DNA marker analysis of adult polycystic kidney disease in Italian families. Italian Cooperative Group on ADPKD.
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Adult↗
Biomedical subjects
G Novelli
Publications and source records attributed to G Novelli.
[Construction of an apparatus for pulsed field electrophoresis for the analysis of high molecular weight DNA].
An inexpensive apparatus for Pulsed Field Gel Electrophoresis constructed by the Authors, is described. According to our experimental condition it was found that this apparatus is capable to resolve DNA fragments in order of 50-1500 Kb. This range of analysis is adequate for the molecular studies adjacent regions of prokaryotes, primitive eukaryotes and mammals.
DNA↗
Multilocus analysis of the fragile X syndrome.
A multilocus analysis of the fragile X (fra(X] syndrome was conducted with 147 families. Two proximal loci, DXS51 and F9, and two distal loci, DXS52 and DXS15, were studied. Overall, the best multipoint distances were found to be DXS51-F9, 6.9%, F9-fra(X), 22.4%; fra(X)-DXS52, 12.7%; DXS52-DXS15, 2.2%. These distances can be used for multipoint mapping of new probes, carrier testing and counseling of fra(X) families. Consistent with several previous studies, the families as a whole showed genetic heterogeneity for linkage between F9 and fra(X).
Chromosome Mapping↗
Deletion 2q31.3----2q33.3: gene dosage effect of ribulose 5-phosphate 3-epimerase.
In a new case of interstitial del(2q), measurements of ribulose 5-phosphate 3-epimerase activity suggested that the locus for this enzyme might be localized to the subregion 2q32----q33.3.
Carbohydrate Epimerases↗
Gene dosage studies regionally assign the phosphoserine phosphatase gene to 7p15.1 or 2.
Gene dosage studies in subjects with imbalances of the short arm of chromosome 7 have suggested the localization of the phosphoserine phosphatase (PSP) structural gene to p22.1----pter or p15----p21.2. We have found normal enzyme activity in the fibroblasts from a patient with dup (7) (p15.3----pter). This result refines the assignment of the PSP gene to the 7p15.1----p15.2 sub-bands.
Chromosome Mapping↗
Linkage disequilibrium between cystic fibrosis and linked DNA polymorphisms in Italian families: a collaborative study.
The locus D7S23 includes a CpG-enriched methylation-free island that maps midway between the markers J3.11 and met and is genetically very close to the mutation causing cystic fibrosis (CF). We have studied the linkage disequilibrium between four polymorphic markers from this locus (KM.19, CS.7, XV-2c, and PT-3) and the CF mutation (CF) in 127 Italian families. Strong linkage disequilibrium is found between KM.19, CS.7, and CF, and weaker but significant disequilibrium is found between XV-2c, PT-3, and CF. The disequilibrium between markers and CF for the Italian population provides additional information on the origin and homogeneity of the CF defect. This panel of probes is sufficiently informative to permit accurate prenatal diagnosis of CF in most families with an affected person, and the disequilibrium also allows indirect carrier detection/exclusion in some cases.
Alleles↗
Protocol for prenatal diagnosis of cystic fibrosis based on studies of alkaline phosphatase isoenzymes.
Amniotic fluid analysis of microvillar enzymes, including alkaline phosphatase (ALP) total activity and ALP isoenzymes, has been widely experimented with and used for the prenatal diagnosis of cystic fibrosis in the second trimester of gestation. Since the development of cystic fibrosis molecular analysis, interest in these biochemical tests has been maintained for those instances in which the pregnancy is not fully informative by restriction fragment length polymorphism analysis or DNA is not available from the index-affected child. However, recommended biochemical protocols do not provide clear-cut diagnostic results in a minority of cases. We have tested the reliability of cystic fibrosis biochemical prediction by ALP high-resolution electrophoresis and ALP kinetic studies after inactivation by urea. With this approach, all the amniotic fluid samples that had not been unambiguously classified as affected or unaffected by standard microvillar enzymes analysis were definitely categorized. The proposed method seems to improve the diagnostic accuracy in pregnancies with a one in four risk of resulting in a child with cystic fibrosis.
Alkaline Phosphatase↗
First trimester studies of a fetus at risk for triose phosphate isomerase deficiency.
A first trimester prenatal diagnosis was offered to a mother whose child had died of haemolytic anaemia and multisystem disease caused by TPI deficiency. The deficiency state was characterized by greatly reduced TPI activity in both erythrocytes and peripheral lymphocytes. Specific activity of TPI in trophoblast homogenates from the index fetus was about 30 per cent less than in the controls, but the heat stability test showed overlap. These data were confirmed in uncultured and cultured amniotic cells, where glycolytic intermediate concentrations DHAP, GAP and FDP fell in the range of controls. These results suggested that the fetus was a TPI heterozygote. This prenatal prediction was confirmed by RBC and haematological studies at birth.
Amniocentesis↗
Alkaline phosphatase expression in human chorionic villi.
The physicochemical properties and electrophoretic mobility of different isoforms of alkaline phosphatase were studied in chorionic villi. Based on selective inactivation and inhibition studies (thermal stability, inactivation by urea, EDTA and L(+)ascorbic acid and L-amino acid inhibition), evidence was obtained for the existence of two distinct types of alkaline phosphatase in trophoblast cells. One type is peculiar to chorionic villi while the other is also found in term placenta. Both show two isoforms. These two isoforms were observed with polyacrylamide gel electrophoresis, carried out at pH 6.0 and 9.5. It is suggested that the qualitative and quantitative methods of alkaline phosphatase analysis could be used for first trimester fetal diagnosis of severe infantile hypophosphatasia and for understanding genetic control during early fetal development.
Alkaline Phosphatase↗
Red blood cell glucose metabolism in Down's syndrome.
The specific activity of red blood cell glycolytic enzymes was determined in 20 Down's syndrome patients and compared with 20 normal controls. According to previous evidence, a 50% increase of phosphofructokinase and a 30% increase of glucose-6-phosphate dehydrogenase and glutathione peroxidase activity was found. Metabolic studies of the patients' erythrocytes revealed a decrease in fructose-6-phosphate and 2, 3-diphosphoglycerate concentrations, while fructose-1, 6-diphosphate and ADP both increased. Glucose utilization by intact erythrocytes from Down's syndrome patients did not differ from that of normal controls. However, addition of methylene blue or inorganic phosphate produced a higher stimulation of erythrocyte glycolysis in patients with Down's syndrome compared to controls. These metabolic abnormalities could be, at least in part, ascribed to the increased phosphofructokinase activity which is due to a gene-dosage effect.
Blood Glucose↗
Polymorphic DNA markers linked to cystic fibrosis locus in 20 Italian nuclear families.
We report the haplotype characterization of 20 cystic fibrosis (CF) families for the allelic systems MET-H, D7S16, D7S13 and D7S8. Linkage disequilibrium between MET-H and CF was found in the population investigated. Twelve of the examined families were fully informative, 6 partially informative and 2 completely uninformative.
Alleles↗
The interaction of rifamycin-SV with the hepatic transport and sulfation of taurolithocolic acid in rats.
Sulfated and non sulfated lithocholic acid were analyzed in serum, bile and urine of bile-fistula Sprague-Dawley female rats by gas-liquid chromatography and thin-layer chromatography. In rats (group A) bile was collected during a 3 h saline infusion followed by a 3 h intravenous infusion of taurolithocholate; animals of group B and C were infused with both taurolithocholate and rifamycin-SV, but in the animals of group C the renal pedicles were ligated. Serum bile acids showed a significant increase only in animals of group C, during the infusion of rifamycin-SV. In bile of animals of group A sulfated lithocholic acid represented 18% of total, but was not found in bile of animals of group B and C, while a significant increase of sulfated lithocholic acid was evident in urine of animals of group B. The results indicate that rifamycin-SV probably interferes with the hepatic secretion of sulfated lithocholic acid in the rat.
Animals↗
Increased erythrocyte adenosine deaminase activity without haemolytic anaemia.
A 4-fold increase of red blood cell adenosine deaminase (ADA) activity was found in a patient without haemolytic anaemia, but with mild anisopoikilocytosis. High-performance liquid chromatography showed a 40% reduction of adenosine-5'-triphosphate (ATP) while all the other nucleotides were in normal ranges. The patient's parents (first cousins) and a brother displayed the same enzyme activities as the controls. This observation suggests that mild increases of ADA activity is neither a marker for congenital hypoplastic anaemia as previously reported nor associated with haemolytic anaemia.
Adenosine Deaminase↗
Isoenzymic analysis of fifteen non-lysosomal enzymes in human chorionic villi.
The electrophoretic pattern of 15 non-lysosomal enzymes was established in human chorionic villi sampled at 8-12 weeks of gestation from 30 women requesting legal abortion. The product of 23 genes was detected at this stage of development. Trophoblast cells are mitotic derivates from the zygote. Thus, chorionic villi have the same genetic constitution as the embryo. Establishing the normal isoenzymatic patterns in fetal life is of wide theoretical and biological interest. In particular, it contributes to the understanding of gene modulation and the differential activity of alleloenzymes during early embryogenesis and development. In addition, the possibility is suggested that in the future electrophoresis analysis can be used in conjunction with quantitative enzyme assay for the fetal diagnosis of specific metabolic disorders involving one or more isoenzymes.
Chorionic Villi↗
Effect of chenodeoxycholic acid on the alterations of biliary lipid secretion induced by ethynylestradiol in the rat.
The role of chenodeoxycholic acid (CDCA) in modifying the biliary secretory alterations induced by ethynylestradiol (EE) was evaluated in male Sprague-Dawley rats. Bile flow and bile acid output were both decreased in rats given EE either alone or with CDCA (EE + CDCA) whereas cholesterol output showed a significant decrease only in animals given the combined treatment. The output of phospholipids remained unchanged in all groups. As a result the lithogenic index was significantly decreased in EE + CDCA-treated rats. In urine, the total bile acid excretion was significantly increased in EE + CDCA-treated animals. The data indicate that in the rat, biliary secretory failure induced by EE is not modified by CDCA, but that CDCA may prevent the increase of biliary cholesterol saturation caused by EE.
Animals↗
Increased rate of superoxide ion generation in Fanconi anemia erythrocytes.
The rate of generation of superoxide ion, the concentration of Cu, Zn superoxide dismutase and the hematological parameters were measured in red blood cells obtained from Fanconi anemia patients and from healthy individuals. No significative difference in the superoxide dismutase concentration was found, while the rate of generation of the superoxide ion doubled in Fanconi anemia patients. The steady-state concentration of the superoxide ion was calculated from these data and was found to be 2.3 times higher in Fanconi anemia erythrocytes than in controls. The possible consequences with respect to the alterations in FA are discussed.
Anemia, Aplastic↗
Red blood cell adenine nucleotides abnormalities in Down syndrome.
The red blood cell adenine nucleotides of 20 Down Syndrome patients and 20 healthy controls were determined by a new high-performance liquid chromatography method. All patients showed increased concentrations of adenosine 5'-diphosphate (ADP) and adenosine 5'-monophosphate (AMP), while adenosine 5'-triphosphate (ATP), nicotinamide adenine dinucleotide (NAD+) and nicotinamide adenine dinucleotide phosphate (NADP+) were within normal ranges. This alternation of the energetic charge could be partly responsible for the impairment of glucose metabolism in these patients.
Adenine Nucleotides↗
Hexokinase in human chorionic villi.
The level, intracellular distribution, and isozymic pattern of hexokinase (EC 2.7.1.1) were determined on human chorionic villi obtained by trophoblast biopsy in the first trimester of pregnancy. About 50% of total hexokinase activity was found to be particle-bound and 96% of this in the overt form (i.e. assayable without the addition of detergents). Both soluble and particulate hexokinase show the same affinities for glucose but differ in the affinity for MgATP2- and in their sensitivity to glucose 1,6-diphosphate inhibition. By chromatographic and kinetic methods the soluble hexokinase was found to be represented by isozymes I, II and traces of hexokinase III while most of the bound enzyme was hexokinase type I. These results provide evidence for the expression of at least three hexokinase isozymes in early human development and can partially explain the high rate of glucose utilization of the placenta.
Adenosine Triphosphate↗