Use of Lewis X antigen and deoxyribonucleic acid image cytometry to increase sensitivity of urinary cytology in transitional cell carcinoma of the bladder.

PURPOSE: To improve sensitivity and specificity of urinary cytology for bladder cancer cell detection we used deoxyribonucleic acid (DNA) image cytometry and the monoclonal anti-Lewis X antibody P12. MATERIALS AND METHODS: Spontaneously voided urine and additional barbotage bladder washings of 25 patients with transitional cell carcinomas and 25 patients with benign diseases of the bladder were analyzed by conventional cytology, immunocytology and DNA image cytometry. The DNA content was determined in specimens stained with Feulgen according to the European Society for Analytical Cellular Pathology consensus report on standardization of DNA image cytometry. For the immunocytological examination we used the avidin-biotin-complex immunoperoxidase method with the monoclonal antibody P12 directed against the Lewis X determinant. RESULTS: The cytological examination revealed a sensitivity of 72% and a specificity of 100%. By using DNA image cytometry with Kolmogoroff-Smirnow test sensitivity increased up to 84% with a specificity of 100%. The immunocytological examination with Lewis X showed a sensitivity of 84% and a specificity of 80%. CONCLUSIONS: Combining cytology and DNA cytometry the overall sensitivity increased to 92% and with the additional application of immunocytology for Lewis X antigen sensitivity it increased to 96% but was accompanied by a decrease in specificity to 80%. DNA image cytometry and Lewis X antigen detection are not suitable for screening but suspicious urothelial cells in urinary cytology specimens can be evaluated specifically by DNA measurements. In the future it needs to be clarified whether DNA image cytometry in combination with Lewis X antigen detection can help to signal relapse and progression of transitional cell carcinoma of the bladder.

Inflammatory pseudotumor of the epididymis.

We report the case of a 73-year-old patient who presented clinically with a palpable left scrotal mass after a 3-month history of therapy-resistant epidiymitis. He underwent epidiymectomy, and the following histopathologic and immunohistochemical evaluation revealed an inflammatory pseudotumor. We present the second case of an inflammatory pseudotumor of the epididymis being reported in the literature and give a brief review of the literature concerning this very rare neoplastic entity.

DNA-aneuploidy as marker for neoplasia in G1-urothelial carcinomas.

Biopsies from 60 grade-1 urothelial carcinomas and 50 normal bladder mucosae have been investigated by DNA-image-cytometry after enzymatic cell separation to determine the diagnostic sensitivity of DNA-aneuploidy for the identification of low grade urothelial neoplasia. Two different modes to detect DNA-aneuploidy were compared: the conventional application of a threshold at 2.2c for the modal DNA value and the use of the Kolmogoroff-Smirnow (KS) test to compare GO/1 phase fractions of the cells under analysis and the internal reference cells. Whereas the specificity for both types of analysis was 100%, the sentitivity of the latter method was 73.3% compared to 48.3% of the conventional procedure. Therefore, the application of the KS-test for the identification of DNA- stemlines as aneuploid results in an increased detection rate was compared with the conventional threshold procedure. The high prevalence of DNA- stemline aneuploidy even in low grade urothelial cancers may qualify this marker as a possible aid in the cytologic evaluation of urine and bladder washings.

Correlation of histology, cytogenetics and proliferation fraction (Ki-67 and PCNA) quantitated by image analysis in meningiomas.

Sixty meningiomas were classified histologically according to the WHO criteria and analyzed cytogenetically. The Ki-67 and PCNA proliferation fractions (PRFs) were assessed by immunhistochemistry. The staining results were quantified by TV-image analysis (Miamed, Leica, Germany). Within meningiomas of WHO-grade II/III and those with complete or partial deletion of # 1p, we found a significantly higher mean and maximal Ki-67 PRFs than in those of WHO-grade I and those with all other karyotypes. This was not the case for PCNA PRFs. No differences in PRFs were detectable between histological subtypes. Although these results were obtained after measurement of five high power fields (HPFs) only, the heterogeneity of PRF distribution within the tumors was high. Even after the measurement of 100 HPFs, the 95% confidence intervals were in a range of 18% to 34.3% PRF. This finding seems to be responsible for the moderate interobserver reproducibility of image analytical determination of PRF (r = 0.74). Nevertheless, there was a good correlation between subjective and objective image analytical assessment of PRF (r = 0.83). The significance of the maximum Ki-67 PRF per specimen implies the possibility of selecting those areas which show the highest fraction of positively stained nuclei for measurement, avoiding the problem of intraslide heterogeneity. Thus the time needed for image analytical quantification may be reduced.

Intratumorous heterogeneity of chromosome 10 and 17 in meningiomas using non-radioactive in situ hybridization.

Contrary to classical cytogenetics non-radioactive chromosomal in situ hybridization (CISH) may be performed within 24 hours while the morphological structure of paraffin-embedded tumor material is preserved. Slides of 34 formalin fixed and paraffin-embedded meningiomas were hybridized with a biotinylate alpha-satellite DNA-probe for chromosome 10 and 17. According to the distribution of hybridization signals per nucleus we found five meningiomas with signs of trisomy 17 whereby all of them exhibited intratumorous heterogeneity. Trisomy 10 was found in five tumors. Monosomy 17 was assumed in two cases and monosomy 10 in one meningioma. As a control six meningiomas were karyotyped by G-banding. Formalin fixed and paraffin-embedded tissue of the same tumors was hybridized against chromosome X and Y. In five of six cases the result of conventional karyotyping could be confirmed by in situ hybridization. CISH of paraffin-embedded tissue is new tool for the analysis of intratumorous cytogenetical heterogeneity with potential prognostic significance.

Quantitative analysis of the neu oncogene in normal and transformed epithelial breast cells by fluorescence in situ hybridization and laser scanning microscopy.

In this study we evaluated indirect fluorescence in situ hybridization of the neu oncogene in combination with laser scanning microscopy for the detection and quantification of fluorescence signals in single cells. Cell lines from human tumor tissue with neu oncogene amplification and one nonneoplastic human epithelial cell line from a lactating breast with a neu single copy as the reference were used. After quantitative fluorescence analysis of three mixed cell populations (neoplastic cells and reference cells), we evaluated the cell morphology and location of the fluorescence signal by laser scanning microscopy. Finally, the number of oncogene copies in the tumor cell population could be estimated from the ratio of its mean background-corrected integrated fluorescence intensities to those of the reference cells. In all mixed cultures the ratio of fluorescence intensities was within the limits of oncogene copy numbers known from the literature and evaluated by southern blot analysis. The lowest oncogene copy number possible to quantify was four to eight genomic copies.

[18FDG-PET in intracranial meningiomas versus grading, proliferation index, cellular density and cytogenetic analysis]. / 18FDG-PET bei intrakraniellen Meningeomen versus Grading, Proliferationsindex, Zelldichte und zytogenetische Analyse.

62 intracranial meningiomas in 60 patients were studied with 18FDG-PET prior to neurosurgery in order to evaluate the relationship between 18FDG uptake and biological aggressiveness of the tumors. Histopathological grading, cellular density, Ki-67 proliferation index and evidence of chromosomal aberrations were used to assess tumor aggressiveness. Significantly elevated 18FDG uptake was found in grade 2- and 3- compared to grade 1-meningiomas, in tumors of high cellularity compared with those of low cellularity, and in meningiomas with an elevated Ki-67 proliferation index (above 2%). The two meningiomas with the most pronounced chromosomal aberrations revealed the highest 18FDG uptake of all cytogenetically studied meningiomas. We conclude that 18FDG-PET is useful for estimating the biological aggressiveness of intracranial meningiomas.

[Pseudo-sulfur granules (pseudo-actinomycotic granules) in intrauterine device patients]. / Pseudosulfurgranula (Pseudoaktinomyzesdrusen) bei Intrauterinpessar-Trägerinnen.

In 14 tissues from uterine curettages from women with intrauterine contraceptive devices (IUD) we found structures were discovered, which could easily be mistaken for actinomycotic sulphur granules. These were characterised by radial filamentous eosinophilic structures with peripheral club-like swellings. The dissimilarities to a true sulphur granule are evident with Gram staining which shows single filaments substantially wider than those of the true actinomycotic granules. Until now the nature of the described structures is not clear. They are possibly deposits on the copper thread of the IUD.