Impact of different sectioning of excision biopsies of the cervical transformation zone for detection of invasive cervical cancer in 2 European centers related to national cervical screening programs.

BACKGROUND: The impact of the number of sections used in cervical excision biopsies of transformation zone on sensitivity of histological detection of cervical cancer is poorly documented. AIM: To assess whether different techniques of sectioning cervical excision biopsies in 2 European laboratories influence the finding of cervical cancer by histopathological examination. MATERIALS AND METHODS: The routine assessment at the Department of Histopathology and Cytology, Sheffield Teaching Hospitals NHS Foundation Trust, Sheffield, UK (Sheffield), encompassed sectioning a cone in 3-mm tissue blocks and cutting 3 levels per block. At the Institute of Pathology, Faculty of Medicine, University of Ljubljana, Slovenia (Ljubljana), cones were cut in a 3- to 4-mm-thick tissue block, and 10 levels were cut per block, spaced every 100 microm. The number of blocks and levels per block were assessed for each cone. Histopathological detection of cervical carcinoma between the laboratories was compared. RESULTS: Among 820 cones in the Sheffield laboratory, we detected 35 invasive carcinomas, whereas 6 invasive carcinomas were detected among 94 cones in the Ljubljana laboratory. Although the Slovenian laboratory examined a significantly larger number of levels per cone biopsy (64 vs 24 in Sheffield), this was not associated with a larger proportion of invasive cervical carcinoma, especially foci of stage IA1 or high-grade cervical intraepithelial neoplasia diagnosed on excision biopsies: 6.4% of invasive cervical carcinomas (5 [83.3%] of stage IA1) in Ljubljana and 4.3% (30 [85.7%] of stage IA1) in Sheffield laboratory (P > 0.05); 71.2% of high-grade cervical intraepithelial neoplasia in Ljubljana and 75.5% in Sheffield (P > 0.05). CONCLUSIONS: Our assessment showed that a method with a large number of levels per cervical cone or large loop excision of the transformation zone biopsy did not increase sensitivity for the detection of cervical cancer. However, extensive sectioning substantially affects the pathologist's workload, and this may need to be reconsidered.

Digital pathology for intraoperative frozen section diagnosis of thoracic specimens: an evaluation of a system using remote sampling and whole slide imaging diagnosis.

BACKGROUND: Digital pathology is now used for primary diagnostic work as well as teaching, research and consultation. In our multisite institution service reorganisation led to histopathology being located in a separate hospital from some surgical specialities. We implemented remotely supervised specimen sampling and frozen section diagnosis using digital pathology. In this study we assessed the concordance of glass and digital slide diagnosis using this system. METHODS: We reviewed cases from the first 2 years of digital frozen section reporting at our institution. Cases with potential digital to glass slide discordance were reviewed by three experienced thoracic histopathologists. The reasons for discordance were determined and common themes identified. We also reviewed critical incidents relating to digital pathology during the study period. RESULTS: The study population comprised 211 cases. Frozen section to final diagnosis concordance between digital and glass slide diagnosis was found in 196 (92.6%) cases. The 15 potentially discordant cases were reviewed. Intraobserver concordance between glass and digital slide review ranged from 9/15 to 12/15 cases across the three pathologists. Glass slide review diagnosis showed better concordance with ground truth in two cases; digital slide review was more accurate in two cases. One relevant critical incident was identified during the study period. DISCUSSION: This is the largest study to examine digital pathology for thoracic frozen section diagnosis and shows that this is a safe and feasible alternative to glass slide diagnosis. Discordance between digital and glass slide diagnoses were unrelated to the processes of whole slide imaging and digital microscopy.

Simultaneous evaluation of maspin and CXCR4 in patients with breast cancer.

AIM: To study simultaneously the actions of maspin and CXCR4, which share several similar pathways in cancer, including apoptosis and angiogenesis. METHODS: Our material consisted of 151 invasive breast carcinomas arranged in a tissue microarray setting. Maspin and CXCR4 expression was evaluated by immunohistochemistry. Microvessel density was assessed by CD34 immunodetection and apoptosis by the Tdt-mediated dUTP nick end labelling assay. RESULTS: Maspin expression was related to CXCR4 expression, apoptosis, patient age and the Nottingham prognostic index. The expression of both maspin and CXCR4 progressively increased in high-grade tumours. In patients with lymph node negative breast cancer, maspin overexpression was associated with increased risk of death. High CXCR4 expression was associated with prolonged survival of patients with high maspin expression. CONCLUSIONS: Our results show that maspin overexpression could prove to be a potentially useful marker, especially for the clinically important group of patients with lymph node negative breast cancer. The expression of CXCR4 is of less significance in our study, but may be informative for specific patient subsets or in a longer time frame.

Expression of mos in astrocytic tumors and its potential role in neoplastic progression.

The c-mos gene and its protein product mos, components of the mitogen-activated protein kinase transduction pathway, are known to be involved in the control of meiosis and mitosis. Apart from a study on lung carcinomas, there is little information about its role in human neoplasia. The aim of this study was to investigate expression of mos in astrocytic tumors and to correlate it with accumulation of p53. We studied expression of mos in 62 cases of supratentorial astrocytic tumor. Intracytoplasmic immunostaining for mos was found in 28 (45%) cases: 3 of 20 (15%) grade 2 astrocytomas, 9 of 20 (45%) grade 3 anaplastic astrocytomas, and 16 of 22 (73%) glioblastomas. Immunopositivity for mos correlated significantly (P < 0.01) with tumor grade but not with p53 expression. In contrast to the findings in relation to lung tumors, immunopositivity for mos in astrocytic tumors did not predict recurrence-free or overall survival time. Cytoplasmic immunostaining was observed in scattered large cortical neurons adjacent to tumors, possibly due to stress-induced abortive entry into the cell cycle. The correlation of mos immunopositivity with tumor grade may reflect the expansion of more malignant mos-positive clones. This study provides evidence that mos may be involved in the neoplastic progression of a proportion of astrocytic tumors.

Expression of mos in ependymal gliomas.

The c-mos gene and its protein product mos, components of the mitogen-activated protein kinase transduction pathway, are known to be involved in the control of meiosis and mitosis. Apart from our previous studies on lung carcinomas and astrocytic gliomas, little has been published about its role in human neoplasia. The aim of this study was to investigate the expression of mos in ependymal neoplasms and to correlate it with tumor grade, proliferative fraction, and clinical behavior. We studied mos expression in biopsy specimens from 34 patients with ependymomas. Intracytoplasmic immunopositivity for mos was found in 16 (47%) and was associated significantly with tumor grade: 5 (24%) of 21 grade II ependymomas; 11 (85%) of 13 grade III anaplastic ependymomas (P < .01). Tumors with an MIB-1 labeling index of more than 4% were significantly more likely than those with a lower proliferative fraction to be immunopositive for mos (P = .012). Expression of mos showed a significant negative association with recurrence-free interval (P = .05) but not with overall survival. Our results suggest that overexpression of mos identifies a biologically aggressive subgroup of ependymal tumors and may be involved in their neoplastic progression.

The complement inhibitor CD59 and the lymphocyte function-associated antigen-3 (LFA-3, CD58) genes possess functional binding sites for the p53 tumor suppressor protein.

p53 is an oncosuppressor protein, which acts via transcriptional and non-transcriptional mechanisms. The transcriptional function of p53 is mediated by specific responsive elements. In the present study we found active responsive elements, specific for the p53 within the 5'flanking region and within the first intron of the gene encoding for the CD59 membrane inhibitor of reactive lysis, and within the first intron of the gene encoding for the CD58 membrane protein (LFA-3). The results suggest that p53 may enhance the transcription of both CD59 and CD58 and imply a novel role for p53 as a direct regulator of the immune response.

Lactate dehydrogenase-B is silenced by promoter methylation in a high frequency of human breast cancers.

OBJECTIVE: Under normoxia, non-malignant cells rely on oxidative phosphorylation for their ATP production, whereas cancer cells rely on Glycolysis; a phenomenon known as the Warburg effect. We aimed to elucidate the mechanisms contributing to the Warburg effect in human breast cancer. EXPERIMENTAL DESIGN: Lactate Dehydrogenase (LDH) isoenzymes were profiled using zymography. LDH-B subunit expression was assessed by reverse transcription PCR in cells, and by Immunohistochemistry in breast tissues. LDH-B promoter methylation was assessed by sequencing bisulfite modified DNA. RESULTS: Absent or decreased expression of LDH isoenzymes 1-4, were seen in T-47D and MCF7 cells. Absence of LDH-B mRNA was seen in T-47D cells, and its expression was restored following treatment with the demethylating agent 5'Azacytadine. LDH-B promoter methylation was identified in T-47D and MCF7 cells, and in 25/25 cases of breast cancer tissues, but not in 5/5 cases of normal breast tissues. Absent immuno-expression of LDH-B protein (<10% cells stained), was seen in 23/26 (88%) breast cancer cases, and in 4/8 cases of adjacent ductal carcinoma in situ lesions. Exposure of breast cancer cells to hypoxia (1% O(2)), for 48 hours resulted in significant increases in lactate levels in both MCF7 (14.0 fold, p = 0.002), and T-47D cells (2.9 fold, p = 0.009), but not in MDA-MB-436 (-0.9 fold, p = 0.229), or MCF10AT (1.2 fold, p = 0.09) cells. CONCLUSIONS: Loss of LDH-B expression is an early and frequent event in human breast cancer occurring due to promoter methylation, and is likely to contribute to an enhanced glycolysis of cancer cells under hypoxia.