Human papillomavirus and p16 expression in inverted papillomas of the urinary bladder.

Human Papillomaviruses (HPVs) have been found in association with benign and malignant growth of epithelia. The cell cycle inhibitor p16(Ink4a) has been shown to be overexpressed in HPV-positive cervical pre-malignant and malignant lesions, probably as a result of pRB targeting by the viral E7 protein. Inverted papillomas of the urinary bladder are epithelial tumors considered to be of benign nature. In this report we analyze the expression of p16(Ink4a) and the presence of HPV sequences in inverted papillomas and in non-tumoral bladder controls. Our results show no association of HPV infection and inverted papillomas. Further, no correlation between p16 overexpression and HPV positivity was found. We conclude that HPV does not play an indispensable role in the development of urinary bladder inverted papillomas and that overexpression of p16(Ink4a) does not correlate with HPV infection in these tumors.

Prostate cancer detected by uPM3: radical prostatectomy findings.

uPM3 is the first urine-based genetic test that is highly specific for detecting prostate cancer. The histopathologic characteristics of uPM3-detected cancer in radical prostatectomy specimens have not been previously described. We evaluated a consecutive series of radical prostatectomies to determine the extent, zonal distribution and other features of prostate cancer following uPM3 detection. A total of 24 whole-mounted, totally embedded radical prostatectomy specimens were evaluated. All patients had clinically localized cancer and none received preoperative therapy. Zonal location of cancer, distance to the urethra, cancer volume, Gleason grade and multicentricity were recorded; volume of cancer was measured using the grid-counting method. Patients ranged in age from 43 to 75 years (mean 65 years). In 21/24 cases, cancer involved the transition and peripheral zones and was multicentric (87%). Mean cancer volume was 3.96 cm3 (range 0.08-16.86 cm3). Mean distance to the urethra was 0.50 cm for the closest cancer and 0.61 cm for the most distant cancer. Mean Gleason score was 7 (range 5-9); pathologic stage was pT2 for 17 cases and pT3 for seven cases. The uPM3 is an independent and specific biomarker that detects prostate cancers of similar volume, location, extent and grade as other tests for early detection. uPM3 does not preferentially identify large or aggressive prostate cancers.

The phosphorylated form of connexin43 is up-regulated in breast hyperplasias and carcinomas and in their neoformed capillaries.

We applied an antiserum (SA226P) specifically recognizing the phosphorylated form of connexin43 (P-Cx43) to human breast samples including normal breast samples, with fibrocystic disease (FCD), fibroadenomas (FA), in situ and infiltrating carcinomas of all major types, and miscellaneous extramammary tumors. The findings were compared with those obtained with commercial antisera recognizing all Cx43 forms (pan-Cx43). A subset of samples was stained for Her2-neu and p44/42 to mitogen-activated protein kinase. Paraffin step sections were used. Immunoblots were performed on frozen samples of a representative subset of cases. In the normal breast, FCD, and FA, SA226P stained strongly and extensively most myoepithelial cells (MECs); luminal cells remained unstained. In proliferative FCD and some cellular FA, SA226P stained MEC and the capillary endothelium (CE). In ductal and lobular in situ carcinomas, SA226P reacted strongly and diffusely with the remaining MEC, the CE, and the transformed luminal cells. SA226P stained all infiltrating carcinomas except the tubular variant. In all breast carcinomas, the CE within and adjacent to tumors and some myofibroblasts stained with SA226P. By contrast, pan-Cx43 stained weakly and sporadically the MEC and rare samples of invasive carcinomas. Notably, Mab p44/42 reacted in parallel with the samples stained with SA226P, whereas reactions with Her2 were negative. Immunoblot findings paralleled those obtained immunohistochemically. We conclude that P-Cx43, restricted to MEC in the normal breast, is up-regulated in the same cells in hyperplasias and dysplasias and FA and is strongly up-regulated in invasive carcinomas. Notably, in some proliferative FCD and in most in situ and infiltrating carcinomas, P-Cx43 is strongly expressed in CE within and adjacent to the lesions but not away from them. These findings were paralleled by the strong nuclear reactions noted with Mab p44/42. These phenomena, although not exclusive to malignancy, are particularly conspicuous in breast carcinomas and seemingly reflect active proliferation associated with abnormal gap junctional intercellular communication.

Immunolocalization of the Epstein-Barr nuclear antigen-1 in conjunctival squamous carcinomas and dysplasias.

Epstein-Barr virus (EBV) has been linked etiologically to infectious mononucleosis, some non-Hodgkin as well as Hodgkin lymphomas, and lymphoepithelioma-like carcinomas. Moreover, various EBV antigens have been identified by a variety of techniques in a number of visceral carcinomas including breast, prostate, colon and lung primaries. We have now demonstrated by immunohistochemistry the presence of EBV nuclear antigen-1 (EBNA-1) in 4 of 15 cases of conjuntival squamous carcinomas and related dysplasias. At present, there is no significant evidence linking etiologically EVB to this type of tumor and dysplasia. However, our findings merit further investigation given the growing evidence that EBV may enhance proliferation and aggressiveness of tumor systems as well as the immortalization of non-neoplastic cells.

Ethnic difference of Helicobacter pylori gastritis: Korean and Japanese gastritis is characterized by male- and antrum-predominant acute foveolitis in comparison with American gastritis.

AIM: To investigate the clinicopathological factors underlying the ethnic differences of Helicobacter pylori gastritis and cancer. METHODS: We analyzed clinicopathological parameters of gastric biopsies having H pylori infection that were randomly selected from different ethnic populations including 147 Americans, 149 Japanese, and 181 Koreans. RESULTS: Males were predominant in Japanese and Korean populations (77.9 and 67.4% respectively) in comparison with Americans (48.3%) (P<0.001). H pylori gastritis in Koreans and Japanese was characterized by the predominant antral involvement. In the antrum, neutrophilic infiltration into the proliferative zone of pit, i.e. acute foveolitis, was more frequent in Koreans (82%) than in Japanese (71%) (P<0.05) and Americans (61%) (P<0.001). Interstitial neutrophilic infiltration, intestinal metaplasia and atrophy were also frequent in Koreans and Japanese. In the body, the prevalence of acute foveolitis was not significantly different among the populations while chronic interstitial inflammation and lymphoid follicles were more pronounced in the body of Americans than in the body of others (P<0.01). CONCLUSION: The male-, and antrum-predominant H pylori gastritis in Koreans and Japanese is compatible with the pattern of sex and topographical distribution of gastric cancer incidence. Our data suggest that persistent acute foveolitis at the proliferative zone is a crucial step in the gastric carcinogenesis.

Cytologic-histologic correlations in the diagnosis of inverted sinonasal papilloma.

We present cytologic data from multiple samples from two cases of inverted sinonasal papilloma (ISP). These samples displayed the entire spectrum of squamous cell changes, including benign squamous papilloma, variable degrees of dysplasia, and invasive squamous cell carcinoma. In all instances, the cytologic impression coincided with the final diagnosis based on frozen and/or permanent histologic sections from the same samples. We suggest that cytologic examination be viewed as a useful initial approach in the diagnosis of ISP, and in the differential diagnosis of other tumors that occur in the same sites.

Assessment of Her-2/Neu status by immunohistochemistry and fluorescence in situ hybridization in mammary Paget disease and underlying carcinoma.

HER-2/Neu overexpression is seen in 20% to 30% of invasive breast carcinomas and has been reported in as many as 80% of high-grade infiltrating carcinomas. Earlier studies have suggested that 100% of the tumor cells in mammary Paget disease show overexpression of HER-2 protein. We undertook this study to assess HER-2 status of mammary Paget disease and of the underlying breast carcinoma, when present, by immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH). Formalin-fixed, paraffin-embedded tissue from 20 cases of mammary Paget disease were analyzed for HER-2 status by IHC and FISH. IHC for estrogen receptor (ER) was also performed. The patients ranged in age from 34 to 88 years, with a mean age of 62 years. Eighty percent of the cases showed strong overexpression (3+) of HER-2 protein by IHC, and all of these cases showed more than 5-fold amplification of the HER-2 gene by FISH. The remaining 4 cases, which were negative for HER-2/Neu by IHC, showed no amplification by FISH. All of the latter cases expressed ER, whereas no case that overexpressed HER-2 expressed ER. Sixteen cases had an underlying tumor, which was in situ in 6 cases. The underlying tumors were identical to the Paget disease with respect to their HER-2/Neu overexpression by both IHC and FISH. HER-2 overexpression was identified in 80% of our cases of Paget disease. There was 100% concordance between HER-2 protein overexpression by immunohistochemistry and gene amplification in both the Paget and the underlying tumor. Moreover, all of the cases negative for HER-2 overexpression expressed ER, whereas those positive for HER-2 did not.

Malgun (clear) cell change in Helicobacter pylori gastritis reflects epithelial genomic damage and repair.

Cancers may develop in the background of genomic instability with accumulated mutations. Helicobacter pylori gastritis is characterized by acute foveolitis of the proliferative zone, which is found in any stage of the gastritis as long as the infection persists. Because acute foveolitis targets specifically the proliferative zone of pits, the proliferating epithelial cells are under severe and persistent mutagenic pressure. In H. pylori gastritis, a characteristic morphological change of epithelial cells, the malgun (clear) cell change is frequently present in association with acute foveolitis. Malgun cells have enlarged euchromatic nuclei and abundant cytoplasm. The expression of proliferating cell nuclear antigen and cytokeratin 8 are typically up-regulated in them indicating that they are mitotically and metabolically active. Here, we report evidence for DNA damage and repair in malgun cells. Significant double-strand DNA breaks were shown by the consistent terminal dUTP nick-end labeling in the nuclei of malgun cells. Proteins related to DNA damage and repair, such as Ku, poly(ADP-ribosyl) polymerase, OGG1, and MSH2 were selectively up-regulated in malgun cells. Inducible nitric oxide synthase was also up-regulated. There were occasional bcl2- and p53-expressing cells suggesting that further steps of carcinogenesis took place at the single cell level. Our results suggest that the malgun cell change represents a characteristic morphological sign of cellular genomic damage and repair, and may be implicated in an early stage of carcinogenesis. It is suggested that acute foveolitis of the proliferative zone is a major pathogenetic step of gastric carcinogenesis in H. pylori gastritis.

Glycoprotein A-80 in the human prostate: immunolocalization in prostatic intraepithelial neoplasia, carcinoma, radiation failure, and after neoadjuvant hormonal therapy.

OBJECTIVES: To review the expression of A-80 in prostate cancer and prostatic intraepithelial neoplasia and compare it with that of normal and hyperplastic prostatic tissue. The ability to recognize cancer after androgen deprivation therapy and residual and/or recurrent cancer after radiotherapy using A-80 staining was also examined. A-80 is a glycoprotein linked to exocrine differentiation that shows little or no expression in normal exocrine cells, but is selectively overexpressed in dysplasias and adenocarcinomas. METHODS: We studied 277 prostate samples with a monoclonal antibody (MAb) to A-80. We applied this MAb to paraffin sections of specimens of fetal (n = 12), benign prostatic hyperplasia (n = 26, from transurethral prostate resection specimens), atypical adenomatous hyperplasia (n = 11), prostate cancer (n = 103, from radical prostatectomy specimens), and autopsy (n = 7) tissue. In addition, 54 prostatectomy specimens after androgen-deprivation therapy and 64 specimens after radiotherapy were similarly studied. RESULTS: MAb A-80 stained the epithelial component of all 12 prostate specimens from fetal tissue; no staining was seen in normal adult prostatic tissue (0 of 7). In benign prostatic hyperplasia, sporadic cells reacted in 13% of cases (4 of 30); the atypical adenomatous hyperplasia samples were all negative (0 of 11). In patients with prostate cancer, more than 99% (102 of 103) stained positive, regardless of the grade or stage of cancer. Low and high-grade prostatic intraepithelial neoplasia reacted in 73% (38 of 52) and 92% (77 of 84) of cases, respectively. All 64 (100%) salvage prostatectomy samples after external beam radiotherapy stained positive for A-80. Carcinoma subsequent to neoadjuvant hormonal therapy stained positive in 98% (53 of 54) of cases. CONCLUSIONS: A-80 is useful in differentiating benign prostatic hyperplasia and atypical adenomatous hyperplasia from prostate cancer. Also, the strong A-80 reactions in most high-grade prostatic intraepithelial neoplasia provide strong molecular support to the precancerous nature of the lesion. In particular, A-80 staining of biopsies may be useful in detecting residual and/or recurrent prostate carcinoma after radiation or hormonal therapy.

Predictors of lymph node metastasis in T1 breast carcinoma, stratified by patient age.

It is important to identify T1-substage breast carcinomas (BCs) which are inherently aggressive, so that these can be managed more assertively. The purpose of this study was to distinguish those T1 BCs with the potential to metastasize to axillary lymph nodes from those lacking that ability by multiparametric analysis of several clinicopathologic features. The authors studied 197 patients with invasive BC who had undergone modified radical mastectomy; 161 tumors were ductal and 26 were lobular BCs. The study group was stratified by age into two groups:

Neuroendocrine tumors of the bronchopulmonary tract: a reappraisal of their classification after 20 years.

This article is an overview of the classification of pulmonary neuroendocrine neoplasms, their presentation, their pathologic appearance, and their clinical management. In addition, the original classification, based on histologic features, is reassessed in the light of newer areas in study, including neurosecretory products, neuroendocrine markers, ultrastructural studies, ploidy analysis, cell adhesion markers, apoptosis, oncogene mutation analysis, and genetic alterations. The histologic classification proposed in 1983 remains the single most valuable factor in establishing the diagnosis and, together with the TNM status, the prognosis of this group of interesting neoplasms.

Demonstration of Epstein-Barr virus in carcinomas of various sites.

EBV is an etiological agent in infectious mononucleosis, is implicated in some malignant lymphomas and lymphoepithelioma-like carcinomas, and has been sporadically reported in carcinomas of the breast, lung, and other sites. We studied immunohistochemically benign and malignant tumors of the breast, lung, colon, and prostate and found EBV in some carcinomas of those sites. Also, EBV reactions were noted in hyperplasias and dysplasias, e.g., breast carcinomas in situ and prostatic intraepithelial neoplasia. Benign tumor counterparts were negative. PCR analysis of selected cases confirmed the presence of EBV. Our results suggest that EBV is not restricted to lymphoepithelioma-like carcinomas but may play an oncogenic role in frequent epithelial cancers and possibly also in hyperplasias and certain dysplasias preceding carcinomas.

Application of adjunct techniques in cytologic material in the diagnosis of rhabdomyosarcoma: case report and review of the literature.

A 4(1/2)-yr-old female presented with right-sided pleural effusion and a retroperitoneal mass. Cytologic analysis of the pleural fluid yielded malignant small round blue cells, which were noncohesive, 3-4 times the size of lymphocytes. The malignant cells had hyperchromatic, pleomorphic nuclei with moderate amounts of vacuolated cytoplasm. A few fiber-shaped cells were also seen. Immunostains for desmin, muscle-specific actin were positive; ultrastructural findings of thick and thin actin-myosin filaments confirmed the diagnosis of embryonal rhabdomyosarcoma. This case illustrates the importance of performing appropriate immunohistochemical stains and ultrastructural studies on cytological material to arrive at a definitive diagnosis.

Nup88 (karyoporin) in human malignant neoplasms and dysplasias: correlations of immunostaining of tissue sections, cytologic smears, and immunoblot analysis.

Nuclear pore complexes (NPCs) are elaborate macromolecular structures that regulate the bidirectional nucleocytoplasmic traffic system. In vertebrate cells, NPCs include a family of 50 to 100 proteins termed nucleoporins (Nups). The 88-kD Nup has been found to be linked in a dynamic subcomplex with the oncogenic CAN/Nup214. Applying a polyclonal antiserum to Nup88 on paraffin sections, we found that it immunoreacts with numerous malignant neoplasms. All carcinomas reacted irrespective of site, type, or degree of differentiation; often, high-grade carcinomas stained more strongly and extensively. Some sarcomas (e.g., fibrosarcomas, leiomyosarcomas, liposarcomas, and rhabdomyosarcomas) reacted intensely; melanomas, gliomas, mesotheliomas, and malignant lymphomas also stained. In situ carcinomas of the colon, stomach, breast, and prostate stained convincingly, as did in situ melanomas; some samples of fetal tissues also reacted. Cytologic smears of some of the aforementioned tumors also stained. In selected samples, enhanced immunostaining of tissue sections and cytologic smears correlated strongly and consistently with immunoblot data. Immunoblots of the same tumors with antibodies to 2 other Nups (Nup214 and Nup153) showed no comparable enhancement. Therefore, it seems that in some malignant tumors, Nup88 overexpression is not parallelled by an overexpression of other Nups. Benign tumors, hyperplasias, and normal tissues showed weak and sporadic staining or absence of staining; immunoblots of the same samples yielded weak signals. Occasional highly proliferative hyperplastic-reactive processes showed focal staining. Thus, our correlative histologic, cytologic, and molecular data indicate that Nup88 may be viewed as a potentially useful, broadly based histodiagnostic and molecular marker of many malignancies and premalignant dysplasias, and further suggest that in some malignant tumors, Nup88 may be selectively overexpressed as compared with other Nups. Thus, we propose that Nup88 be designated as karyoporin.

Role of expression of cell cycle inhibitor p27 and MIB-1 in predicting lymph node metastasis in male breast carcinoma.

Tumor expression of the proliferation marker (MIB-1) and the cell cycle-related protein (p27) may predict the biologic behavior of various human tumors. The purpose of this study was to evaluate the role of p27 and MIB-1 expression in predicting lymph node metastasis in male breast carcinomas (MBCs). We studied 67 patients with invasive MBC who had undergone modified radical mastectomy. Pathologic lymph node status was correlated with the p27 protein and the MIB-1 proliferation index. These factors were studied immunohistologically by standard methods. Men in this study ranged from 36 to 92 years of age (mean, 63 years); 43 (64%) were T1 lesions, and 24 (36%) were T2 lesions. Twenty-nine patients (43%) had positive nodes. p27 was expressed in 43 tumors (64%) and MIB-1 in 13 tumors (19.4%). Tumors with positive p27 showed positive lymph nodes in 10 cases (23%). In contrast, p27-negative tumors had positive lymph nodes in 18 cases (75%). Tumors positive for MIB-1 show positive lymph nodes in 11 cases (85%). However, when MIB-1 was negative, only 16 patients (30%) had positive lymph nodes. Multivariate logistic regression analysis confirmed the utility of MIB-1 overexpression in predicting lymph node metastasis ( p < 0.0006). Also, decreased p27 protein expression strongly correlates with lymph node metastasis ( p < or = 0.0001). Furthermore, when p27 was negative and MIB-1 was positive, 100% of the patients had positive lymph nodes. In contrast, when p27 was positive and MIB-1 was negative, only 12% of patients had positive lymph nodes. The reduced expression of the p27 protein and the overexpression of the MIB-1 proliferation index in this study show a significant correlation in predicting lymph nodes metastasis in MBCs.