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Biomedical subjects

E Kucera

Publications and source records attributed to E Kucera.

At least 37 records · Page 2Linked to original sources
PubMed2000

p53 polymorphism at codon 72--does it constitute a risk for squamous intraepithelial lesions and invasive cancer of the cervix in Central Europeans?

BACKGROUND: Polymorphisms of the tumour suppresser gene p53 especially at codon 72 are suspected to be associated with an increased risk for malignant transformation. In invasive cervical cancer, the arginine form of the p53 gene is estimated to be more susceptible to degradation mediated by tumour-associated human papilloma viruses (HPV) than the proline form. METHODS: To test the prevalence of p53 polymorphism at codon 72 in 133 healthy women, 50 patients suffering from squamous intraepithelial lesions of the cervix (SIL), and 105 patients with invasive cervical cancer, we developed a polymerase chain reaction (PCR) and microtiter plate-based hybridisation assay. Furthermore, we tested whether the two p53 isoforms increased the risk of developing cervical cancer. RESULTS: The proportions of individual homozygous for arginine, homozygous for proline and heterozygous for arginine and proline in the investigated patient groups did not significantly deviate from the Hardy-Weinberg equilibrium. We found no increased risk of developing cervical cancer in respect to p53 polymorphism, independent of histological diagnosis. DISCUSSION: In conformity with other study groups, our findings do not support the hypothesis that the p53 polymorphism at codon 72 is important in determining an increased risk of developing HPV-associated SIL or invasive cervical cancer in Central Europeans.

Adolescent↗
PubMed2000

Immunohistochemical expression of matrix metalloproteinases 1 and 2 (MMP-1 and MMP-2) and tissue inhibitor of metalloproteinase 2 (TIMP-2) in ruptured and non-ruptured tubal ectopic pregnancies.

OBJECTIVE: Recent evidence suggests that matrix metalloproteinases (MMPs) and their inhibitors (TIMPs) are crucial for trophoblast implantation in normal pregnancy. To evaluate the expression of MMP-1, MMP2, and the tissue inhibitor of MMP-2 (TIMP-2) along the invasive pathway of trophoblast in ruptured and non-ruptured tubal ectopic pregnancies, we performed a retrospective immunohistochemical study. METHODS: In 15 tissue specimens of patients with ruptured (N = 7) and non-ruptured (N = 8) first trimester tubal ectopic pregnancies who underwent laparoscopic salpingectomy, immunohistochemical staining against MMP-1, MMP-2, and TIMP-2 was performed. Serial paraffin sections were photographed and digitized for a computerized quantitative image analysis. Mean percentages of positive stained areas by MMP-1, MMP-2, and TIMP-2 antibodies in the extravillous trophoblast were determined for ruptured and non-ruptured tubal ectopic pregnancies and compared. RESULTS: In our 15 tissue specimens of ectopic pregnancies MMP-1 and TIMP-2 were found to be more prominent in the immunohistochemical distribution pattern than MMP-2. However, no statistically significant difference could be detected between the mean percentages of positive stained area by MMP-1, MMP-2, and TIMP-2 antibodies in ruptured and non-ruptured tubal pregnancies. DISCUSSION: For the first time, we measured the comparative immunohistochemical expression of MMP-1, MMP-2, and TIMP-2 in ruptured and non-ruptured tubal ectopic pregnancies. Although our results did not show any statistically significant difference between ruptured and non-ruptured tubal ectopic pregnancies, we conclude that MMP-1, MMP-2, and TIMP-2 are functionally involved in the highly proliferative early first part of ectopic implantation.

Adult↗
PubMed2000

Systemic methotrexate treatment of interstitial pregnancy--magnetic resonance imaging (MRI) as a valuable tool for monitoring treatment.

OBJECTIVE: Interstitial pregnancy occurs in 2-4% of ectopic pregnancies and is defined as implantation of the trophoblast in the interstitial part of the tuba uterina. Therefore the term intramural pregnancy can also be found in the literature. In 20% of the cases that progress beyond 12 weeks of amenorrhea a potentially life-threatening rupture of the uterus occurs, leading to a maternal mortality rate of 2.5%. According to the literature until a few years ago diagnosis was mainly made intraoperatively, and resulted in cornual resection or hysterectomy per laparotomy. Better methods of diagnosis and treatment of interstitial pregnancy can help to decrease morbidity and mortality associated with this condition. PATIENTS: We describe two cases of interstitial pregnancies that were eventually diagnosed and also monitored by magnetic resonance imaging (MRI) after systemic methotrexate treatment. Both patients were uniparous and experienced their second spontaneous pregnancy. METHODS: Treatment consisted of four doses (50 mg/m2 body surface area) of systemic intramuscular methotrexate alternating with four doses (6 mg) of intramuscular folic acid. When beta-hCG levels were undetectable, MRI results were compared with pre-therapeutic MRI findings. RESULTS: In patients A and B, beta-hCG levels were undetectable 64 and 88 days after initiation of methotrexate treatment, while magnetic resonance imaging revealed nearly equally persisting interstitial pregnancies. They initially presented as hyperintense lesions with hypointense zones and changed into a hypointense lesion with a central hyperintense area for patient A, and a completely hyperintense lesion for patient B at the time of negative beta-hCG levels in follow-up MRI. CONCLUSION: Systemic methotrexate treatment with an intramuscular regimen is effective in the treatment of interstitial pregnancy. MRI has the ability of correct tissue differentiation and objective three-dimensional measuring of interstitial pregnancy. We therefore propose this imaging modality as a valuable tool for monitoring systemic methotrexate treatment of interstitial pregnancy that should be used additionally to beta-hCG clearance curves.

Adult↗
PubMed2000

Ectopic pregnancy.

Ectopic pregnancy is a implantation occurring elsewhere than in the cavity of the uterus, whereas ninety-nine percent of extrauterine pregnancies occur in the fallopian tube. The incidence of extrauterine pregnancy has increased from 0.5% thirty years ago, to a present day 1-2%. The most frequent cause of tubal pregnancy is previous salpingitis. Mortality rates for tubal pregnancies used to be approximately 1.7% in the 1970s but dropped to 0.3% in 1980s. DIAGNOSIS: Using transvaginal ultrasound it is possible to obtain positive evidence of an ectopic pregnancy at a very early stage. In cases of hCG titers>2,000 IU/l, intrauterine pregnancy can be diagnosed with certainty. The most important differential diagnosis of ectopic pregnancy is early intrauterine pregnancy. CLINICAL MANAGEMENT AND THERAPY: Regardless of the therapeutic strategy selected by the physician, informing the patient is a major aspect of the management of ectopic pregnancy. If surgery is considered appropriate, the patient must be informed about the nature, side effects and complications of the procedure. However, it should be remembered that in some cases, the actual chances of cure first become apparent at surgery. In asymptomatic patients with a serum hCG titer <1,000 IU/l that is falling, it is appropriate to wait and watch. In clinically stable patients with an unruptured tubal pregnancy and steady hCG levels, systemic treatment with methotrexate might also be considered. In unruptured tubal pregnancy with a hCG titer between 1,000 and 2,500, a further therapeutic alternative is intratubal injection of prostaglandins, hyperosmolar glucose of NaCl. Generally speaking, the currently widespread laparoscopic surgical treatment of the fallopian tube hardly influences the risk of recurrence. If the gestational mass is larger, the serum hCG titer higher than the approximate limit of 2,500 mU/ml and/or the tube already ruptured, surgery is usually required. PREVENTION: The most effective prevention is to avoid tubal inflammation or, in cases of preexisting inflammation, to administer effective therapy.

Chorionic Gonadotropin↗
PubMed2000

Human papillomavirus in the cervix and placenta.

OBJECTIVE: To determine the prevalence and association of human papillomavirus (HPV) infection in the cervices and placentas of pregnant women. METHODS: Cervical samples were taken from 179 of 226 women who had placental biopsies because of abnormal ultrasound findings or were older than 35 years, to detect HPV infections with hybrid capture II tests. Polymerase chain reaction (PCR) was done on placental tissue of 147 of the 226 women to detect HPV DNA. RESULTS: We found 44 of 179 women (24.6%, 95% confidence interval 18.3, 31.0) to test positive for HPV in their cervices. Logistic regression analyses showed decreased prevalence of HPV infection with increased maternal age (P =.039). The HPV DNA E6 PCR from the villus tissue was negative in the 147 cases examined. However, a significant contingency coefficient between low-risk HPV infection and elevated risk of chromosome aberration was found (&phi; = V = 0.15, P =.050). CONCLUSION: The infection rate of 24.6% in women without clinical symptoms of HPV infection was high, but there seemed to be no virus transmission to the placenta in women with subclinical infections. Low-risk cervical HPV infection might be associated with a slightly higher risk of abnormal fetal karyotype.

Adult↗
PubMed2000

Could we treat more unruptured ectopic pregnancies with intramuscular methotrexate?

The main reason for the restricted use of methotrexate in the treatment of ectopic pregnancy (EP) obviously is the fear of tubal rupture in patients with lower abdominal pain after the administration of methotrexate. Therefore, we wanted to find out if patient characteristics at first presentation, such as age, pretreatment beta-hCG level, adnexal mass as visualized by transvaginal ultrasonography, or history of prior EP, would identify patients at risk for tubal rupture if they were hemodynamically stable and showed no signs of peritoneal irritation. We examined whether more patients could have been treated medically with methotrexate, because tubal rupture was unforeseeable at first presentation and inclusion criteria for methotrexate treatment were fulfilled. From January 1996 to August 1998, 122 patients diagnosed as having EP were treated at the Gynecologic Department of the University Hospital of Vienna. Inclusion criteria for medical treatment with intramuscular methotrexate (50 mg/ m(2) body surface area) were (1) hemodynamic stability, (2) an unruptured ectopic mass < or = 5 cm at the greatest dimension demonstrated at transvaginal ultrasonography; (3) beta-hCG level < or = 5,000 mIU/ml; (4) no cardiac activity of the extrauterine embryo; (5) wish of future fertility, and (6) informed consent. Patients with hemodynamic instability, severe abdominal pain, an ectopic mass > or = 5 cm at the greatest dimension, beta-hCG levels > or = 5,000 mIU/ml, cardiac activity of the extrauterine embryo, and no wish of future fertility, or disagreement with methotrexate treatment, primarily underwent surgery. Despite the fact that none of the above patient characteristics at first presentation identified patients at risk for tubal rupture, only 60/122 patients (49%) actually underwent medical treatment whereas our inclusion criteria would have granted medical treatment in 101/122 patients (83%). We determined the actual and maximal possible percentages of patients with unruptured EP eligible for medical treatment of EP with intramuscular single-dose methotrexate 50 mg/m(2) body surface area. Our data show that tubal rupture in hemodynamically stable patients is not foreseeable and should not lead to a restricted use of medical treatment in patients preferring methotrexate.

Adult↗
PubMed2000

Accuracy of intraoperative frozen-section diagnosis in stage I endometrial adenocarcinoma.

The purpose of our study was to determine if frozen-section diagnosis accurately identified patients suffering from endometrial adenocarcinoma FIGO stage I for surgical staging consisting of total abdominal hysterectomy, bilateral salpingo-oophorectomy, peritoneal cytology, and complete bilateral pelvic lymphadenectomy in moderately differentiated tumors with myometrial invasion. In all poorly differentiated tumors, and in all tumors with deep myometrial invasion (more than 50%) surgical staging included additional para-aortic lymphadenectomy. We performed a retrospective study including 70 patients. Frozen-section diagnosis of myometrial invasion and tumor grade was compared with permanent-section diagnosis. The accuracy rates were determined, and compared with accuracy rates of frozen-section diagnosis in the literature, and a total accuracy rate for 624 patients suffering from stage I endometrial adenocarcinoma was evaluated. In our patient collective, the overall accuracy rate of frozen-section diagnosis for myometrial invasion and tumor grade was 80 and 84%, respectively. In the five comparable studies, the mean accuracy rate for myometrial invasion and tumor grade was 89 and 84%, respectively. In combination with the five comparable studies our recent study produced an accuracy rate of frozen-section diagnosis for myometrial invasion and tumor grade of 88 and 84% in 624 patients, respectively. Despite an accuracy level of frozen-section diagnosis for myometrial invasion of 80 and 84% for tumor grade in our patient collective, all patients who required surgical staging were accurately identified.

Adenocarcinoma↗
PubMed1999

Prognostic significance of mutations in the p53 gene, particularly in the zinc-binding domains, in lymph node- and steroid receptor positive breast cancer patients. Austrian Breast Cancer Study Group.

The aim of our study was to evaluate if p53 mutations, especially those in the L2/L3 domains of the p53 gene, add prognostic information for node-positive and steroid receptor positive breast cancer patients. Two hundred and five tumour samples from a randomised clinical trial of 596 lymph node- and steroid receptor positive breast cancer patients were included. All patients had been randomly allocated to receive 20 mg of adjuvant tamoxifen (TAM) daily for 2 years or TAM plus one cycle of low-dose, short-term chemotherapy. For detection of p53 mutations we used in vitro amplification by polymerase chain reaction and consecutively performed temperature gradient gel electrophoresis (PCR-TGGE) and direct sequencing. We found p53 mutations in 42/205 (20%) cases: 16/42 (38%) p53 mutations occurred within the L2/L3 domains of the p53 gene, and 26/42 (62%) outside the L2/L3 domains. p53 mutation served as a statistically significant parameter in predicting disease-free survival in univariate (P = 0.02) and multivariate (P = 0.009) analysis. For overall survival, no significant differences were observed. Patients with tumours that had p53 mutations within the L2/L3 domains of the gene showed no significant difference to those with mutations outside the L2/L3 domains for disease-free survival. For overall survival, mutations in the L2/L3 domains showed a marginally significant difference (P = 0.05) in multivariate analysis, but not in univariate analysis (P = 0.13). We conclude that mutation in the L2/L3 domains of the p53 gene is not an independent prognostic indicator of disease outcome for patients suffering from breast cancer with lymph node metastases and positive steroid receptors.

Adult↗
PubMed1999

Relationship of serum levels of endogenous relaxin to cervical size in the second trimester and to cervical ripening at term.

OBJECTIVE: To assess whether cervical size in the early second trimester and cervical ripening at term may be related to serum levels of endogenous relaxin, 17beta-oestradiol or progesterone. DESIGN: A cross-sectional study investigating the relationship between hormone concentrations and cervical parameters as measured by ultrasound and Bishop score, respectively. PARTICIPANTS: Uncomplicated human pregnancies with normal fetal outcome, 72 women in the second trimester and 40 women at term. METHODS: Vaginal ultrasound and palpation were used to estimate cervical parameters. Hormones were analysed either by dissociation-enhanced fluoroimmunoassay (relaxin) or by automated electrochemiluminescent immunoassays. RESULTS: Cervical length and diameter correlated positively during mid-gestation and negatively at term. During mid-gestation, but not at term, relaxin was significantly associated with cervical length and volume, and with progesterone. Bishop score only correlated inversely with progesterone at term. CONCLUSION: Corpus luteum function is reflected by progesterone and relaxin in the early second trimester. An impact of relaxin on cervical growth, previously demonstrated by animal models and in vitro experiments, was confirmed during human mid-gestation. In contrast to many other species, human cervical ripening was not associated with endogenous relaxin at term, but with decreased progesterone.

Adult↗
PubMed1999

Messenger RNA determination of estrogen receptor, progesterone receptor, pS2, and plasminogen activator inhibitor-1 by competitive reverse transcription-polymerase chain reaction in human breast cancer.

Estrogen receptor (ER), progesterone receptor (PR), the estrogen-inducible protein pS2, and plasminogen activator inhibitor-1 (PAI-1) are important prognostic factors in primary breast cancer. The protein concentrations of these factors in breast tumors have been well documented. However, few data about the mRNA expression of ER, PR, pS2, and PAI-1 in breast cancer are available, which is mostly due to the limitations of conventional techniques for mRNA analysis. We have described a competitive reverse transcription-PCR system for the simultaneous quantification of ER, PR, pS2, and PAI-1 mRNA in tumor samples. Here, we evaluated 100 tumor biopsies from breast cancer patients for the mRNA expression of ER, PR, pS2, and PAI-1. The results were analyzed for correlations with protein status and with clinical data. Significant correlations between mRNA expression levels and protein concentrations of all tested markers were found. In only a few cases was there an obvious discordance between the measurable amounts of mRNA and protein, especially for ER and PR. In addition, ER, PR, and pS2 mRNA levels correlated significantly with each other. No correlation between PAI-1 mRNA amount and the expression of the other markers was found. With respect to clinical data, ER and PR mRNA levels were found to be inversely correlated to tumor size and histological grade but not to the lymph node status. pS2 and PAI-1 mRNA expression were not correlated with tumor size, grade, or lymph node involvement. In conclusion, competitive reverse transcription-PCR may be used as an alternative for the study of prognostic factors in human breast cancer and other malignancies. However, before mRNA expression is measured for diagnostics, a presumed concordance of mRNA and protein expression must be evaluated very carefully for every gene.

Adult↗
PubMed1998

Cytosol concentrations of CD44 isoforms in breast cancer tissue.

The role of the adhesion molecule CD44 in the natural history of breast cancer is controversial. We investigated the CD44 isoform CD44v5 and CD44v6 concentrations in the cytosol of 90 breast cancer specimens, 9 fibroadenomas and 22 normal breast tissue specimens by means of ELISA. CD44v5 and CD44v6 cytosol concentrations were statistically significantly higher in breast cancer compared with fibroadenoma and normal breast tissue (Mann-Whitney U-test, p = 0.009 and p < 0.001, respectively). When CD44 isoforms were correlated with lymph node involvement, histological grading, histological type, tumor stage and age at diagnosis, we found no statistically significant correlation with any of the investigated clinico-pathological parameters. In univariate and multivariate analyses, CD44v5 and CD44v6 were of no prognostic relevance regarding disease-free survival in breast cancer patients (log-rank test, p = 0.16 and p = 0.08, respectively). Our results indicate that CD44 isoforms are increased in samples from tumors relative to normal tissue. Our data show that CD44v5 and CD44v6 isoform expression, although up-regulated by malignant transformation, is not required to acquire a metastatic phenotype in breast cancer. Furthermore, our data support the assumption that cytosolic CD44 isoforms are of no prognostic relevance for disease-free survival of breast cancer patients.

Adult↗
PubMed1998

Amplification and expression of the c-erbB-2 oncogene in Müllerian-derived genital-tract tumors.

This study focused on 18 uterine corpus tumors and 7 ovarian endometrioid tumors, all of them malignant and Müllerian derived. Differential polymerase chain reaction (DPCR), dot blot hybridization, and immunohistochemical technique were employed to determine c-erbB-2 amplification and expression. Of 25 Müllerian-derived tumors, 17 (68.0%) demonstrated amplified c-erbB-2 (two to eight copies) by DPCR. These 25 samples were reexamined by dot blot and immunohistochemical technique, revealing c-erbB-2 amplification and expression of to be 52.0 and 40.0%, respectively. There seemed to be a slight correlation between the amplification and expression of c-erbB-2 and patient survival. Although c-erbB-2 was frequently present in Müllerian-derived genital-tract tumors, it is uncertain whether this oncogene may serve as their sole prognostic marker. The question remains whether c-erbB-2 alone, or in conjunction with other oncogenes or suppressor genes, accounts for the pathogenesis of Müllerian-derived tumors. However, these results suggest for the first time in the literature that DPCR is a sensitive enough technique for detecting c-erbB-2 amplification in M ullerian-derived tumors.

Adult↗
PubMed1998

Treatment of endometrial carcinoma with high-dose-rate brachytherapy alone in medically inoperable stage I patients.

PURPOSE: To review the results of treatment with high-dose-rate brachytherapy alone in 228 patients with stage I endometrial carcinoma who are unfit for surgery. METHODS: All patients received an exclusive radiation therapy by means of high-dose-rate Iridium 192 intracavitary brachytherapy without additional external beam radiation. RESULTS: At 5 years, the overall survival rate was 59.7% and disease specific survival 85.4% at 10 years 30.2% and 75.1%. In clinical stage Ia disease specific survival was 88.6% at 5 years and 82.7% at 10 years, in stage Ib 80.2% and 63.4%, respectively (p<0.02). Disease specific survival was not affected by tumor grade or age. The rates of local control are related to the size of the uterus but not to the tumor grading. Intrauterine recurrence occurred in 17.5% but extrauterine pelvic relapse in only 0.4% of patients. The calculated probability of severe complications was 4.6% at 5 years. CONCLUSION: HDR brachytherapy alone achieves excellent disease specific survival rates in patients with medically inoperable stage I endometrial carcinoma.

Aged↗
PubMed1997

Characterisation of genotoxic properties of 2',2'-difluorodeoxycytidine.

The genotoxic properties of 2',2'-difluorodeoxycytidine (dFdC) were characterised using diploid, mortal low-passage fibroblasts (LPF cells) and the spontaneously transformed fibroblast cell line V79. In both cell types, incorporation of dFdC into the DNA led to an increase of DNA single-strand breaks evaluated by an in situ nick translation assay and to an accumulation of cells in the S-phase of the cell cycle. At concentrations below those leading to cell cycle arrest, dFdC neither induced sister chromatid exchange (SCE) nor structural chromosome aberrations in LPF cells, whereas V79 cells accumulated SCEs as well as chromosome breaks over a broad dose range. In LPF cells treated with dFdC, chromosomal alterations were detected by the micronucleus assay within a narrow concentration range, whereas in V79 cells, a dose-dependent increase in the appearance of micronuclei was seen up to cytotoxic concentrations. In addition, V79 cells went into apoptosis, as evaluated by nuclear fragmentation and condensation, whereas this phenomenon was not detectable in LPF cells.

Antimetabolites, Antineoplastic↗