Clinico-morphological analysis of the neuroendocrine neoplasms of the gastroenteropancreatic system.

Gastroenteropancreatic neuroendocrine neoplasms (GEP-NENs) are unusual and rare epithelial tumors for which the diagnosis of the grade of malignancy and prognostic assessment on the basis ofhistology represent considerable challenges for the pathologist. In this study we conducted a retrospective analysis of the cell proliferation (Ki-67 nuclear antigen) in primary and metastatic GEP NENs of 137 patients as well as the assessment of keratin 19 (CKI9) and progesterone receptors (PR) expression in pancreatic NENs of 57 patients. In 19 (27,1%) GEP NEN metastases was found I,5-4,5-fold increase of Ki-67 index compared with their primaries. Consequently, 6 (21,4%) cases of NETGI and 4 (7,0%) cases of NET G2 were up-graded Pancreatic NETs G2 with Ki-67 index >5% were significantly associated with presence of distant metastases (p =0,007) and decreased survival (p=0,03). Decreased survival also was found in the group of gastrointestinal NET G2 with Ki-67 index > 15% (p=0,005). Further analysis of immunomorphological features and proliferative activity allowed to separate a rare group of tumors - ("NET G3", characterized by decreased survival comparing to NET G2. Expression of CKI9 in pancreatic NETs was significantly associated with higher proliferative activity of primary tumor (p =0,04) and adverse outcome (p=0,003). On other hand, PR expression correlated with lower Ki-67 index (p=0,006), absence of metastases (p=0,004) and favorable outcome (p=0,000). Our results show that Ki67 index is a key parameter of morphological diagnosis of GEP NENs. Thus, the studied markers are important parameters of the morphological diagnostic of GEP NENs, which allow more accurately assess the degree of malignancy, prognosis and treatment of the disease.

[Expression of estrogen receptors beta and beta1 in tissue of human non-small cell lung cancer].

Comparability of the level and intensity of estrogen receptors beta (ERbeta) expression in non-small cell lung cancer tissue of 32 patients was analyzed by flow cytometry using various antibodies--to the total fraction of ERbeta (clone 14C8) as well to the full-length ERbeta1 isoform (clone EMRO2). The differences in the ER expression indexes detected by anti-ERbeta or anti-ERbeta1 antibodies were revealed in some patients, but it had no influence on average indexes of the ERbeta expression in the patient groups investigated. It was confirmed by the findings on more frequent and more intensive expression of ERbeta in the non-small cell lung cancer tissue of female patients vs. the males irrespective of antibody type - anti-ER/ or anti-ERbeta1. Therefore, in comparative analysis of ERbeta expression in the groups of the patients with different clinicomorphologic characteristics of the disease it is possible to use both the antibodies. For individual disease prognosis in the routine clinical practice it is recommended to use the antibodies to the total fraction of ERbeta, since there are individual differences between the ERbeta expression indexes revealing by various types of antibodies.

Squamous cell carcinoma of the esophagus: evaluation of the status of epidermal growth factor receptors (EGFR and HER-2) by immunohistochemistry and in situ hybridization.

We performed a parallel evaluation of the status of epidermal growth factor receptors EGFR and HER-2 in tumor samples from 31 patients with squamous cell carcinoma of the esophagus. Hyperexpression of proteins was detected by immunohistochemical methods and gene amplification and other chromosome abnormalities were studied using FISH reaction. Evaluation of EGFR status showed that amplification of EGFR gene was present in 25% cases and chromosome 7 polysomy was detected in 29.2% cases positive by protein expression (2+/3+). Immunohistochemically positive EGFR status was confirmed by the results of FISH reaction for gene amplification and chromosome 7 polysomy in 54.2% cases (p=0.002). During evaluation of HER-2 status in the tumor, hyperexpression of the protein detected histochemically was not confirmed by FISH reaction for detection of amplification of the corresponding gene in 16.1% cases. In 22.6% patients, chromosome 7 polysomy was detected; it was not accompanied by amplification of HER-2 gene, but was related to immunohistochemically positive status of the tumor. Hyperexpression of EGFR protein significantly correlated with the presence of intravascular invasion (p=0.006) and increased depth of invasion (p=0.044), while amplification of EGFR gene (≥2.2) correlated with low differentiation degree of the tumor (p=0.006). The outcome of the disease was not associated with EGFR status at the gene and protein levels, whereas clinical course of the disease in patients with immunohistochemically negative expression of HER-2 protein was more favorable than in patients with positive expression (p=0.004). The results of this study suggest that hyperexpression/amplification of EGFR and hyperexpression of HER-2 are important clinical markers for evaluation of disease prognosis and development of new regimens of targeted therapy for patients with squamous cells carcinoma of the esophagus.

[Immunohistochemical study of epidermal growth factor receptor expression in esophageal squamous cell carcinoma].

Primary tumors from 31 patients with esophageal squamous cell carcinoma (ESCC) were immunohistochemically studied for the expression of markers of ESCC in order to define the clinical value of the levels of EGFR and HER-2 in the tumors. EGFR and HER-2 hyperexpression in the tumors of patients with ESCC was ascertained to be an important marker for the analysis of the clinical features of ESCC. There was an association of the elevated levels of EGFR and HER-2 in the tumors of ESCC patients with the presence of vascular tumor invasion (p = 0.006) and that with the poor outcome of the disease (p = 0.004). The findings suggest that estimation of changes found in EGRF and HER-2 expression in the tumors of patient with ESCC is of great interest for the individual prediction of the disease course and for the development of new approaches to treating these tumors, including targeted therapy against these tyrosine kinase receptors.

[Actins and keratins in the diagnosis of human basal-like breast cancer].

The most common forms of luminal breast cancer (BC) were compared with basal-like and Her2/neu3+ BC. Their primary classification was based on morphological diagnosis and the expression of estrogen receptor (ER), progesterone receptor (PR), and Her2/neu receptors. Monoclonal antibodies to actins and keratins were used for the study. Basal-like BC cells (ER/PR/Her2/ neu-) were regularly stained with antibodies to basal keratins 5/6 and 17, smooth muscle alpha-actin, and p63. Luminal keratin 8 staining was reduced. The solid regions had beta-actin staining with disappearance in the scirrhous component. beta-actin and basal keratins were also observed in metaplastic BC with ER/PR/Her2/neu3+. Immunomorphology using cytoskeletal markers along with the expression of steroid hormone and Her2/neu receptors may be used in the diagnosis of basal-like forms of BC.

Clinical significance of hyperexpression of epidermal growth factor receptors (EGFR and HER-2) in esophageal squamous cell carcinoma.

Immunohistochemical study of marker expression in primary tumors of patients with esophageal squamous-cell carcinoma was carried out in order to evaluate prognostic significance of EGFR and HER-2 levels in the tumors. Hyperexpression of EGFR and HER-2 in the tumors is an important marker for the analysis of prognosis and clinical course of the disease. A relationship between high levels of EGFR and HER-2 in the tumors of patients with esophageal squamous-cell carcinoma and intravascular tumor invasion (p=0.038) and poor outcome of the disease (p=0.019) was detected. The results indicate that evaluation of changes in the expression of EGFR and HER-2 in tumors is essential for individual prediction of the disease course and development of new approaches to the treatment of these tumors, including target therapy aimed at these tyrosine kinase receptors.

Epidemiological, clinical and viral determinants of the increased prevalence of high-risk human papillomavirus (HPV) infections in elderly women.

BACKGROUND: Population-based studies have reported a second peak of human papillomavirus (HPV) prevalence among women > 55 years, but reasons for this U-shaped HPV prevalence curve are poorly understood. OBJECTIVES: To analyse determinants of high-risk HPV (HR-HPV) infections among postmenopausal women. STUDY DESIGN AND METHODS: A cohort of 3,187 women was stratified into three age categories: i) youngest age group < 25 years (n = 1.103); ii) women between 26-55 years (n = 2.004), and iii) women > 55 years (n = 80), analysed for epidemiological, clinical and virological determinants of their HR-HPV infections. Real-time PCR was used for HPV genotyping, analysis of viral loads for HPV16, 18/45, 31, 33/52/58, 35 and 39, and load of integrated HPV16. RESULTS: Age-standardised prevalence of HR-HPV infections showed a second peak among women > 55 years, with a perfect U-shaped curve (R2 = 0.966). The factors explaining this increased HR-HPV prevalence among older women include: i) cohort effect, ii) higher viral loads for HR-HPV types with cubic model curve (R2 = 0.714) for HPV 16, iii) distinct shift (p = 0.0001) from multiple-type infections to single HR-HPV types, iv) transition from episomal to integrated HPV16 (p = 0.009), v) higher load of integrated HPV16 (p = 0.009), and, vi) higher proportion of incident infections, higher rate of viral persistence, and lower rate of HR-HPV clearance. CONCLUSIONS: These data suggest that in women who fail to eradicate their HR-HPV infection until menopause, selection of integrated viral clone has taken place, driving the process towards progressing disease. Consequent to this, most of the HR-HPV infections in women > 55 years were associated with high-grade CIN or invasive carcinoma.

[Female genital tract polyneoplasia: primary multiple neoplasms or metastases?].

The analysis of 49 cases of synchronous and metachronous malignant mucinous tumors of the colon (rectum) and ovaries in the patients treated in 1990 to 2004 again has confirmed the data that metastatic ovarian cancer occurs from a primary focus in the colorectal region. Immunohistochemical studies (using cytokeratin 7 and cytokeratin 20) may be used in the differential diagnosis of ovarian mucinous ovarian carcinoma from metastatic colonic mucinous tumors.

Clearance of high-risk human papillomavirus (HPV) DNA and PAP smear abnormalities in a cohort of women subjected to HPV screening in the New Independent States of the former Soviet Union (the NIS cohort study).

BACKGROUND: We analysed the temporal relationships of the clearance of human papillomavirus (HPV) DNA and cytological abnormalities in women participating in a screening study in three NIS countries. METHODS: The 274 patients included in this analysis were prospectively followed-up for 21.6 months (range: 0.5-42.9). All 274 women had abnormal PAP test (ASC-US or higher) and high-risk HPV-positive test (HCII) at baseline. Two groups were compared: 132 women who cleared both tests (Group 1), and 142 women who cleared either HPV or abnormal PAP test (Group 2). The first clearance during the follow-up, and the last visit clearance were modeled using life-table techniques, and the predictive factors were analysed using univariate (Kaplan-Meier) and multivariate (Cox) survival analysis. RESULTS: There was no difference in the mean clearance time for the abnormal PAP test (14.4 months; 0.7-40.5 and 12.6 months; 0.5-35.0) and high-risk HPV DNA (12.67 months; 0.6-33.5 and 10.8 months; 0.7-33.4) in Group 1 and Group 2 (Mann-Whitney: P = 0.107 and P = 0.082, respectively). Clearance times for HPV DNA and abnormal PAP test did not deviate from each other in either groups (Wilcoxon: P = 0.063 and P = 0.088). The monthly clearance rates for the abnormal PAP test are 1.32 and 1.38%, and those for the HPV DNA 1.62 and 1.61%, in Groups 1 and 2, respectively. Of the factors predicting the last visit clearance, the issues related to smoking are of particular interest. CONCLUSIONS: The clearance of high-risk HPV type and abnormal PAP test shows a close temporal relationship, the former preceding the latter, however, by an interval of 1.0-2.0 months.

Factors predicting persistence of high-risk human papillomavirus (HPV) infections in women prospectively followed-up in three New Independent States (NIS) of the former Soviet Union.

BACKGROUND: We completed an analysis of the factors predicting the persistence of high risk (HR) HPV infections in women participating in a multicenter screening trial in three NIS countries. METHODS: The 543 baseline HR HPV-positive women included in this analysis are derived from a sub-cohort of 887 women who were prospectively followed-up for a mean of 21.6 months (range: 0.5-42.9) as a part of a multi-center screening study in three NIS countries (the NIS cohort study; n = 3,187 women). Of these 543 women, 273 showed persistent HR-HPV in serial Hybrid Capture II (HCII) testing during the follow-up (Group 1), whereas 270 women cleared their infection (Group 2). These two groups were compared with their epidemiological, clinical, and virological data (HCII, PCR) to disclose the factors predicting persistent HR-HPV infection. RESULTS: Women with persistent HR-HPV infections were significantly younger (27.3 yrs) than those who cleared their infection (29.1 yrs) (p = 0.006), and their follow-up time was shorter; 14.1 and 21 months, respectively (p = 0.0001). Both variables were treated as confounders in the multivariate analyses. Of the 66 recorded epidemiological variables, only being a current smoker proved to be an independent predictor (OR 1.693; 95% CI 1.114-2.573; p=0.014). Baseline colposcopy, biopsy or Pap smear did not predict HPV persistence, whereas an incident or persistent abnormal Pap during the follow-up were independent predictors in a multivariate model (p = 0.005), together with the high viral load (HCII RLU/CO at 100 pg/ml cut-off), and HR HPV positive PCR test (p = 0.0001). When all significant variables were entered in the regression model, only the follow-up time (OR 0.950, 95% CI 0.924-0.976; p = 0.0001) and HR-HPV positive PCR (OR 4.169, 95% CI 1.741-9.987; p = 0.001), remained independent predictors. CONCLUSIONS: While several factors were related to HR-HPV persistence in univariate analysis and when adjusted for age and follow-up time as confounders, the only independent predictors in the multivariate regression model were follow-up time and HR-HPV positive PCR. Clearly more data are needed on type-specific persistence and HPV integration as its predictors.

[Infiltrating lobular breast cancer: histological and immunohistochemical characteristics].

Lobular carcinoma ranks second among most frequent tumors of the breast. Metastases to the lymph nodes and distant organs are one of the most important prognostic factors. 60% regional lymph node metastases were morphologically diagnosed during surgical removal of the tumor. Distant metastases rate was 1% at the beginning of the treatment and reached 10% within 5-year follow-up. Cases of secondary uterus and epiploon lobular carcinoma are rare and tumor location in the breast should be proved.

Elevated content of p53 protein in the absence of p53 gene mutations as a possible prognostic marker for human renal cell tumors.

p53 tumour suppressor gene expression was estimated immunohistochemically using DO-1 monoclonal antibody (recognising both wild-type and mutant p53 in 88 human renal tumours. Single strand conformation polymorphism (SSCP) analysis of possible mutations within exons 4-8 of the p53 gene was performed in 29 of the tumours (mostly immunostaining-positive cases). Obviously elevated p53 content was detected with DO-1 antibody in chromophobic cell carcinomas and most clear/chromophilic cell tumours (in chromophilic cell populations). In contrast, clear cell carcinomas demonstrated either complete absence of p53 expression or the presence of single immunopositive nuclei. Oncocytomas were completely negative. Additional immunostaining of the positive samples with mutant p53-specific Pab240 monoclonal antibody failed to detect immunopositive material. No p53 mutation was found in any of the samples analysed by SSCP. Our results suggest that the elevated p53 content in human renal cell carcinomas does not result from gene mutation and the p53 gene alterations are probably not an important mechanism in the development of human renal cell carcinomas. Accumulation of the wild-type p53 protein may be a useful prognostic marker indicating neoplastic progression malignancy.

[Effect of sex hormones on the induction of renal capsule angiosarcomas in mice]. / Vliianie polovykh gormonov na induktsiiu angiosarkom pochechnoi kapsuly u myshei.

Pararenal angiosarcomas appear in CBA male mice treated with 1,2-dimethylhydrazine (DMH). No such tumours appeared in the intact females treated with DMH alone. Testosterone propionate (TP) given in combination with DMH (28%) was efficient in the incidence of DMH-induced pararenal tumours in females. Pararenal angiosarcomas appeared in 92% of castrated CBA female mice under combined treatment with DMH and TP. No such tumours appeared in C57B1 males treated under the same experimental conditions.

[1,2-dimethylhydrazine induction of epithelial tumors of the kidneys in CBA strain mice]. / Induktsiia 1,2-dimetilgidrazinom épitelial'nykh opukholei pochek u myshei linii CBA.

The results of experiments on the 1,2-dimethylhydrazine (DMH) induction of epithelial renal tumours in CBA male mice are presented. The dose-response study shows a sharp increase (from 5 to 75%) of the epithelial renal tumour incidence in the range of 2, 4 and 8 injections of DMH. Higher doses induce a decrease of the tumour incidence due to the early death caused by other tumours. DMH is shown to be the most powerful renal carcinogen in mice. Serial sacrifice of mice after 5 injections of DMH is a convenient model for the study of renal carcinogenesis in mice. Main histological types of epithelial renal tumours are illustrated.

[Histogenesis of experimental renal tumors in mice]. / O histogeneze éksperimental'nykh opukholei pochek myshei.

Epithelial kidney tumours induced in CBA male mice by 1,2-dimethylhydrazine were studied histochemically and immunohistochemically. Histologically, 48 tumours studied were diagnosed as clear-cell, acidophilic or mixed adenomas located in the renal cortex. Gamma-glutamyl transpeptidase (GGT) was strongly positive in normal proximal convoluted tubules, slightly positive in the cells of Bowman capsule and negative in 47 of 48 tumours examined. Antibodies against the new antigen obtained from the mouse liver oval cells and called A6 antigen were also used. This antigen in normal kidney is negative in proximal tubules but always positive in distal tubules and collective ducts. It was also positive in all 47 GGT-negative tumours. One tumour was GGT-positive and A6 antigen-negative. The conclusion is made that the majority of renal cell adenomas in mice most likely originate from the collective duct system and not from the proximal tubules.

[Combination of pulmonary lymphangioleiomyomatosis (LAM) with angiomyolipoma (AML) of the kidney and multilocular cystic nephroma]. / Sochetanie limfangioleiomiomatoza, angiolipomy pochki i mul'tilokuliarnoi kistoznoi nefromy.

A rare case of pulmonary LAM in combination with AML and multilocular cystic nephroma of the kidney in a 18-year-old female is described. Granules of the melanosomic type in AML, HMB-45 antigen and steroid receptors expression in LAM and AML were observed.

[Effect of castration and mouse strain on 1, 2-dimethylhydrazine induction of pararenal angiosarcomas and kidney adenomas]. / Vliiania kastratsii i linii myshei na induktsiiu pararenal'nykh angiosarkom i adenom pochek 1, 2-dimetilgidrazinom.

Castration preceding the administration of 1, 2-dimethylhydrazine (DMH) to CBA male mice inhibits the induction of pararenal angiosarcomas (from 76% to naught) and renal adenomas (from 89 to 57%). Male mice of different strains treated with DMH developed the following incidences of pararenal angiosarcomas and renal adenomas: C3H - 35.5 and 13%, respectively, CBA - 97 and 78.8%,m (CBA X C57B1) F1-36.1 and 30%. C57B1-4.2 and 23.4%, BALB/c-13 and 43.4%, C3HA -7.1 and 14.3%. Pararenal angiosarcoma develops most likely from vascular bed of renal capsula. The mechanisms of androgen stimulation of renal adenoma and pararenal angiosarcoma induction are discussed.

[Modifying effect of DDT and of its various metabolites on the toxic effect of 7,12-dimethylbenz(alpha)anthracene]. / Modifitsiruiushchee deistvie DDT i nekotorykh ego metabolitov na toksicheskii effekt 7,12-dimetilbenz(alpha)antratsena.

In rats a preliminary three times injection of DDT in doses of 10 and 100 mg/Kg eliminated completely DMBA (30 mg/per 100 g of weight) lethal effect and prevented or decreased the development of adrenal necrosis. Among the metabolites under study (DDT, DDON, DDMU, DDE, DDA, DVR) DDE was found to render the strongest protective action in its single administration in a dose of 10 mg/Kg a day before the exposure to DMBA.

[Morphogenesis and histogenesis of experimental malignant hemangioendothelioma]. / Morfogenez i gistogenez éksperimental'noi zlokachestvennoi gemangioéndoteliomy.

Film preparations, various histological stains, autohistoradiography and electron microscopy were used to study morphologic changes occurring in the course of formation of vascular kidney tumors in male CBA mice after subcutaneous injection of 1.2-dimethylhydrazine. Twenty-thirty animals were examined every 4 weeks up to 45-50 weeks. In mice, renal capsule (site of malignant hemangioendothelioma growth) is avascular. Blood vessels developed due to "formation of lumen" in bands of anastomosed cells. Endothelium of the newly formed vessels originated from multipotent mesenchymal cells. Also, intracytoplasmic formation of vascular lumens was observed. The initial neoplastic vascular structure (telangiectases) were registered after 20-30 weeks. They were composed of markedly atypical cells characterized by columnar shape, high nucleo-cytoplasmic ratio and high proliferative activity. Further progression was associated with the development of cavernous hemangioma-like tumors bearing cytologic signs of malignancy as well as well- and poorly-differentiated hemangioendotheliomas.

[Morphology of experimental malignant hemangioendothelioma]. / Morfologiia éksperimental'noi zlokachestvennoi gemangioéndoteliomy.

Various histological stains, histoautoradiography and electron microscopy were used to study the morphologic features of 77 malignant hemangioendotheliomas induced in CBA mice with subcutaneously injected 1,2-dimethylhydrazine. The histologic and cytologic structure of those tumors proved similar to those of malignant hemangioendothelioma in man. The ability of tumor cells to form vascular structures and degree of their neoplastic atypia were used to identify three types of neoplasia: "cavernous hemangioma" with cytologic signs of malignity and well- and poorly--differentiated malignant hemangioendotheliomas. Due to high rate of tumor development and inevitable growth in the renal capsule which makes tumor easy to detect at early stage, the model offers promise for evaluating morphogenesis of malignant hemangioendothelioma.