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Biomedical subjects

M De Rosa

Publications and source records attributed to M De Rosa.

18 recordsLinked to original sources
PubMed1979

5'-Methylthioadenosine phosphorylase from Caldariella acidophila. Purification and properties.

The occurrence of 5'-methylthioadenosine phosphorylase in Caldariella acidophila, a thermophilic bacterium growing optimally at 87 degrees C, is reported. It represents the first example in prokaryotes of a phosphoryolytic cleavage of the thioether. The reaction products, purified by ion-exchange chromatography, have been identified as 5-methylthioribose-1-phosphate and adenine by several analytical procedures. The enzyme has been purified to homogeneity in 32% yield by using DEAE-cellulose and hydroxyapatite chromatography, gel filtration and isoelectric focusing. The enzyme shows a high degree of thermophilicity, its temperature optimum being at 93 degrees C; furthermore no loss of activity is observable after exposure for 1 h at 100 degrees C. The kinetic data indicate a sequential mechanism of the reaction. The apparent Km values are 0.095 mM for 5'-methylthioadenosine and 6.1 mM for phosphate. The specificity of the reaction is rather strict. Experiments performed with analogues of the substrate, i.e. 5'-methylthioinosine, 5'-dimethylthioadenosine sulfonium salt, 5'-n-butylthioadenosine, 5'-isobutylthioadenosine, 5'-isobutylthioinosine, adenosylhomocysteine, 5'-thioethanoladenosine, adenosine, indicate the relevance of the adenine amino group and the sulfur in thioether form in the binding to the enzyme protein.

Adenosine↗
PubMed1979

[Determination of umbilical cord blood Ig and its correlation with prenatal infections (author's transl)].

IgG, IgM and IgA were measured in 120 funicular blood samples. At the same time, these determinations were assayed in the mothers. In the serum samples of the newborn, IgA and IgM were found to have levels over 20 mg/100 ml. Antibodies against rubella virus, cytomegalus virus, Listeria monocytogenes and Toxoplasma gondii were measured in order to investigate a possible intrauterine infection. All Ig determinations were performed through the radial immunodiffusion method.

Adult↗
PubMed1979

[Determination of alpha-1-fetoprotein during pregnancy].

218 samples of serum of pregnant women from 144 normal and 12 risky pregnancy, are tested for AFP RIA, radial immunodiffusion and slide agglutination. The results, according to thet of other AA., as regards un normal pregnancy AFP behaviour, emphasize the necssity to have more data from the same subjects during pregnancy, in order to plot a significant curve for the interpretation of the single values. As regards the technique, the delicacy and the specificity of the research, suggest the use of very sensible methods, which leave simplicity of execution and immediacy of results, out of consideration.

Adult↗
PubMed1978

[5'-methylthioadenosine phosphorylase from Caldariella acidophila. 1. Purification and partial characterization].

5'-Methylthioadenosine phosphorylase has been isolated from C.acidophila, a thermophilic bacterium living in acid hot springs at temperatures ranging from 63 to 89 degrees C. The enzyme has been purified to homogeneity in 32% yield. The enzyme shows a high degree of thermophilicity, its temperature optimum being 93 degrees C in the in vitro assay. The enzyme is exceptionally stable; no loss of activity was observable after exposure for 1 h at 100 degrees C. The optimum pH is about 7,2, with one-half of the maximal activity occurring at pH 6 and 9. The apparent Km for the substrates are: 8,3 x 10(-5) M for MTA and 4,3 x 10(-4) M for phosphate ions.

Adenosine↗
PubMed1978

[Lipid content of aorta, spleen and kidneys in rats on a high fat diet and treated with a chromatographic thymus gland fraction].

A chromatographic fraction obtained from a thymus homogenate of young calf, was administered by intraperitoneal means to a distinct group of rats previously fed on a diet with high content of dietary fat for 50 days. The serum values of cholesterol and triglycerides and of total lipids in spleen, kidneys and aorta which were obtained from these animals were compared with those of rats fed on the same diet and not injected with the thymus fraction. The considerable decrease of the seric values was not followed by conparable results of the lipidic content in the organs.

Animals↗
PubMed1978

The biosynthetic pathway of new polyamines in Caldariella acidophila.

1. Spermidine and sym-nor-spermine (1,11-diamino-4,8-diazaundecane) were identified as the major components of the polyamine pool in Caldariella acidophila, an extreme thermoacidophilic bacterium. A minor component, a new polyamine, sym-nor-spermidine (1,7-diamino-4-azaheptane) was isolated and characterized. 2. To elucidate the biosynthetic pathway, labelled methionine, putrescine, spermidine and spermine were fed to Caldariella acidophila. Incubation of the bacterium in the presence of putrescine or spermidine labelled in the tetramethylene moiety gave unlabelled sym-nor-spermidine and sym-nor-spermine, whereas the radioactivity of propylamine-labelled methionine or spermidine was incorporated into these molecules. No radioactivity was recovered in the polyamines pool when spermine was fed to Caldariella acidophila. 3. S-Adenosylmethionine and S-(5'-adenosyl)-3-methylthiopropylamine were identified as intermediates of the biosynthetic pathway; the cellular contents of the two sulphonium compounds, measured with a new isotope-dilution technique, are 60 and 15nmol/g wet wt. of cells respectively. 4. The above results are indicative of a new pathway characterized by three propylamine-transfer reactions, decarboxylated S-adenosylmethionine being the common donor of the propylamine moiety. The reactions yielding sym-nor-spermidine and sym-nor-spermine are reported for the first time. 5. The probable intermediates related to the catabolism of the tetramethylene moiety of spermidine, gamma-aminobutyraldehyde, gamma-aminobutyric acid or Delta(1)-pyrroline were not detectable. Experiments with [3-aminopropyl-3(n)-(3)H]spermidine trihydrochloride plus [tetramethylene-1,4-(14)C]spermidine trihydrochloride gave rise to an amount of labelled CO(2) equivalent to the spermidine catabolized.

Bacteria↗
PubMed1976

DNA-dependent RNA polymerase from the thermophilic bacterium Caldariella acidophila. Purification and basic properties of the enzyme.

A DNA-dependent RNA polymerase has been isolated from Caldariella acidophila, a thermophilic bacterium living in acidic hot springs at temperatures ranging from 63 to 89 degrees C. The enzyme was purified 180-fold and is composed of five different subunits having the following molecular weights: a = 127000, b = 120000, c = 72000, d = 65000, and e = 38000. The enzyme is activated by Mn2+ and Mg2+ and exhibits optimal activity in the presence of 0.5 mM Mn2+. The activity depends on ionic strength, with a maximum at 0.25 M KCl, and exhibits a pH optimum at 7.8 in the presence of Tris-HCl buffer. The enzyme shows a high degree of thermophilicity, its temperature optimum being 80 degrees C in the in vitro assay. The thermophilicity of C. acidophila RNA polymerase allows studies on enzyme-template interactions to be performed in a temperature range where many templates are close to their Tm.

Bacteria↗
PubMed1976

Stable, inducible thermoacidophilic alpha-amylase from Bacillus acidocaldarius.

Bacillus acidocaldarius Agnano 101 produces an inducible thermoacidophilic alpha-amylase. The enzyme production occurs during the stationary phase of growth in the presence of compounds with alpha-1,4-glucosidic linkages. The enzymatic activity is both present in the culture medium and associated with the cells; the enzymes purified from both sources show identical molecular and catalytic properties. The purified amylase has a single polypeptide chain of molecular weight 68,000 and behaves like an alpha-amylase with affinity constants for starch and related substances of 0.8 to 0.9 mg/ml. The pH and temperature optima for activity are 3.5 and 75degreesC, respectively. The amylase is stable at acidic pH (below 4.5). Its thermal stability is strictly dependent upon protein concentration; the half-life at 60degreesC of the amylase in a 70-mug/ml solution is about 5 days.

Amylases↗