Otto Gröntoft

Cytometric characterization and clinical course of breast cancer diagnosed in a population‐based screening program

A randomized controlled trial evaluating mammographic screening was started in two Swedish counties in 1977. In one of these, Óstergótland county, the authors performed static cytofluorometry on 161 cancers detected at the second and third screening rounds, 50 interval cancers, and 219 cancers appearing in the nonscreened control group during the same time period. The median follow‐up time was 42 months. No difference in mean S‐phase was found between screening and control group cancers, but interval cancers, appearing between two screenings, had increased mean S‐phase levels (P = 0.01) compared to both of the other groups. A high S‐phase fraction was associated with distant recurrence in both node‐negative and node‐positive tumors. Aneuploid tumors were more often found in the control group (67%) and among interval cancers (72%) than among screening detected cancers (55%, P = 0.02). In Coxs multivariate analysis, including all patients, the lymph node status, tumor size, estrogen receptor content, and S‐phase all contributed independent prognostic information about the clinical course. DNA ploidy predicted the outcome in simple but not in multivariate Coxs analysis. When analyzing screening‐detected cancers separately, only the S‐phase significantly predicted distant recurrence in multivariate analysis. In tumors with local recurrence, a high S‐phase implicated an increased, although not statistically significant, risk for distant recurrence. Survival with metastatic disease was significantly influenced by the S‐phase level (P = 0.002). The authors conclude that S‐phase fraction provides valuable kinetic information related to the clinical outcome for all stages of the disease and serves as a prognostic factor in screened populations, which have tumors predominantly in early stages.

Carcinoma in Situ and Severe Dysplasia of the Vocal Cords: A Clinic pathological and Photometric Investigation

Premalignant lesions of the vocal cords vary in their gross appearance. and diagnosis invariably relies on microscopic examination. This retrospective clinicopath-ological and photometric study of 20 cases of severe dysplasia and carcinoma in situ showed that various degrees of squarnous cell differentiation may he recognized for laryngeal premalignant lesions and that such distinction may be of prognostic significance. Carcinoma in situ is widely accepted as a premalignant lesion. while there has been an uncertainty about the histological criteria and the biological nature of severe dyasplasia. None of the cases of carcinoma in situ of classic type recurred. whereas well differentiated severe dysplasia had a high recurrence rate and development of invasice carcinoma. A classitlcation of severe dysplasia into well and moderately, to poorly differentiated is proposed. Photometric examination proved to be of value in the assessment of nuclear atypia. which was sometinies masked. and especislly in the case o...

A fast and reliable system for microcomputerized DNA cytofluorometry in tumour pathology

SummaryA microcomputerized cytofluorometry system based on a Leitz MPV 3 cytophotometer, and intended for DNA measurements in tumour pathology is described. The system has been equipped with a reference channel for correction of excitation light instability. The importance of the adjustment of the epi-illumination for optimal performance of the reference channel is stressed, and a detailed description is provided. An apparatus variability well below 1% CV is obtained even during periods of marked instability of the arc-lamp. Software for conventional cytofluorometry with which about 200 cells can be measured in 20 min is presented. In addition a measuring technique where cells are not positioned, but are just passed through the excitation light spot is described. Preliminary results on cytocentrifuged specimens of human cancers indicate that this system is at least three times faster than conventional fluorometry. The increased speed of measurements considerably extends the possibilities to evaluate cell proliferation using static cytofluorometry. The measuring capacity of course is dependent upon the quality of the specimen. In well prepared cytocentrifuged specimens 1000 cells have been measured in 35 min.

The DNA content and nuclear size in normal, dysplastic and carcinomatous laryngeal epithelium. A spectrophotometric study.

The aim of the present study was to obtain a more objective evaluation of nuclear hyperchromasia and polymorphism in laryngeal epithelium. The method is based on a photometric assay of nuclear size and DNA content in Feulgen stained slides. The DNA content of hyperchromatic nuclei should exceed the mean for nuclei of normal epithelium by more than twice the standard deviation. In comparison with lymphocytes (2N) the DNA content in hyperchromatic nuclei corresponds to 3.3 N. The mean DNA contents of normal, dysplastic and carcinomatous laryngeal nuclei were 69, 71 and 118 A.U. The mean nuclear area for normal, dysplastic and carcinomatous epithelial cells were 48, 41 and 73 micrometer2. The higher the degree of atypia displayed by the cells the greater was the variability of the DNA content and the nuclear size. The investigation shows that the method may provide a more objective basis for evaluating hyperchromasia and polymorphism.

HISTOSCAN: Computer program for cytophotometry in tissue sections and its application in the evaluation of nuclear atypia

SummaryThe grading of nuclear atypia has a great and well recognized value when predicting the malignancy of neoplastic processes. Although the basic criteria for the grading are generally agreed upon, in the individual case, the final judgement is to some extent still a matter of subjectivity, which naturally impairs reproducibility. The present paper describes a method for quantification of variability of nuclear size and Feulgen-stainability. By plotting the mean optical density against the area value in a scatterdiagram, a cluster is obtained, the size of which reflects the degree of nuclear atypia. The measurements are performed in tissue sections using stage scanning cytophotometry. A computer program — HISTOSCAN — has been developed which enables measurements in highly cellular tissues. The system is also insensitive to the influence of light scattering, a factor of importance when measuring in tissue sections. The performance of the program is tested in both imprints and tissue sections.

Cytometric and histopathologic features of tumors detected in a randomized mammography screening program: correlation and relative prognostic influence

SummaryCytometric determination of S-phase fraction and ploidy type was performed on 430 tumors detected within a randomized trial of mammographic screening. The results were compared to several histopathologic features. A high S-phase fraction was estimated in tumors with a high grade of malignancy and other histopathologic findings related to rapid tumor progression, including lack of tubule formation, a high mitotic index, marked nuclear pleomorphism, multifocal cancer growth, tumor emboli in lymphatic and blood vessels, tumor necrosis, and inflammatory reaction. DNA aneuploidy was correlated with a high malignancy grade, frequent mitoses, a high degree of nuclear pleomorphism, vascular invasion, necrosis, and the presence of noninvasive ductal carcinoma. Both cytometric variables were inversely related to the degree of elastosis. Positive nodes, large tumor size, DNA aneuploidy, a high S-phase fraction, high grade of malignancy, lack of tubule formation, as well as high mitotic index and pleomorphism, presence of multifocal cancer, and vascular invasion, predicted a significantly shorter distant recurrence-free interval after a median follow-up time of 46.6 months. Elastosis and the presence of estrogen and progesterone receptors indicated favorable prognosis. In the multivariate analysis, only lymph node status, tumor size S-phase fraction, and multifocal growth pattern had independent prognostic value.

Chondrosarcoma of the larynx.

A chondrosarcoma arising in the posterior cricoid plate is presented. The tumour gave rise to increasing inspiratory stridor: laryngectomy was performed. The tumour consisted of loose cartilaginous tissue with great predominance of highly differentiated cartilage cells and only small areas with nuclear polymorphism. This complicated the differential diagnosis between chondroma and highly differentiated chondrosarcoma. From the fact that the patient died from massive pulmonary metastases 3 1/2 years later, it is evident that the degree of malignancy of cartilaginous tumours should be determined even on small polymorphic foci. The DNA histogram for the foci of the laryngeal tumour with atypia differed distinctly from those for benign chondroma and normal cartilage, but resembled those of the pulmonary and splenic metastases and of a nasal chondrosarcoma. Photometric examination may provide an aid in the difficult differential diagnosis between chondroma and highly differentiated chondrosarcoma.

Cytophotometric characterization of the WHO grades of transitional cell neoplasms.

The WHO classification of urothelial bladder tumours is widely used, but does not seem to be entirely satisfactory. The main problem is the unhomogeneous WHO II group, and borderline cases in particular are difficult to evaluate histopathologically. Objective assessment of atypia would allow more reliable correlation between morphology and biological behaviour of the tumours to be made. We describe measurements of atypia in 3 characteristic cases of papillary bladder tumour of each WHO grade I, II, and III using Feulgen-DNA-cytophotometry. The mean optical density and nuclear area is determined in 5 micron thick tissue sections. The optical density and nuclear area for each specimen are plotted in a scatter diagram, and the position and variability of the cluster are evaluated. Cytophotometric characterization resulted in defined groups of WHO-graded tumours, and due to this encouraging result the method will be applied in a more comprehensive study of WHO II carcinomas.

Histopathology grading in small brast cancers ≤ 10mm – results from an area with mammography screening

Histopathological malignancy grading ysing the Bloom-Ricardson classification of ductal cancers was performed for 248 invasive ductal breast cancers ≤10mm operated 1978-1985. There weresignificantly more grade 1 lesions in the prevalence screening round. Grade 3 was correlated with aneuploidy, higher S-phase (SPF), and more receptor negative tumours. There were also significantly more positive lymph nodes in grade 3 lesions, 18% compared to 5% and 12% respecrively for grades 1 and 2 (p<0.05). In life table analysis for survival, when the high risk group of grade 3 lesions was compared to the grade 1 and 2 lesions combined, five-year disease-free survival was 84.6% vs. 99.1% (p<0.001).With good training and care from the pathologist, malignancy grading seems useful for prognostication of eventual recurrence and death. In umours 10mm or smaller only grade 3 lesions need to be included in follow-up systems and should probably have adjuvant treatmentMalignancy grading is especially good in small ductal brast cancers whre grading can always be performed while other prognostic determinations are hampered by shortage of material. Lymph node positivity is also low in this groupHistopathological malignancy grading ysing the Bloom-Ricardson classification of ductal cancers was performed for 248 invasive ductal breast cancers ≤10mm operated 1978-1985. There weresignificantly more grade 1 lesions in the prevalence screening round. Grade 3 was correlated with aneuploidy, higher S-phase (SPF), and more receptor negative tumours. There were also significantly more positive lymph nodes in grade 3 lesions, 18% compared to 5% and 12% respecrively for grades 1 and 2 (p<0.05). In life table analysis for survival, when the high risk group of grade 3 lesions was compared to the grade 1 and 2 lesions combined, five-year disease-free survival was 84.6% vs. 99.1% (p<0.001). With good training and care from the pathologist, malignancy grading seems useful for prognostication of eventual recurrence and death. In umours 10mm or smaller only grade 3 lesions need to be included in follow-up systems and should probably have adjuvant treatment Malignancy grading is especially good in small ductal brast cancers whre grading can always be performed while other prognostic determinations are hampered by shortage of material. Lymph node positivity is also low in this group

Influence of S-phase fraction on metastatic pattern and post-recurrence survival in a randomized mammography screening trial

SummaryUsing static cytofluorometry, S-phase was determined on the primary tumors of 421 patients with breast carcinomas in stages I–III diagnosed 1981–85 during the second and third screening rounds of a randomized trial evaluating the effect of mammographic screening. Through December 1988, 82 patients had developed local and/or distant recurrence, 51 of whom had died of cancer during the same period. The distribution among sites of recurrence differed between patients with tumors detected by mammography screening and cancers diagnosed due to clinical symptoms. The mean S-phase fraction was highest in patients with liver or brain metastases and lowest in patients with metastases in subcutaneous and cutaneous tissue and lymph nodes only. In univariate analysis, survival after first recurrence was significantly associated with the site of primary recurrence, the disease-free interval, and node status and tumor size at diagnosis, as well as the S-phase level. The median survival period was 31.3. months for patients with a S-phase fraction below 6%, and 10.7 months in cases with S-phase exceeding 10%. Neither ploidy nor the estrogen receptor content had significant influence on post-recurrence survival. In Coxs multiple regression analysis, only metastatic site, disease-free interval, and S-phase fraction showed significantly independent prognostic value.