Treatment of locally advanced prostatic carcinoma with LHRH analogues: cytological, DNA-cytophotometrical, and clinical results.

From June 1, 1981 to December 31, 1985, 122 patients aged 54 to 83 years, with locally advanced prostatic carcinoma, were treated with buserelin. Nineteen of the patients received combined therapy with buserelin and androcur for the first 3 months. To control the response of the primary tumor to therapy, fine-needle aspiration biopsy of the prostate was made in all patients at 3-month intervals. Fifty-eight (76.3%) of 76 patients with locally advanced prostatic carcinoma, with or without bone metastases, who underwent buserelin therapy for periods of 12-54 months showed good to satisfactory regression grades in the primary tumor. Eighteen patients (23.7%) showed poor regression or none, established by cytological findings and the measure of DNA by means of single cell-scanning cytophotometry. In three of the 58 patients, tumor progression or bone metastases occurred despite favorable regression grade; these were the only cases in which there was a discrepancy between the clinical course of the disease and the grade of regression in the primary tumor. According to TNM classification, 68 of the 78 patients treated for 12-54 months were in stage T3 NX M0; eight were in stage T3/T4 NX M1. On the basis of our long-term studies, it can be stated that buserelin therapy induces positive therapy response in more than 75% of locally advanced, inoperable, primary prostatic carcinoma. The clinical castration caused by buserelin through selective suppression of gonadotrophic secretion in the pituitary gland is, as the term implies, no more effective than surgical castration. However, the gonadotrophin suppression induced by buserelin is reversible and spares the patient the psychic stress of orchiectomy. This is a decisive advantage in light of the fact that in 20-40% of patients with locally advanced primary prostatic carcinoma, the primary tumor is hormone-refractory, and surgical castration would prove unnecessary after all.

Fine needle aspiration biopsy vs. ultrasound-guided transrectal random core biopsy of the prostate. Comparative investigations in 246 cases.

OBJECTIVE: To compare fine needle aspiration biopsy (FNAB) and ultrasound-guided transrectal core biopsy of the prostate. STUDY DESIGN: FNAB and the random core biopsy were performed simultaneously on 246 patients who either had striking palpable signs or increased levels of prostate-specific antigen, at least 5 micrograms/mL, on two separate examinations over a six-week period, RESULTS: The histologic and cytologic evaluations showed very close conformity between the two methods. Fifty-eight percent of the patients had benign prostate hyperplasia. Of the total number of patients, 103 (42%) had prostate carcinoma. This figure was confirmed in 101 (98%) of cases using FNAB. In two cases (0.8%) atypical prostate hyperplasia was diagnosed. In five cases the examination had to be repeated since insufficient prostate cell material was collected. Ultrasound-guided random core biopsy showed the presence of carcinoma in 96 of 103 patients (93%). It was necessary to repeat the examination in one patient due to a lack of adequate cell material. CONCLUSION: In our hands, FNAB method has a sensitivity of 98%, whereas core biopsy attained 91%. Both methods showed 100% concurrence in specificity.

[Lavage-cytology of the renal pelvis and ureter with special reference to tumors (author's transl)]. / Lavage-Zytologie von Nierenbecken und Harnleiter unter besonderer Berücksichtigung von Tumoren.

On 138 patients with radiologic filling defects in the ureter and renal pelvis lavage cytology was done. With this method it was possible to recognize clearly 39 of 47 urothelial tumors of renal pelvis and ureter (81,3%). Lavage cytology failed in patients with hypernephroma. Severe cellular atypias could be seen in patients with urolithiasis and inflammation. The diagnostic reliability of lavage cytology with urothelial tumors of renal pelvis and ureter is far greater than that of exfoliative urinary cytology and reaches results as good as those from brush biopsy.

The prognostic significance of ploidy and DNA-heterogeneity in the primary diagnosis and monitoring of patients with locally advanced prostatic carcinoma.

Single-cell DNA cytophotometry was employed to analyze the tumors of 271 patients with locally advanced prostatic carcinoma as to DNA ploidy and heterogeneity and the distribution of the phases of the cell cycle before and during therapy, with the intention of establishing prognostic factors apart from those already known (stage, grade). Follow-up periods ranged from 1 to 9 years. One hundred and ninety-eight (73%) of the 271 patients had carcinoma stage T3 NO MO, and 73 (27%) of them had carcinoma state T3/T4 N+ M1. The tumors were evaluated cytologically to establish the grades of malignancy. 11.8% were grade-1 carcinoma, 64.3% were grade-II and 23.8% were grade-III carcinoma. Single-cell DNA cytophotometry demonstrated aneuploidy rates of up to 73% and diploidy rates of up to 23.8% for the higher grades of malignancy, whereas the diploidy rate established for grade I carcinoma was 71% and the respective aneuploidy rate was 15.2%. These differences are significant (p less than 0.001). There was a significant correlation between the results of DNA cytometry and the clinical course of the disease. Patients with diploid tumor-cell nuclei developed no metastases and no local tumor progression during the follow-up of 9 years, whereas patients with aneuploid tumor-cell nuclei showed metastases and local tumor progression within 8-22 months, despite changes in therapy. These patients died of carcinoma after an average 18 months following primary diagnosis.

[Renal cell carcinoma: DNA cytometry and its clinical significance. An 8 year survival study]. / Nierenzellkarzinom: DNS-Zytometrie und ihre klinische Bedeutung. Eine 8-Jahres-Uberlebenszeit-Studie.

In 157 patients with renal cell carcinoma, the ploidy, DNA heterogeneity and the phases of the cell cycle occurring in the tumors were determined by means of single-cell DNA cytophotometry, in order to establish further prognostic factors in addition to the ones known so far (stage/grade). Patients with aneuploid tumors or tumors with more than one DNA frequency peak were found to have lymph node metastases intraoperatively and died earlier than patients with diploid tumors. Patients who had tumors with low proliferative activity survived longer than patients with highly proliferative tumor activity (p = 0.001).

Prognostic relevance of ploidy and proliferative activity of renal cell carcinoma.

In 112 patients with renal cell carcinoma, the ploidy, DNA heterogeneity and the phases of the cell cycle occurring in the tumors were determined by means of single-cell DNA cytophotometry, in order to establish further prognostic factors in addition to the ones known so far (stage/grade). Patients with aneuploid tumors or tumors with more than one DNA frequency peak were found to have lymph node metastases intraoperatively and died earlier than patients with diploid tumors. Patients who had tumors with low proliferative activity survived longer than patients with highly proliferative tumor activity (p = 0.001).

Nuclear DNA analysis: DNA heterogeneity in the monitoring of patients with locally advanced prostatic carcinoma.

Single-cell DNA cytophotometry was employed to analyze the tumors of 271 patients with locally advanced prostatic carcinoma as to DNA ploidy and heterogeneity and the distribution of the phases of the cell cycle before and during therapy, with the intention of establishing prognostic factors apart from those already known (stage, grade). Follow-up periods ranged from 1 to 9 years. 198 (73%) of the 271 patients had carcinoma stage T3 N0 M0, and 73 (27%) of them had carcinoma stage T3/T4 N+M1. The tumors were evaluated cytologically to establish the grades of malignancy. 11.8% were grade I carcinoma, 64.3% were grade II and 23.9% were grade III carcinoma. Single-cell DNA cytophotometry demonstrated aneuploidy rates of up to 73% and diploidy rates of up to 23.8% for the higher grades of malignancy, whereas the diploidy rate established for grade I carcinoma was 71% and the respective aneuploidy rate was 15.2%. These differences are significant (p < 0.001). There was a significant correlation between the results of DNA cytophotometry and the clinical course of the disease. Patients with diploid tumor cell nuclei developed no metastases and no local tumor progression during the follow-up period of 9 years, whereas patients with aneuploid tumor cell nuclei showed metastases and local tumor progression within 8-22 months, despite changes in therapy. These patients died of carcinoma after an average 18 months following primary diagnosis.

Transitional cell carcinoma of the renal pelvis and ureter: prognostic relevance of nuclear deoxyribonucleic acid ploidy studied by slide cytometry: an 8-year survival time study.

In 72 patients with urothelial carcinoma of the renal pelvis or ureter the ploidy, deoxyribonucleic acid (DNA) heterogeneity and counts of cell cycle phases in the tumor were analyzed by means of single cell DNA cytophotometry with the intention of finding new prognostic factors in addition to those already known (stage and grade). Followup ranged from 1 to 8 years. The results of the DNA analyses were related to the tumor categories, histopathological grading of the tumors and clinical course. Malignancy grade 1 tumors showed DNA frequency peaks in the diploid range, while tumors assessed as malignancy grade 2 showed heterogeneous DNA distribution patterns. Malignancy grade 3 tumors exhibited 71% aneuploid and 29% tetraploid DNA values. The proliferation rate of the tumor cells was statistically significantly higher in malignancy grades 2 and 3 than in malignancy grade 1. The prognosis for grade 1 tumors is good, whereas it is unfavorable in the case of grade 3 tumors. For these 2 groups (patients with grades 1 and 3 tumors) DNA ploidy affords no additional prognostic information. Grade 2 tumors, on the other hand, are heterogeneous in respect to DNA ploidy although they exhibit the same histomorphological degree of differentiation. These tumors can be subclassified as aneuploid (biologically aggressive) and diploid or tetraploid (biologically less aggressive) tumors. There was also a positive correlation between T category and DNA ploidy. The cell lines were aneuploid in 38% of the patients with stage T1 tumors, 56% with stage T2 tumors and almost 85% with stage T3, N+ tumors. A significant correlation was found between the results of DNA cytophotometry and the clinical course of the disease. Patients with diploid tumor cell nuclei had no metastases and no local tumor progression for up to 8 years, whereas patients with aneuploid tumor cell nuclei suffered metastasis and local tumor progression within 24 to 36 months. The patients died of the tumor 36 months after primary diagnosis on the average. The determination of DNA ploidy, tumor heterogeneity and tumor cell proliferation by means of DNA cytophotometry affords valuable clues as to prognosis.

Nuclear DNA analysis: the relevance of ploidy, DNA heterogeneity and phases of the cell cycle in 329 patients with prostatic carcinoma. A study on a follow-up of eight years.

The ploidy, DNA heterogeneity and the phases of the cell cycle of the tumor were analyzed, by means of single-cell DNA cytophotometry, in 329 patients with locally advanced prostatic carcinoma to find out and establish prognostic factors apart from those already known (stage, grade). Follow-up periods ranged from 1 to 9 years. 253 (76.8%) of the 329 patients had carcinoma stage T3 Nx M0, and 76 of them (23.1%) had carcinoma stage T3/T4 N2-4 M1. 11.8% of the patients showed a cytological grade of malignancy I, while 64.3% had grade II carcinoma and 23.8% had grade III carcinoma. Single-cell DNA cytophotometry demonstrated aneuploidy rates of up to 71% and diploidy rates of up to 23.8% for the higher grades of malignancy, i.e. grades II and III, whereas the diploidy rate established for grade I was 68% and the respective aneuploidy rate was 21%. These differences are significant (p less than 0.001). There was a significant correlation between the results of DNA cytophotometry and the clinical course of the disease. Only 3 (3.7%) of the patients with diploid tumor cell nuclei developed metastases and local tumor progression within 8 years, whereas patients with aneuploid tumor cell nuclei showed metastases and local tumor progression within 8-22 months. These patients died of carcinoma after an average 18 months following primary diagnosis.

Deoxyribonucleic acid content and survival rates of patients with transitional cell carcinoma of the bladder.

In 127 patients with urothelial carcinoma of the bladder the ploidy, deoxyribonucleic acid (DNA) heterogeneity and counts of cell cycle phases in the tumor were analyzed by means of single cell DNA cytophotometry with the intention of finding new prognostic factors in addition to those already known (stage and grade). Patients were followed for 1 to 9 years. The results of the DNA analyses were related to the tumor categories, histopathological grading of the tumor and clinical course. Tumors were histologically classified as grade 1--DNA frequency peaks in the diploid range, grade 2--heterogenous DNA distribution patterns, and grade 3-73% aneuploid and 27% tetraploid DNA values. The proliferation rate of the tumor cells was statistically greater in cases of histological grades 2 and 3 malignancy than in grade 1 malignancy. There was also a positive correlation between tumor stage and DNA ploidy. The cell lines were aneuploid in 38% of the patients with stage pT1, 64% with stage pT2 and almost 85% with stage pT3 tumors. A significant correlation was found between the results of DNA cytophotometry and the clinical course of the disease. Patients with diploid tumor cell lines had no metastases and no local tumor progression for up to 9 years, whereas patients with multiple aneuploid tumor cell lines suffered recurrence and local tumor progression within 6 to 36 months. On the average, the patients died of the tumors 26 months after primary diagnosis. The difference in tumor recurrence and in tumor progression between patients with aneuploid and diploid tumors was highly significant (p < 0.001). The prognosis for patients with grade 1 tumors is good, whereas it is unfavorable in the case of grade 3 tumors. For these 2 groups DNA ploidy affords no additional prognostic information. Grade 2 tumors, on the other hand, are heterogeneous in respect to DNA ploidy, although they exhibit the same degree of histomorphological differentiation. These tumors can be subclassified as aneuploid (biologically aggressive) and diploid or tetraploid (biologically less aggressive). In terms of multivariate Cox regression analysis, DNA ploidy compared with grade and tumor stage was the strongest predictor of survival.

Prognostic relevance of DNA ploidy and proliferative activity in urothelial carcinoma of the renal pelvis and ureter. A study on a follow-up period of 6 years.

In 55 patients with urothelial carcinoma of the renal pelvis or ureter, the ploidy, the DNA heterogeneity and the counts of cell cycle phases in the tumor were examined by means of single-cell DNA cytophotometry in order to find more prognostic factors than those already known (stage and grade). Follow-up periods ranged from 1 to 6 years. At the time of first diagnosis, 42 (76%) of the patients had tumors of the renal pelvis, 13 (24%) of them had ureteral tumors. 23 (42%) patients were in stage pT 1 N 0, 15 (27%) in stage pT 2 N 0, 12 (22%) in stage pT 3 N 0, and 5 (9%) were in stage pT 3 N+. The histological malignancy grade most frequently seen in the patients examined--i.e. in 51% of cases--was malignancy grade II. 25% of the patients had grade III tumors whereas only 24% had grade I tumors. With malignancy grade I, DNA cytophotometry showed DNA frequency peaks to be in the diploid range while tumors with malignancy grade II showed heterogenous DNA patterns. 71% of the patients with malignancy grade III showed aneuploid DNA values; 29% of them had polyploid DNA values. For malignancy grades II and III, the proliferation rate of the tumor cells was statistically significantly higher than for malignancy grade I. The determination of tumor heterogeneity and tumor cell proliferation by means of DNA cytophotometry gives valuable clues regarding prognosis.

Effect of different local and systemic therapy upon urinary bladder cytology.

1,900 cytological analyses of urine and bladder washings were made in 127 patients with urothelial bladder carcinomas before, during and after therapy. Following transurethral resection, all patients were treated by intravesical instillation of mitomycin C or thiotepa. Because of a locally advanced bladder carcinoma, 26 patients who were not candidates for radical cystectomy were given an integrated treatment of radiotherapy and chemotherapy. Intravesical administration of mitomycin C and thiotepa as well as integrated radiotherapy and chemotherapy induce a variety of cytological effects (toxic and/or metabolical) which may lead to cytological misinterpretations in the follow-up. DNA measurements by means of single-cell spectrocytophotometry show that the cytological effects induced by the above-mentioned therapies are not accompanied by an increase in the nuclear DNA content. It is concluded that the knowledge of these induced effects is mandatory for a correct interpretation of urinary cytology in the follow-up. Considering these effects and the clinical history, bladder carcinoma recurrences during and after intravesical chemotherapy or integrated radiotherapy and chemotherapy may be detected early by urinary cytology in the hands of an experienced cytopathologist or urologist. Furthermore, alterations of the urinary cytology occur after systemic application of cyclophosphamide and under immunosuppressive therapy.

[Decrease of disaccharidase activity following application of bencyclan-hydrogenfumarate (author's transl)]. / Depression der Disaccharidaseaktivitäten unter Bencyclan-Hydrogenfumarat

The disaccharidase activity of the small bowel mucosa of the rat is decreased by intraperitoneal application of Bencyclan-hydrogenfumarate. The doses applied were 10, 50 and 100 mg/kg body weight. The decrease rate after 100 mg/kg attained up to 50% of the control values. Furthermore, a reduction of villous height and an increase of the crypt depth has been observed in correlation with the enzyme activity decrease. Though functional and morphological alterations show a correlation, they cannot be explained by an antimetabolic-like effect because of the fact that the enzyme function recovers 72 h after application of the drug. Whilst our results do not sustain a cytostatic effect of Bencyclan-hydrogenfumarate, there is no doubt that the intraperitoneally applied drug influences the disaccharidase activity. These facts have to be considered in clinical therapy.

Immunohistochemical detection of p53 protein in transitional cell carcinoma of the bladder in correlation to DNA ploidy and pathohistological stage and grade.

Mutations of p53 gene have been found in a variety of human malignancies; however, the impact of immunohistological detection of p53 expression in the development and progression of TCC of the bladder is still uncertain. In the present study, we investigated the p53 oncoprotein expression and compared the findings to DNA ploidy and pathohistological stage and grade. The study included 147 patients with transitional cell carcinoma of the bladder investigated between February 1981 and September 1994. The average age of the 55 women and 92 men was 67 years (range: 20-71 years). A total of 76 patients (52%) had stage pTa to pT1, 35 (24%) stage pT2, 25 (17%) stage pT3, and 11 (7%) stage pT4 disease. Frozen sections of tumor biopsies obtained by transurethral resection were immunohistochemically stained using the monoclonal antibody clone D0-7 (DAKO), which recognized two different epitopes for mutant and wild-type p53 protein. Tumors expressing p53 in more than 10% of the tumor nuclei were regarded as positive. The DNA ploidy was determined by image analysis. Immunohistochemical detection of p53 expression was found in 84 (57%) of the 147 tumors examined. Positive p53 staining was seen in grade I tumors in 10 to 25%, in grade II tumors 25 to 75%, and in grade III up to 58% of the tumor nuclei. There was a positive correlation between p53 expression and pathological stage (28% in pTa, 73% in pT1-2, and 68% in pT3-4 tumors). There was no appreciable relationship between DNA Ploidy and p53. Although carcinomas with p53 expression had a slight tendency to be more prevalent among higher disease stages and poorly differentiated transitional cell carcinoma, immunohistochemical detection of p53 is not a valuable tool for predicting the outcome of patients with TCC or for identifying subgroups of patients that may be at a higher risk for tumor progression.

[Morphometric studies on epithelial cells of the rat stomach under chronic effect of alcohol]. / Morphometrische Untersuchungen an Belegzellen des Rattenmagens unter chronischem Alkoholeinfluss

In studying the effect of ethanol on the size of parietal cells 60 Wistar rats received 15% ethanol ad libitum and were killed after 8 days, 2, 4, 8, and 12 weeks. For the control group 24 untreated animals were used. The parietal cells were stained with Kongo-red for the histological preparations and with a Papanicolaou modification for the cytological smear. With two measuring-methods -- the Integration-disc and the Mikrovideomat -- the cell and nucleus surface were determined and the nucleus-plasma-relation was calculated. After 8 days of alcohol administration a significant increase of parietal cell surface appeared in the histological and cytological preparation. The cytoplasma surface area increased continuously until the 8th week. With further application of alcohol until the 12th week the zytoplasma surface area decreased again but remained above the starting size. The calculation of the cell surface areas produced nonuniform results. Further the sizes of parietal cells (cytoplasma and nucleus surface areas) in the cytological smear and after being embedded in the histological preparation were compared. In the applied measuring methods a comparison was also made.

[1st clinical results of the treatment of locally advanced prostatic cancer with a powerful LH-RH analog: buserelin acetate]. / Premiers résultats cliniques concernant le traitement du cancer de la prostate à un stade local avancé au moyen d'un puissant analogue de la LH-RH: l'acétate de buséréline.

From a series of 39 patients presenting with a locally advanced cancer of the prostate (Stage C), 26 were treated with an LH-RH analogue, Buserelin, for a period ranging between 7 and 22 months. After an initial period of subcutaneous injections, the treatment was administered by intranasal spray (400 mcg three times a day). This resulted in levels of testosterone similar to those of castrated men. In order to evaluate the effects of buserelin on the primary tumour, a degree of cytological regression was established for all of the patients, using fine-needle aspiration biopsy which was performed every three months. The cytological results correspond with the DNA analyses performed by cytophotometry of isolated cells and reveal a statistically significant reduction in the degree of anaploidy or polyploidy in the cases in which the prostatic cancer had responded favourably to treatment with buserelin. 21 of the 26 patients treated with this potent LH-RH analogue showed a good therapeutic response. 5 patients who did not show any signs of cytological regression were secondarily treated with estramustine phosphate because of their resistance to hormones. One patient received cyclophosphamide as a tertiary treatment because of progression of his symptoms after 15 months. No appreciable side effects were observed.(ABSTRACT TRUNCATED AT 250 WORDS)

Treatment of prostatic cancer with LH-RH analogues.

Twenty-one of 32 patients with locally advanced prostatic cancer (stage C) were treated with the LH-RH analogue Buserelin for 7-19 months. After an initial sequence of subcutaneous injections, treatment was continued with intranasal spray application (three daily doses of 400 micrograms each) which ensured maintenance of serum testosterone within the range seen in castrated men. To evaluate the response of the primary tumor to Buserelin, cytological regression was established for all patients by fine-needle aspiration biopsy every 3 months. The cytological results corresponded with those of DNA analyses of single-cell cytophotometry showing a statistically significant drop of the grade of aneuploidy or polyploidy when the prostatic carcinoma responded positively to Buserelin therapy. Seventeen of 21 patients treated with the potent LH-RH analogue showed good therapy response. Four patients with no cytological signs of tumor regression received secondary treatment with estramustine phosphate because of hormone resistence. One patient had to be crossed over to cyclophosphamide, the third drug, for clinical progression after 15 months. Essential side effects have not been observed. Continuous treatment of locally advanced prostatic cancer with Buserelin, combined with close control of the patient, offers not only a real alternative to surgical castration--as the patient is spared the psychical stress of orchiectomy--but also to estrogen therapy with its risk of cardiovascular side effects.

DNA ploidy is a valuable predictor for prognosis of patients with resected renal cell carcinoma.

BACKGROUND: Renal cell carcinomas (RCCs) are heterogeneous and include several distinct entities with a range of biologic and clinical behaviors from relatively favorable to extremely aggressive. The heterogeneity leads to unpredictable outcome and survival. DNA ploidy is a relatively new predictor differentiating diploid from aneuploid tumor cells according to regular or irregular DNA content. The authors evaluated the predictive value of DNA ploidy in patients who underwent resection because of RCC. METHODS: In a prospective study, 180 patients who underwent resection because of RCC were investigated. DNA cytometry was conducted on each resected tumor to determine DNA ploidy. Patients were completely followed up until death or up to 12 years. RESULTS: Survival analysis showed that patients who underwent resection because of RCC in tumor classifications pT1, pT2, and pT3 survived 10 years in 85%, 53%, and 8% of cases, respectively. Patients suffering from small tumors (pT1 and pT2, n = 44) with diploid nuclei survived 10 years in 94% but only in 8% if the tumor was aneuploid (n = 55). In addition, 91% of patients who underwent resection of large tumors (pT3, n = 12) with diploid nuclei survived 10 years, but no patient with large and aneuploid tumor (n = 51) survived more than 3 years. Furthermore, 92% of all patients afflicted from diploid RCC survived 10 years. This finding was independent of tumor stage. CONCLUSIONS: The results of this study suggest that DNA ploidy is a significant and independent predictor for survival of patients afflicted from RCC and superior to tumor classification and grade. DNA ploidy is a reliable prognostic factor for RCC and yields considerable information for patient management and predicting clinical outcome.