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C102 SURVIVIN EXPRESSION IN THE CYTOPLASM AND NUCLEUS OF HAL FLUORESCENT CANCER CELLS
< 0.01). Compared to the incidence rates of industrialized countries of the western world, this risk-adapted strategy represents an increase in effectiveness of 377. Conclusions: Risk-adapted screening in bladder cancer delivers a reasonable approach to diagnose bladder cancer before emerging symptoms. We could demonstrate that risk-adapted screening in BC is work effective, cost effecitve and aim achieving in an asymptomatic population. The questionnaire RCBC integrates evidence based bladder cancer inductors, is easy in use, calculates risk for BC automatically and is as an open-access tool available in 10 languages via internet www.riskcheck-bladder-cancer.info. In consequence the tool is ideal for European urology to be used in daily routine.
C100
BCGitis: A purely clinical picture?
Götz Mg., Pavlik M., Ikic M., Hamidov Z., Schneider S., Atanassov G., Hauschild E., Krah X. Helios-Kliniken Blankenhain, Dept. of Urology, Blankenhain, Germany Introduction & Objectives: The adjuvant intravesical instillation of Bacillus Calmette-Guérin (BCG) is the gold standard in preventing recurrence of superficial bladder tumors and carcinoma in situ (CIS) according to the EAU Guidelines. Severe side effects in terms of a BCGitis are described in less than 5% of cases and a BCG sepsis in less than 0.5% of cases. Material & Methods: A 71-year-old patient presents with fever up to 39.8 °C and poor general condition. A week earlier, a BCG instillation was performed after a cystoscopy and gross hematuria. Since then, he has been suffering from fever, increasing fatigue and voiding problems. Already earlier he had been treated with a 6-fold BCG instillation therapy because of recurrence of urothelial carcinoma of the bladder.Primarily, he had an acute cystitis with coagulate-negative Staphylococcus spp. In a situation with an unclear initial focus of infection and lack of detection of Mycobacterium bovis BCG in the blood cultures, we started a calculated antibiotic therapy with piperacillin-tazobactam. After this therapy mainly the pulmonary situation deteriorated. A thoracic CT showed a generalized pneumonitis with an incipient pulmonary fibrosis. Then we started - because of the clinical picture of a BCGitis - a tuberculostatic therapy (isoniazid, rifampicin, ethambutol). In the meantime an ARDS necessitated an intensive care therapy with medical monitoring and a tracheostomy. A marked improvement of general condition and pulmonary situation was reached in the course. The antimycobacterial therapy was administered for 6 months because of the severity of the initial clinical picture. Results: BCGitis is caused by Mycobacterium bovis BCG entering the blood stream. Clinically, it presents as miliary tuberculosis. Early treatment with tuberculostatic drugs is indicated, even if no pathogen is detected because blood cultures as well as the latest PCR technologies show poor sensitivity in this context. According to the literature complications after BCG instillations often occur after a history of tuberculosis, if the instillation is performed early post-interventionally, and if gross hematuria or acute cystitis were present. BCG instillation should not be performed in such cases. As far as therapy of BCGitis is concerned, the literature names various options: 2-week therapy with quinolones, 6-month triple anti-mycobacterial therapy, etc. Since the complication is caused by a defined, industrially produced Mycobacterium bovis BCG, there is no drug resistance besides natural resistance against pyrazinamide. The attenuated bacterium can’t persist in an immunocompetent patient; therefore we could also consider a shortterm therapy. Conclusions: BCGitis is a serious complication after instillation therapy. Usually, treatment is sufficient and successful even in severe cases. Still, if the abovementioned risk factors or contraindications are observed, BCG instillation should be continued as a prophylaxis of recurrence in superficial bladder tumors and carcinoma in situ taking into account the encouraging available data of this therapy.
C101
Vascular Endothelial Growth Factor expression in the PDD-positive urothelium
Czech A., Glazar B., Dobrowolski Z., Lipczyński W., Okoń K., Maliszewski T. Jagiellonian University, Clinic and Dept. of Urology, Cracow, Poland Introduction & Objectives: One of the prominent features of the neoplastic process is the vascular network formation. The regulation of the formation of new blood vessels depends on a number of soluble factors. Particularly important are vascular endothelial growth factors-VEGF-A. The aim of this study was to evaluate HAL fluorescent urothelium in PPD endoscopy using VEGF-A expression. Material & Methods: 96 patients with non-muscle-invasive bladder cancer underwent PDD endoscopy using Hexvix. The bladder was inspected two hours after photosensitizer instillation by white light cystoscopy first. Lesions or suspicious areas were mapped onto bladder chart. Then HAL fluorescence cystoscopy was performed. Lesions and suspicious areas were also mapped onto bladder chart. From all the lesions mapped specimens were collected with forceps for histopathological examination and lesions were bipolar loop resected. Specimen were fixed in formalin and embedded in paraffin. EDTA buffer antigen retrieval in water bath pH 8.0 for 60 minutes was performed. A-20 (Santa Cruz) primary antibody and Ultra Vision LP Value (LabVision) detection system were used.
Eur Urol Suppl 2012;11(4):106
Reaction intensity was microscopically evaluated (ranging 0-3). Results: 410 HAL fluorescent specimens from 96 patients with non-muscleinvasive bladder cancer were obtained. 354 specimens were analyzed, the remaining 56 were rejected due to the artifacts. Estimation of 354 HAL – positive specimens using VEGF.In normal urothelium 102 (28,8%) HAL-positive specimens the expression of VEGF was demonstrated in 80 (78,43%) specimens and no expression of VEGF in 22 (21,27%) specimens.In cancer tissue 105 (29,7%) HAL-positive specimens the expression of VEGF was demonstrated in 69 (65,7%) specimens and no expression of VEGF in 36 (34,3%) specimens. In 54 (15%) atypia HAL-positive specimens the expression of VEGF was demonstrated in 42 (77,77%) specimens and no expression of VEGF in 12 (22,23%) specimens. In 93 (26,6%) hyperplasia HAL-positive specimens the expression of VEGF was demonstrated in 71 (76,34%) specimens and no expression of VEGF in 22 (23,66%) specimens. Conclusions: There was no expression of VEGF-A, could be used for differential diagnosis of changes in bladder epithelium. This may result from the fact that the onset of neoangiogenesis is not an early phenomenon in cancer formation. The appearance of VEGF expression in inflammatory lesions may be related to the participation of this factor in the inflammatory and repair processes.
C102
Survivin expression in the cytoplasm and nucleus of HAL fluorescent cancer cells
Zembrzuski M.1, Glazar B.2, Dobrowolski Z.2, Lipczyński W.2, Okoń K.2, Bąk M.2, Maliszewski T.2 1 Jagiellonian University, Clinic and Dept. of Urology, Kraków, Poland, 2 Jagiellonian University, Clinic and Dept. of Urology, Cracow, Poland Introduction & Objectives: The main element of carcinogenesis is to obtain tumor cell resistance to apoptosis. One of the factors regulating apoptosis is survivin. Survivin is a protein product of the BIRC5 gene, under physiological conditions present in fetal cells, is not present in good differentiated cells. Reexpression of survivinu observed in a number of malignancies, it is associated with cell proliferation and to protect against cancer cell apoptosis.The aim of the study was to evaluate the expression of survivin on the nuclei and cytoplasm urothelial cells, which in condition of PDD in the endoscopy under ultraviolet light showed fluorescence. Material & Methods: 96 patients with non-muscle-invasive bladder cancer underwent PDD endoscopy using Hexvix. From all the lesions specimens were collected with forceps for histopathological examination and lesions were bipolar loop resected.Specimens were fixed in 10% formalin solution. Reaction in the direction of survivin was performed in a typical way available antigen in a water bath in EDTA, pH 8.0. Primary antibody was used AB-1 8B2 (Lab Vision). Using a detection system Volu LP Ultra Vision (Lab Vision), DAB as chromogen and Mayer hematoxylin as a contrast. Examining microscopically the cytoplasm of urothelial cells rated power on a scale pH from 0 to 3.Examining the contents of survivin in the urothelial nuclei evalueted microscopically percentage of positive nuclei. Results: Specimens from 96 patients with non-muscle-invasive bladder cancer were obtained. 363 specimens were analyzed, the remaining 47 were rejected due to the artifacts.Estimation of 354 HAL – positive specimens using expression of cytoplasmic Survivin in urothelium .In all 354 HAL –positive specimens the expression of Survivin C was demonstrated in 328 (92,6%) specimens and no expression of Survivin C in 26 (7,4%) specimensIn normal urothelium 102 specimens the expression of Survivin C was demonstrated in 91 (89,21%) specimens and no expression of Survivin C in 11 (10,79%) specimensIn cancer tissue 105 HALpositive specimens the expression of Survivin C was demonstrated in 93 (88,6%) specimens and no expression of Survivin C in 12 (11,4%) specimens. In 54 atypia HAL-positive specimens the expression of Survivin C was demonstrated in 51 (94,44%) specimens and no expression of Survivin C in 3 (5,565%) specimens. Estimation of 354 HAL – positive specimens using expression of nuclear Survivin in urothelium .In all 354 HAL –positive specimens the expression of Survivin was demonstrated in 218 (61,5%) specimens and no expression of Survivin C in 136 (38,5%) specimensIn normal urothelium 102 specimens the expression of Survivin was demonstrated in 69 (67,64%) specimens and no expression of Survivin C in 33 (32,36%) specimensIn cancer tissue 105 HAL-positive specimens the expression of Survivin was demonstrated in 68 (64,8%) specimens and no expression of Survivin in 37 (35,2%) specimens. In 54 atypia HAL-positive specimens the expression of Survivin was demonstrated in 31 (57,4%) specimens and no expression of Survivin in 23 (42,6%) specimens. Conclusions: In 92,6% HAL–positive specimens the expression of Survivin was demonstrated. There was no statistical differences between Survivin expresion between normal, cancer and also atypical urothelium.
C100
BCGitis: A purely clinical picture?
Götz Mg., Pavlik M., Ikic M., Hamidov Z., Schneider S., Atanassov G., Hauschild E., Krah X. Helios-Kliniken Blankenhain, Dept. of Urology, Blankenhain, Germany Introduction & Objectives: The adjuvant intravesical instillation of Bacillus Calmette-Guérin (BCG) is the gold standard in preventing recurrence of superficial bladder tumors and carcinoma in situ (CIS) according to the EAU Guidelines. Severe side effects in terms of a BCGitis are described in less than 5% of cases and a BCG sepsis in less than 0.5% of cases. Material & Methods: A 71-year-old patient presents with fever up to 39.8 °C and poor general condition. A week earlier, a BCG instillation was performed after a cystoscopy and gross hematuria. Since then, he has been suffering from fever, increasing fatigue and voiding problems. Already earlier he had been treated with a 6-fold BCG instillation therapy because of recurrence of urothelial carcinoma of the bladder.Primarily, he had an acute cystitis with coagulate-negative Staphylococcus spp. In a situation with an unclear initial focus of infection and lack of detection of Mycobacterium bovis BCG in the blood cultures, we started a calculated antibiotic therapy with piperacillin-tazobactam. After this therapy mainly the pulmonary situation deteriorated. A thoracic CT showed a generalized pneumonitis with an incipient pulmonary fibrosis. Then we started - because of the clinical picture of a BCGitis - a tuberculostatic therapy (isoniazid, rifampicin, ethambutol). In the meantime an ARDS necessitated an intensive care therapy with medical monitoring and a tracheostomy. A marked improvement of general condition and pulmonary situation was reached in the course. The antimycobacterial therapy was administered for 6 months because of the severity of the initial clinical picture. Results: BCGitis is caused by Mycobacterium bovis BCG entering the blood stream. Clinically, it presents as miliary tuberculosis. Early treatment with tuberculostatic drugs is indicated, even if no pathogen is detected because blood cultures as well as the latest PCR technologies show poor sensitivity in this context. According to the literature complications after BCG instillations often occur after a history of tuberculosis, if the instillation is performed early post-interventionally, and if gross hematuria or acute cystitis were present. BCG instillation should not be performed in such cases. As far as therapy of BCGitis is concerned, the literature names various options: 2-week therapy with quinolones, 6-month triple anti-mycobacterial therapy, etc. Since the complication is caused by a defined, industrially produced Mycobacterium bovis BCG, there is no drug resistance besides natural resistance against pyrazinamide. The attenuated bacterium can’t persist in an immunocompetent patient; therefore we could also consider a shortterm therapy. Conclusions: BCGitis is a serious complication after instillation therapy. Usually, treatment is sufficient and successful even in severe cases. Still, if the abovementioned risk factors or contraindications are observed, BCG instillation should be continued as a prophylaxis of recurrence in superficial bladder tumors and carcinoma in situ taking into account the encouraging available data of this therapy.
C101
Vascular Endothelial Growth Factor expression in the PDD-positive urothelium
Czech A., Glazar B., Dobrowolski Z., Lipczyński W., Okoń K., Maliszewski T. Jagiellonian University, Clinic and Dept. of Urology, Cracow, Poland Introduction & Objectives: One of the prominent features of the neoplastic process is the vascular network formation. The regulation of the formation of new blood vessels depends on a number of soluble factors. Particularly important are vascular endothelial growth factors-VEGF-A. The aim of this study was to evaluate HAL fluorescent urothelium in PPD endoscopy using VEGF-A expression. Material & Methods: 96 patients with non-muscle-invasive bladder cancer underwent PDD endoscopy using Hexvix. The bladder was inspected two hours after photosensitizer instillation by white light cystoscopy first. Lesions or suspicious areas were mapped onto bladder chart. Then HAL fluorescence cystoscopy was performed. Lesions and suspicious areas were also mapped onto bladder chart. From all the lesions mapped specimens were collected with forceps for histopathological examination and lesions were bipolar loop resected. Specimen were fixed in formalin and embedded in paraffin. EDTA buffer antigen retrieval in water bath pH 8.0 for 60 minutes was performed. A-20 (Santa Cruz) primary antibody and Ultra Vision LP Value (LabVision) detection system were used.
Eur Urol Suppl 2012;11(4):106
Reaction intensity was microscopically evaluated (ranging 0-3). Results: 410 HAL fluorescent specimens from 96 patients with non-muscleinvasive bladder cancer were obtained. 354 specimens were analyzed, the remaining 56 were rejected due to the artifacts. Estimation of 354 HAL – positive specimens using VEGF.In normal urothelium 102 (28,8%) HAL-positive specimens the expression of VEGF was demonstrated in 80 (78,43%) specimens and no expression of VEGF in 22 (21,27%) specimens.In cancer tissue 105 (29,7%) HAL-positive specimens the expression of VEGF was demonstrated in 69 (65,7%) specimens and no expression of VEGF in 36 (34,3%) specimens. In 54 (15%) atypia HAL-positive specimens the expression of VEGF was demonstrated in 42 (77,77%) specimens and no expression of VEGF in 12 (22,23%) specimens. In 93 (26,6%) hyperplasia HAL-positive specimens the expression of VEGF was demonstrated in 71 (76,34%) specimens and no expression of VEGF in 22 (23,66%) specimens. Conclusions: There was no expression of VEGF-A, could be used for differential diagnosis of changes in bladder epithelium. This may result from the fact that the onset of neoangiogenesis is not an early phenomenon in cancer formation. The appearance of VEGF expression in inflammatory lesions may be related to the participation of this factor in the inflammatory and repair processes.
C102
Survivin expression in the cytoplasm and nucleus of HAL fluorescent cancer cells
Zembrzuski M.1, Glazar B.2, Dobrowolski Z.2, Lipczyński W.2, Okoń K.2, Bąk M.2, Maliszewski T.2 1 Jagiellonian University, Clinic and Dept. of Urology, Kraków, Poland, 2 Jagiellonian University, Clinic and Dept. of Urology, Cracow, Poland Introduction & Objectives: The main element of carcinogenesis is to obtain tumor cell resistance to apoptosis. One of the factors regulating apoptosis is survivin. Survivin is a protein product of the BIRC5 gene, under physiological conditions present in fetal cells, is not present in good differentiated cells. Reexpression of survivinu observed in a number of malignancies, it is associated with cell proliferation and to protect against cancer cell apoptosis.The aim of the study was to evaluate the expression of survivin on the nuclei and cytoplasm urothelial cells, which in condition of PDD in the endoscopy under ultraviolet light showed fluorescence. Material & Methods: 96 patients with non-muscle-invasive bladder cancer underwent PDD endoscopy using Hexvix. From all the lesions specimens were collected with forceps for histopathological examination and lesions were bipolar loop resected.Specimens were fixed in 10% formalin solution. Reaction in the direction of survivin was performed in a typical way available antigen in a water bath in EDTA, pH 8.0. Primary antibody was used AB-1 8B2 (Lab Vision). Using a detection system Volu LP Ultra Vision (Lab Vision), DAB as chromogen and Mayer hematoxylin as a contrast. Examining microscopically the cytoplasm of urothelial cells rated power on a scale pH from 0 to 3.Examining the contents of survivin in the urothelial nuclei evalueted microscopically percentage of positive nuclei. Results: Specimens from 96 patients with non-muscle-invasive bladder cancer were obtained. 363 specimens were analyzed, the remaining 47 were rejected due to the artifacts.Estimation of 354 HAL – positive specimens using expression of cytoplasmic Survivin in urothelium .In all 354 HAL –positive specimens the expression of Survivin C was demonstrated in 328 (92,6%) specimens and no expression of Survivin C in 26 (7,4%) specimensIn normal urothelium 102 specimens the expression of Survivin C was demonstrated in 91 (89,21%) specimens and no expression of Survivin C in 11 (10,79%) specimensIn cancer tissue 105 HALpositive specimens the expression of Survivin C was demonstrated in 93 (88,6%) specimens and no expression of Survivin C in 12 (11,4%) specimens. In 54 atypia HAL-positive specimens the expression of Survivin C was demonstrated in 51 (94,44%) specimens and no expression of Survivin C in 3 (5,565%) specimens. Estimation of 354 HAL – positive specimens using expression of nuclear Survivin in urothelium .In all 354 HAL –positive specimens the expression of Survivin was demonstrated in 218 (61,5%) specimens and no expression of Survivin C in 136 (38,5%) specimensIn normal urothelium 102 specimens the expression of Survivin was demonstrated in 69 (67,64%) specimens and no expression of Survivin C in 33 (32,36%) specimensIn cancer tissue 105 HAL-positive specimens the expression of Survivin was demonstrated in 68 (64,8%) specimens and no expression of Survivin in 37 (35,2%) specimens. In 54 atypia HAL-positive specimens the expression of Survivin was demonstrated in 31 (57,4%) specimens and no expression of Survivin in 23 (42,6%) specimens. Conclusions: In 92,6% HAL–positive specimens the expression of Survivin was demonstrated. There was no statistical differences between Survivin expresion between normal, cancer and also atypical urothelium.