[Genetic screening: the contribution of 2-dimensional DNA electrophoresis].
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Biomedical subjects
Publications and source records attributed to V Romano-Spica.
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The use of antiblastic drugs has opened up new perspectives in improvement of therapy and life quality for cancer patients. The widespread clinical application of cytostatic drugs implies risks for exposed hospital personnel, due to genotoxic and toxic-reproductive effects. Biological monitoring is fundamental to identify individuals at risk but is limited by the long latency of chronic effects, absence of unique cellular targets and low sensitivity of available laboratory tests. The objective of this study was to investigate toxic mechanisms by a molecular biology approach, searching for biomarkers potentially useful in monitoring programs. The proposed experimental model consisted of cell line exposure to cyclophosphamide, an alkylating agent of wide clinical use. Cellular response has been investigated focusing on potential targets at RNA level, through reverse transcription polymerase chain reaction (RT-PCR) and differential display analysis. We studied the expression of several genes involved in differentiation, apoptosis and chemoresistance: ets1, bax, bcl-2, bag-1, bcl-X, mdr1 and mrp. Specific patterns of mRNA modulations were observed. Differential display analysis revealed candidate genes induced or repressed following exposure: their characterization is in progress. Besides improving the understanding of toxic mechanisms, identification of modulated molecular targets opens up new perspectives in exposure risk assessment, biomonitoring and preventive strategies at occupational level.
BACKGROUND/AIMS: Several epidemiological studies have shown the existence of other routes of transmission of the hepatitis C virus besides the parenteral one, but the mechanisms involved are not yet understood. The general aim of this study was to evaluate the prevalence of hepatitis C virus infection in family contacts of infected patients and to analyze the possible risk factors and alternative transmission routes. METHODOLOGY: One hundred and thirty-eight relatives of 45 patients (index cases) affected by C virus-related chronic hepatitis were studied. The relatives were 45 spouses, 89 children and 4 cohabitants who underwent detection of serum anti-HCV antibodies; the anti-HCV-positive subjects were tested for serum HCV-RNA. The index cases, all the spouses and only other infected relatives were tested for the presence of HCV-RNA in saliva RESULTS: Antibodies to hepatitis C virus were detected in 5.7% of the family members while 11.1% of the analyzed spouses were serum HCV-RNA-positive. HCV-RNA was found in 44% of the examined saliva and 39% of these were found serum HCV-RNA-negative. The prevalence of hepatitis C virus among household contacts, excluding cases with previous parenteral exposure, was 3.6%. CONCLUSIONS: The epidemiological data on the intrafamilial spread of hepatitis C virus may be underestimated owing to the existence of infected relatives serum-negative but saliva-positive for the presence of the virus. The whole of these observations suggests a possible role of biological fluids in intrafamilial spread of hepatitis C virus.
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We have previously shown that electromagnetic field (EMF) exposure induces ETS1 oncogene overexpression in different cell lines. In order to investigate in vivo EMF effects, BALB/c mice were exposed at different times to 50 MHz radiation, modulated (80%) at 16 Hz. The exposed and control animals were sacrificed and the spleen excised for rt-pcr and western blot analysis. We observed an increase in ETS1 mRNA and protein expression, but a decrease in ETS2 protein levels. Preliminary results from this experimental model show in vivo evidence of the effect of EMF on ETS oncogene expression.
Ets genes are transcription factors expressed in different tissues. Biochemical pathways interfering with the intracellular calcium ion concentration are known to influence members of this gene family. Considering experimental observations and previous reports, we proposed a hypothetical model of interaction between EMF and Ets, based on possible interference in pathways involving calcium as a second messenger. This approach comprehend different hypothesis and may contribute to clarify EMF biological effects, providing an experimental model and a procedure based on gene expression analysis. Additional knowledge on biological mechanisms is required to support these promising results. Further experiments are in progress to assess the role of EMF in modulating gene expression and its biological meaning. Identification of early endpoints is needed for mechanism understanding and development of sensitive biomarkers for molecular epidemiology studies.
A multicentric Italian investigation on legionnaires' disease is in course to clarify host factors as well as pathogen associated characteristics involved in the infection/disease. The main goal of the research plan is to account for some critical aspects concerning identification and prevention of legionellosis. To improve knowledge on factors associated with Legionella spp colonisation in hot waters, to detect cases and to characterize risk factors in subjects which develop pneumonia are specific objectives of the research programme. Preliminary results show that hot waters of houses and hotels are frequently contaminated (22.6% and 54.6%, respectively), mainly by L. pneumophila. Microbial concentrations were low in domestic waters (<1.000 ufc/l), but higher in samples from the hotels (geom. mean 1.85 x 10(3) ufc/l). Warming system, age of the plant, type of building were risk factors significantly associated with Legionella spp positivity. The active surveillance on patients affected by pneumonia with search for Legionella urinary antigen allowed the identification of 34 cases, 3 of which of nosocomial origin, corresponding to 4.2% of the screened pneumonia. After informed consent, 26 subjects were recruited for a case-control-study to clarify risk factors for the disease.
Management of genetic data needs the fulfillment of technical requirements, involving ethics, law, and social issues. Introduction of genetic information in preventive medicine implies complex consequences and specialized expertise. Genetic screenings represent a promising tool, but further advances are needed to provide their effective and appropriate use in the respect of the privacy. Application areas, tests validation procedures and availability of effective laboratory and qualified service networks are considered. The Authors present main laws and bioethics guidelines in the field.
In Italy controls on the hygienic quality of swimming pools are fixed by the Accordo tra il Ministero della Salute, le regioni e le province autonome di Trento e di Bolzano, come into force in 2003. In the present study swimming pools were investigated from the microbiological point of view on the basis of the new legislation. Contemporaneously, for the first time in Italy, analytical controls on the occurrence of the protozoa Cryptosporidium and Giardia were performed with both techniques of immunofluorescence and polymerase chain reaction. Results evidenced a good water quality when the bacterial parameters stated by law were considered. However in some swimming pools, the protozoa were also recovered. Factors affecting the use of molecular techniques on the analysis of environmental samples are also put into evidence.