G.F. Sterrett

Histological grading of breast carcinomas: A study of interobserver agreement

Interobserver variation in the histological grading of breast carcinoma was investigated using the hypothesis that optimal fixation, more precise grading guidelines, some experience, the use of training and test sets, and a comparison of results with an expert group might allow higher levels of agreement. For the training sets sections from 50 consecutive cases of breast carcinoma received at the Sir Charles Gairdner Hospital (SCGH) and fixed in both B5 and buffered formal saline (BFS) were graded by consensus of three pathologists at the SCGH and independently by consensus of two pathologists at the Nottingham City Hospital (NCH) using a modified Scarff-Bloom-Richardson histological grading system with guidelines as suggested by NCH pathologists. The section quality and degree of preservation of nuclear morphology were judged by NCH pathologists to be superior for B5-fixed material. Complete agreement in grade between SCGH and NCH results was achieved for 83.3% of B5-fixed cases and 73.5% of BFS-fixed cases (P = .05) with relative disagreement rates (RDRs) of 0.15 and 0.29 and kappa statistic values of 0.73 and 0.58, respectively. Approximately 80% complete agreement was achieved for tubule formation, nuclear score, and mitotic count, with RDRs ranging from 0.19 to 0.27 and kappa values from 0.46 to 0.69. There was a consistent bias in the SCGH results toward a higher tubule score in both B5- and BFS-fixed material because of a difference in interpretation of cribriform or complex gland patterns and a consistent bias in SCGH results toward a lower nuclear size/pleomorphism score for B5 and BFS material. For the test set sections from 50 further consecutive cases of breast cancer fixed in B5 were examined using similar criteria but taking into account the sources of error shown by the training set. Approximately 80% complete agreement was again achieved for grade components and grade (RDRs, 0.18 and 0.72). Systematic bias was reduced in the test set, but no other improvement was observed. Of the tumors designated as grade I by NCH, 87.5% were called grade I tumors by SCGH in the B5 training set, 84.6% in the B5 test set, and 66.6% in the BFS training set. The levels of agreement shown in both the training and test sets were satisfactory and represented a significant improvement over our previous study, suggesting that experience and precise grading guidelines are of value. The similar levels of agreement in training and test sets suggest that reasonable results can be achieved without direct training by expert groups.(ABSTRACT TRUNCATED AT 400 WORDS)

Detection of tissue CEA-like substance as an aid in the differential diagnosis of malignant mesothelioma

Summary Sections of various adenocarcinomas and malignant mesotheliomas were tested for carcinoembryonic antigen (CEA) localized in tissues by the immunoperoxidase technique; epithelial mucin was demonstrated with the PAS technique. While CEA and mucin were found in many adenocarcinomas, both were absent in the 43 cases of malignant mesothelioma we investigated. In the problem of distinguishing between adenocarcinoma and mesothelioma, the CEA‐test in combination with conventional stains for mucin is a useful technique and clearly identifies most adenocarcinomas. A dual negative result for CEA and mucin, although not proving that a given lesion is a mesothelioma, adds considerable support to this histological diagnosis.

Routine analysis of p53 mutation in clinical breast tumor specimens using fluorescence-based polymerase chain reaction and single strand conformation polymorphism.

Improved prognostic and predictive markers in breast cancer management would help considerably in therapeutic decision making, particularly in patients with early-stage breast cancer. Tumor factors currently used for prognostication and management decisions are tumor size, histologic type and grade, axillary lymph node status, and estrogen receptor content. The discovery of various somatic genetic alterations in breast cancer has raised the possibility that these may provide additional and independent prognostic and predictive information. Alterations of the p53 tumor suppressor gene in particular have received the most attention as potential prognostic and predictive factors. In multivariate analysis, p53 gene mutation is consistently associated with a two- to threefold increased risk of relapse and death from breast cancer. One of the major reasons preventing the introduction of p53 mutation as a routine marker to assist in therapeutic decision making is the lack of a simple, reproducible, and inexpensive assay. In the present study the authors optimized a polymerase chain reaction-based mutation screening method, fluorescence-single strand conformation polymorphism (F-SSCP), that allows p53 status to be assessed accurately and reproducibly in routinely handled, formalin-fixed and paraffin-embedded tumor specimens. The frequency of p53 mutation observed using F-SSCP in a consecutive series of invasive ductal breast carcinomas was 17% (28/164). The authors propose that the prognostic and predictive values of p53 mutation in breast cancer should be further evaluated in prospective, randomized studies using this standardized technique.

Pulmonary lymphomatoid granulomatosis with immunodeficiency terminating as malignant lymphoma

Lymphomatoid granulomatosis was diagnosed in a 60-yr-old woman 2 yr after presentation with a multi-system disorder resembling sarcoidosis. Five months later autopsy revealed malignant lymphoma. Large aggregates of intracytoplasmic tubular structures resembling nucleocapsid material of the paramyxovirus group were found within cells of lymphoma deposits in the liver. Sequential immunological studies over more than 2 years demonstrated a relatively stable T-cell deficiency associated with variable B-cell dysfunction. The latter was characterized by the production of immunoglobulins of restricted electrophoretic mobility. Intermittent hypercalcaemia was associated with increases in serum IgG and appeared to be due to the presence of Ca-binding paraproteins. It is suggested that lymphomatoid granulomatosis may be a pre-malignant lymphoproliferation, with immune deficiency as a predisposing cause. The pattern of immunological abnormalities suggests that the lymphoma may have been due to B-cell malignant transformation.

Detection of estradiol-induced messenger RNA (pS2) in uninvolved breast tissue from mastectomies for breast cancer

SummarypS2 mRNA was estimated in uninvolved breast tissue and breast carcinoma from the same patients. pS2 mRNA was clearly detected in 14 of 59 uninvolved breast tissues and in 30 of 58 breast carcinomas. pS2 mRNA was found more frequently in uninvolved breast tissue of premenopausal women than in that of post-menopausal women.

Detection of DNA of human papillomavirus types 6/11 and 16/18 in cell scrapings of the uterine cervix by filter in situ hybridisation. Correlation with cytology, colposcopy and histology

The application of filter in situ hybridisation (FISH) to detect the presence of the DNA of human papillomavirus genotypes 6/11 and/or 16/18 in cell scrapings of the uterine cervix of 248 women in Western Australia is described. The results obtained by FISH are related to cervical dysplasia as assessed by cytology, colposcopy and histology.The detection of HPV infection was more sensitive and specific by FISH than by either histological/cytological evidence of an HPV cytopathic effect (koilocytosis) or immunohistochemical staining for HPV capsid antigen using antiserum against genus specific antigen of the bovine papillomavirus. Viral DNA was detected by FISH in 65% of women with atypical cytology and of the HPV positive cases, 68% were HPV 16/18, 22% HPV 6/11 and 10% of mixed types.HPV-DNA was detected in the cervical smears of 16 women who also had HPV capsid antigen (HPV-Ag) in their cervical biopsies; HPV 6/11 was found in 4 cases and HPV 16/18 in 12 cases. The relative frequency of the HPV-Ag positive cases decreased markedly from 44% to 49% with an increase in the severity of cervical dysplasia. By comparison, the percentage of HPV-DNA positive cases remained relatively constant between 70% and 80% for all 3 categories of dysplasia. Of these, the percentage of HPV 6/11 positives decreased slightly with an increase in the severity of dysplasia as assessed by either cytology or histology, whereas the percentage of HPV 16/18 positive cases was relatively constant.HPV-DNA was found in cervical smears of 11 of 48 (23%) women who were diagnosed colposcopically to have a typical transformation zone of the cervix. In 138 women with an atypical transformation zone, 96 (70%) were found to have HPV-DNA -xvith HPV 16/18 contributing to 83% of the HPV positive cases. All four genotypes of HPV were found to be associated with the colposcopy morphology of mosaicism (26 cases) but only HPV 16/18 was found in 12 HPV-DNA positive cases associated colposcopically with punctation and an atypical transformation zone.The detection and typing of HPV-DNA in cell scrapings by FISH is a relatively fast and non-invasive procedure which complements cytology, colposcopy and histology and should be useful in further studies of the natural history of different HPV infections and their role in cervical cancer.

Malakoplakia of the prostate: a morphological and biochemical study

Summary A case of malakoplakia of the prostate is presented. Electron microscopic appearances support the origin of the Michaelis‐Gutmann bodies from phagolysosomes in the histiocytes characteristic of the lesion. Biochemical analysis revealed the presence of muramic acid in the prostate with malakoplakia. This amino sugar is characteristic of bacterial cell walls and despite the absence of demonstrable bacteria in the affected tissues indicates the involvement of bacteria in the disease process.

Comparison of Peroxidase-Antiperoxidase and Avidin-Biotin Complex Methods for the Detection of Papillomavirus in Histological Sections of the Cervix Uteri

&NA; Summary A study was undertaken to determine the relative sensitivities of the peroxidase‐antiperoxidase (PAP) and avidin‐biotin complex (ABC) methods for the detection of human papillomavirus (HPV) antigens in acetic acid‐ethanol fixed paraffin‐embedded cervical tissue. Tissue sections prepared from 14 women suspected to have HPV infections with either atypia or dysplasia were stained immunohistochemically using an antiserum against genus‐specific (common) antigen of bovine papillomavirus. Detection of HPV antigen was approximately twice as frequent by the ABC method as by the PAP method. Of the 14 cases studied, 43% were found to be HPV positive by the PAP method whereas 79% were HPV positive by the ABC method. In addition, the number of cells found to be HPV positive by the ABC method was approximately double the number by the PAP method.

Assessing the effectiveness of a mammography screening service

Background:  Trials have shown that mammography screening reduces mortality and probably decreases morbidity related to breast cancer.

Histological grading of breast cancer: a study of reproducibility of consensus grading

Summary There has been a resurgence of interest in the use of histological grade in predicting prognosis in patients with breast cancer, although several studies have questioned the reproducibility of the techniques used. Interobserver agreement rates of 90% have been reported by the Nottingham group, recognized experts in the field, but a previous study from this institution showed only a 70% agreement rate between two patho- logists independently reviewing histological sections. This study reports the results of an attempt to improve internal consistency in reporting histological grade by using established, well defined criteria, and a consensus opinion by pathologists ‘trained’ in the method. Criteria established by the Nottingham group were used by three pathologists to obtain a consensus histological grade in 98 cases of breast cancer, assessed on two occasions at an interval of 6 months. The intra-observer agreement rate of 86 % showed an improvement on the previous interobserver agreement rate, and is comparable to agreement rates of 90% reported by the Nottingham group. Relative disagreement rate (RDR) of 0.10 (95% CI 0.05–0.19) and kappa statistic 0.77 (95% CI 0.66–0.88) also showed improvement over previous results, and compares favourably with other studies. The results suggest ‘training’ of pathologists in assessing histological grade, strict adherence to guidelines, and a consensus opinion, may all contribute to greater reproducibility.