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Chronic obstructive pulmonary disease (COPD) is a relatively common disease, affecting mainly males in the western world. Although substantial data are available as regards the clinicopathological characterization of COPD, little is known of the molecular basis of the disease. In the present study we analysed the incidence of microsatellite instability (MI) in cytological specimens from patients with COPD. MI reflects increased mutational rate and is associated with decreased accuracy in the DNA repair, resulting in the accumulation of somatic mutations in cells manifesting this genetic alteration. Among 31 specimens tested, 7 (23%) exhibited MI in at least one among 6 microsatellite markers tested. 5 cases were affected in only one marker while the remaining two cases exhibited evidence of MI in two microsatellite markers. These data suggest that an elevated mutational rate as reflected by the increased incidence of MI is associated with the development of the disease.

Journal URL: http://content.karger.com/ProdukteDB/produkte.asp?Aktion=JournalHome&ProduktNr=223857

Journal URL: http://www.spandidos-publications.com/ijo/ Compatible with the epidemiology and natural history of cervical cancer and with experimental findings that HPV-immortalized nontumorigenic cells are similar to cervical carcinoma cells, in terms of the quantity and type of viral RNA expressed, it is believed that additional cellular genetic events are necessary for tumorigenic conversion. In the current study we detected codon 12 point mutations of the K-ras oncogene with an incidence of 28.2% in malignant lesions of the cervix, as well as HPV 18 at a higher rate than HPV16 (30.2% vs 27.9%) in genital lesions, by PCR and RFLP analysis. Codon 12 point mutations of K-, H- and Nras were also found in benign lesions of the cervix, in endometrial and in ovarian carcinomas, although at a lower frequency. It is suggested that the mutationally activated ras oncogenes cooperate with HPV in the early stages of carcinogenesis of the human female reproductive tract.

Journal URL: http://jmm.sgmjournals.org/

Journal URL: http://www.sciencedirect.com/science/journal/03043835

Journal URL: http://www.spandidos-publications.com/or/ TGF-ß1 belongs to a family of pluripotent growth factors (TGFßs) and has been implicated in the development and progression of human breast cancer. There are conflicting data though, suggesting that TGF-ß has the pontency both to promote and inhibit the progression of mammary neoplasia. We examined the expression of TGF-ß1 mRNA in 24 breast carcinomas using the technique of the reverse transcription polymerase chain reaction (RT-PCR) to obtain quantitative results. Overexpression of TGF-ß1 gene was found in 75% of the cases. We also correlated the overexpression of the TGF-ß1 gene with clinicopathological parameters including histological grade, tumour cellularity, oestrogen receptor status (ER), progesterone receptor status (PR) and lymph node involvement. The results led us to the conclusion that the increasing rado of overexpression related to the stage of cancer in an analogous way (P~1). No significant association was identified between the ratio of overexpression and the grade, ER, PR, or lymph node involvement (rs=0.5, 0.2, 0.1, 0.1 respectively; P

Journal URL: http://www3.interscience.wiley.com/journal/29331/home

Journal URL: http://www.sciencedirect.com/science/journal/03043835

Journal URL: http://www.spandidos-publications.com/or/ Infections caused by Human Cytomegalovirus (HCMV) are very common in patients who undergo immunosuppression or immunocompromisation. The techniques used for routine HCMV detection are timeconsuming and lack specificity and sensitivity. The ability of the Polymerase Chain Reaction (PCR) to amplify HCMV DNA from clinical samples of the patients is a valuable diagnostic tool for the detection of HCMV in the early stages of the infection. We used a pair of primers to amplify a 435 bp region of the immediate early-1 gene, to detect HCMV DNA in clinical samples from patients at high risk for HCMV infection. We found HCMV in the following type of patients: 6 out of 20 in immunosuppressed, 11 out of 31 in immunocompromised, 5 out of 8 in pregnant women, 4 out of 25 in patients with high anti-CMV IgM and IgG titres, 1 out of 2 in patients with kidney failure, and 6 out of 14 in patients with opthalmic disorders. Sixty-seven specimens, which were found to be negative for CMV by the PCR technique, were used to inoculate human fibroblast monolayer cultures and PCR was performed to the DNA extracted from the cultured cells. Only in 1 out of the 67 cases HCMV DNA was detected.

Journal URL: http://www.journalarchive.jst.go.jp/english/jnltop_en.php?cdjournal=cancersci1985

Journal URL: http://www.sciencedirect.com/science/journal/0006291X

Journal URL: http://stemcells.alphamedpress.org/

Journal URL: http://www.spandidos-publications.com/or/ Investigation of the nucleotide sequence of the HIV-1 LTR showed the presence of four novel short DNA regions which are homologous to the recognition site for the cellular transcription factor AP-1. Four short oligonucleotide hybrids containing these potential AP-1 sites were constructed and used in gel retardation assays and in competition experiments in order to determine the role of the AP-1 protein in the regulation of HIV-1 expression. The breast MDA MB 468 and cervical HeLa tumor cell lines, which are known to overexpress the AP-1 protein were used in a gel retardation assay as a control to study the affinity of the AP-1 to synthesized oligonucleotide sequences. We have observed specific binding of nuclear factor AP-1 to three of these oligonucleotide hybrids. These results demonstrate the presence of three novel AP-1 binding sites on HIV-1 LTR, one of which was found within the TAR element and in the Tat protein binding region. Moreover, they suggest that AP-1 could be contributing to HIV-1 transcriptional regulation through its interaction with the AP-1 binding sites of HIV-1 LTR.

Journal URL: http://www.spandidos-publications.com/or/ Chronic obstructive pulmonary disease (COPD) is a relatively common disease, affecting mainly males in the western world. Although substantial data are available as regards the clinicopathological characterization of COPD, little is known of the molecular basis of the disease. In the present study we analysed the incidence of microsatellite instability (MI) in cytological specimens from patients with COPD. MI reflects increased mutational rate and is associated with decreased accuracy in the DNA repair, resulting in the accumulation of somatic mutations in cells manifesting this genetic alteration. Among 31 specimens tested, 7 (23%) exhibited MI in at least one among 6 microsatellite markers tested. 5 cases were affected in only one marker while the remaining two cases exhibited evidence of MI in two microsatellite markers. These data suggest that an elevated mutational rate as reflected by the increased incidence of MI is associated with the development of the disease.

Journal URL: http://www.spandidos-publications.com/or/ DNA extracted from the blood of immunosuppressed and immunocompromised individuals and from patients with infectious mononucleosis, leukaemias and lymphomas were studied using the Polymerase Chain Reaction (PCR) technique. The oligonucleotide primers used for the detection of the Epstein-Barr virus (EBV) amplify a 375bp sequence from the EcoRI Β fragment of the viral genome. EBV specific sequences were amplified from the blood samples of 18 out of 65 patients, most of which were transplant patients (9 out of 31 ). The results confirmed the association of EBV with clinical disorders in immunodeficient and immunocompromised patients and the importance of PCR method in routine diagnosis.

Journal URL: http://www.spandidos-publications.com/or/ Infections caused by Human Cytomegalovirus (HCMV) and Epstein-Barr virus (EBV) are common in multiple transfused patients, such as patients with ßthalassaemia. The ability of the Polymerase Chain Reaction (PCR) to amplify HCMV and EBV DNA from blood and other samples makes this technique a valuable diagnostic tool for the detection of both viruses in the early stages of the infection. PCR was used for the amplification of a 435 bp region of the immediate early-1 (IE-1) gene of HCMV and a 375 bp sequence from the EcoRI Β fragment of EBV genome. Blood samples from 80 patients with ß-thalassaemia were examined. HCMV was found in 14 and EBV in 12 patients. The results obtained confirm the implications of HCMV and EBV in the diagnosis of viral infections in multiple transfused patients as well as the importance of PCR technique as a valuable diagnostic tool.

Journal URL: Istituto nazionale per lo studio e lo cura dei tumori di Milano

The purpose of this study was to assess the extent of involvement of the ras oncogene activation by point mutations in endometrial carcinoma in the Greek population. The PCR technique was employed, followed by RFLP analysis to identify the point mutations in codon 12 of the K-ras, H-ras and N-ras genes. K-ras gene point mutations were detected in 8 of the 55 cases (15%) of primary endometrial carcinoma, H-ras in 4 (7.3%), while no mutations were found for the N-ras gene. No correlation was found between the presence of ras gene mutations and the clinicopathological parameters, or patient survival. The only association found was between H-ras mutations and the FIGO stage of the tumor (Fisher’s exact test, p = 0.011). These results indicate a possible role of ras gene activation in a small subset of endometrial carcinomas. © 1996 S. Karger AG, Basel.

We used the PCR technique to detect the Epstein-Barr virus (EBV) and human papillomavirus (HPV) DNA in paraffin-embedded tissues from Greek patients with nasopharyngeal carcinoma (NPC). The oligonucleotide primers used for the detection of EBV amplify a 375-bp long sequence from the EcoRI B fragment of the viral genome, whereas for HPV the primers amplify a 151-bp long sequence of the viral genome. The PCR products were analysed by agarose gel electrophoresis and visualised by UV illumination after staining with ethidium bromide. Sixty-three specimens were examined. EBV specific sequence was amplified in 20 (32%) and HPV in 12 (19%) out of the 63 samples. There was no co-infection with EBV and HPV. Although there is a high correlation of EBV infection with poorly differentiated NPC in patients from Southern China and South-East Asia, the restricted distribution suggests genetic or environmental cofactors in the development of the neoplasm. Our results confirm this suggestion since there was only a 32% correlation of EBV with NPC in Greece. HPV may also be involved in the carcinogenesis of EBV-negative squamous cell nasopharyngeal carcinomas.

Epstein-Bar virus (EBV) is a B-lymphotropic virus with a tumorigenic potential. EBV infection has been recognized as rite main cause of nasopharyngeal carcinoma and Burkitt’s lymphoma. The aim of our study was to determine the incidence of EBV in squamous cell carcinomas of the larynx. We employed for our analysis a sensitive polymorphism chain reaction (PCR) assay, followed by restriction fragment length polymorphism (RFLP) for further confirmation of the specificity of the PCR-amplification reaction, Our analysis revealed that 9 of 27 (33%) specimens harbored the EBV genome in the tumor tissue while only 4 (15%) specimens from adjacent normal tissue exhibited evidence of EBV infection. Three were EBV positive for both normal and tumor tissue. No association has been found with disease stage, histological differentiation and nodes at pathology. The relatively high incidence of EBV in the tumor tissue (33%) of patients with laryngeal cancer, as compared to the low (15%) incidence of the virus genome detected in the adjacent normal tissue of the patients, indicates a probable role of EBV in the development of the disease.

Cytomegalovirus injection is common in kidney transplanted patients. Viremia is the only marker of active CMV infection, but the use of cell culture for the direct detection of the virus is time-consuming and not very sensitive, while the detection of CMV by measuring the titre of antibodies is difficult due to the immunosuppression these patients undergo. Thus the ability to amplify CMV DNA by the polymerase chain reaction from blood or urine samples of the patients becomes a valuable diagnostic tool for the detection of CMV in the early stages of the infection. Using a set of primers specific for the amplification of a 435 bp region of the IE-1 gene, we detected CMV DNA in blood leucocytes of a kidney transplanted patient who received the transplant from a CMV-seropositive donor, 45 days after the operation, while the antibody titre showed no evidence of active CMV infection.