Superior performance of liquid-based versus conventional cytology in a population-based cervical cancer screening program.

OBJECTIVE: Liquid-based cytology may offer improvements over conventional cytology for cervical cancer screening. The two cytology techniques were compared in a group of 86,469 women who participated in a population-based screening program. Using a nation-wide pathology database containing both cervical cytology and histology records for all patients, we compared the outcome of the two screenings methods with regard to the detection rate of histological proven abnormalities and the determination of the true false-negative rates for both methods. METHODS: Two cohorts of women living in the same geographical region were used. Cohort 1 (n=51,154 women) was analysed using conventional cytology (conventional cohort) and cohort 2 (liquid cohort) (n=35,315 women) was analysed using liquid-based cytology (SurePath). The samples were processed in one laboratory. The results of histological follow up were available via a central database. RESULTS: The rate of unsatisfactory slides was significantly lower using liquid-based cytology (0.13% vs. 0.89%, p<0.0001). Detection of ASCUS+ (Atypical squamous cells of unknown significance or higher abnormalities) was significantly higher using liquid-based cytology (2.97% vs. 1.64%, p<0.0001), mainly due to the increase in the ASCUS category. The percentage of histological abnormalities within the ASCUS samples was approximately equal in both cohorts, indicating that more true abnormal cases were detected using liquid-based cytology. The sensitivity for detection of a histological proven lesion is significantly higher in the liquid cohort compared to the conventional cohort (96.2% vs. 92.0%), with only a slight difference in specificity (97.8% vs. 98.2%). CONCLUSION: This population study confirmed previous institution-based reports of decreased numbers of unsatisfactory samples based on liquid-based cytology and showed an increased sensitivity for the detection of cytological abnormalities that was validated by subsequent histological investigation.

Ultrasound-guided fine needle aspiration cytology of axillary lymph nodes in breast cancer patients. A preoperative staging procedure.

Currently, axillary lymph node dissection is increasingly being replaced by the sentinel node procedure. This method is time-consuming and the full immunohistochemical evaluation is usually only first known postoperatively. This study was designed to evaluate the accuracy of preoperative ultrasound-guided fine needle aspirations (FNAs) for the detection of non-palpable lymph node metastases in primary breast cancer patients. We evaluated the material of 183 ultrasound-guided FNAs of non-palpable axillary lymph nodes of primary breast cancer patients. The cytological results were compared with the final histological diagnosis. Ultrasound-guided FNA detected metastases in 44% (37/85) of histologically node-positive patients, in 20% of the total patient population studied. These pecentages are likely to be higher when women with palpable nodes are included. Cytologically false-negative and false-positive nodes were seen in 28 (15%) and three cases (1.6%), respectively. Interestingly 25% (n=7) of the false-negative nodes, revealed micrometastases on postoperative histology. The sensitivity was 57%, the specificity 96%. We conclude that ultrasound-guided FNA of the axillary lymph nodes is an effective procedure that should be included in the preoperative staging of all primary breast cancer patients. Whether lymph nodes are palpable or not, it will save considerable operating time by selecting those who need a complete axillary lymph node dissection at primary surgery and would save a significant number of sentinel lymph node dissections (SLNDs).

Increased transplant-related morbidity and mortality in CMV-seropositive patients despite highly effective prevention of CMV disease after allogeneic T-cell-depleted stem cell transplantation.

We evaluated the efficacy, toxicity, and outcome of preemptive ganciclovir (GCV) therapy in 80 cytomegalovirus (CMV)-seropositive patients allografted between 1991 and 1996 and compared their outcome to 35 seronegative patients allografted during the same period. Both cohorts were comparable with respect to diagnosis and distribution of high- versus standard-risk patients. All patients received a stem cell graft from an HLA-identical sibling donor, and grafts were partially depleted of T cells in 109 patients. Patients were monitored for CMV antigenemia by leukocyte expression of the CMV-pp65 antigen. Fifty-two periods of CMV reactivation occurring in 30 patients were treated preemptively with GCV. A favorable response was observed in 48 of 50 periods, and only 2 patients developed CMV disease: 1 with esophagitis and 1 with pneumonia. Ten of 30 treated patients developed GCV-related neutropenia (less than 0.5 x 10(9)/L), which was associated with a high bilirubin at the start of GCV therapy. Overall survival at 5 years was 64% in the CMV-seronegative cohort and 40% in the CMV-seropositive cohort (P =.01). Increased treatment-related mortality accounted for inferior survival. CMV seropositivity proved an independent risk factor for developing acute graft-versus-host disease, and acute graft-versus-host disease predicted for higher treatment-related mortality and worse overall survival in a time-dependent analysis. We conclude that, although CMV disease can effectively be prevented by preemptive GCV therapy, CMV seropositivity remains a strong adverse risk factor for survival following partial T-cell-depleted allogeneic stem cell transplantation.

Immunocytochemical detection of prognostic markers in breast cancer; technical considerations.

The purpose of this study was to establish a good technical procedure for immunocytochemical (IC) staining of prognostic markers in breast cancer specimens. The influence of various preparation, fixation and storage methods on ER, P53 and Ki-67 IC staining was assessed, using cells of two breast cancer cell lines T47D (ER/P53+) and ZR-75-ER (ER+, P53-). In addition we searched for a suitable transport medium. Depending on the technical procedure, great variations in expression of the tested antigens were found. Cytospins fixed and stored according to the Abbott method gave the best results. Histocon appeared to be the medium of choice. A good concordance of IC and immunohistochemical (IH) results was found when the adopted method was tested on material of 10 breast cancers. This study underlines the importance of quality controlled standardization of cell processing, fixation and storage of fine needle aspiration (FNA) aspirates in order to obtain reproducible and consistent IC results.

Altered expression of p53 and its regulated proteins in phyllodes tumours of the breast.

The histological characteristics of phyllodes tumours of the breast are often not related to their clinical outcome. Additional studies must therefore be performed to investigate the possible relationship of cell biological parameters to the biological behaviour of these tumours. The expression of Ki-67, p53, and its regulated proteins has been studied in 19 primary phyllodes tumours, from patients with known follow-up, using immunohistochemical and molecular biological techniques. Overexpression of the p53 protein was observed in four cases and mutation in two cases. In only one case, the sequence alteration, at codon 273, was associated with overexpression of p53 protein and with strong expression of Ki-67 (30 per cent). This alteration was found in the primary, the recurrent, and the metastatic tumour samples. Moreover, the same p53 gene mutation, Arg273Cys, was detected in all tumour samples. No mutation was found in adjacent normal breast tissue, indicating that this was an acquired mutation. Unexpectedly, strong BAX expression was observed in the primary tumour. The patient died during the follow-up period. It is concluded that p53 gene status and an accumulation of BAX, both involved in the same apoptosis-controlling pathway, may be of prognostic relevance in phyllodes tumours.

Immunocytochemical staining of effusions; an external quality control study in The Netherlands.

In The Netherlands an external quality control study of immunocytochemical (IC) staining of effusions was initiated, consisting of three test rounds. The 12 participating laboratories received samples of malignant effusions (runs 1, 2 and 3), and five unstained control specimens prepared from the same material in runs 2 and 3. The laboratories used their own protocols to prepare and stain the samples ('in-house' specimens). Two persons viewed and scored the slides following preset criteria concerning number and morphology of diagnostic cells, background staining and staining specificity. Better scoring results were found for control specimens, compared with 'in-house' specimens, primarily caused by cell loss in the latter. This finding underlines the view that high quality IC needs well organized processing and staining procedures, and warrants external quality control systems.

The clinical significance of androgen receptors in breast cancer and their relation to histological and cell biological parameters.

To analyse the clinical significance of the presence of androgen receptors (AR) in breast carcinomas, clinical and histological parameters of 153 primary breast carcinomas (median follow-up 46 months) were examined. Oestrogen (ER) and progesterone receptor (PR) levels were determined in cytosol preparations using enzyme immunoassay assays and in cryostat sections by immunohistochemistry. AR and Ki-67 levels were only determined immunohistochemically. Data were analysed by uni- and multivariate models. 94/153 (61%) breast carcinomas were ER+ PR+ AR+, while 14 cases were only positive for AR. All grade III tumours (n = 17) were steroid receptor negative and 14 (76%) of these cases demonstrated high Ki-67 values suggestive of more aggressive behaviour. Strikingly, 14 ductal carcinomas negative for ER and PR were positive for AR. In univariate analysis, AR as well as ER, tumour size, lymph node status, grade and Ki-67 proved to be significant prognostic factors for disease-free survival (DFS). Multivariate analysis, however, showed lymph node status, tumour size and ER status to be the only independent prognostic factors for DFS within this model. We conclude that simple histological and cell biological parameters, including AR, can be used to select high- and low-risk patients at the time of primary surgery and can provide valuable information on treatment options.

Quality control of immunocytochemical staining of effusions using a standardized method of cell processing.

OBJECTIVE: To improve the quality and reproducibility of immunocytochemical staining of effusions by using a standardized method of cell processing. STUDY DESIGN: The study included the specimens of 108 effusions (44 benign, 56 adenocarcinoma metastases and 8 mesotheliomas). Hemorrhagic effusions were lysed using isotonic ammonium chloride. All pellets were fixed in 1% paraformaldehyde dissolved in phosphate-buffered saline (PBS), pH 7.4, and washed in PBS. A Burker counting chamber was used to adjust the pellets to a standard cell concentration. The panel of monoclonal antibodies (MAbs) included MOC-31, Ber-EP4 and anticarcinoembryonic antigen (anti-CEA). The alkaline phosphatase/anti-alkaline phosphatase technique was applied. RESULTS: Standardized material processing resulted in reproducible specimens with good preservation of cell morphology, reduction of nonspecific interaction and good immunostain intensity. MAbs MOC-31 and Ber-EP4 gave similar results: both were positive in all adenocarcinomas. Anti-CEA was positive in 73%. Benign effusions showed no expression. In contrast to the literature, seven mesotheliomas showed variable membranous expression of MOC-31 and Ber-EP4. CONCLUSION: High-quality immunostaining results were obtained by using a standardized method of cell processing. MAbs MOC-31 and Ber-EP4 cannot be used as differentiation markers between mesotheliomas and adenocarcinomas. Discrepancies in immunocytochemical staining results may be caused partly by differences in cell preparation.

[Good results in periodic screening in women with increased risk of breast cancer; retrospective study]. / Gunstige resultaten van periodieke controle bij vrouwen met verhoogd risico van borstkanker; retrospectief onderzoek.

OBJECTIVE: To determine the efficiency of surveillance among women with a high risk of breast cancer by semiannual breast examination and annual mammography. SETTING: 'Dr. Daniel den Hoed' Cancer Centre, Rotterdam, the Netherlands. DESIGN: Retrospective study. METHOD: In 1994 breast cancer was diagnosed in 92 patients: in 19 patients during surveillance (SU), in 48 referred because of symptoms (SY) and in 25 during the national screening programme (SC). The contributions of physical examination, mammography, ultrasound and fine needle aspiration cytology to the diagnosis were evaluated and the tumour stages in the 3 groups were compared. RESULTS: The mean age was 53 years in the SU group, 51 years in the SY group and 64 years in the SC group. Breast cancer was detected by mammography only in 6%, 40% and 42% in the SY, SC and SU groups, respectively. 42% of patients had a family history of breast cancer in the SU group. Mammography failed to reveal malignancy in 25% of patients in the SU and SY groups combined. In women in the SU group 53% of breast cancers were: carcinoma in situ (Tis) or minimal invasive carcinoma (< 0.5 cm diameter; T1a). In patients with a family history of breast cancer 75% (6/8) were diagnosed in stages Tis or T1a, by surveillance. In the SY group 4% were Tis or T1a and 40% N1 + 2, in the SC group 8% and 20% respectively. 15.7% of the SU group had axillary lymph node metastases (N1) but only one of the 8 patients with a family history of breast cancer. CONCLUSIONS: Surveillance by palpation and mammography in women with a high risk of breast cancer is useful, because it detects this disease frequently at an early stage, when the prognosis is still good.

Immunohistochemical determination of androgen receptors in relation to oestrogen and progesterone receptors in female breast cancer.

The expression of oestrogen (ER), progesterone (PR) and androgen (AR) receptors in female breast cancer was investigated by immunohistochemistry on snap-frozen tissue specimens of a series of 100 breast cancers. For detection of the AR we used a recently developed mouse monoclonal antibody specific for the N-terminal domain of the human AR. Expression of AR was compared with that of ER and PR as well as with tumour grade and age. Of the breast cancers investigated, 76% were AR-positive. This high percentage corresponds well with previous data on AR expression in breast cancer determined with ligand-binding assays. In 53% of the tumours AR, ER and PR were present, while 9% of the tumours were positive for AR and negative for ER and PR. In 13% of the tumours no ER, PR or AR expression was seen; these were all grade-III tumours. A positive correlation was found between age and ER expression, but no correlation was seen between age and PR or AR. Future studies should establish the prognostic value of the combination ER, PR and AR determinations on female breast cancer with regard to biological behaviour and response rate to hormonal therapy.

Ki-67 staining in histological subtypes of breast carcinoma and fine needle aspiration smears.

Thirty four cases of invasive breast carcinoma were analysed for heterogeneity of Ki-67 reactivity in a tumour, and proliferative activity in various histological subtypes was compared. The growth factions determined in areas of central and peripheral tumour were the same. Mucinous and lobular carcinoma showed lower Ki-67 activity than ductal carcinomas. When ductal carcinomas were subdivided according to their dominant growth pattern, the carcinomas with a solid or comedo growth pattern showed the highest proliferative activity. These results largely confirm data from previous cell kinetic studies on the incorporation of radioactively labelled thymidine. A correlation between the growth fraction determined by Ki-67 in fine needle aspiration smears and cryostat sections of corresponding tumours was shown, implying that the immunostaining of cytological smears gives a reliable impression of the growth fraction of a tumour and may therefore be used in prospective studies.

Prognostic scoring using cytomorphometry and lymph node status of patients with breast carcinoma.

Applying morphometry on FNA smears may lead to meaningful prognostic subgrouping of breast carcinomas. A statistical analysis of the cytomorphometric and clinical data of 73 patients with breast carcinoma was performed. By multivariate analysis, taking into account various prognostic factors, it was shown that variations in nuclear area and the presence of axillary metastases were the most important prognosticators. By weighing these factors a prognostic score was obtained. According to that score the patients were classified into 4 score groups: the decreasing survival with increasing score was evident. The prognostic score was also related to the metastasis-free period. This prognostic scoring is relatively easy to perform, and can be done in routine pathology laboratories.