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The GCT is a primary bone tumor of intermedia dignity, whereby the forecast of the biological behaviour and outcome is impossible on the base of the conventional histological diagnosis. The authors observed in one of their GCT-patients a malignant transformation of the formerly benign tumor. Through announcement of their case the authors want to call the attention to the importance of the DNA-Cytophotometry and to their possible future additional role in the diagnostics of the GCT and in the prediction of their biological behaviour.

DNA-cytophotometry is one of the methods that may contribute to a more precise evaluation of the biological behaviour of tumours in addition to the WHO-classification. In this study 121 tumour specimens of 50 patients suffering from gliomas with one or up to three recurrencies were investigated. In all cases the histological type and WHO-grade and the Ki-67/MIB1 labeling index were determined. DNA cytophotometry was performed after single cell preparation on Feulgen-stained preparations, and the following parameters were calculated: stemline ploidy, 5c-exceeding rate, and 2c-deviation index. Statistical evaluation revealed a highly significant correlation between recurrence-free interval and WHO-grade only. The DNA parameters, however, furnished additional information about increasing genetic instability in the majority of the recurrencies independently of changes in the WHO-grade. They thus seem to be useful as additional parameters for the determination of glioma progression.

Intension of our examination is to investigate changes in ploidy during radiotherapy and to find out radiosensitive cloni of tumors.In November 1993 a 83-year old patient with a carcinoma of the vulva was introduced to the Strahlenklinik der Städtischen Kliniken Offenbach. It was a stage T3N0M0 tumor (UICC 1990). We performed daily smears of the tumor surface during radiotherapy. The smears became air-dried before fixation in PBS/formaldehyde and staining with the Feulgen procedure. The DNA-analyse took place at an image-analysis-system (Ahrens Cytometry Analyse System).Nine and 16 weeks after radiotherapy the patient showed clinically complete remission. Macroscopically and histopathologically there was no tumor found. The ploidy-analysis shows the take off of the hyperpentaploid cells on radiotherapy. Nine and 16 weeks after finished therapy there are no more hyperpentaploid cells detectable.The DNA-determination is a senseful completition to estimate the success of therapy. It is possible to earn informations about the behaviour of different tumor cell populations during the course of radiotherapy. The hyperpentaploid cells are a radiosensitive part of the tumor. For being able to judge about the route of this cells, if they are proliferating cells of a peritriploid tumor population, or if they proliferate themselves, are some more examinations and additional investigations necessary.

Histiocytosis X is very rare in late adulthood, usually taking a benign course with a good prognosis. A 56-year-old patient developed acute disseminated histiocytosis X with multiorgan involvement. Despite extensive diagnostic work-up the diagnosis could only be established on the grounds of a skin biopsy. Histiocytosis X cells are S 100 protein-positive and express the membrane antigens CD 1, CD 4 and HLA-DR. Electron microscopy reveals Birbeck granules. On static cytophotometry, histiocytosis X cells exhibited normal DNS content without aneuploidy. One year after the onset of disease the patient's condition deteriorated rapidly despite aggressive polychemotherapy (CHOP), and he died of multiorgan failure due to histiocytosis X.

Carcinomas were induced to the thyroid gland of female rats, using a method originally proposed by Thomas and Bollmann (metachronous application of nitrosomethylurea and methylthiouracil), to establish cytomorphological, histomorphological, cytochemical, and flow-cytophotometric criteria for diagnosis of thyroid carcinoma. Verification was also intended of the diagnostic value of each of the methods involved for differentiation of nodular goitre. Another purpose of the study was to find out, whether chemically induced thyroid tumours in rat were comparable to thyroid neoplasms in man. This provided to the examiners genetically coherent and morphologically comparable biological material at various stages of thyroid tumour growth which included diffuse and adenomatous hyperplasias, adenomas, and, from the 18th to 42nd experimental weeks, papillary as well as follicular carcinomas in 31 to 100% of all experimental animals involved. Cytomorphological comparability of rat thyroid material (imprint specimens) with human material (fine-needle aspiration cytology) was ensured for normal as well as for hyperplastically altered thyroid glands, including adenomatous and carcinomatous changes. Hence, group typing of thyroid cytology, originally devised for human specimens, could be easily adapted to material obtained from rat. Assessment of cytological samples by microscopic criteria yielded an accuracy of 89% in malignoma diagnosis and proved to be an approach of highly informative potential also in the context of rat experiments. Use of additional cytochemical techniques (PAS, toluidine-blue, peroxidase, alkaline and acid phosphatases, gamma-glutamyl transpeptidase) as well as quantitative DNA determination by means of flow-cytophotometry was helpful in casting light at some scattered trends of change from normal for certain stages of proliferation, but it failed to enhance information in cytomorphological diagnosis of the individual case.

The DNA-content of mononuclear cells of the peripheral blood of infantile and juvenile ALL patients was investigated using Pulse Cytophotometry. The fraction of cells in S- and G2 + M-phase is significantly increased in comparison with samples of healthy probands. The fraction of DNA-synthesising cells (S-phase) of both peripheral blood (mononuclear cells) and bone marrow of leukemia patients cannot be significantly distinguished by mathematical methods. On the other hand, the fraction of cells in later phases of cell cycle (G2 + M-phase) is significant enhanced in the bone marrow in comparison with the peripheral blood. A high correlation was found between the number of leukocytes and fraction of G2 + M-phase cells in the peripheral blood of SR- and MR-patients. No correlation was found between the number of leukocytes and S-phase-fraction. The occurrence of aneuploid cell populations in the mononuclear fraction of peripheral blood in the acute state of ALL could be of importance for prognosis and regime of therapy.

The development of cancer in humans is considered as a multistep process ultimately leading to clinically detectable disease and further progression of the tumour. By use of cytometric techniques it has been possible to perform quantitative measurements and detect qualitative alterations in malignant tumours. This mainly applies to DNA aberrations and deviations in nuclear size and morphology. Such alterations may also be present for a long period prior to malignant, invasive growth, i.e. at a preneoplastic stage. A survey of available data on such preneoplastic changes is given. Since the type of aberrations is both dependent on the tissue of origin and the tumour type, this must always be taken into consideration. A particularly rapid expansion has taken place in the field of flow cytometry, where high resolution measurements of cellular DNA content as well as cell cycle distribution and other parameters can be performed shortly after sampling of the tissue. It is expected that cytometric methods will be used in future diagnostics of early cancerous growth in several organ systems, including the skin, the gastrointestinal tract, the urinary bladder, the breast and female genital organs.

The aim of the study was to describe aging changes in the functional nuclear edema and the classification of human hepatocyte nuclei, by determining three parameters--the size of the nucleus, the relative DNA quantity and the number of chromocenters. For this purpose, karyometry and DNA cytophotometry were performed on 10 human liver preparations. The data obtained was subjected to correlation, cluster and discriminance analysis. The results indicated a reduction in the capacity of liver cells for functional nuclear edema as aging progressed. Whereas at a young age there is only a loose correlation between nuclear size and DNA content, it becomes much stricter later on, partly caused by polyploidization. Cluster analysis, followed by discriminance analysis, is well suited for dividing the nuclei of human hepatocytes into two or three statistical populations provided the nuclear area, DNA quantity and number of chromocenters are all used as characteristics simultaneously. When allowance is made for functional edema, the biological interpretation of clusters from young liver preparations permits meaningful conclusions, but it appears problematic for old preparations. Here it might be more practical to analyze the mixed distributions resulting from a determination of the DNA quantity or nuclear size alone.

Macroscopically and microscopically precancerous lesions of the oral mucosa cannot be diagnosed with sufficient accuracy. Dysplasias represent a particular diagnostic difficulty. The reproducibility of grading of dysplasias may be increased by the quantification of morphological parameters using image analysis systems. However an unequivocal identification of obligatory precancerous lesions cannot be attained by this method. Measurement of the nuclear DNA content represents a non-morphological method for the identification of obligatory precancerous lesions. The diagnostic criterion of malignancy is aneuploidy. Structural or numerical chromosomal aberrations that are absent in benign tumours and normal cells typify aneuploidy. Aneuploidy is manifested by an atypical nuclear DNA distribution that can be identified objectively by suitable algorithms. Dysplasias of the squamous epithelium with aneuploid nuclear DNA contents are regarded as obligatory precancerous lesions. DNA measurements, computation of data and printout of a 'DNA diagnosis' are possible with a TV-image analysis system and an automated microscope within 30 min.

We summarize our own cytophotometric and flow cytometric measurements from malignant tumors and premalignant lesions. DNA, non-histoprotein and dry mass (by interference microscopy) have been assessed. Ploidy and the compartments of the cell cycle were evaluated. For most tumors (cervix, breast, ovary, melanoma, larynx, oro-facial, bladder) a bad prognosis was suggested by the following cytophotometric findings: aneuploidy, polyploidy, and/or a high proportion of S-G2/M nuclei. This has been statistically confirmed for breast cancer by case studies conducted over a period of five years. Among the changes associated with lung cancer were alterations in protein content. Cytophotometric findings can provide useful additional information concerning a patient's prognosis.

Antigens A, B, Lea, Leb, CA 12-5, CA 19-9 and CEA were prepared immunohistologically, a DNA-cytophotometry was carried out and the value of a new in vitro proliferation assay to test cytostatic drugs investigated. Material was as follows: 20 normal ovaries and 105 ovarian tumors. Blood group antigens were expressed at a high percentage from well-differentiated epithelial cells, CEA in 50% and CA 12-5 in 70% of the ovarian carcinoma. Predictions about the dignity of borderline tumors can be made with DNA-cytophotometry. The efficacy of 6 different cytostatic agents was investigated in a new in vitro proliferation assay.

The diagnostic importance of a new approach for the detection of N. go. by 1,2-Propanediol: NAD+-Oxidoreductase (1,2-PDOR) counting was demonstrated. The advantages are established in supporting of the culture diagnosis of cervical and urethral swabs and in the shorting of the diagnostic way by isolation of N. go. colonies and biochemical differentiation.

Cytophotometric determination of nuclear DNA content was performed in 121 cases by examining endometrial preparations. The endometrial cells were obtained by means of a MiMark cell sampler before curetting. A variety of groups (normal endometrium, adenomatous hyperplasia, endometrial carcinoma) were compared with each other and examined under statistical conditions. The differences were statistically significant according to the 3rd quartile (75% of DNA-population in a histogram) between the group of secretory transformed and atrophic endometrium and the group of well and moderately differentiated adenocarcinoma. Determination of the median and deviation index of the DNA population yielded similar results. The group of adenomatous hyperplasia did not differ from the normal endometrium according to all statistical parameters. Obviously it is impossible to distinguish precancerous endometrium from normal endometrium by cytophotometry. Differences between proliferative endometrium and well differentiated adenocarcinoma were seen in individual cases only. On the other hand, moderately differentiated adenocarcinoma could obviously be differentiated and delimitated.

1. Compared with the peripheral blood lymphocytes of healthy children the cell fractions in the S- and G2 + M-phase are significantly higher in the bone-marrow of those children affected with ALL. This increase was proved in the SR- and MR-group irrespective of the cytomorphological subtype and cytochemical reaction. In patients with relapses the percentage of S-phase cells is below 6%. 2. In about 30% of our patients (mainly in the SR-group with L1-morphology) an initial DNA-aneuploidy was identified. As the risk of relapse is higher in children without DNA-aneuploidy, this symptom has a pretherapeutical-prognostic significance. 3. In the phase of hematological full remission, DNA-frequency distribution correlates with the proliferative activity of normal hematopoiesis. It provides no additional information about the pretherapeutical risk.

DNA ploidy and the proportion of DNA-synthesized cells (number of S-phase fractions) were measured by DNA impulse cytophotometry in 39 patients with untreated carcinomas of the oral cavity and the oropharynx. 84.7% of the tumours of the oral cavity and 80.8% of the tumours of the oropharynx were DNA-aneuploid. The mean value of S-phase fractions of the carcinomas of the oral cavity was 9.18 +/- 4.82%, that of the oropharynx carcinomas was 9.16 +/- 5.85%. There was no correlation between the type of DNA ploidy or the number of S-phase fractions and the presence of regional metastases and the clinical outcome of the tumour diseases. A case report suggests that determination of the number of S-phase fractions may give important information on the prognosticable effect of cytostatic therapy. So pretherapeutic ICP determination might become helpful with regard to the indications for cytostatic therapy of oro-pharyngeal tumours.

1. As compared with the peripheral blood lymphocytes of healthy subjects the cells in the mononuclear fraction of peripheral blood and in the bone marrow of children with ANLL show a significant higher S- and G2 + M-phase. 2. A high proliferative activity correlate with a bad prognosis or with reaching no haematological remission. 3. Frequently, aneuploid cell populations will occur in the morphological subtypes M 4 and M 5.

With the help of the scanning microscope photometry DNA content and size of Feulgen-stained cytoblasts may be measured. On smears of 28 tumours of the urinary bladder of all degrees of differentiation in each case 200 nuclei were investigated, the DNA and surface values were applied in histograms and these were compared one with another. With increasing dedifferentiation the mean nucleus-DNA content and the spreading of DNA and surface values significantly increased. The mean size of the nuclei did not increase. Increasingly ploidy deviations appeared--clinically significant for the recidivation. With the help of the proportion of non-diploid nuclei the histo-pathological graduation could be repeatedly performed and statements could be made as to the proliferative behaviour of tumours of the same classification. The "chromatin density" of non-diploid nuclei allowed the exact separation of the papillomas (Gx2) from the carcinoma preparations. The results are discussed on the basis of the literature.

The single-cell fluorescence cytometric data presented in this study basing on measurements on 6 osseous Ewing's-Sarcoma and 17 classical osteosarcoma are showing a mirrorlike behaviour to cytological and histological appearance. The cellular monotony in Ewing's-Sarcoma and the polymorphic appearance of osteosarcoma are represented by typical aneuploid DNA-histograms. Ewing's-Sarcoma histogram's show a large peak between the diploid and tetraploid region and little deviation to the octoploid value. The conventional osteosarcomas are characterised by a very polyploid and changeable DNA-histogram with variable blocks and peaks and deviation up to 34c. Special trends in the subtypes are remarkable, but depend on further investigations. DNA-measurements support objectivization of histological appearance on the one hand and the controlling of therapeutical responses on the other hand.

In koilocytotic dysplasia of the uterine cervix the DNA content in squamous cells was quantitated by cytophotometry in histological preparations stained according to Feulgen. Three patterns of DNA distribution in the squamous cells were found. In type one (21.4%) the cells had DNA content in the diploid and paradiploid zone of the histogram. In type two (35.7%) cells with triploid and tetraploid DNA values were found, but with a conspicuous modal class of cells. In type three no modal class cells were found (42.9%). The quantitative DNA changes in squamous cells show that some of the HPV induced alterations may be regarded as precancerous.

Cytologic slides from 11 women with clinical, cytological and histological signs of a papillomavirus infection of the cervix uteri are investigated by cytophotometry and cytomorphometry. The mean values of DNA content, chromatin density, nuclear area and nuclear circumference of koilocytes were notably higher than for atypical or normal squamous cells. In those patients with concurrent cervical intraepithelial neoplasia the DNA content of koilocytes was inversely related to the degree of atypia. Squamous cells with HPV-induced lesions can be characterized by cytophotometry and cytomorphometry as a distinct cell population forming a continuum with the preneoplastic epithelium.

A retrospective study of 95 follicular thyroid carcinomas was conducted to evaluate the prognostic value of different morphological and clinical features. The biological behaviour of these tumours was primarily influenced by presence or absence of a capsule type of confinement: the frequency of lethal outcome among widely invasive carcinomas was six times higher than among encapsulated neoplasms. Furthermore, dismal prognosis could be demonstrated for tumours occurring in older patients (with a sharp break in the prognosis at the age of 52 years) and for those lesions which displayed oxyphilic metaplasia. The same effect was shown for presence of lymph node metastasis, tumour invasion of the cervical soft tissue and, for the case of encapsulated carcinomas, distant haematogenous spread. Conversely, the degree of differentiation and the patients' sex proved to have no significant influence on prognosis. In 22 carcinomas, cytophotometric and flow-cytometric determinations of DNA values were performed. These procedures revealed to have only limited diagnostic value, since for the majority of the tumours, benignancy or malignancy could not be judged from the DNA histograms. However, DNA measurements proved to contribute valuable information for the prognosis in individual cases of widely invasive follicular carcinomas. The discussion focuses on the diagnostic, therapeutic and prognostic relevance of these findings and on their impact on subclassification of follicular thyroid carcinomas.

The average relative DNA-content, the status of ploidy, the area of the nucleus, the compactness of chromatin and the variability of the shape of the nucleus were determined in 33 experimental induced malignant neoplasms and 5 benign lesions of gastric and intestinal mucosa of rats. The results were compared with diploid controls and divided into 3 types of DNA-histograms. Normal and malignant cells differed significantly in their DNA-content, in the area of the nucleus and in the percentage of hypertetraploid cells. 27 tumors belonged to the aneuploid type of histogram. The dignity was not clear in 6 cases. Diploid carcinomas did not occur. The DNA-cytophotometry gave the decisive hint to diagnosis of malignancy in 8 carcinomas. The various types of tumors differed not in the determined parameters. The problem of differential diagnosis between benign lesions with preneoplastic potential and malignant tumors is discussed.