- Email: [email protected]
224Dynamic activity of rhabdosphincter and urethra in continent and incontinent women
221
222
INNERVATION INDUCED BY CYSTITIS. COMPARISON OF EXPERIMENTAL CYSTITIS M O D E L IN PIGS VERSUS INTERSTITIAL CYSTITIS IN HUMANS Radziszewski P.I, Bossowska A. 2, Borkowski A.I, Majewski M ?
COMPARISON OF MESH-TO-TISSUE ATTACHMENT CAPACITY AND DETACHMENT STRENGTH OF F O U R DIFFERENT SLING MATERIALS USED IN THE TREATMENT OF STRESS URINARY INCONTINENCE IN AN IN VIVO RABBIT MODEL
IWarsaw School of Medicine, Urology, Warsaw, Poland, 2University of Warmia arid Mazury, Division of Clinical Physiology, Olsztyn, Poland
Yildirim A. Gulpinar T., Basok E., Gurbuz C., Tokuc R.
INTRODUCTION & OBJECTIVES: The etiology of interstitial cystitis (IC) is still far from being understood. The disease has different clinical manifestations and different treatment modalities are tried to treat it. The main problem with studying pathogenesis of IC is the lack of an appropriate animal model. Therefore in our present study we wanted to compare innervation changes induced by IC in humans with that observed in a recently established by us animal model of bladder inflmrmaatinn. MATERIAL & METHODS: In our studies we included n=5 female patients suffering from interstitial cystitis. During the cystoacopy, cold-cup bladder biopsies from the posterior bladder wall were collected. Control tissue samples n-7 were obtained during cystectomy performed for invasive transitional cell carcinoma. Samples were collected from unchanged region of the posterior bladder wall and processed for immunohistofiuorescence. In sexually immature female pigs (n-4) cystitis was achieved by combining acute bladder overdistension (pressure 100 cm H20 ) and 30 rain chemical irritation with 50% acetone solution (7 days after overdistension). Afterwards animals were kept on normal chow conditions for 30 days and then sacrificed. Bladder scraps from the ventral bladder wall were processed for inmmnohistofluorescence. Antisera against following putative neurotransmitters were used: SP-substance P, CGRP- calcitonine gene related peptide, VIP-vasoactive intestinal polypeptide, PACAP-27-pituitary adenylate cyclase polypetide 27, NPYneuropeptide Y, 5-HT-serotonine, DbH-dopamine-[3-hydroxylase. RESULTS: Mean patients age was 63.8 years~:6.3, mean disease duration was 3.3 years±2.1. No neoplasmatic cells were found in the obtained specimens. In patients with IC an increase in the expression of CGRP immunoreactive (IR) nerve fibers, NPY-IR nerve fibers and PACAP-IR nerve fibers was observed, while fibers being SP-IR, VIP-IR and DbH-IR remained unchanged and 5-HT-IR nerve fibers were wanished. Similar findings were observed in specimen from experimental animals, however no increase in PACAP-IR nerve fibers was observed. Details are presented in Tab. 1 Substance
Pig
Human Control
SP
+
CGRP
+
IC
Contml
Cystitis
+
DbH 5-HT
+
NPY
INTRODUCTION & OBJECTIVES: The aim of the study was to compare mesh-to-tissue detachment strengths of four different sling mesh materials used in Tension-Free Vaginal Tape (TVT), Intravaginal Slingplasty (IVS), polypropylene mesh hernia-repair, and Suprapubic approach to suburethral polypropylene tape (SPARC) procedures at second, 7th, 15th and 30 ~hpost-implantation day. MATERIAL & METHODS: Twenty female New Zealand white rabbits were
randomized to four groups as group 1 (2 days), group 2 (7 days), group 3 (15 days) and group 4 (30 days). After rabbits were anesthetized, four subfascial tunnels 1.5 cm in length, 2 in the medial surface of each lower extremities were prepared, and 1.0 x 0.5 cm strips of TVT, IVS, SPARC and polypropylene mesh were implanted in each tunnel. At the second, 7th, 15th and 30 *h day after implantation, all animals were re-anesthetized, and a mechanical testing model was performed to define the detachment strength by using weights as grams. RESULTS: The mean detachment strengths (the minimum weight needed to move the mesh) of TVT, IVS, SPARC and polypropylene mesh were 291.6 gr./ 92.8 gr./178.4 gr./188.4 gr. at 2 nd day, 840.0 gr./280.0 gr./860.0 gr./510.0 gr. at 7th day, 1930.0 gr./1070.0 gr./1390.0 gr./1290.0 gr. at 15th day, and 2390.0 gr./ 1510.0 gr./2160.0 gr./1850.0 gr. at 30 th day respectively.
V1P PACAP-27
SSK Goztepe Teaching Hospital, Urology, Istanbul, Turkey
+++
Expression of neuropeptides within the urothelium. -: no nerve fibers; +: few fibres; ++: moderate number of fibres; +++: numerous nerve terminals CONCLUSIONS: Our findings demons'a-ate that the population of CGRP and SP-IR nerve fibers underwent same changes in IC and experimentally induced cystitis in pigs. This makes the proposed model and interesting option to study interstitial cystitis.
CONCLUSIONS: The attachment capacity of TVT was superior and IVS was the least among the four, this difference was slightly statistically significant only at the second and 7th days. Surgical hernia mesh gave similar results as TVT and SPARC.
223
224
W H I C H MUSCARINIC RECEPTOR SUBTYPE PREDOMINATES IN THE UROTHELIUM?
DYNAMIC ACTIVITY OF RHABDOSPHINCTER AND URETHRA IN CONTINENT AND INCONTINENT W O M E N
Scott R. t, Manandar A. 2, Chess-Williams R. ~, Chapple C.3
Strasser H. t, Mitterberger M. J, Rehder p.1, Pinggera G.M. 1, Herwig R. 1, Klauser A. 2, Bartsch G?, Frauscher F.z
~University of Sheffield, Biomedical Science, Sheffield, United Kingdom, 2University of Varna, Urology, Varna, Bulgaria, 3Royal Hallamshire Hospital, Urology, Sheffield, United Kingdom
IUniversity of Iunsbruck, Department of Urology, Innsbrnck, Austria, 2University of Innsbruck, Department of Radiology 2, Innsbruck, Austria
INTRODUCTION & OBJECTIVES: Stimulation of the muscarinic receptors in the bladder evokes the release of a diffusible inhibitory factor from the urothelium in both human and pig that suppresses contractile responses of the tmderlying detrusor muscle. In previous radioligand binding studies, a population of mnscarinic receptors with a density twice that found in the detrusor smooth muscle has been identified. The aim of this study was to identify which muscarinic receptor subtypes make up this receptor population in the pig urothelium. MATERIAL & METHODS: Urothelial cell membranes were prepared from
female pig bladders. Competition experiments using a range of concentrations of muscarinic receptor subtype antagonists and the radioligand [3H] QNB (0.6nM) were performed (n=4-10) to identify the muscarinic receptor subtype population present in this tissue.
INTRODUCTION & OBJECTIVES: The goal of our study was to assess the
dynamic changes of the lower urinary tract in response to contractions of the rhabdosphincter in females with and without urinary stress incontinence by means of three-dimensional trasurethral ultrasound. MATERIAL & METHODS: In 29 female patients with clinically proven stress urinary incontinence and in 6 continent females transurethral ultrasound was performed using special intraurethral ultrasound probes (B&K, Denmark; Hitachi, Japan). The ultrasound probes were mounted on a mechanically driven puller system and pulled back slowly with a speed of 1 mm/sec. The cross-sectional images were transferred to a workstation. 3-D reconsmactions were calculated using specialized software packages. Contractility of the rhabdosphincter was assessed by measuring the distance between the inner aspects of the rhabdosphincter and the ultrasound transducer during voluntary muscle contraction. Changes in length and diameter of the urethra and the rhabdosphincter in response to voluntary contractions of the rhabdospbincter were measured.
RESULTS: The M2-receptor selective antagonist methoctramine had a high
affinity (PKi=7.7+0.1) for the receptors of the urothelium, whilst pirenzipine (Ml-selective) and oxybutynin (M3-selective) had relatively low affinities (PKi=6.2~0.1 and 6.9±0.1 respectively). Atropine, a non-subtype selective antagonist had a high affinity for the urothelial muscarinic receptor (pKi=8.5±0.3). These antagonists bound to a single binding site with Hill slopes of unity. Similarly, 4-DAMP (M3-selective) bound to a single receptor in 6 experiments, which had a low affinity for this antagonist (pK=7.1±0.l), but in 4 other experiments it identified an additional minor population (approximately 20%) of receptors with a high affinity for this antagonist. Correlation plots of the affinities obtained at the urothelial receptor against values published in the literature for the various cloned receptors yielded poor correlations for the M1 (r2=0.07), M3 (r2=0.16), M4 (r2=0.41) and MS-receptors (r2-0.31), but correlated closely with the cloned M2-receptor (r2=0.87). CONCLUSIONS: The urothelium possesses a population of muscarinic receptors that appears to consist predominately of the M2 muscarinic receptor subtype.
European Urology Supplements 4 (2005) No. 3, pp. 58
RESULTS: Transurethral ultrasound and subsequent 3D-reconstrnction were
successfully performed in all patients. It could be shown that during voluntary contraction of the rhabdosphincter the urethra was compressed. The cranio-caudal length of the urethra as well as the rhabdosphincter was substantially increased in continent patients (up to 2 ram) in response to compression of the urethra. In patients with stress urinary incontinence a statistically significantly reduced contractility of the rbabdosphincter was observed compared to continent females. Increase of urethral length during contraction of the rhabdosphincter was statistically significantly lower in these patients. Furthermore, in patients with stress urinary incontinence atrophies of the rhabdospbincter could be seen. 3-D reconstruction provided detailed images which allowed excellent orientation in all dimensions. CONCLUSIONS: Investigation of the female urethra with transurethral ultrasound and 3-D reconstruction proved to be a simple and easy procedure which provided excellent anatomic and physiologic information about the urethra and the rhabdosphincter. It could be shown that contraction of the rhabdosphincter not only compresses but also increases the cranio-caudal length of the urethra.
222
INNERVATION INDUCED BY CYSTITIS. COMPARISON OF EXPERIMENTAL CYSTITIS M O D E L IN PIGS VERSUS INTERSTITIAL CYSTITIS IN HUMANS Radziszewski P.I, Bossowska A. 2, Borkowski A.I, Majewski M ?
COMPARISON OF MESH-TO-TISSUE ATTACHMENT CAPACITY AND DETACHMENT STRENGTH OF F O U R DIFFERENT SLING MATERIALS USED IN THE TREATMENT OF STRESS URINARY INCONTINENCE IN AN IN VIVO RABBIT MODEL
IWarsaw School of Medicine, Urology, Warsaw, Poland, 2University of Warmia arid Mazury, Division of Clinical Physiology, Olsztyn, Poland
Yildirim A. Gulpinar T., Basok E., Gurbuz C., Tokuc R.
INTRODUCTION & OBJECTIVES: The etiology of interstitial cystitis (IC) is still far from being understood. The disease has different clinical manifestations and different treatment modalities are tried to treat it. The main problem with studying pathogenesis of IC is the lack of an appropriate animal model. Therefore in our present study we wanted to compare innervation changes induced by IC in humans with that observed in a recently established by us animal model of bladder inflmrmaatinn. MATERIAL & METHODS: In our studies we included n=5 female patients suffering from interstitial cystitis. During the cystoacopy, cold-cup bladder biopsies from the posterior bladder wall were collected. Control tissue samples n-7 were obtained during cystectomy performed for invasive transitional cell carcinoma. Samples were collected from unchanged region of the posterior bladder wall and processed for immunohistofiuorescence. In sexually immature female pigs (n-4) cystitis was achieved by combining acute bladder overdistension (pressure 100 cm H20 ) and 30 rain chemical irritation with 50% acetone solution (7 days after overdistension). Afterwards animals were kept on normal chow conditions for 30 days and then sacrificed. Bladder scraps from the ventral bladder wall were processed for inmmnohistofluorescence. Antisera against following putative neurotransmitters were used: SP-substance P, CGRP- calcitonine gene related peptide, VIP-vasoactive intestinal polypeptide, PACAP-27-pituitary adenylate cyclase polypetide 27, NPYneuropeptide Y, 5-HT-serotonine, DbH-dopamine-[3-hydroxylase. RESULTS: Mean patients age was 63.8 years~:6.3, mean disease duration was 3.3 years±2.1. No neoplasmatic cells were found in the obtained specimens. In patients with IC an increase in the expression of CGRP immunoreactive (IR) nerve fibers, NPY-IR nerve fibers and PACAP-IR nerve fibers was observed, while fibers being SP-IR, VIP-IR and DbH-IR remained unchanged and 5-HT-IR nerve fibers were wanished. Similar findings were observed in specimen from experimental animals, however no increase in PACAP-IR nerve fibers was observed. Details are presented in Tab. 1 Substance
Pig
Human Control
SP
+
CGRP
+
IC
Contml
Cystitis
+
DbH 5-HT
+
NPY
INTRODUCTION & OBJECTIVES: The aim of the study was to compare mesh-to-tissue detachment strengths of four different sling mesh materials used in Tension-Free Vaginal Tape (TVT), Intravaginal Slingplasty (IVS), polypropylene mesh hernia-repair, and Suprapubic approach to suburethral polypropylene tape (SPARC) procedures at second, 7th, 15th and 30 ~hpost-implantation day. MATERIAL & METHODS: Twenty female New Zealand white rabbits were
randomized to four groups as group 1 (2 days), group 2 (7 days), group 3 (15 days) and group 4 (30 days). After rabbits were anesthetized, four subfascial tunnels 1.5 cm in length, 2 in the medial surface of each lower extremities were prepared, and 1.0 x 0.5 cm strips of TVT, IVS, SPARC and polypropylene mesh were implanted in each tunnel. At the second, 7th, 15th and 30 *h day after implantation, all animals were re-anesthetized, and a mechanical testing model was performed to define the detachment strength by using weights as grams. RESULTS: The mean detachment strengths (the minimum weight needed to move the mesh) of TVT, IVS, SPARC and polypropylene mesh were 291.6 gr./ 92.8 gr./178.4 gr./188.4 gr. at 2 nd day, 840.0 gr./280.0 gr./860.0 gr./510.0 gr. at 7th day, 1930.0 gr./1070.0 gr./1390.0 gr./1290.0 gr. at 15th day, and 2390.0 gr./ 1510.0 gr./2160.0 gr./1850.0 gr. at 30 th day respectively.
V1P PACAP-27
SSK Goztepe Teaching Hospital, Urology, Istanbul, Turkey
+++
Expression of neuropeptides within the urothelium. -: no nerve fibers; +: few fibres; ++: moderate number of fibres; +++: numerous nerve terminals CONCLUSIONS: Our findings demons'a-ate that the population of CGRP and SP-IR nerve fibers underwent same changes in IC and experimentally induced cystitis in pigs. This makes the proposed model and interesting option to study interstitial cystitis.
CONCLUSIONS: The attachment capacity of TVT was superior and IVS was the least among the four, this difference was slightly statistically significant only at the second and 7th days. Surgical hernia mesh gave similar results as TVT and SPARC.
223
224
W H I C H MUSCARINIC RECEPTOR SUBTYPE PREDOMINATES IN THE UROTHELIUM?
DYNAMIC ACTIVITY OF RHABDOSPHINCTER AND URETHRA IN CONTINENT AND INCONTINENT W O M E N
Scott R. t, Manandar A. 2, Chess-Williams R. ~, Chapple C.3
Strasser H. t, Mitterberger M. J, Rehder p.1, Pinggera G.M. 1, Herwig R. 1, Klauser A. 2, Bartsch G?, Frauscher F.z
~University of Sheffield, Biomedical Science, Sheffield, United Kingdom, 2University of Varna, Urology, Varna, Bulgaria, 3Royal Hallamshire Hospital, Urology, Sheffield, United Kingdom
IUniversity of Iunsbruck, Department of Urology, Innsbrnck, Austria, 2University of Innsbruck, Department of Radiology 2, Innsbruck, Austria
INTRODUCTION & OBJECTIVES: Stimulation of the muscarinic receptors in the bladder evokes the release of a diffusible inhibitory factor from the urothelium in both human and pig that suppresses contractile responses of the tmderlying detrusor muscle. In previous radioligand binding studies, a population of mnscarinic receptors with a density twice that found in the detrusor smooth muscle has been identified. The aim of this study was to identify which muscarinic receptor subtypes make up this receptor population in the pig urothelium. MATERIAL & METHODS: Urothelial cell membranes were prepared from
female pig bladders. Competition experiments using a range of concentrations of muscarinic receptor subtype antagonists and the radioligand [3H] QNB (0.6nM) were performed (n=4-10) to identify the muscarinic receptor subtype population present in this tissue.
INTRODUCTION & OBJECTIVES: The goal of our study was to assess the
dynamic changes of the lower urinary tract in response to contractions of the rhabdosphincter in females with and without urinary stress incontinence by means of three-dimensional trasurethral ultrasound. MATERIAL & METHODS: In 29 female patients with clinically proven stress urinary incontinence and in 6 continent females transurethral ultrasound was performed using special intraurethral ultrasound probes (B&K, Denmark; Hitachi, Japan). The ultrasound probes were mounted on a mechanically driven puller system and pulled back slowly with a speed of 1 mm/sec. The cross-sectional images were transferred to a workstation. 3-D reconsmactions were calculated using specialized software packages. Contractility of the rhabdosphincter was assessed by measuring the distance between the inner aspects of the rhabdosphincter and the ultrasound transducer during voluntary muscle contraction. Changes in length and diameter of the urethra and the rhabdosphincter in response to voluntary contractions of the rhabdospbincter were measured.
RESULTS: The M2-receptor selective antagonist methoctramine had a high
affinity (PKi=7.7+0.1) for the receptors of the urothelium, whilst pirenzipine (Ml-selective) and oxybutynin (M3-selective) had relatively low affinities (PKi=6.2~0.1 and 6.9±0.1 respectively). Atropine, a non-subtype selective antagonist had a high affinity for the urothelial muscarinic receptor (pKi=8.5±0.3). These antagonists bound to a single binding site with Hill slopes of unity. Similarly, 4-DAMP (M3-selective) bound to a single receptor in 6 experiments, which had a low affinity for this antagonist (pK=7.1±0.l), but in 4 other experiments it identified an additional minor population (approximately 20%) of receptors with a high affinity for this antagonist. Correlation plots of the affinities obtained at the urothelial receptor against values published in the literature for the various cloned receptors yielded poor correlations for the M1 (r2=0.07), M3 (r2=0.16), M4 (r2=0.41) and MS-receptors (r2-0.31), but correlated closely with the cloned M2-receptor (r2=0.87). CONCLUSIONS: The urothelium possesses a population of muscarinic receptors that appears to consist predominately of the M2 muscarinic receptor subtype.
European Urology Supplements 4 (2005) No. 3, pp. 58
RESULTS: Transurethral ultrasound and subsequent 3D-reconstrnction were
successfully performed in all patients. It could be shown that during voluntary contraction of the rhabdosphincter the urethra was compressed. The cranio-caudal length of the urethra as well as the rhabdosphincter was substantially increased in continent patients (up to 2 ram) in response to compression of the urethra. In patients with stress urinary incontinence a statistically significantly reduced contractility of the rbabdosphincter was observed compared to continent females. Increase of urethral length during contraction of the rhabdosphincter was statistically significantly lower in these patients. Furthermore, in patients with stress urinary incontinence atrophies of the rhabdospbincter could be seen. 3-D reconstruction provided detailed images which allowed excellent orientation in all dimensions. CONCLUSIONS: Investigation of the female urethra with transurethral ultrasound and 3-D reconstruction proved to be a simple and easy procedure which provided excellent anatomic and physiologic information about the urethra and the rhabdosphincter. It could be shown that contraction of the rhabdosphincter not only compresses but also increases the cranio-caudal length of the urethra.