Showing papers by "Francesco Salvatore published in 1997"

Genetic history of cystic fibrosis mutations in Italy. I. Regional distribution

Abstract: Earlier analysis of the Italian population showed patterns of genetic differentiation that were interpreted as being the result of population settlements going back to pre-Roman times. DNA disease mutations may be a powerful tool in further testing this hypothesis since the analysis of diseased individuals can detect variants too rare to be resolved in normal individuals. We present data on the relative frequencies of 60 cystic fibrosis (CF) mutations in Italy and the geographical distribution of the 12 most frequent CF mutations screened in 3492 CF chromosomes originating in 13 Italian regions. The 12 most frequent mutations characterize about 73% of the Italian CF chromosomes. The most common mutation, delta F508, has an average frequency of 51%, followed by N1303K and G542X, both with average frequencies around 5%. Multivariate analyses show that the relative frequencies of CF mutations are heterogeneous among Italian regions, and that this heterogeneity is weakly correlated with the geographical pattern of non-DNA 'classical' genetic markers. The northern regions are well differentiated from the central-southern regions and within the former group the western and eastern regions are remarkably distinct. Moreover, Sardinia shows the presence of mutation T338I, which seems absent in any other European CF chromosome. The north-western regions of Italy, characterized by the mutation 1717-1G-->A, were under Celtic influence, while the north-east regions, characterized by the mutations R1162X, 2183AA-->G and 711 + 5G-->A, were under the influence of the Venetic culture.

Lung cancer metastatic cells detected in blood by reverse transcriptase-polymerase chain reaction and dot-blot analysis.

Abstract: PURPOSEWe analyzed the blood of patients with lung cancer at different stages of presentation for the presence of carcinoembryonic antigen (CEA) mRNA detected by reverse transcriptase-polymerase chain reaction (RT-PCR) combined with the dot-blot procedure as an indicator of micrometastatic malignant cells.PATIENTS AND METHODSWe studied 24 lung cancer patients (10 with distant metastases and 14 with no evidence of distant metastases), eight age- and sex-matched patients affected by nonneoplastic respiratory diseases (four smokers), and eight healthy subjects. We used immunohistochemistry and RT-PCR dot-blot analysis to evaluate CEA expression in the neoplastic tissue, and the RT-PCR dot-blot procedure to analyze CEA mRNA in circulating cells.RESULTSThe RT-PCR dot-blot procedure was highly sensitive aspecific: it detected CEA mRNA in samples of RNA from lung cancer diluted 10(6)-fold with RNA extracted from normal blood cells, and sequence analysis confirmed that the amplified product was CEA. CEA mRNA was ...

JURL-MK1 (c-kit high /CD30 − /CD40 − ) and JURL-MK2 (c-kit low /CD30 + /CD40 + ) cell lines: ‘two-sided’ model for investigating leukemic megakaryocytopoiesis

Abstract: Two novel cell lines (JURL-MK1 and JURL-MK2) have been established from the peripheral blood of a patient in the blastic phase of chronic myelogenous leukemia. The cells grow in a single cell suspension with doubling times of 48 h (JURL-MK1) and 72 h (JURL-MK2). Cytogenetic analysis has shown that JURL-MK1 is hypodiploid whereas JURL-MK2 is near triploid and that both cell lines retain t(9;22). Moreover, JURL-MK1 and JURL-MK2 have a bcr/abl-fused gene with the same junction found in the patient's fresh cells, and both cell lines express the b3/a2 type of hybrid bcr/abl mRNA. The morphology and immunophenotype of these cell lines are reminiscent of megakaryoblasts. In both lines, a limited but consistent percentage of cells expresses gpIIbIIIa (CD41a), gpIIIa (CD61) and CD36, with no expression of gplb (CD42b), glycophorin A, hemoglobin and CD34. Both cell lines are clearly positive for CD33, CD43, CD45RO and CD63, while CD13, CD44, CD54, CD30 and CD40 are specific features of JURL-MK2. Among cytokine receptors, CD117/SCF-R is strongly displayed by a large fraction of JURL-MK1 cells but is hardly detectable on about 20% JURL-MK2 cells. Both cell lines are clearly positive for CD25/IL2R alpha, while a marked expression of CD116/GM-CSF-R and CDw123/IL3R alpha is restricted to JURL-MK2. Induction of cell differentiation in vitro has demonstrated that TPA is able to modulate the JURL-MK1 phenotype, causing an increased expression of platelet-associated antigens. The JURL-MK2 phenotype is easily modulated by both TPA and DMSO, which cause an increased expression of CD41a and CD117 accompanied by a decreased expression of CD30. Proliferation studies demonstrated that JURL-MK1 cell growth is enhanced by stem cell factor, while JURL-MK2 proliferation is unaffected by this cytokine. JURL-MK1 and JURL-MK2 are two novel cell lines with divergent biological features, representing a 'two-sided' model for investigating new aspects of megakaryocytopoiesis.

Rapid identification of HLA DQA1*0501, DQB1*0201, and DRB1*04 alleles in celiac disease by a PCR-based methodology

Abstract: Celiac disease (CD) is an autoimmune disorder associated with a small bowel lesion induced by toxic gliadin components [1 , 2] . In this condition, an antigen peptide from α-gliadin, corresponding to the amino acid sequence between residues 31 and 49, initiates the cellular immune response, which is mediated by gliadin-specific T lymphocytes [3] . Antigen recognition by T lymphocytes in CD mainly occurs if the gliadin-derived peptides are carried by the HLA class II molecules HLA-DQ2 or HLA-DR4 [4 , 5] . Thus, the genetic susceptibility towards CD derives from inheriting certain HLA class II alleles encoding for the above-mentioned specific molecules. Among the CD-HLA associations that have been described so far, the one caused by the presence of alleles DQA10501/DQB10201 (encoding for the DQ2 molecule) is present in most cases, whereas the DRB104 alleles (encoding for the DR4 molecule) occurs almost invariably in the other cases [5 , 6] . In fact, these two associations characterized >95% of the affected celiac patients in European populations [7] . The aim of our study was to improve a PCR-based methodology for the rapid typing of the DQA10501, DQB10201, and DRB104 alleles, and thus provide an additive simple tool for the diagnosis of CD. DNA was extracted from 5 mL of fresh whole blood in EDTA by proteinase K …

Severe liver impairment in a cystic fibrosis‐affected child homozygous for the G542X mutation

Abstract: The clinical and laboratory findings of a cystic fibrosis (CF) patient homozygous for the G542X mutation are described. This is the first case, among the 7 G542X homozygous CF subjects described so far who shows severe liver involvement, associated pancreatic insufficiency, and moderate pulmonary expression of the disease, as demonstrated by laboratory and imaging data. This case adds to the conclusion that genotype/phenotype correlation in cystic fibrosis is more complex than formerly suspected.

The transcription of the human fructose-bisphosphate aldolase C gene is activated by nerve-growth-factor-induced B factor in human neuroblastoma cells.

Abstract: A DNA region located at around -200 bp in the 5' flanking region (region D) of the human brain-type fructose-bisphosphate aldolase (aldolase C) gene has been analysed. We show by transient transfection assay and electrophoretic-mobility-shift assay (EMSA) that the binding of transcriptional activators to region D is much more efficient (80% versus 30%) in human neuroblastoma cells (SKNBE) than in the non-neuronal cell line A1251, which contains low levels of aldolase C mRNA. The sequence of region D, CAAGGTCA, is very similar to the AAAGGTCA motif present in the mouse steroid 21-hydroxylase gene; the latter motif binds nerve-growth-factor-induced B factor (NGFI-B), which is a member of the thyroid/steroid/retinoid nuclear receptor gene family. Competition experiments in EMSA and antibody-directed supershift experiments showed that NGFI-B is involved in the binding to region D of the human aldolase C gene. Furthermore, the regulation of the aldolase C gene (which is the second known target of NGFI-B) expression during development parallels that of NGFI-B.

Three Novel Germline Mutations in the Adenomatous Polyposis Coli Gene

Abstract: Maria 1. Scarano, Marina De Rosa, Maurizio Gentile, Luigi Bucci, Giuseppe P. Ferulano, Nicola Carlomagno, Andrea Renda, Ginevra Guanti, Francesco Salvatore*, and Paola Izzo Dipartimento di Biochirnica e Biotecnologie Mediche, CEINGE-Biotechnologie Avanzate, Naples, Italy; Fax: + 39-81-746-3650 II Divisione di Chirurgia Generale III Divisione di Chirurgia Generale Instituto di Chirurgia Generale e Trapiand d’Organo, Medical School, University of Naples Federico 11, 80131 Naples Cattedra di Genetica Medica, Istituto di Medicina del Lavoro, Policlinico, Bari, Italy