Molecular epidemiology of hepatitis C virus genotypes and subtypes among injecting drug users in Hungary.

The aim of this study was to determine the geographical distribution of hepatitis C virus genotypes/subtypes among people who inject drugs (PWID) recruited at 22 needle exchange sites and drug outpatient services in all seven Planning and Statistical Regions of Hungary. Of 198 such PWID, 147 (74.2%), 45 (22.7%) and six (3.0%) carried genotype 1, 3 or 4, respectively, and 31 (72.1%) of the 43 genotype 1 sequences were of subtype 1a. Genotype 3 was significantly more prevalent in provincial towns than in the capital, Budapest. Injecting for a longer period and an older age both correlated with a higher prevalence of genotype 3, suggesting possible future changes in genotype distribution. The distributions of hepatitis C virus genotypes/ subtypes differed significantly between the tested PWID and the general population. The identification of genotype 3 reflected its worldwide occurrence among PWID. Our results underline the importance of genotyping before treatment, especially among people who have ever injected drugs in Hungary.

Time-lapse cleavage rating predicts human embryo viability.

Chronology of three consecutive mitotic events in human pre-implantation embryos was examined by time-lapse imaging. In zygotes producing well-formed and pregnancy-yielding expanded blastocysts, uniform time-patterning of cleavage clusters (c) and interphases (i) was revealed: i2=11+/-1, i3=15+/-1, i4=23+/-1 h / c2=15+/-5, c3=40+/-10, c4=55+/-15 min. Oppositely, shortened or prolonged durations of one or more cell cycles were strongly predictive of poor implantation and development. Furthermore, trichotomic mitosis was discovered in 17 % of cases - zygotes cleaved into 3 blastomeres and 2-cell embryos into 5-6 cells (instead of normal 2 and 4). During conventional clinical assessment, such embryos are indistinguishable from normal, often considered just-in-course of the next cell cycle. Only detailed time-lapse monitoring paced at 10-minute intervals had proven all these embryos to be absolutely unviable, even in rare cases when they reduced their hypercellularity to normal cell counts via cell-cell fusion. Overall, we demonstrate that time-lapse embryo cleavage rating (ECR) as a standalone diagnostic procedure allows for effective identification of viable early embryos with 90 % specificity, while elimination of good-looking but unviable embryos can be assumed with a specificity of 100 %. Thus, making this non-invasive and contactless approach worth of addition to routine embryo screening in clinical IVF programs.

[Non-invasive monitoring of the timing of early embryo cleavages--objectively measurable predictor of human embryo viability]. / Neinvazívne meranie dlzky bunkového cyklu v prvých dnoch embryonálneho vývoja--objektívne meratel'ný ukazovatel' zivotaschopnosti l'udských embryí.

OBJECTIVE: The evaluation of the developmental abilities of human embryos according to the timing of their early mitotic cleavages. DESIGN: Retrospective study. SETTING: Prague Fertility Centre and Institute for Care of Mother and Child, CAR, Prague. METHODS: The embryos obtained in IVF program were used for further observations and subjected to automated time-lapse monitoring (PrimoVision, Cryo-Innovation, 1 picture/10 min, intermittent white-light illumination) under standard cultivation conditions (37.0 degrees C, 5% CO2 in humid air). Image sequences were digitally recorded for later use. For intravital spindle detection we used polaryzing microscopy (Oosight, Research Instruments) and Hoechst 33342 fluorescent dye for intravital chromatin visualization. A total number of 180 human embryos which gave a vital pregnancies (FHB, fetal heart beat) were analysed retrospectively for timing of early cleavages. In our study, the exact timing of the four interphases (IP) and synchrony of sister cell divisions (ID, interval division) occurring after fertilization were identified and manually recorded. Interphases: IP1 was defined as the period from fertilization till 2 cell stage. IP2 between 2 and 3 cells stages, IP3 between 3 and 5 and IP4 between 5 and 9 cells embryo. INTERVAL DIVISION: ID2 was recorded as a time interval between 3 and 4 cells, ID3 between 5 and 8 cells and ID4 between 9 and 16 cells stage embryos. RESULTS: In the embryos giving viable pregnancies, the durations of IP1 was 20-26 hrs. IP2 was 10-12 hrs, IP3 was 14-16 hrs and IP4 was 20-26 hrs. In these embryos, the sister blastomeres cleaved in a very synchronous manner. The duration of ID1 was recorded to varry from 120 to 210 min. ID2 from 20 to 60 min., ID3 from 120 to 240 min. and ID4 from 230 to 360 min. CONCLUSION: The viable embryos cleave in a very similar time pattern which can be defined and applied as referencial value. Non-invasive monitoring of the timing of early embryo cleavages can be used as an objectively measurable predictor of human embryo.

Malignant changes in adenomyosis in patients with endometrioid adenocarcinoma.

OBJECTIVE: The aim of our retrospective study was to evaluate pathological changes in adenomyotic foci in hysterectomy specimens, and point out a possible mechanism of carcinogenesis in adenomyotic foci inside the myometrium. METHODS: Retrospective analysis of clinical data; 219 patients were operated at our departments from 2003-2008 with the diagnosis of early endometrial cancer. Standard staging operation was used in all cases and all hysterectomy specimens were afterwards routinely analyzed. RESULTS: Adenomyosis was found in 88 of a total of 219 hysterectomy specimens, while 205 of these 219 were affected by endometrioid adenocarcinoma, ten with clear cell carcinoma and four with papillary serous carcinoma. Within these subgroups adenomyosis was documented in 87 of 205 specimens with endometrioid adenocarcinoma (42.4%) and in one specimen of ten with clear cell carcinoma (2.2%), all found in the eutopic endometrium. All cases of malignant changes (n = 6) in adenomyosis were found exclusively with coexisting endometrioid adenocarcinoma: adenocarcinoma in adenomyosis was well or moderately differentiated in five cases, and poorly differentiated in just one case. Differentiation of the tumor in adenomyosis correlated with differentiation of the eutopic endometrial cancer in 50%. Hyperplastic changes like benign glandular hyperplasia, or atypical complex hyperplasia (ACH) were identified simultaneously in all cancer-positive adenomyotic foci. CONCLUSION: Malignant changes in adenomyosis were present in 6.8% of patients with endometrial cancer. All malignancy-positive cases of adenomyosis were associated with endometrioid adenocarcinoma of the eutopic endometrium. Interestingly, in all these cases, different stages of hyperplastic changes were also simultaneously identified. This observation suggests a similar pathway of carcinogenesis in adenomyosis as is known in estrogen-responsive endometrial cancer type I.

Altered patterns of RB expression define groups of soft tissue sarcoma patients with distinct biological and clinical behavior.

BACKGROUND: Function of the retinoblastoma tumor suppressor protein (pRB) may be compromised at a genetic level by gene loss or mutation or at a post-translational level by hyperphosphorylation. In this study, we examined adult soft tissue sarcomas (ASTS) to determine if alterations of pRB were associated with distinct patterns of pRB expression and clinical outcome. DESIGN: We investigated 86 ASTS patients using monoclonal antibodies that distinguish between hyperphosphorylated and underphosphorylated pRB products. We also used microsatellite analysis to investigate the genetic status of the RB locus. We correlated pRB alterations with proliferative activity, and with clinicopathological outcomes. RESULTS: Altered patterns of pRB expression are common in ASTS occurring in 84% of cases, and it is significantly associated with proliferative activity (p<0.001). Patients whose tumors either lack expression of pRB, or express hyperphosphorylated forms of pRB, have poor survivals compared to patients whose tumors exhibit a normal, underphosphorylated pattern of pRB expression (p=0.03). In addition, 63% of cases lacking expression of pRB showed loss-of-heterozygosity at the locus. CONCLUSIONS: Inactivation of pRB is common in adult STS, which may be due to either gene loss or post-translational modification, namely hyper-phosphorylation. Both mechanisms are associated with tumor cell proliferation and poor survival.

Transformations of elemental mercury to inorganic and organic forms in mercury and hydrocarbon co-contaminated soils.

There are many industrial sites, such as gas processing plants, that are contaminated with both mercury and hydrocarbons. These sites tend to be localized but can have very high concentrations of mercury in the soil and heterogeneous distribution of hydrocarbons. The original form of mercury in many cases was elemental mercury from broken manometers. Over time the mercury has become redistributed within soil and has undergone chemical transformations into new forms. The forms of mercury will govern the chemical behavior and the availability of the mercury to biological receptors. The availability of the mercury is important as it will govern the risk associated with the contaminated soil and will also determine the effectiveness of any attempts at remediation. In the present study a chemical extraction protocol was used to determine the forms of mercury in soil originally contaminated by spillage of elemental mercury and petroleum hydrocarbons. Chemical extractions have been used in the past to determine the forms of mercury in uncontaminated soils and several researchers have used them to study contaminated soils. However, to date, no researchers have studied the forms of mercury in soils following years of weathering of elemental mercury after a spill. This study shows that decades after the original spill the elemental mercury has transformed and is dominantly (up to 85%) associated with soil organic matter, and to a lesser extent the mineral fraction of soil.

Sorption of 1-naphthol to soil and geologic samples with varying diagenetic properties.

Remediation of contaminated land requires a firm understanding of the processes that occur between xenobiotics and soil colloids. It is currently accepted that the extent of xenobiotic uptake is proportional to the carbon quantity and character of the soil or geologic sample. Previous studies have developed empirical equations to predict the extent of sorption based on the aromatic carbon content. We examined these relationships with an independent set of soil and geologic samples and 1-naphthol. The 1-naphthol sorption coefficients varied significantly (P < 0.01) among sorbents and are consistent with the diagenetic properties of the organic matter in these samples. The cross-polarization magic angle spinning (CPMAS) 13C nuclear magnetic resonance (NMR) and elemental data did not concur with the sorption data for most of the soil samples. We suggest that this contradiction may be due to a third variable, the physical organization of the organic matter. Chemical methods measure the whole sample, whereas short-term sorption occurs on the surface; therefore, only some organic matter domains in the soil are available for interaction with 1-naphthol. Hence, chemical data alone may be insufficient for predicting the sorption behavior of xenobiotics in soil and geologic samples.

Alterations of cell cycle regulators affecting the RB pathway in nonfamilial retinoblastoma.

We undertook the present study to examine alterations affecting the RB pathway in the G1 checkpoint and to determine their potential clinical significance in children affected with nonfamilial retinoblastoma. Using immunohistochemistry, patterns of expression of pRB, p16/INK4A, and E2F1 were analyzed in tissue from a cohort of 86 well-characterized patients with nonfamilial retinoblastoma diagnosed at the "Instituto Nacional de Pediatria" in Mexico City. The relationship of these phenotypes to proliferative index was assessed by analysis of Ki67 antigen expression. pRB expression was found in 11 (13%) cases. Using a hypophosphorylated specific pRB antibody, we observed low levels of underphosphorylated pRB expression in only 1 of 9 evaluable positive cases. These data suggest that the detected pRB products were hyperphosphorylated and thus had decreased functional activity. Increased p16 nuclear expression was found in only 6 tumors. No tumors showed deletions or mobility shifts of the INK4A gene. Undetectable pRB levels were significantly associated with undetectable p16 expression (odds ratio, 10.8; 95% confidence interval, 1.4-81.3; P =.03). All tumors showed nuclear immunoreactivities for E2F1 and Ki67. Increased Ki67 proliferative index was associated with increased staining for E2F1 (r =.44; P =.008) and increasing clinical stage (P =.03). Among children with unilateral disease, the mean Ki67 proliferative index was significantly higher in children with advanced clinical disease (stages 3 and 4) (mean 81.25; SD 6.78) than in those with earlier stage disease (mean 69.50; SD 9.45) (P = 0.001). Among children with bilateral disease, however, the mean proliferative index was not significantly higher for children with advanced clinical stage. When examining all cases together, there was a significant trend toward increasing proliferative index with increasing clinical stage (P =.03). In unilateral tumors, we also found that presence of detectable pRB was associated with a lower percentage of cells expressing E2F1 (46.7% v 70.8%) (P = 0.05), whereas there was no association between presence of pRB and E2F1 among bilateral tumors. We have found that expression of some of the cell cycle markers examined varies according to laterality, suggesting underlying differences in the capacity for cell cycle regulation between these 2 forms of the disease. Differences in capacities for cell cycle regulation may account for some differences in clinical behavior. Thus, the inclusion of molecular markers may become useful adjuncts to clinicopathological staging and subsequent determination of therapy.

Experimental attempts to extend the current preimplantation genetic diagnosis with individual karyotypization of human blastomeres.

The drug-induced chromosome condensation using okadaic acid, a potent protein phosphatase inhibitor, was studied in day 1 to day 4 (D1-D4) spare human preimplantation embryos. In order to obtain cells for genetic tests, we developed a modified displacement blastomere biopsy method. During the okadaic acid treatment, approximately 40% of biopsied cells were lost due to heavy changes of the plasma membrane; this detrimental effect of okadaic acid differed markedly with the respect to the age of embryos. In comparison with the natural embryonic mitotic index, day 1 and day 2 embryonic cells gave increased yields of chromosome spreads (up to 51% of the initial D1-D2 cell number); on days 3 and 4 we were not able to obtain from surviving cells more than 31% blastomeres with condensed chromosomes (9% of total D3-D4 cell number). All chromosome spreads were successfully used for recycling in G-banding and subsequent FISH analysis.

Characterization of molecular abnormalities in human fibroblastic neoplasms: a model for genotype-phenotype association in soft tissue tumors.

Desmoid tumors and fibrosarcomas (FS) are part of a wide spectrum of disordered fibroblastic growth that display striking clinical and phenotypic differences. This study was designed to characterize molecular abnormalities that are associated with these differences and to determine their clinical relevance. A cohort of 24 desmoid tumors and 25 low-grade (LG) and 14 high-grade (HG) FS that were clinically and pathologically well characterized was analyzed for alterations in expression of Ki-67, Bcl-2, retinoblastoma gene product (pRB), and p53 by immunohistochemistry. LG-FS and HG-FS showed abnormal expression of Ki-67 (32 versus 86%), Bcl-2 (48 versus 57%), and pRB (56 versus 93%). In contrast, desmoid tumors showed a normal phenotype with these markers. p53 overexpression was identified in 20% of LG-FS and in 29% of HG-FS cases but only in 4% of desmoid tumors. There was an increasing trend in the proportion of abnormal expression of Ki-67, Bcl-2, pRB, and p53 with the increase of tumor aggressiveness from desmoid tumors to LG-FS to HG-FS. The molecular differences between tumor entities were highly statistically significant (P < 0.01). Significant associations between abnormal expression of pRB and recurrence-free survival of LG-FS patients (P = 0.05) and between Ki-67 overexpression and recurrence-free survival for tumors of >5 cm were observed (P = 0.02). The demonstrated differences of molecular alterations in HG-FS, LG-FS, and desmoids appear to be related to biological aggressiveness of such tumors, and they might be useful to differentiate between histologically similar cases of desmoid tumors and LG-FS. pRB and Ki-67 status may be useful to predict recurrence in certain subsets of patients.

Validation of tissue microarrays for immunohistochemical profiling of cancer specimens using the example of human fibroblastic tumors.

Tissue microarrays allow high-throughput molecular profiling of cancer specimens by immunohistochemistry. Phenotype information of sections from arrayed biopsies on a multitissue block needs to be representative of full sections, as protein expression varies throughout the entire tumor specimen. To validate the use of tissue microarrays for immunophenotyping, we studied a group of 59 fibroblastic tumors with variable protein expression patterns by immunohistochemistry for Ki-67, p53, and the retinoblastoma protein (pRB). Data on full tissue sections were compared to the results of one, two, and three 0.6-mm core biopsies per tumor on a tissue array. Ki-67 and p53 staining was read as two categories (positive or negative). Concordance for this staining between tissue arrays with triplicate cores per tumor and full sections were 96 and 98%, respectively. For pRB staining was read as three categories (high, moderate, or negative), where concordance was 91%. The use of three cores per tumor resulted in lower numbers of lost cases and lower nonconcordance with standard full sections as compared to one or two cores per tumor. Correlations between phenotypes and clinical outcome were not significantly different between full section and array-based analysis. Triplicate 0.6-mm core biopsies sampled on tissue arrays provide a reliable system for high-throughput expression profiling by immunohistochemistry when compared to standard full sections. Triplicate cores offer a higher rate of assessable cases and a lower rate of nonconcordant readings than one or two cores. Concordance of triplicate cores is high (96 to 98%) for two category distinction and decreases with the complexity of the phenotypes being analyzed (91%).

Prognostic role of cyclin D1 in retroperitoneal sarcomas.

PURPOSE: Cyclin D1 regulates the G1 checkpoint of the cell cycle and is overexpressed in a number of cancers. This study was designed to determine if cyclin D1 overexpression had prognostic value in patients undergoing surgery with curative intent for primary retroperitoneal soft-tissue sarcoma. METHODS: Tissue was available for analysis on 79 patients who underwent resection between September 1983 and May 1997. Clinicopathologic data and follow-up was obtained from a prospective sarcoma database and a patient and family interview. Immunohistochemical analysis was used to determine overexpression (> or = 5% of nuclei labeled). Survival was analyzed using the Kaplan-Meier method, and statistical analysis was performed by using log rank testing and the Cox regression model. RESULTS: Median follow-up was 3.5 years. On univariate analysis of disease-specific mortality, significant prognostic factors were high grade (n = 42, 53%), positive microscopic margins (n = 36, 46%) or macroscopic margins (n = 15, 19%), and cyclin D1 overexpression (n = 37, 47%). On multivariate analysis, macroscopically positive margins (P = 0.02) and the combination of high grade and cyclin D1 overexpression (P = 0.04) both were associated independently with poor survival. CONCLUSION: High grade retroperitoneal sarcomas demonstrating cyclin D1 overexpression have had an extremely poor prognosis. Continued study of multiple biological markers, exemplified by cyclin D1, may aid characterization of tumor behavior and response to treatment in this diverse group of tumors.

Alterations of cell cycle regulators in localized synovial sarcoma: A multifactorial study with prognostic implications.

Genetic alterations of cell cycle regulators are thought to represent uncommon and possible secondary events in sarcomas characterized by recurrent chromosomal translocations. The present study investigates this hypothesis on synovial sarcoma (SS), assessing the frequency of expression and possible clinical implications of detecting alterations in critical cell cycle regulatory proteins. A homogeneous cohort of 49 patients with localized SS, restricted to the extremity and with available long-term follow-up information, was selected from our files. We focused our study on molecules involved in the G1 checkpoint and G1-S transition, including cyclins D1 and E, p21(WAF1), p27(Kip1), mdm2, p53, and Ki67. A cutoff point of 10% immunoreactive tumor cell nuclei was selected to define a positive phenotype for any given marker, except for Ki67. High Ki67 proliferative index was considered when >/=20% tumor cells displayed nuclear immunoreactivity. Biphasic SS were analyzed, taking into account separately the expression of these proteins in the spindle and glandular components. Disease specific survival was modeled using the Kaplan-Meier method with log rank test and Cox regression. The cohort of patients analyzed included 23 females and 26 males, and the histological type distribution was 35 monophasic and 14 biphasic SS. The median follow-up for survivors was 53 months, with a 5-year disease-specific survival of 63% and a metastatic disease-free survival of 40%. The positive phenotypes identified for the different markers studied were as follows: cyclin D1, 59%; cyclin E, 29%; p21, 51%; p27, 69%; mdm2, 59%; p53, 16%; and Ki67, 59%. We observed that positive p53, cyclin E, and high Ki67 proliferative index were correlated with survival, but only Ki67 and p53 were independent variables for prognostication. The present study suggests that alterations of cell cycle regulators are more common events in SS than originally thought. p53 overexpression could be of use as a marker together with a high Ki67 proliferative index, in identifying a subset of SS patients with increased risk of tumor relapse.

[Genetic polymorphism of enzymes metabolizing xenobiotics as a factor in susceptibility to tumor development]. / Genetický polymorfizmus enzýmov metabolizmu xenobiotík ako faktor susceptibility pri vzniku nádorových ochorení.

Organisms are exposed to a large number of xenobiotics (compounds foreign to the body), such as drugs, pesticides, natural food constituents and so on. To deal with these, usually lipophilic substances, a range of biotransformation enzyme systems are available. Experimental data have shown that metabolic activation and detoxification play an important role in chemical carcinogenesis. There is a considerable interindividual genetic variability in these pathways which might explain the differences in cancer susceptibility and might ultimately enable the identification of people at increased risk and to offer individualized cancer prevention. Recently, molecular genetic bases of many polymorphic enzyme activities involved in xenobiotics metabolism have been elucidated. The purpose of this article is to provide a brief review of recent findings concerning the association of genetically determined metabolic variants with different risks of environmentally induced cancer. (Tab. 6, Fig. 1, Ref. 30.)

Expression of p27(kip) and other cell cycle regulators in malignant peripheral nerve sheath tumors and neurofibromas: the emerging role of p27(kip) in malignant transformation of neurofibromas.

There is little information regarding the status of cell cycle regulators in malignant peripheral nerve sheath tumors (MPNSTs) and neurofibromas (NFs). In this study, we investigated patterns of expression of p53 and pRB, cyclin-dependent kinase inhibitors (CKIs) p21 and p27, as well as cyclins D1 and E, in a cohort of 35 well-characterized MPNSTs and 16 NFs. These phenotypes were correlated with proliferative index, as assessed by Ki-67, as well as clinicopathological parameters of poor outcome. p53 nuclear overexpression was found in 10 of 35 (29%) MPNSTs, and it was lacking in NFs (P = 0.02). There were no differences in the patterns of expression of pRB, cyclin D1, and p21 between MPNSTs and NFs. However, p27 nuclear expression was present in most NFs, but it was absent in the majority of MPNSTs, which displayed cytoplasmic staining (P < 0.001). Nuclear cyclin E expression was more pronounced in MPNSTs than in NFs. We observed inverse patterns of expression for nuclear p27 and nuclear cyclin E expression. The staining profiles of cytoplasmic p27 and nuclear cyclin E expression were found to be statistically associated (P = 0.01). High Ki-67 expression was found in 20 of 34 (59%) MPNSTs but was absent in NFs (P < 0.001). Furthermore, detection of cytoplasmic p27 expression was found to be a prognostic factor for poor survival in MPNSTs (P = 0.03, relative risk = 2.4).

Involvement of the Ink4a gene (p16 and p19arf) in murine tumorigenesis.

The INK4A and INK4B genes map to 9p21, with the INK4A gene encoding two products, p16 and p19ARF. Many neoplasms in which INK4A and INK4B genes are altered show deletions involving both genes. Mice carrying a targeted Ink4a deletion develop tumors at an early age. In the present study we examined the genetic alterations affecting the remaining Ink4a allele and the Ink4b gene in tumors arising in heterozygous Ink4a mice. We identified deletion of the remaining Ink4a allele in 7 of 18 (39%) tumors. We also observed deletion of the exon 1beta in 3 cases, one of them presenting this deletion as a unique alteration. In conclusion, the deletion of the remaining Ink4a allele was the alteration most frequently observed, representing the inactivation of two proteins capable of arresting the cell cycle through different pathways that involve the tumor suppressors pRB and p53.

Prognostic significance of transcription factor E2F-1 in bladder cancer: genotypic and phenotypic characterization.

BACKGROUND: We sought to identify and characterize potential alterations in E2F-1, a transcription factor that binds to the retinoblastoma protein (pRB), in bladder neoplasms and to elucidate a possible role for E2F-1 as an oncogene or a tumor suppressor gene. METHODS: Tumor samples from 133 evaluable patients with bladder cancer were analyzed for E2F-1 gene mutations by use of polymerase chain reaction-single-strand conformational polymorphism (PCR-SSCP) analysis and DNA sequencing. In addition, tumors were studied for E2F-1 and pRB protein expression by use of immunohistochemistry. Results from the above analyses were correlated with clinicopathologic parameters and outcome. All P values are two-sided. RESULTS: A polymorphism, consisting of a nucleotide change at amino acid codon 393 in exon 7 (GGC-->AGC [Gly-->Ser]), was identified in seven of 133 case patients, being present in both tumor and corresponding normal tissues. No bandshifts were identified in the nuclear-localization or DNA-binding domains on PCR-SSCP analysis. On immunohistochemical analysis, E2F-1 nuclear reactivity was observed in less than 5% of the cells from 53 tumors and in 5%-75% of the cells from the remaining 80 tumors. The pattern of E2F-1 protein expression was not altered in relation to the identified polymorphism. pRB nuclear reactivity greater than 20% (of tumor cells stained) was present in 66% of the samples. E2F-1 nuclear reactivity correlated inversely with the percentage of cells showing pRB reactivity (Kendall tau(b) = -0.18; P = .019). On multivariate analysis, patients with lower E2F-1 reactivity had statistically significantly increased risks of progression to metastases (P = .001) and death (P = .02). CONCLUSIONS: E2F-1 alterations occur at the phenotypic level, rather than at the genotypic level, in bladder cancer. The adverse outcome for patients whose tumors exhibit low E2F-1 nuclear expression suggests a possible tumor suppressor role for E2F-1 in bladder cancer.

[Clinical and pathologic description of stroke patients (1990-1994)]. / Stroke betegek klinikai és patológiai jellemzói (felmérés 1990-1994-ból).

UNLABELLED: Data relating to stroke patients at a department of internal medicine (50 beds) in a county teaching hospital were studied in period 1990-1994. In this five-year period, 1184 patients were admitted because of some forms of stroke. The mortality due to the stroke was 16.8% (199 patients, deceased group). Autopsy was performed on 159 of these 199 patients (autopsy rate: 79.8%). Age- and sex-matched controls were selected from the survivors (n = 159). The main risk factors of stroke were analyzed in both groups: hypertension, cardiac events (decompensation, atrial fibrillation, and old myocardial infarction), previously verified diabetes, and recurrent stroke in the history. The mean hospital nursing time for the survivors was 12.3 +/- 6.3 days, while that in the deceased 7.2 +/- 7.6 days. RESULTS: 1. Hypertension was present to similar extents in both groups (survivors: 82.1%, deceased group: 77.8%) 2. Decompensation occurred in 5% vs 18.2% fibrillation in 11.3% vs 13.8%, and old myocardial infarction in 5.6% vs 18.2% 3. Diabetes was observed in 21.3% vs 36.4% and 4. Recurrent stroke in 22.6% vs 39.6%. These risk factors strongly predicted the outcome of the stroke. Other recently observed factors (haemorrhagic form, conscious state, time of hospital admission, seasonal variation, higher erythrocyte sedimentation rate, hyperglycaemia, proteinuria, early deep vein trombosis) revealed also significant differences between survivors and deceased patients. Since pulmonary thromboembolism was twice as frequent in the deceased patients as in the survivors, early heparin prevention is necessary immediately after computer tomography which excluded the haemorrhagic type of cerebrovascular diseases.

Overexpression of cyclin D1 is associated with poor prognosis in extremity soft-tissue sarcomas.

Binding of G1 cyclins to cyclin-dependent kinases leads to phosphorylation of the retinoblastoma protein and progression through G1 and S phases of the cell cycle. Overexpression of cyclins is thought to deregulate this process and has been noted in many human malignancies. This study was conducted to assess patterns of expression and potential gene amplification of the G1 cyclins in 84 patients affected with extremity soft-tissue sarcomas. Sixty cases were primary tumors, whereas the remaining 24 cases were locally recurrent lesions. There were 58 high-grade and 26 low-grade tumors. Immunohistochemical analyses were conducted using antibodies to cyclins D1, E, and A. Southern blot analysis was performed on DNA available from 53 of 84 patients with a cyclin D1-specific probe. Cyclin D1 overexpression was noted in 23 of 79 informative cases (29%), whereas cyclin E was found overexpressed in 26 of 80 cases (33%) and cyclin A overexpression was observed in 9 of 81 cases (11%). Overexpression of cyclins D1, E, or A each correlated significantly with high tumor grade (P <0.05). On multivariate analysis, neither cyclin E nor cyclin A were significant predictors of outcome. However, overexpression of cyclin D1 was significantly associated with worse overall survival for the entire group as well as in the subset of high-grade lesions (P <0.05), notwithstanding the relatively short follow-up time (mean, 2.4 years). Nevertheless, the presence of a significant association between laboratory data and outcome implies that the study is adequately powered. Furthermore, none of the cases demonstrated CCND1 gene amplification. These data support the concept that cyclin D1 overexpression determines the evolution of a particularly aggressive subset of these lesions.