A Cytophotometric Study of Neoplastic Field Change in the Human Urinary Bladder

Accepted 85

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A CYTOPHOTOMETRIC STUDY OF NEOPLASTIC FIELD CHANGE IN THE HUMAN URINARY BLADDER. *Aidan O'Brien, Michael R. Butler,

MIAC: A MODULAR IMAGE ANALYSIS COMPUTER FOR AUTOMATED

John M. Fitzpatrick, Dublin, Ireland. (Presentation to be made by Mr. A. O'Brien)

Leiden, The Netherlands (Presentation to be made by Dr. Tanke). A new modular image analysis computer (MIAC) (Leitz, FRG)

Hofstaedter

et

al

(1984) have shown that the

Diploid Deviation Quotient (DDQ) is an accurate indicator of the biological behaviour of bladder cancer, irrespective of

histological

grade

or

stage, in that the survival of

patients with tumours with a DDQ less than 1.2 does not differ from that of controls, whilst a DDQ greater than 1.2 is

was

associated with a poor prognosis. The aim of this study

to

extent

change.

use and

the

cytophotometric

prognostic

technique to assess the

implications

of

neoplastic field

Cold cup biopsies of endoscopically normal mucosa

were taken from 7 predetermined, disparate sites in each of 10 bladders containing tumour, as well as a biopsy of the

tumour itself. Ten sex- and age-matched patients were used as controls. After fixation in Bohm 1 s medium and paraffin embedding, adjacent 5um cuts were stained with H&E for

histopathology

and with a Feulgen stain for cytophotometry

using a scanning microdensitometer. Histological examination of the endoscopically normal mucosal biopsies revealed that 4 (6%) contained

frank tumour, 6 (9%) carcinoma-j_n-situ (CIS), 13 (19%) varying types of cystitis, and 44 (66%) entirely normal. Two of those with frank tumour, all of those with CIS and 4 of those with cystitis had a DDQ greater than 1. 2. It was surprising to find that 20 (45%) of the histologically normal biopsies had a DDQ greater than 1.2. Moreover, in 10 of these cases, the DDQ of the associated tumour was less than 1.2. This study would indicate that it may, not only, be possible to identify histologically normal mucosa already involved in the oncogenic process, but that such mucosa may have even greater prognostic implications than the associated tumour itself.

CYTOLOGY. H.J. Tanke*, J.S. Ploem*, U. Jonas, University of

has been incorporated in the Leyden Television Analysis

System (LEYTAS) for applications in the field of automated cytology. The new system consists of a multiparameter

microscope equi.pped with a special 40X objective lens (N.A. 1.30), enabling the analysis of microscopic fields of 500x 500 µm with a pixel resolution of about I µm. Due to demagnification, the total microscope magnification reaching the TV camera is only 20X, allowing rapid analysis of a large number of cells. The same objective also provides a 80x magnification for more detailed analysis. The microscope is connected to a MIAC, an image analyzer

constructed around a Versa Module Europe (VME) bus and a high-speed TV bus. The instrument contains a Motorola 68,000 microprocessor and combines rapid image analysis based on image transformations with conventional pattern recognition performed in the Motorola microprocessor. With

the MIAC 512x512 pixels, as detected by a plumbicon tube, are stored directly in the computer memory using fast AD

conversion (20 msec per field). This system has been used for automated classification of

Feulgen-DNA stained cytology preparations from the bladder and the prostate, based on the presence of cells with abnormal DNA content. Furthermore, it has been applied for automatic recording of DNA histograms of single cells only

for grading of carcinoma on the basis of DNA ploidy. Screening of cytological samples takes about 5 min with the

new MIAC system. Typical results of the automated analysis of bladder and prostate samples will be presented,

Reference: Hofstaedter et al: British Journal of Urology, (1984): 56, 289 - 295.

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ND-YAG LASER NORMOGRAM DOSIMETRY SCALE FOR THE BLADDER.

CHARACTERIZATION OF HUMAN BLADDER CARCINOMA BY MONOCLONAL ANTIBODIES. Bernd J. Schmitz-Drager*, Thomas Ebert*, Christoph Peschkes*, Matthias Uhlig* and Rolf Ackermann, Dilsseldorf, FRG (Presentation to be made by Dr. Schmitz-Drager) Today the diagnosis of bladder carcinoma (BC) is based on cystoscopy and cytological evaluation of urine. Both methods are time consuming or of low specificity. Urine analysis with monoclonal antibodies (mAb) directed against BC might become an valuable tool in the early detection of BC. We report on two recently developed mAb Due aBC 1 and Due aBC 2 directed against human BC. MAb were generated by immunization of Balb/c mice with BC cell line SW 1710. Screening and specificity analysis was performed by an indirect immunoperoxidase method on frozen sections of bladder tumors, normal bladder wall, and various other tissues of benign or malignant origine. Due aBC 1 and Due aBC 2 were found to react with 5/6 and 6/9 BC specimen tested. As demonstrated by light microscopy Due aBC 1 did react with an antigen present on nearly all carcinoma cells. The lack of binding in one case might be due to thermic damage caused by electroresection. Due aBC 2 did bind to another antigen mainly expressed on low grade BC. Both mAb were found to react with some other malignant but not with normal tissues. Immunocytochemical examination of long term BC cell lines showed positive results in 6/8 cell lines with antibody Due aBC 1. Due aBC 2 did react with 4/8 BC cell lines. Characterization of the antigens and the investigation of the question if the results of cytological urine evaluation might improve by the use of these 2 antibodies are subject of current research.

Louis R. Cos and *P. Anthony diSant'Agnese, Rochester, NY

(presentation to be made by Dr. Cos) Bladder perforation following Nd-YAG Laser irradiation has been documented. To prevent unwanted damage to the bladder and vicinity a normogram dosimetry scale was attempted. Under general anesthesia 25 female pigs under-

went cystoscopy. Multiple pulses at different settings of Laser energy were delivered to several areas in the bladder. One week following treatment, exploratory laparotomy and cystectomy were performed. Microscopic analysis of the

depth of Laser irradiation completed the study. It was observed that although the larger the energy delivered, the deeper the tissue damage, this was not uniform. Using 60

watts (w) of power for 10 pulses (p) of one second each, with a total of 600 joules (j), the depth of penetration varied from half the bladder wall thickness to transmural,

with or without perforation and/or damage to bowel. If the same amount of energy was delivered by giving 30 w for 20 p only deep penetration was observed. A temperature summation

factor was demonstrated by giving 200 j in 3 different forms: 30 w x 10 s, 13 w x 15 sand 10 w x 20 s. In the first group the maximum penetration was 2 mm, in the second,

3.5 mm, and in the third transmural. When large energy delivery or high number of pulses were given massive ·necrosis of the bladder wall and injury of surrounding organs were observed. The density and thermal conditions of tissue, type of Laser and wave length, optimal focusing, and power and time settings are deciding factors. With all these variables a normogram Laser dosimetry scale is

probably impossible to set. When treating a pedunculated tumor, it is probably safe to use settings of 40 w for several hundred pulses of 1 or 2 second duration using visual blanching and shrinking as parameters. However, when treating a sessile, flat lesion a setting of 20 w for no

more than 10 pis advisable. A solid knowledge of Laser physics and tissue interaction is mandatory for all surgeons using this form of treatment.

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