M. Koffa

Evaluation of Parvo B19, CMV and HPV viruses in human aborted material using the polymerase chain reaction technique

OBJECTIVE To investigate the role of human parvovirus B19 (Parvo B19), cytomegalovirus (CMV) and human papilloma virus (HPV) viruses in the aetiopathogenesis of spontaneous abortions. STUDY DESIGN Abortion material from 102 cases of women with spontaneous abortions were analysed for the presence of Parvo B19, CMV and HPV DNA using the polymerase chain reaction (PCR) technique. Serological assays were used for the detection of specific IgM and IgG antibodies against Parvo B19 virus and CMV in the maternal sera. RESULTS Parvo B19 virus genome was detected in two cases of spontaneous abortion, by PCR amplification, while CMV and HPV genomes were not observed. Serological markers were indicative for Parvo B19 virus and CMV infection in ten and four cases, respectively. CONCLUSIONS PCR is a useful method for investigating the viral contribution to the aetiopathogenesis of spontaneous abortions and for detecting the viral genome in the abortion material. This study of 102 cases of spontaneous abortion does not implicate CMV and HPV in the aetiopathogenesis of spontaneous abortion, although it indicates a possible abortional role for Parvo B19 virus.

Codon 12 ras mutations in patients with myelodysplastic syndrome: incidence and prognostic value

Abstract To determine the prevalence of activated ras-oncogenes (N-ras, Harvey-ras Kirsten-ras), DNA derived from peripheral blood of 51 patients with myelodysplastic syndrome (MDS) was investigated. The method was based on the polymerase chain reaction (PCR) technique to amplify DNA, followed by restriction fragment length polymorphism (RFLP) analysis. Among the French-American-British (FAB) subtypes, N-ras mutations were found in two patients with refractory anemia with excess of blasts (RAEB), in one patient with refractory anemia with excess of blasts in transformation (RAEB-t), and in two patients with chronic myelomonocytic leukemia (CMML). MDS patients with a mutation at codon 12 of the N-ras gene showed shorter survival duration than other MDS patients of the same FAB subtypes, although these findings proved to be not statistically significant (P>0.1). Interestingly, all but one patient with N-ras mutation developed acute myelogenous leukemia (AML). In conclusion, the presence of mutation at codon 12 of the N-ras gene might serve as a negative prognostic factor at diagnosis of MDS.

ras Gene Mutations in Human Endometrial Carcinoma

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 (Fishers exact test, p = 0.011). These results indicate a possible role of ras gene activation in a small subset of endometrial carcinomas.

Glucocorticoid and Estrogen Receptors Have Elevated Activity in Human Endometrial and Ovarian Tumors as Compared to the Adjacent Normal Tissues and Recognize Sequence Elements of the H‐ras Proto‐oncogene

We examined the level of receptor binding in H‐ras elements, using nuclear extracts derived from human endometrial and ovarian lesions and from adjacent normal tissue in gel retardation assays. We found increased binding of the glucocorticoid receptor (GR) to the H‐ras GR element in more than 90% of endometrial tumors and in all ovarian tumors tested, as compared to the corresponding adjacent normal tissue. Additionally, we found elevated binding of the estrogen receptor (ER) in H‐ras ER element in all pairs of ovarian tumor/normal tissue tested, whereas in ER‐negative control breast tumor/normal tissue pairs, no differences in ER DNA‐binding levels were observed. These results suggest that steroid hormone receptor binding could directly activate the H‐ras oncogenic potency in human endometrial and ovarian lesions, providing additional evidence for the role of H‐ras expression in hormonally responsive human cancers.

Detection of Herpes simplex Virus (HSV) in Aborted Material Using the Polymerase Chain Reaction Technique

Objective: To investigate the contribution of HSV to the aetiopathogenesis of spontaneous abortion. Design: A hospital-based, case-control study. Setting: Department of Obstetrics and Gynecology, University Hospital and Laboratory of Clinical Virology, Medical School, University of Crete, Heraklion, Crete, Greece. Population and Methods: Abortion material from 102 cases of women with spontaneous abortion was analysed for the presence of HSV DNA applying the PCR technique. Serological assays were used for the detection of specific IgM and IgG antibodies in the maternal sera of 90 pregnant women with successful outcome of their pregnancy while 70 non-pregnant women at reproductive age were also examined as control. Results: The HSV genome was detected by PCR amplification in 3 cases of spontaneous abortion, 2 of them exhibited serological markers of virus reactivation while the 3rd showed a past infection. There were no obvious clinical manifestations indicating a current herpes infection. Both groups of pregnant women, either with spontaneous abortion or with a successful outcome of pregnancy, displayed serological markers of HSV reactivation at higher rates compared with non-pregnant women (χ2, p < 0.05). Conclusions: Using the PCR technique we were able to detect the HSV genome in gestational tissues of spontaneous abortions, even in cases without any clinical symptoms or seropositivity for a primary infection. Serological assays were not very useful for the elucidation of the role of HSV in inducing spontaneous abortions, although they indicate that the state of pregnancy predisposes to HSV reactivation. However, the detection of HSV in 3 out of a total number of 102 cases does not support HSV infection as a major abortion-related factor.