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M Montagna

Publications and source records attributed to M Montagna.

At least 55 records · Page 3Linked to original sources

A 100-kb physical and transcriptional map around the EDH17B2 gene: identification of three novel genes and a pseudogene of a human homologue of the rat PRL-1 tyrosine phosphatase.

In this paper, we describe the physical map and transcriptional organisation of a 100-kb region with the BRCA1 locus at 17q12-21. Using the cDNA of the EDH17B2 gene as a probe, we screened a human genomic cosmid library. Positive cosmid clones were aligned and a contig around the EDH17B2 gene was established, expanding the previously reported map. In order to identify genes located in this region, we used the cosmid inserts to select cDNAs from a human ovarian cDNA library. Among the clones identified, cDNA OV-1 corresponds to a human homologue of a rat PRL-1 tyrosine phosphatase gene that shows enhanced expression during hepatic regeneration and in some tumour cell lines. Neither the OV-1 nor the PRL-1 protein shares strong homology with any previously characterised phosphotyrosine phosphatase, suggesting that they probably belong to a new phosphatase family. In an attempt to characterise the OV-1 gene, we found that the genomic sequence present on chromosome 17 probably corresponds to a nonfunctional copy of the gene, as it contains several sequence changes that disrupt the potential coding information of the gene. Three other cDNAs, corresponding to unrelated genes, were also identified and characterised. They did not reveal striking homologies in database sequence comparison and therefore represent new genes localised on chromosome 17q, in a region that frequently shows loss of heterozigosity in sporadic breast and ovarian cancers.

Amino Acid Sequence↗

TP53 gene mutations in gastric carcinoma detected by polymerase chain reaction/single-strand conformation polymorphism analysis of archival material.

TP53 gene mutations, one of the most common alterations described in human tumors, have also been detected in gastric carcinoma, and shown to occur rather late in disease progression. A better assessment of the prognostic value of TP53 gene mutations can be obtained by examining archival material, as this allows stored cases with well-defined histories to be monitored. We performed immunohistochemical and polymerase chain reaction/single-strand conformation polymorphism (PCR-SSCP) analyses of formalin-fixed paraffin-embedded material from nine selected cases of gastric carcinoma at different pathological stages. PCR-SSCP analysis of TP53 exons 5-8 detected missense point mutations in two out of five immunostain(PAb1801)-positive tumors, and a deletion (allowing for a premature stop codon) in one of the remaining four immunostain-negative tumors. Thus, PCR-SSCP analysis represents a feasible strategy for the detection of TP53 alterations in archival material of gastric carcinoma cases.

Aged↗

Dominance of a single Epstein-Barr virus strain in SCID-mouse tumors induced by injection of peripheral blood mononuclear cells from healthy human donors.

Severe Combined Immune Deficiency mouse tumors, induced by inoculating peripheral blood mononuclear cells from 11 healthy human donors (hu-PBMC-SCID tumors), were used to analyse Epstein-Barr virus (EBV) type and strain variations. PCR analysis of EBNA 2- and EBNA 3C-specific sequences showed that EBV type A was present in SCID-mouse tumors induced by PBMC from all donors but one, while, using amplimers for a highly polymorphic region within the latent membrane protein (LMP) coding sequence, 5 different strains could be detected among the samples examined. The same LMP fragment was present in different tumors arising in the same animal, as well as in different mice injected with PBMC from any donor. Compared to B95.8 and AG876 prototype viruses, sequence analysis of LMP variants disclosed a higher homology to the latter, with 33 bp additional repetitions and a few point mutations in specific sites. This study confirms and extends previous data on the presence of a single EBV type and strain in the peripheral blood of most normal healthy subjects using the SCID-mouse system.

Amino Acid Sequence↗

Toxic PCB congeners and organochlorine pesticides in Italian human milk.

Human milk from four major Italian cities was analyzed for individual congeners of polychlorinated biphenyls (PCB), DDT, DDE, hexachlorobenzene, and beta-hexachlorocyclohexane. Minimum and maximum concentrations in milk from individual mothers for most compounds ranged between one order of magnitude below and above the mean value of all mothers. Good agreements were found between results from pooled samples and mean values of individual samples. No statistically significant difference between cities was found and the levels in milk from Italian mothers did not differ significantly from published levels from other parts of the world. Principal component analyses revealed that the PCB congener distribution pattern was very similar in all mothers, independent of location. Average concentrations in milk from the four cities were 19(+/- 5) micrograms liter-1 total PCB or 3.2(+/- 0.8) ng liter-1 toxicity equivalents according to the most conservative TCDD toxicity equivalent factors of PCBs proposed in the literature, 70(+/- 18) micrograms liter-1 DDE, 4.5(+/- 1.2) micrograms liter-1 DDT, 5.6(+/- 1.9) micrograms liter-1 HCB, and 4.4(+/- 1.7) micrograms liter-1 beta-hexachlorocyclohexane.

Female↗

Occasional loss of constitutive heterozygosity at 11p15.5 and imprinting relaxation of the IGFII maternal allele in hepatoblastoma.

The 11p15.5 chromosomal region contains one or more loci involved in congenital developmental abnormalities and in the genesis of embryonal tumors, such as Wilms' tumor, embryonal rhabdomyosarcoma, and hepatoblastoma. In these tumors, a loss of constitutive heterozygosity, selectively involving a specific parental allele, suggests both the presence of onco-suppressor genes and a phenomenon of genomic imprinting. We present evidence that both genetic events could be occasionally involved in hepatoblastoma. In fact, loss of heterozygosity at 11p15.5 could be documented in 3 of 13 patients with hepatoblastoma, and in 2 cases the paternal origin of the residual allele in the tumor was assessed. Moreover, imprinting of the paternal IGFII allele and the maternal H19 allele was confirmed in normal tissues of 5 informative patients. Finally, imprinting relaxation of IGFII was detected in the tumor tissue of 1 patient.

Alleles↗

A fatal case of trichlorofluoromethane (Freon 11) poisoning. Tissue distribution study by gas chromatography-mass spectrometry.

A case of lethal poisoning due to trichlorofluoromethane (FC11) inhalation is described. The fluorocarbon was determined in biological tissues by headspace gas chromatography-mass spectrometry. FC11 was detected in all the examined tissues, with decreasing levels in heart, lung, brain, liver, blood, kidney, and spleen. The highest concentration measured in heart could be related to the mode of toxic action of fluorocarbons postulated by many authors, characterized by the sensitization of the myocardium to the catecholamines producing arrhythmia and cardiac arrest. Nevertheless the aspecific picture of the anatomo-pathological and histological findings does not exclude that the described accidental fatality may have been caused by the combination of direct from toxicity with hypoxemic asphyxiation, due to the saturation of the atmosphere by FC11 in the closed environment in which the intoxication occurred.

Adult↗

Properties of tumors arising in SCID mice injected with PBMC from EBV-positive donors.

Groups of SCID mice were injected with different PBMC sub-populations, and established LCL cells. In about 80% of PBMC-injected animals, tumors developed in association with high levels of human Ig in mouse serum and detectable IL-6 levels. The tumors showed a histopathologic pattern reminiscent of large cell immunoblastic non-Hodgkin's lymphoma; in situ hybridization invariably evidenced EBV sequences in a minority of cells. Genotypic analysis of tumors arising in PBMC-injected mice showed the presence of different oligoclonal B cell populations in different tumor sites. Southern blot analysis disclosed the presence of both linear (replicating) and episomal (latent) EBV DNA forms; sequential analysis of LCL cells serially passaged into animals revealed the progressive selection of clonal cells with only the latent episomal form. Attempts to dissect the events underlying tumor development revealed that the presence of T cells within the injected population was essential for tumor generation; however, the putative T cell-derived factors involved are unclear, and IL-6 seems to play a minor role.

Animals↗

Gas chromatographic/electron impact mass spectrometric selective confirmatory analysis of clenbuterol in human and bovine urine.

A method for the confirmatory analysis of clenbuterol in human and bovine urine was developed. After a double washing of the acidified sample with tert-butyl methyl ether (only for bovine urine), and a solid-phase extraction with Bond-Elut Certify columns, derivatization was performed with trimethylboroxine. The extract was then analysed by gas chromatography/electron impact mass spectrometry. The advantages of the methylboronate derivative with respect to other derivatives are discussed in terms of chromatographic properties and, above all, of specificity of the electron impact mass spectrum. The method was successfully used for the confirmatory analyses of clenbuterol during the XXV Olympic Games of Barcelona.

Animals↗

Rapid and highly selective GC/MS/MS detection of heroin and its metabolites in hair.

A direct treatment of methanol-washed hair with a silylating solution is proposed to extract heroin, O-6-monoacetylmorphine, morphine, acetylcodeine, and codeine, obtaining the simultaneous derivatization of the hydroxylated metabolites and reducing potential sample contamination. Analysis is performed by capillary gas chromatography-tandem mass spectrometry (GC/MS/MS) using multiple selected reaction monitoring. Owing to the selectivity and sensitivity of the GC/MS/MS analysis, and to the extremely simple treatment of the sample, the method fulfils the requirements of both clinical and forensic diagnosis of heroin use.

Codeine↗

[Correlations between membrane integrins and granulocyte defects in myelodysplastic syndromes].

The aim of the present study was to evaluate some functions of neutrophil granulocytes (PMNs), such as aggregation, superoxide production, chemotaxis and adhesion molecules involved in these processes, in 22 patients suffering from Myelodysplastic Syndrome (MDS), to clarify if granulocytes alterations described in this syndrome is really correlated with the expression of surface membrane integrins. Several patients suffering from MDS present granulocytopenia and/or absolute monocytoses; neutrophil granulocytes can have typical nuclear and cytoplasmatic alterations. These granulocytic anomalies are valuable in about 90% of patients suffering from MDS. The granulocytes showed a significant deficit in chemotaxis stimulated by serum activated with E. Coli, casein and formyl-methionyl-leucylphenylalanine (fMLP) (p < 0.01) and in superoxide production stimulated by phorbol-myristate-acetate (PMA). We also studied the role of membrane integrin CD11/CD18 using specific monoclonal antibodies (MoAb). The cytofluorimetric analysis demonstrated a significant inhibition in expression of CD11b/CD18 receptors in patients suffering from MDS (p < 0.001), while the expression of CD11a/CD18 and CD11c/CD18 receptors was normal. In conclusion we found specific alterations in PMNs functions in MDS and a correlation of these anomalies with membrane integrins of PMNs is therefore possible.

Adult↗

Determination of clenbuterol in urine as its cyclic boronate derivative by gas chromatography-mass spectrometry.

A rapid and reliable gas chromatographic-mass spectrometric method for the determination of clenbuterol in urine is described. Penbutolol was used as internal standard. Four derivatization procedures have been tested, of which 1-butaneboronic acid gave the best results. The method includes extraction of the alkalinized urine (3 ml) with tert.-butyl methyl ether-n-butanol (9:1), derivatization with 1-butaneboronic acid (15 min at room temperature), and analysis in the selected-ion monitoring mode of the derivatives of clenbuterol at m/z 243, 327 and 342 and of penbutolol at m/z 342 and 357. The detection limit is 0.5 ng/ml and the recovery better than 90%.

Boronic Acids↗