Use of a mucolytic agent (Cytoclair) in the preparation of cell material for the detection of malignant cells in sputum.

A mucolytic agent, methyl cysteine HC1 (Cytoclair), has been used to obtain a centrifuged cell deposit for the detection of malignant cells in sputum. In 178 sputum samples from 52 patients with primary lung cancer there was a slightly higher rate of detection with the mucolytic method than with the routine method: 54 positives from 27 patients compared with 48 positives from 24 patients. Morphological studies using the mucolytic agent indicated that a millipore filter with fixation of cells in suspension produces a satisfactory cell preparation and is valuable in certain problem cases.

Cross contamination of cytological smears, with automated staining machines and bulk manual staining procedures. With a specific study of the problems of the Cytotek and the Shandon Elliott staining machines.

Further development of an individual staining machine is to be strongly encouraged but meanwhile, using bulk stainers, frequent changing of wash fluids and staining solutions, particularly leading up to and following the haematoxylin pot, is essential to reduce the risk of cross contamination. Certain smears, such as from semen or from serous fluids where malignancy is suspected or known, must be stained on separate racks. In some laboratories it is the rule not to stain semen or serous fluids in bulk staining machines at all and this may have to become the rule everywhere until we are provided with safe individual slide stainers.

Feulgen-hydrolysis profiles in cells exfoliated from the cervix uteri: a potential aid in the diagnosis of malignancy.

By varying the time of hydrolysis for the Feulgen reaction, done under conditions that protect the backbone of the DNA, it is possible to distinguish three species of DNA that are characterised by their lability to acid hydrolysis. The most labile DNA was found, in greatest proportions, in malignant cells; this may be helpful in diagnostic cytology. The fact that the cytologically normal cells, in grade V smears, also show this labile DNA may well facilitate cytological screening even in those smears that contain very few neoplastic cells.

A modified tetrazolium reaction for identifying malignant cells from gastric and colonic cancer.

When non-malignant cells were reacted for glucose-6-phosphate dehydrogenase activity, with neotetrazolium chloride as the indicator of the activity, oxygen competed with the neotetrazolium and nullified the reaction. In contrast, about 30% of the activity was retained in malignant cells, in sections and in smears, from cancer of the stomach or colon. This could provide the basis of a qualitative (black-or-white) functional test for distinguishing malignant cells in these conditions.

Monoclonal immunofluorescence compared with silver stain for investigating Pneumocystis carinii pneumonia.

Two hundred and eighty two specimens from 220 patients positive for HIV with respiratory tract symptoms, or febrile illness, or both, were examined for the presence of Pneumocystis carinii. Specimens were either induced sputum samples or bronchoalveolar lavage fluids. To establish the optimal method for laboratory diagnosis a comparison was made of detection of the organism by use of monoclonal antibody and immunofluorescence with conventional silver staining methods. Three commercially available reagents for immunofluorescence were also compared. Immunofluorescence was significantly more sensitive than the silver stain and the best results for immunofluorescence were obtained using. Northumbria Biologicals Ltd reagents.

Quality control in cervical cytology.

From surveys conducted by the authors it is concluded that the best and most acceptable quality control methods in cytology are those from within the laboratory. Most of these have results which can be reported centrally. Where the overall control and codes of practice are high, there the results are the most reliable, as sources of error from whatever cause are quickly brought to light. These conclusions are illustrated by data from the five centres and correlated in the tables.

Interobserver variation in the diagnosis and grading of dyskaryosis in cervical smears: specialist cytopathologists compared with non-specialists.

AIMS: To compare the assessment of dyskaryosis in cervical smears made by specialist consultant cytopathologists and consultant general histopathologists. METHODS: One hundred and ten cervical smears were circulated to 10 observers from five district general hospital histopathology departments and five major departments of cytopathology. Their responses were analysed by five consultant general histopathologists and five consultant specialist cytopathologists. In 54 of the 110 cases, the histology of a corresponding cervical biopsy specimen was compared with the smear assessments. RESULTS: Specialist cytopathologists were more consistent than non-specialists when diagnosing and grading dyskaryosis. They chose the higher grades of dyskaryosis more frequently than the non-specialists. The cytopathologists recommended referral for colposcopy more frequently, but if they asked for a repeat smear, they wanted it done within three months more frequently than the histopathologists. The specialists were more frequently in agreement with the biopsy grade of intra-epithelial neoplasia than the non-specialists, whose smear diagnoses tended to underestimate the severity of the histopathological abnormality. CONCLUSIONS: This study has shown major differences between specialist and non-specialist cytopathologists in the diagnosis and grading of cervical smears and in the recommended management of patients with abnormal smears. These differences may result in uneven clinical management of women with smear abnormalities. It is therefore important to explore possible strategies for standardising the reporting of cervical smears, such as centralisation of screening services, accreditation in cytopathology for non-specialist consultants, and the value of participation in external quality assessment schemes.

Quantitative deoxyribonucleic acid analysis of patients with mild cervical atypia: a potentially malignant lesion?

Quantitative deoxyribonucleic acid (DNA) analysis of cervical biopsy material from 32 women with cytologic, colposcopic, and histologic evidence of mild cervical atypia consistent with cervical intraepithelial neoplasia I, reactive atypia, or human papillomavirus infection was carried out using flow cytometry and Feulgen microspectrophotometry. Evidence of aneuploidy, ie, neoplastic transformation, was demonstrated in all cases of cervical intraepithelial neoplasia I and 68% of cases with human papillomavirus-induced atypia. These results support the growing impression that human papillomavirus-induced cervical atypia should be regarded as a true precursor of cervical neoplasia, and emphasize the necessity to refer patients with mild atypia for definitive diagnosis and management.

Cytopathology in the United Kingdom: 1854 to the present.

The paper traces the history of cytopathology in the U.K. from the time of the pioneers in the last century to the 1930s and continues with the development of cervical and breast screening, with reference to training and quality control, to the present time. Diagn. Cytopathol. 2000;22:203-206.

Consistency of a double PAPNET scan of cervical smears.

OBJECTIVE: To determine the reproducibility and accuracy of the PAPNET system in finding and presenting abnormal and atypical cells. STUDY DESIGN: To assess the potential variation of a double PAPNET scan on the same cervical smear, 516 cervical smears from women with abnormal histologic diagnoses were scanned twice via PAPNET and reviewed by two independent examiners in a double-blind trial. RESULTS: The false negative rate of 5.7% in conventional screening was reduced to 0.4% and 0.8%, respectively, by PAPNET testing. The resulting concurrence of the first and the second PAPNET reviews was 99.2%, indicating the accuracy, sensitivity and consistency of this supplemental test. CONCLUSION: The results obtained demonstrate the reproducibility and accuracy of the system in finding and presenting abnormal and atypical cells.

PAPNET. The human and other dimensions.

OBJECTIVE: To study the effect of specimen preparation and observer variation on the accuracy of interpretation of PAPNET images. STUDY DESIGN: The PAPNET scanner makes use of conventional Papanicolaou-stained smears and presents the cytologist with pictures of selected "abnormal" cells. This minitrial was designed to investigate the human variability in different grades of personnel and to consider any problems caused by the use of conventional smears. RESULTS: The personnel involved consisted of a consultant cytopathologist, a senior chief medical laboratory scientific officer and two senior cytotechnologists, one from another laboratory. Experience with the system ranged from six months to a few years. The best interobserver agreement was found between the senior chief and the senior technologist, who worked in the same laboratory. Majority agreement with the "gold standard" (at least three out of four) was highest for the extremes of "negative" and "high grade" smears. Three high grade smears were "missed" due to the quality of the smears and scantiness of "abnormal" cells. CONCLUSION: Although fatigue in screening is reduced by the interactive mode, it does not overcome the subjectivity of human examination both in review of the dat tape and in microscopic examination of selected cells on the slides. Experience with the method is important, and different background experiences will have an effect. The technical quality of the smear is important since the automated scanner cannot be expected to select abnormal cells if they are absent or obscured for any reason. A monolayer may be the solution, although at greater cost.

A sample preparation for automated cervical cancer screening.

A procedure for cluster disruption, cell dispersal and the production of a monolayer of cells from fluid suspensions for automated cell-scanning machines in an interactive system has been developed. The combination of 0.1 per cent dithiothreitol and 40 per cent alcohol in a balanced salt solution provides a self-limiting form of mucolysis which is completed by a controlled syringing regime to produce an optimally dispersed cell sample without the production of cell debris from the more effete cells. The induction of a positive charge on a microscope slide to attract the negatively charged cells has been achieved by coating the slides with the cationic polymer, polylysine. The cells appear to be sufficiently well attached to permit wet fixation and subsequent stain processing without loss. Quantitative data is given to demonstrate the superior cell presentation that is achieved by this new technique for automated scanning.

Cell preparation methods and criteria for sample adequacy. International Academy of Cytology Task Force summary. Diagnostic Cytology Towards the 21st Century: An International Expert Conference and Tutorial.

ISSUES: Cell Preparation Methods Standardized fixation and optimal staining Sampling of cervix, sampling error, homogenization of sample, subsampling Assessment of liquid-based preparations: efficacy and economic impact Training and transitional procedures before full implementation of new technologies Criteria for Sample Adequacy Clinician responsibility for collecting and providing representative sample to laboratory Collection instruments, number of slides Cellular content of samples: evidence of transformation zone (TZ) sampling, number of squamous cells present, obscuring factors Screening issues CONSENSUS POSITION The conventional cervical smear remains the standard method of cervical cancer screening but has limitations in individual test sensitivity and specificity. Sample takers should: (1) receive appropriate training in sample collection, (2) be held responsible for providing the laboratory with appropriate samples, and (3) have their performance monitored. The instruments used for sampling should collect cells from both the ectocervix and endocervix; optimally, TZ sampling, represented by the presence of endocervical or squamous metaplastic cells, should be identifiable in samples other than atrophic specimens. The adequacy of a specimen (as judged microscopically) does not guarantee that it is representative of the cervix. Each cytology report should include a comment on cellular content/adequacy of the specimen. Liquid-based preparations may overcome many of the inherent problems with the conventional cervical smear. ONGOING ISSUES: We need further data on the cost-effectiveness of making two slides from cervical specimens and/or using two samplers rather than a single one. Do we have enough information to make recommendations as to the appropriate type of sampler to be used in particular situations, such as routine screening? What is the best method of screening for/detecting endocervical glandular neoplasia? How are such terms as unsatisfactory and inadequate defined in cervical cytology classifications other than the Bethesda System? What number and types of epithelial cells should be present (visualized) in a cervical smear or liquid-based preparation for it to be considered adequate? Do we need to have evidence of TZ sampling in specimens taken during the follow-up period after treatment of squamous intraepithelial lesion or after detection of endocervical glandular neoplasia? What criteria for obscuring factors, such as blood and inflammation, should be used in assessing adequacy? Cost-benefit analyses of utilizing liquid-based preparations are needed. Should we inform women about the technical details of the test methods available or chosen by the laboratory? Are women in a position to decide which method is the most appropriate to assess their cervical scrape sample? We need to obtain more information about the properties of proprietary liquid fixative/transport media with respect to inactivation of viral pathogens, tuberculosis and other bacterial pathogens and suitability for immunobiologic and molecular tests, etc. We need to obtain more information on the use of stoichiometric stains and the limitations of Papanicolaou stain for image analysis systems. The use of liquid-based preparations for nongynecologic cytopathology and ancillary tests must be considered, including criteria for adequacy. We need to obtain more information on the time required for and best methods of training experienced cytotechnologists to become competent at assessing liquid-based cervical preparations.

Health and cancer prevention: knowledge and beliefs of children and young people.

OBJECTIVE: To collect information from children and young people about their knowledge of and attitudes towards cancer and their understanding of health and health related behaviours to inform future health promotion work. DESIGN: Questionnaire survey of 15-16 year olds, and interviews with play materials with 9-10 year old children. SETTING: Six inner city, suburban, and rural schools. SUBJECTS: 226 children aged 15-16 years and 100 aged 9-10 years. MAIN OUTCOME MEASURES: Knowledge about different types of cancer; beliefs about health; sources of information; quality of research data obtainable from young children about cancer and health. RESULTS: Both samples knew most about lung cancer, but there was also some knowledge of breast and skin cancer and leukaemia. Smoking, together with pollution and other environmental factors, were seen as the dominant causes of cancer. Environmental factors were mentioned more often by the inner city samples. Television and the media were the most important sources of information. Young people were more worried about unemployment than about ill health. More than half the young people did not describe their health as good, and most said they did not have a healthy lifestyle. Children were able to provide detailed information about their knowledge and understanding by using drawings as well as interviews. CONCLUSIONS: Children and young people possess considerable knowledge about cancer, especially about lung cancer and smoking, and show considerable awareness of predominant health education messages. Despite this knowledge, many lead less than healthy lifestyles. Health is not seen as the most important goal in life by many young people; the circumstances in which many children and young people live are not experienced as health promoting.