Search

Analyses of the cellular DNA content were carried out in 226 patients with acute lymphoblastic leukemia (ALL) and in 61 children with acute myeloid leukemia (AML) to assess the incidence and clinical significance of DNA aneuploidies. All children were treated within the BFM studies ALL 79/81 and 81/83 as well as AML 78 und 83. DNA aneuploidies were identified in ALL in 39,8% and in AML in 36,1% of cases. Within the ALL group a significantly higher rate of aneuploid DNA stemlines was observed for non-T/non-B ALL with 44,1% as compared to T-ALL with 14,3% (p less than 0,01). In AML the morphologic subgroups M 1/2 revealed a lower frequency of 21,7% DNA aneuploidies than M 4/5 leukemias with 44,4%. The rates of complete remissions were not different between patients with and without DNA aneuploidy neither in ALL nor in AML. In the study ALL 79/81, however, a tendency towards longer remissions for patients with DNA aneuploidy was found (p = 0.053) which could not be confirmed at present by the study ALL 81/83. In both ALL trials patients with the lowest pretherapeutic risk-score revealed the highest rate of aneuploid DNA stemlines.

Analysis of the cellular DNA content was carried out in 162 children with ALL and 34 children with AML admitted to the university children's hospital Münster between 1979 and 1984. DNA aneuploidies were identified at a similar frequency in ALL (40%) and AML (44%). However, the degree of DNA aneuploidies (DNA-index) was significantly lower in aneuploid AML (median 1.09) than in aneuploid ALL (median 1.19). We found a significantly lower incidence of DNA-aneuploidies in T-ALL (3/21; 14%) as compared to non-T/non-B-ALL (60/137; 44%). No differences were found between children with and without DNA aneuploidy in PAS score and TdT activity. In non-T/non-B-ALL DNA aneuploidy is highly correlated with a long pretherapeutic history, with a low WBC and blast count and with a low serum LDH. Under the conditions of the ALL protocols BFM-79/81 and 81/83 no difference in the remission rate was found between the two patient groups. However, more relapses occurred so far within the group of children without DNS aneuploidy.

The aim of this prospective and blinded study was to investigate the diagnostic accuracy of conventional cytopathology of oral brush biopsies taken from suspicious oral lesions. In addition we checked slide based DNA image cytometry as an adjuvant diagnostic tool. Our hypothesis is that DNA aneuploidy is a sensitive and specific marker for earliest detection of oral cancer using brush biopsies. Therefore the nuclear DNA contents were measured after Feulgen re-staining using a TV image analysis system. DNA aneuploidy was assumed if abnormal DNA stemlines or cells with DNA content greater 9c were observed. Sensitivity of our cytological diagnosis in oral smears for the detection of cancer cells thus was 91.3%, specificity for the detection of non-neoplastic cells was 95.1%, positive predictive value 94.4% and negative predictive value 92.3%. The adjuvant DNA image cytometry reached a sensitivity of 97.8%, the specificity and the positive predictive value were 100% and negative predictive value was 98.1%, respectively. Smears from oral brush biopsies of all visible oral lesions are an easily practicable, cheap, minimal invasive, painless and safe screening method for detection of oral precancerous lesions and squamous cell carcinomas in all stages. We conclude that DNA image cytometry is a very sensitive and highly specific, objective and reproducible adjuvant tool for identification of neoplastic cells in oral smears.

The aim of this prospective study was to investigate the diagnostic accuracy of DNA image cytometry in combination with non-invasive brush biopsies taken from suspicious oral lesions.Cytological diagnoses obtained from 1328 exfoliative smears of 332 different lesions were compared with histology and/or clinical follow-ups of the respective patients. Additionally, nuclear DNA contents were measured after Feulgen restaining using a TV image analysis system. DNA aneuploidy was assumed if abnormal DNA stemlines or cells with DNA content greater than 9c were observed. RESULTS; The sensitivity of our cytological diagnosis in addition to DNA image cytometry on oral smears for the detection of cancer cells was 97.8%, specificity 100%, positive predictive value 100%, and negative predictive value 98.1%.The application of DNA image cytometry with DNA aneuploidy as a marker for neoplastic transformation in oral smears secures cytologic diagnosis of carcinomas. Smears from brushings of all visible oral lesions are an easily practicable, cheap, noninvasive, painless, and safe screening method for detection of oral precancerous lesions and squamous cell carcinoma in all stages. We conclude that DNA image cytometry is a very sensitive and highly specific, objective, and reproducible adjuvant tool for identification of neoplastic cells in oral smears.

The diagnostic group of dysplasias has been described as "a group of diagnostic impotence". DNA-aneuploidy detected by image cytometry of Feulgen stained pap smears indicates a potentially progressive lesion, representing a high-grade lesion. DNA-cytometry can identify cases of dysplasias which are likely to progress: DNA-aneuploid dysplasias are HSIL. DNA-aneuploidy is the expression of an integral HPV infection. CIN-lesions with episomal HPV infections are DNA-diploid. With the newly developed laserscanning cytometer (LSC) of ThinPrep-Preparation an automatisation of ploidy measurement is possible in combination with HPV-PCR. We recommend DNA-Image-cytometry as a routine method for classification of uterine cervical borderline lesion into regressive and progressive.

Dysplastic epithelia represent potentially precancerous conditions in which the risk of progression to cancer is unknown in the individual case. The positive predictive value of mild and moderate dysplasias of the uterine cervix is only about 13%. Using DNA image cytometry on restained, conventional Papsmears the cytometric equivalent of chromosomal aneuploidy can be detected as marker for neoplastic transformation of cells. The identification of DNA aneuploidy in dysplastic squamous epithelia can increase the predictive value for malignant transformation to over 90%. DNA aneuploidy qualifies squamous intraepithelial lesions as high grade (H-SIL) which have to be treated whereas lack of DNA aneuploidy characterizes low grade squamous intraepithelial lesions (L-SIL) which have only to be controlled. The methodology is meanwhile internationally standardized concerning performance and diagnostic interpretation.

The prognostic relevance of DNA stem line ploidy was ascertained by comparative DNA analysis performed by flow cytometry (FCM) and by Feulgen image cytometry (ICM) of histologic slides on paraffin-embedded resection material from 221 carcinomas of the cardia and stomach. While flow cytometric detection of DNA stem line aneuploidy was rather insecure in smaller or diffuse carcinomas, the methodological restrictions of ICM were seen in a reduced measuring accuracy varying from one case to the other. This resulted in a lower security in distinguishing mere peridiploidy from real hyperdiploid DNA aneuploidy. Despite the difference of methodology, the results were principally concurring. The assessment of DNA stem line ploidy offered no appreciable information about the prognosis of Lauren's diffuse carcinomas. For intestinal carcinomas, however, there was a significant positive correlation between DNA stem line aneuploidy and the evidence of regional nodal metastases, which might explain to a considerable extent the significantly unfavorable clinical course of tumor cases with recorded DNA aneuploidy. The correlation between DNA stem line ploidy, nodal status, and clinical outcome was distinctly less strong in intestinal carcinomas of the cardia than in those of the stomach, because nodal metastatic spread of cardia carcinomas is strongly influenced by local conditions of lymph drainage and the special mechanical strain.

DNA ploidy and cell cycle phases were evaluated by flow cytometry in colonic biopsy specimens from 107 patients with ulcerative colitis in order to analyse the prevalence of DNA aneuploidy as an indicator of numerical chromosomal aberrations and the cell proliferation in all forms of ulcerative colitis. Whereas G2/M-phase fractions in ulcerative colitis and in controls were comparable (2.7 +/- 1.1% vs. 2.8 +/- 1.1%), S-phase fractions in ulcerative colitis exceeded those of controls (7.5 +/- 3.2% vs. 6.5 +/- 2.3%; p0.01). In 28 control patients, only diploid DNA histograms existed. Single or multiple aneuploid stem lines were detected in 10 patients with ulcerative colitis (9.3%). Aneuploidy was nearly exclusively associated with pancolitis. Dysplasia was present in 13 patients (indefinite: 8; low-grade: 5), of whom 5 patients also showed DNA aneuploidy. 5 patients with non-dysplastic mucosa exhibited DNA aneuploidy. Because dysplasia and DNA aneuploidy can be discordant and might therefore identify different subgroups at risk, flow cytometry might play a role as a valuable complement to histological examination in surveillance programs of ulcerative colitis.

Patients with ulcerative colitis carry an increased colorectal cancer risk. The cumulative cancer risk for patients with pancolitis is 0.5-0.7% per patient year. Dysplasia as a histologic marker of a neoplastic transformation is used to identify patients with an increased cancer risk during colonoscopic surveillance. Because the classification of dysplasia is subject to inter- and intraobserver variation new methods for the detection of risk patients have been investigated. DNA analysis by flow cytometry seems to be of value for the detection of DNA aneuploidy and the identification of patients who are at risk for neoplastic progression. The significance of DNA aneuploidy is being evaluated in prospective studies. Surveillance guidelines depend on duration and anatomical extent of the colitis as well as on the detection of dysplasia.

Tumor and lymph node infiltration, and the DNA-ploidy status of a tumor contain prognostic information in addition to the information obtained by histological examination of surgical samples. Specimens from 112 patients with non-small-cell lung carcinoma obtained immediately after surgery were investigated by means of flow cytometry. DNA-aneuploidy was found in 43% of the primary tumors. Independent from tumor stage, patients with DNA-euploid tumors lived significantly longer (p less than 0.01) than with DNA-aneuploid carcinomas. In 29 cases the DNA-ploidy status of the primary tumor (PTU) could be compared with that of the N2 lymph node metastases (LM). 7 samples revealed a change from DNA aneuploidy in the PTU to DNA-euploidy in the LM. Patients with DNA-euploid PTU and DNA-euploid LM lived significantly longer than patients with DNA-aneuploid PTU/DNA-euploid LM, and patients with DNA-aneuploid PTU/DNA-aneuploid LM. In case of lymph node infiltration only the simultaneous measurement of DNA ploidy of PTU and LM offers an accurate prognostic evaluation. Local tumor recurrence exhibited stability of DNA ploidy, showing DNA euploidy in 12 out of 13 PTU and their corresponding recurrent tumor. Thus, the DNA-ploidy status offers additional prognostic informations which is useful for an extended tumor classification.

The nuclear DNA content of 163 colorectal carcinomas was determined by flow-cytometry (FCM) on formalin-fixed and paraffin-embedded tissue. DNA-aneuploidy was found in 97 cases (59.5%), without correlation to sex, mean age, tumor-stage (DUKES and pTNM) and grading. The frequency of aneuploidy was statistically significantly higher in patients younger than 70 years of age (p less than 0.01) and in tumors localized in the left colon and rectum (p less than 0.002). The tumors in which different areas could be analyzed (n = 80) showed a heterogeneous DNA-ploidy pattern in 18%. The comparison of DNA-content in primaries and in lymph-node metastases (n = 49) resulted in a difference of DNA-ploidy in 38% of the DNA-aneuploid tumors, but only in 6% of the DNA-diploid carcinomas (p less than 0.02). Carcinomas with DNA-aneuploidy showed a trend to a higher rate of loco-regional recurrences, a higher S-phase fraction (13.5% +/- 5.9 vs. 8.1 +/- 7.0), and proved to be associated with a poorer prognosis (p = 0.04). The statistically significantly higher mortality of patients with DNA-aneuploid carcinomas in DUKES A and B stages indicates that DNA-aneuploidy could perhaps be regarded as a stage-independent additional risk factor.

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.