HER2 status in advanced gastric carcinoma: A retrospective multicentric analysis from Sicily.

According to the ToGA trial, HER2 has been shown to be predictive for the success of treatment with trastuzumab in advanced gastric cancer (AGC). A number of studies have analyzed HER-2/neu overexpression in gastric carcinoma and identified the rate of HER2 positivity to be markedly varied. To date, the prevalence of HER2 overexpression in Sicilian people with AGC is unknown. Therefore, in the present study, a retrospective immunohistochemical analysis of HER2 was performed in a cohort of 304 AGC samples that were obtained from the archives of 10 Sicilian anatomopathological diagnostic units in order to verify the positive rate of HER2-positive cases. Furthermore, the characteristics of histotype, grade, stage and Ki-67 expression were also analyzed. HER2 overexpression was encountered in 17.43% of all the gastric adenocarcinomas, which was consistent with the results that have been reported elsewhere in the literature. A progressive increase in HER2 overexpression was observed, from the poorly cohesive histotype to the tubular adenocarcinomas and gastric hepatoid adenocarcinomas. HER2 overexpression was significantly associated with a high grade, advanced stage and high Ki-67 labeling index. Further investigations performed jointly by pathologists and oncologists within the geographical area of the present study should confirm that the association of trastuzumab with chemotherapy results in an improvement of survival in patients with AGC.

Polymorphous low-grade adenocarcinoma of the salivary glands: clinicopathological and immunohistochemical study of a case.

Polymorphous low-grade adenocarcinoma (PLGA) is a distinctive salivary gland neoplasm with a predilection for intraoral sites. Histological and immunohistochemical analyses were used to study a case involving the minor salivary glands of the buccal mucosa in a 68-year-old woman. Histologically, the tumor was characterized by unenca-psulated, progressing and focally-infiltrative borders and showed variable growth patterns including solid, tubular, microcystic and cribriform; papillary and papillary-cystic areas of more than focal extent were present. Cytologically the neoplasm was composed of uniform, round to cuboidal cells with bland, round to oval nuclei. Immunohistochemical analysis showed that the tumor cells were positive for cytokeratin, S-100 protein, vimentin, focally-positive for EMA and actin and negative for CEA. The significance of the papillary component and the possible relationship to other salivary gland tumors is discussed.

BCL2 and BAX expression in hyperplastic and dysplastic rectal polyps.

BACKGROUND/AIMS: Members of the gene family that includes BCL2 and BAX are functionally antagonists in the apoptosis process and they have been observed in normal and neoplastic tissues. The aim of this study is to investigate the combined effects of BCL2 and BAX protein in normal mucosa, dysplastic and hyperplastic polyps of the rectum. METHODOLOGY: We studied BCL2 and BAX protein expression in 40 cases of adenomatous polyps all located in the rectum, with different dysplastic gradings, and the mean time in 10 cases of normal rectal mucosa. RESULTS: BCL2 expression was found more frequently in hyperplastic and in low dysplastic polyps with moderate and strong positivity compared to moderate and severe dysplasia. BAX expression was found in normal mucosa in hyperplastic and dysplastic polyps, the immunoreactivity was prevalently moderate and strong. CONCLUSIONS: These preliminary data suggest that BCL2 and BAX confirm a probably different role in apoptosis. Nevertheless, it is important to know the relation between the molecular pathways of apoptosis, the defective mismatch repair and the tumor suppressor genes associated with an increased mutation rate in cancerogenesis of the colorectum.

Fine needle aspiration and core needle biopsy techniques in the diagnosis of nodular thyroid pathologies.

It is widly accepted that Fine Needle Aspiration Biopsy (FNAB) is the main test to distinguish benign from malignant thyroid lesions. Nevertheless, this technique presents some limits such as the possibility of false-negative or inadequate samples and it is unable to cytologically discriminate among adenomathosus goiter, follicular adenoma and well-differentiated follicular carcinoma. The aim of this study was to evaluate the possibility of restricting these limitations using Core Needle biopsy (CN) technique. Therefore we selected for CN, 40 out of 136 patients who underwent FNAB during a one year period; among these patients only 32 agreed a to this technique. Forty-two out of 136 patients underwent surgery; 29 of them were subjected to both biopsies. Sixteen of the diagnostic microbiopsies have been histologically confirmed. We had no discordant cases between cytological and microhistological diagnosis, except for one case which appeared cytologically colloid goiter, microhistologically follicular neoplasm and histologically follicular adenoma. In this case it was not possible to microhistologically discriminate benign from malign follicular lesion. In our experience not all patients accepted CN biopsy as well as FNAB and, moreover, this technique showed no advantage over FNAB diagnosis. On this base we think that actually FNAB should be the main procedure in the diagnosis of the thyroid lesions. It is easily performed, accepted by the patients and has a low cost-benefit ratio. If the sample is not diagnostic it can be easily repeated and false-negative cases could be discovered thanks to an adequate clinical and ultrasonographical follow-up of the patients.

PAPNET-assisted primary screening of conventional cervical smears.

The PAPNET System is the only device with a neural-network-based-artificial intelligence to detect and show the images of abnormal cells on the monitor to be evaluated in an interactive way. We effectively used the PAPNET in rescreening of conventional cervical smears and we detected its advantages and its disadvantages. In this paper, we report our results from PAPNET-assisted primary screening performed on 20,154 conventional smears. The smears were classified as Negative or as Review. The Negative cases were rapidly rescreened mainly near the coverslip edges, which are the slide areas not analyzed by automated devices because of focusing problems. The Review cases were fully reanalyzed by the optic microscope. In summary, 140 positive smears were detected: 57 cases showed changes due to HPV, 63 LSIL, 15 HSIL, and 5 carcinomas. Therefore, the PAPNET System was confirmed as useful in primary screening of conventional cervical samples as well as rescreening.

Immunohistochemical expression of p53, nm23-HI, Ki67 and DNA ploidy: correlation with lymph node status and other clinical pathologic parameters in breast cancer.

BACKGROUND: The different clinical evolution of breast cancer with similar pathological characteristics prompted the authors to investigate the prognostic significance of different biological markers. METHODS: Seventy-one primary breast carcinoma specimens obtained by mastectomy or quadrantectomy were examined for the determination of the p53, nm23 and Ki67 expression, with immunohistochemistry, and the DNA content, with static cytometry. RESULTS: p53 protein was expressed as nuclear staining in 58% of the cases and was associated with high levels of Ki67, non-diploid lesions and lymph-node status. Positive staining for nm23 was significantly correlated only with histologic grading. A predictive role in disease recurrence was demonstrated only in patients with a high Ki67 nuclear expression. CONCLUSIONS: From these data, we may conclude that, besides all the other traditional clinical morphological parameters, a panel of different biological markers, such as Ki67 and the determination of p53 expression, may be utilized to further characterize breast cancer and its biological behavior.

[The PAPNET system in cytological rescreening of cervical smears]. / Il sistema PAPNET per il rescreening citologico dei preparati eso-endocervicali.

BACKGROUND: The Papanicolaou test has reduced mortality from cervical cancer but has not completely eradicated the disease. A reason for the false negative screening errors may be found in one of three steps of the test (obtaining, processing and interpretation of the cervicovaginal smears). The possible false negative primary screening errors may be detected with the help of rescreening. Therefore, an automated quality control by means of a system such as in the PAPNET is necessary. The PAPNET combines the use of algorithmic image analysis with neural networks and allows to conventionally reassess cervicovaginal smears. Nevertheless, the cytopathologist always decides whether to reassess the sample with the optic microscope as suggested by the presence of abnormal cells in the frames selected by the PAPNET system. Therefore the cytopathologist always formulates the conclusive diagnosis. METHODS: In this study, we rescreened 300 cervicovaginal smears. RESULTS: Only 122 smears have been reassessed by the optic microscope. The human papillomavirus infection and the SIL (squamous Intraepithelial Lesion) diagnoses formulated by means of rescreening with the PAPNET have coincided with the primary screening diagnoses. The only exception was a human papillomavirus infection not previously detected. CONCLUSIONS: Our study indicates that the use of the PAPNET system facilitates the quality control measures and has reduced the false negatives.

Epithelial-myoepithelial carcinoma of the salivary glands: a clinico-pathologic, immunohistochemical and DNA flow cytometric study of three cases.

Epithelial-myoepithelial carcinoma is a rare neoplasia of the salivary glands. Histologic, immunohistochemical and flow cytometry analysis were used to study three new cases involving the major salivary glands of two women and one man. The mean age was 66.7 years. Immunohistochemistry confirmed the epithelial-myoepithelial dualism of the tumors: the predominant growth pattern showed ducts made up of an external myoepithelial layer and an internal epithelial layer. Proliferative activity, assessed by means of the PCNA and Ki-67 immunostaining and the study of the S-phase with flow cytometric analysis, proved to be higher in a recurrent case. All three cases were diploid. The use of immunohistochemical and flow cytometric techniques for the assessment of proliferative activity would seem to be an efficient method for the selection of a subgroup of epithelial-myoepithelial carcinomas with unfavorable prognosis.

The PAPNET system for quality control of cervical smears: validation and limits.

The Pap-test may fail to identify false negative cases. Probably only full rescreening assisted by instrument can prevent potential screening and interpretation errors. Therefore, the PAPNET System was used obtain accurate Quality Control (QC) of 3000 conventional cervical smears. Sixteen false negatives were detected with the rescreening assisted by PAPNET. Thirteen positive cases were suspected during evaluation assisted by the PAPNET; two cases were classified as Review for potential inflammatory cellular changes and underwent full optic microscope. In one case both primary screening and the PAPNET did not detect the only abnormal cell cluster located near the coverslip edge, being part of the slide excluded from the scanning process due to focusing problems: only coverslip edge rescreening by optic microscopy allowed the formulation of the correct conclusive diagnosis. In conclusion, the PAPNET System is useful for QC of conventional cervical smears: in fact this instrument allows the identification of false negative cytological errors easily and effectively. Nevertheless, some slide scanning program modifications are needed to detect all abnormal cells present on slide under coverslip.

Image analysis in multisample biopsy after ileal pouch-anal anastomosis.

Pouchitis in ileal anal anastomosis represents an important clinical complication after restorative proctocolectomy. Acute and chronic inflammation of the reservoir is a frequent event sometimes associated with villous atrophy and colonic metaplasia. After ileal pouch anastomosis, twenty-one patients affected by ulcerative colitis were studied. An image analyzer CAS 200 (Becton Dickinson) was utilized to evaluate the DNA intranuclear content in every biopsy. In two cases abnormal DNA distribution was observed, and in one case a poliploid pattern was seen. Abnormal DNA distribution was also present in colonic metaplasia. Therefore, image analysis for the detection of DNA aneuploidy may be of additional value together with histologic parameters in follow up, in order to exclude transformation of the ileal mucosa in neoplastic epithelia.

Pleomorphic adenoma and adenoid-cystic carcinoma of salivary glands: immunohistochemical assessment of proliferative activity in comparison with flow-cytometric study.

In this study, 32 pleomorphic adenomas (PAs) and seven adenoid cystic carcinomas (ACCs) were analysed for the evaluation of proliferating cell nuclear antigen (PCNA) indices and flow cytometric variables. Our aim was to assess any possible relationship between these parameters and the clinico-pathological variables and to clarify their histogenesis and reasons for their biological differences. The tumours were divided into three groups, mainly epithelial (E), myxoid (M) and chondroid (C); PCNA labelling index (LI) and weighted mean index (WI) and the WI/LI ratio were analysed in the predominant components; a single PCNA index, weighted by the percentage of each component, was also calculated. Only WI/LI was found to be significantly different in the three components, while PCNA single index did not show either significant differences by sex, age, site and size, or any correlation with the S phase fraction. A significant difference was found between PAs and ACCs by site (P < 0.01) and DNA ploidy (P < 0.05); furthermore, all PCNA indices (single index) were significantly lower in PAs than in ACCs.

Prognostic significance of cell kinetics in laryngeal squamous cell carcinoma: clinicopathological associations.

A consecutive series of 99 untreated patients undergoing radical surgical resection for stage I-IV laryngeal carcinomas has been studied prospectively. Our purpose was to analyze the predictive relevance of proliferative variables studied [proliferating cell nuclear antigen (PCNA) expression, volume-corrected mitotic (M/V) index, and S-phase fraction (SPF)] on clinical outcome in relation to DNA ploidy and clinicopathological features. All of the patients were followed up for a median of 32 months (range, 5-58 months). A weak, but significant, positive correlation was found between M/V and PCNA indices (except the PCNA weighted mean index:labeling index ratio) or these indices and SPF. At univariate analysis, node positivity (P < 0.05), poor histological grade (P < 0.01), DNA aneuploidy (P < 0.01), a high SPF (P < 0.01), and a high M/V index (P < 0.05) proved to be related significantly to quicker relapse, whereas T4 (P < 0.05), subglottic site (P < 0.05), DNA aneuploidy (P < 0.01) and a high SPF (P < 0.01) were related significantly to shorter overall survival. With multivariate analysis, a high SPF (> 12.1%) and histological grade (G3) were related to the risk of relapse (relative risk, 8.65 and 5.45, respectively), whereas only a high SPF was related independently to the risk of death (relative risk, 7.30). Our study has identified SPF, in addition to histological grade, as an important biological indicator in laryngeal carcinomas.

Vimentin expression, proliferating cell nuclear antigen and flow cytometric factors. Prognostic role in breast cancer.

A series of 71 patients undergoing surgery for primary breast carcinoma was prospectively studied in order to evaluate the relative weight for four biologic factors (intermediate filament vimentin expression, proliferating cell nuclear antigen [PCNA], flow cytometric DNA ploidy and S-phase fraction) and of several clinicopathologic and biologic features in predicting clinical outcome (disease-free interval). In univariate statistical analysis, positivity of axillary nodes, high number of mitoses, high nuclear grade, high histologic grade, positivity of vimentin, high flow cytometric S-phase fraction (FCM-S) value, high PCNA and high silver-stained nuclear organizer regions scores were significantly related to risk of relapse. In multivariate analysis (Cox's logistic regression) only histologic grade (3) and high FCM-S values (> 10.7) were independently related to risk of relapse, with hazard ratios of 9.84 and 7.98, respectively. The results of our preliminary, prospective study suggest that FCM-S, in addition to morphologic criteria (histologic grade), may be an important biologic indicator in determining breast cancer patients' prognosis.

p53 expression in human renal cell carcinoma: an immunohistochemical study and a literature outline of the cytogenetic characterization.

In this study both the incidence and pattern of p53 over-expression in various histological subtypes of a series of 36 cases of renal cell Grawitz carcinoma, partially studied in a previous paper, were analyzed. This series consisted of these histologic subtypes: clear cell non papillary (18 cases), clear cell papillary (2 cases), granular cell (5 cases), mixed (clear and granular cell) (9 cases) and spindle cell (2 cases). At present, our aim was, firstly, to see which were the best technical conditions for detection of p53 in the available paraffin-embedded tumor specimens, using several antibodies, specific for various epitopes; secondly, to investigate if some relation might exist between this expression and the histological features of these tumors. Twenty-five per cent (9/36 cases) resulted p53 immunoreactive, the highest percentage being represented in the papillary clear and granular cell carcinomas; low expression was detected in 11 cases (30%) and no reactivity in 16 cases (44%). Neither technical or dilution modifications proved to transform these latter results; however, detection was maximal using the CM-1 polyclonal rabbit antiserum. Thus, in RCC, expression of p53, analyzed in the light of the cytogenetic characterization through a literature review, resulted at low frequency. This finding means that mutation of the p53 gene are not frequent in the neoplastic transformation in RCC. Nevertheless, in spite of the small number of cases and of the short follow-up period of this study, detection of p53 positivity in tumors with either high grade and stage or high proliferative activity could suggest that p53 mutations lead to tumors of a more aggressive type.(ABSTRACT TRUNCATED AT 250 WORDS)

P53 expression in stage iii-iv squamous-cell carcinoma of the larynx - an immunohistochemical study related to clinicopathological, flow-cytometric DNA analysis and prognosis.

A series of 71 patients undergoing radical surgical resection for stage III and IV laryngeal carcinoma (LC) consecutively diagnosed was prospectively studied in order to evaluate the relative weight of p53 expression in predicting clinical outcome, All the patients taking part in this study were followed up for a median of 18 months (range: 6-41 months). Positive staining for p53 protein was detected in 44 of 71 (62%) of these tumors on paraffin-embedded tissue, even in dysplastic areas. Among the clinico-pathological and biological parameters analyzed, only flow-cytometric S-phase (FCM-S) Values of turners showed a significant relationship to p53 immunostaining (p=0.01). With Kaplan-Meier estimation, in multivariate analysis only high FCM-S (>15.1) was independently related to risk of relapse (RR=5.82), while both FCM-S and site (subglottis) were related to risk of death (RR=6.83 and RR=14.3, respectively). These findings indicate that p53 immunoreactivity, though of no utility as a prognostic indicator, probably plays a role in the early stages of LC tumorigenesis.

Malignant myoepithelioma of the breast - a case-history and review of literature.

A malignant myoepithelioma, mainly intraductally growing, of the breast was studied, emphasizing the immunohistological features of such tumors, in comparison with the more common, architecturally similar, infiltrating ductal carcinoma with intraductal component. Immunohistochemically, cell population of the tumor studied expressed intense reaction to alpha-SM-actin, cytokeratin 14, vimentin, S-100 protein and collagen IV antibodies, confirming myoepithelial differentiation. Since myoepitheliomas of the breast are very rare, complete knowledge of these neoplasms has not been attained. Nevertheless, histogenetic investigations may be useful in differentiating this type of tumor, from the above one, which is known to be characterized by severe clinical and prognostic implications.

[Micro-glandular hyperplasia of the uterine cervix. Histo-cytopathological evaluation, differential diagnosis and review of literature]. / L'iperplasia microghiandolare della cervice uterina. Valutazioni isto-citopatologiche, diagnosi differenziale e revisione della letteratura.

Microglandular hyperplasia is a lesion of the uterine endocervical mucosa, often associated with a story of contraceptive drugs or pregnancy. It was frequently confused with adenocarcinoma or premalignant glandular changes until its benign nature was recognized. In order to ascertain clinical presentation, hormonal or gestational status, histologic patterns with their variants and cytologic counter-part, we collected and reviewed 28 cases of this condition over a period of a decade. The age ranged from 23 to 54, with a mean of 37.2 years. 11 patients had been receiving oral contraceptives and 5 were pregnant. All samples came from uterine cervix. Cervical smears were available from 17 women. Four histologic patterns were identified, including glandular, reticular, trabecular and solid. Cytologic features varied from aspecific, inflammatory changes of the columnar endocervical cells to more particular findings, including clear cells, strips, sheets, papillae, rosettes and corolla-like aggregates. Moreover, differential diagnostic criteria from cervical neoplasms and various pseudoneoplastic conditions of the cervical glandular epithelium are discussed. An extensive review of the literature is also presented.

Evaluation of integrated morpho-biological indicators in breast cancer.

A series of 71 patients undergoing surgery for primary breast carcinoma was prospectively studied in order to verify the relationships between clinical and pathological variables, proliferative indexes and DNA-ploidy. Significant correlations between proliferative indexes were found; conversely, DNA-ploidy showed no correlation with either PCNA/cyclin or AgNORs, but only with flow cytometry S-phase fraction (FCM-S) and IF-vimentin. In the 42 patients with a mean follow-up of 42 months, disease-free rate was best predicted by nuclear grade (p < 0.001), histological grade (p < 0.001), mitoses (p 0.001), IF-vimentin (p 0.002), FCM-S (p 0.002), PCNA (p 0.005) and Ag-NORs (p 0.007).

[Collagenous colitis. Morphologic and immunohistochemical study]. / Colite collagena. Studio morfologico ed immunoistochimico.

Collagenous colitis is a clinico-pathological entity characterized by chronic diarrhoeas and deposition of collagen beneath the epithelium surface of large bowel. We revised 265 endoscopy biopsy specimens of the large bowel from 198 consecutive patients with "aspecific chronic colitis". Morphometric study showed that were not significant differences among various tracts in the same patients regarding to the thickness of basament membrane. It was more than 11.9 +/- 0.49 mu only in 13 pts (6.6%), while it was 3.96 +/- 1.4 mu in the others. Immunohistochemistry study confirmed the normality of subepithelial basement membrane and the below deposition of the large quantity of collagen IV.

Renal cell (Grawitz) carcinoma: an immunohistochemical study in correlation with nuclear grade, histological pattern and proliferative activity of tumor cells.

Since morphological features of renal cell carcinomas may not always predict their clinical behaviour, other biological parameters are more and more searched for, as prognostic indicators. Among those, the Authors evaluate the cytokeratin-vimentin intermediate-sized filaments co-expression in a series of 32 renal carcinomas and analyze this feature in correlation with both traditional criteria and proliferative activity of tumor cells, studied by means of the proliferation cellular nuclear antigen (PCNA) expression as well as of the nucleolar organizer regions (NORs) count, within tumor cells. From this study, nuclear grade stands out as the best parameter which, furthermore, shows a significant relationship with the cytokeratin-vimentin co-expression and the mean NORs count in tumor cell populations. Further investigations about relationships between these parameters and survival of the patients, who, at present, are almost all alive, will be performed afterwards.