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    Immunomodulating activity of heparin: evaluation of structure and physico-chemical properties of heparin

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    Immunomodulating activity of heparin: evaluation of structure and physico-chemical properties of hepari

    Activity of chondroitin ABC lyase and hyaluronidase on free-radical degraded chondroitin sulfate

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    High molecular mass-chondroitin sulfate was characterized for M(r), charge density and constituent disaccharides. This glycosaminoglycan was depolymerized by a controlled free-radical process mediated by hydrogen peroxide in the absence or presence of cupric or ferrous ions. Hydrogen peroxide depolymerizes chondroitin sulfate, and the velocity of the reaction increases in the presence of cupric ions and, further, of ferrous ions. Different low molecular mass-chondroitin sulfate fractions were produced and analyzed by high-performance size-exclusion chromatography and polyacrylamide-gel electrophoresis. This last technique strongly supports the hypothesis that the free-radical process proceeds by the destruction of disaccharide units. The treatment of free-radical chondroitin sulfate samples with chondroitinase ABC and testicular hyaluronidase results in a lower capacity of these enzymes to degrade these glycosaminoglycan derivatives with respect to the natural sample. This was confirmed by polyacrylamide-gel electrophoresis and by the time-courses of enzymatic treatment evaluated by spectrophotometric technique (for treatment with chondroitin ABC lyase)

    QUALITATIVE AND QUANTITATIVE STUDIES OF HEPARIN AND CHONDROITIN SULFATES IN NORMAL HUMAN PLASMA

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    Heparin was extracted and purified from normal human plasma, and full characterization of its structure and physico-chemical properties was achieved for the first time. Plasma was submitted to exhaustive proteolytic treatment with papain, trypsin, chymotrypsin, collagenase and pepsin, anion-exchange chromatography and precipitation with organic solvents. By this procedure, we recovered heparin (about 0.7 mg/100 ml of plasma) and chondroitin sulfate (about 0.1 mg/100 ml of plasma). Chondroitin sulfate has a peak molecular mass of about 15 630, and it is composed of about 60% nonsulfated disaccharide, 3.5% disaccharide 6-monosulfate and about 40% disaccharide 4-monosulfate, with a sulfate-to-carboxyl ratio of 0.41. Heparin, identified by agarose-gel electrophoresis, is constituted by about 40% slow-moving component and about 60% fast-moving species. This glycosaminoglycan had a peak molecular mass of about 7000, and was identified as 'typical' heparin by its constituent disaccharide composition. About 70% of disaccharides were identified as trisulfated disaccharide, and about 18% as disulfated disaccharides, 3% as monosulfated disaccharides and 10% as nonsulfated disaccharide. Heparin extracted from normal human plasma has a high sulfate-to-carboxyl ratio (2.47) and in vitro anticoagulant activity of about 70 I.U. A more quantitative and statistical analysis performed on 10 ml of plasma obtained from 10 human healthy volunteers revealed a heparin level of 0.54 +/- 0.17 mg/100 ml plasma (mean +/- standard deviation) with a coefficient of variation of about +/- 32%. These findings demonstrate for the first time the presence of heparin molecules in normal human plasma and confirm the importance of adequate extraction processes to purify a molecule that strongly interacts with plasma protein components. This is discussed in light of other authors that described a polysaccharide molecule named heparan sulfate in human plasma

    activity of chondroitin ABC lyase on dermatan sulfate partially degraded by cupric-ion-mediated free-radical treatment

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    Dermatan sulfate was extracted and purified from bovine intestinal mucosa, pig intestinal mucosa and pigskin. Small differences in M(r), charge density and constituent disaccharides were detected for the three purified natural dermatan sulfates. Bovine intestinal mucosa dermatan sulfate was depolymerized by a controlled free-radical process mediated by cupric ions in the presence of hydrogen peroxide. Different low-molecular-mass dermatan sulfate fractions were produced and analysed by high-performance size-exclusion chromatography and polyacrylamide gel electrophoresis. The results obtained by this last technique strongly support the hypothesis that the free-radical process proceeds essentially via the destruction of disaccharide units. The partial degradation of dermatan sulfates by cupric-ion-mediated free-radical treatment reduces or even eliminates the capacity of chondroitin ABC lyase to depolymerize these derivatives. This was confirmed by polyacrylamide gel electrophoresis and the time curves of enzymatic treatments evaluated by spectrophotometry

    Studio qualitativo e quantitativo di eparina e condroitin solfato in plasma umano non patologico

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    Studio qualitativo e quantitativo di eparina e condroitin solfato in plasma umano non patologic

    Going Beyond Counting First Authors in Author Co-citation Analysis

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    The present study examines one of the fundamental aspects of author co-citation analysis (ACA) - the way co-citation counts are defined. Co-citation counting provides the data on which all subsequent statistical analyses and mappings are based, and we compare ACA results based on two different types of co-citation counting - the traditional type that only counts the first one among a cited work's authors on the one hand and a non-traditional type that takes into account the first 5 authors of a cited work on the other hand. Results indicate that the picture produced through this non-traditional author co-citation counting contains more coherent author groups and is therefore considerably clearer. However, this picture represents fewer specialties in the research field being studied than that produced through the traditional first-author co-citation counting when the same number of top-ranked authors is selected and analyzed. Reasons for these effects are discussed

    EFFECTS OF MONOCARBOXYLIC AND DICARBOXYLIC-ACIDS ON MYOSIN ATPASE ACTIVITY TESTED BY LUMINOMETRIC PROCEDURE

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    L(+)Lactic acid enhances myosin ATPase in vitro. Different organic acids were tested for activation of myosin ATPase activity. L(+)Lactic is more effective in stimulating ATPase than D(-)Lactic. D(+) and L(-)Malic acids were also effective at the concentration of 2.5 x 10(-2)-5.0 x 10(-2) mmoles/l. At 3.0 x 10(-2) mmoles/l concentration the following acids are activators: acetic, oxalic, malonic, oxaloacetic, pyruvic, glyoxylic, glycolic; succinic is an inhibitor and acetoacetic is without effect. The activation is not in relation with the pKa of these acids. The inhibitory effects of organic acids are evident at the concentration of 5.0 x 10(-2) mmoles/l. This inhibitory effect is linearly increasing with their pKa. The results are discussed in connection with the possible role of these metabolites in controlling not only ATPase activity towards splitting of ATP, but also in controlling the removal of its hydrolytic products

    Effetto di campi elettromagnetici pulsati sulla concentrazione di nucleotidi adenilici in lievito

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    Effetto di campi elettromagnetici pulsati sulla concentrazione di nucleotidi adenilici in lievit

    Effect of low-frequency electromagnetic pulsed field stimulation on yeast fermentation in presence of dicarboxylic and tricarboxylic acids

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    Effect of low-frequency electromagnetic pulsed field stimulation on yeast fermentation in presence of dicarboxylic and tricarboxylic acid
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