanthine phosphoribosyltransferase (HPRT) in lymphocytes which exhibits high sensitivity and requires low quantities of lymphocytes. 5 normal subjects and 4 patients affected by chronic lymphocytic leukemia (CLL) were considered. Human lymphocytes were prepared and treated as previously reported. To the incubation mixtures buffered with 50 mM TRIS-HCl pH 7.4 either 14C-adenine or 14C-hypoxanthine was added: after deproteinization and neutralization we followed the formation of either 14C-adenylic acid (AMP) or 14C-inosinic acid (IMP) by HPLC. A Supelcosil C18 5 microns (250 X 4.5 mm) column was used: IMP was eluted with 20 mM KH2PO4 pH 5.5 while AMP with a linear gradient to 40% B in 20 min., where A was 20 mM KH2PO4 pH 5.5 and B methanol/water 60:40. Evaluation of AMP and IMP formed was carried out by determination of the radioactivity of the collected peaks. The values of APRT in leukemic patients were enhanced when referred to the proteins and those of HGPRT decreased: the Authors propose to complete the study evaluating the intracellular content of adenine and hypoxanthine.

Adenine Phosphoribosyltransferase↗

Double inhibition of L-threonine dehydratase by aminothiols.

The inhibition of highly purified rat liver L-threonine dehydratase (L-threonine hydro-lyase (deaminating), EC 4.2.1.16) by aminothiols (L-cysteine, D-cysteine, cysteamine) has been studied. Single inhibition experiments evaluated by Lineweaver-Burk and Dixon plots showed, in a given concentration range, partially (parabolic) competitive inhibitions, indicating two binding sites for each inhibitor. Double inhibition experiments revealed that the inhibition was antagonistic, the two inhibitors weakening each other's effect. Formation of EI1 and EI2 binary complexes, and ESI1, ESI2 and EI1I2 ternary complexes was demonstrated, while formation of the quaternary complex ESI1I2 was ruled out. It is assumed that one inhibitor-binding site coincides with the substrate-binding center while the second inhibitor-binding (allosteric, regulatory) site may comprise the pyridoxal-phosphate-binding SH group(s). The comparison between Km and Ki values and the evaluation of intracellular concentrations of L-threonine, L-cysteine and cysteamine suggest a possible physiological role of the inhibition.

Animals↗