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PubMed2004

Urodynamic effects of the K+ channel (KCNQ) opener retigabine in freely moving, conscious rats.

PURPOSE: Retigabine is a novel anticonvulsant drug that not only augments gamma-aminobutyric acid mechanisms, but also opens voltage gated K+ channels (KCNQ). In this study we investigated the effects of retigabine on detrusor activity in rats. MATERIALS AND METHODS: To conscious, female Sprague-Dawley rats undergoing continuous cystometry retigabine was given intravenously (0.5, 1 and 5 mg/kg(-1)). The KCNQ channel blocker linopirdine was given intravenously (2 mg/kg(-1)) 5 minutes prior to retigabine (1 mg/kg(-1)). In addition, retigabine was given intracerebroventricularly (1, 5 and 10 microg) and intravesically (100, 500 and 1,000 ng ml(-1)). The effects of the drug (intravesical administration) on capsaicin induced bladder overactivity were also tested. RESULTS: Retigabine given intravenously (1 mg/kg(-1)) decreased baseline and maximal bladder pressures, increased voided and infused volumes, and increased voiding intervals. Retigabine (10 microg) given intracerebroventricularly decreased baseline pressure and increased voided and infused volumes as well as voiding intervals. However, bladder pressures were not significantly affected. Intravesical retigabine (1,000 ng.ml(-1)) decreased maximal bladder pressure, increased voided and infused volumes, and increased voiding intervals. Given intravesically for 30 minutes prior to intravesical capsaicin (30 microM) instillation retigabine (1,000 ng.ml(-1)) decreased the detrusor overactivity induced by capsaicin. The KNCQ channel blocker linopirdine (2 mg/kg(-1)) completely blocked the effects of intravenous retigabine (1 mg/kg(-1)). CONCLUSIONS: Retigabine given intravenously, intracerebroventricularly and intravesically increased micturition volume and voiding intervals and, when given intravesically, it decreased capsaicin induced detrusor overactivity, suggesting that KCNQ channels can be interesting targets for drugs aiming at micturition control. Retigabine may be a candidate to test as a treatment for detrusor overactivity in humans.

Animals↗
PubMed2001

Administration of endocannabinoids prevents a referred hyperalgesia associated with inflammation of the urinary bladder.

BACKGROUND: Referred hyperalgesia to a somatopically appropriate superficial site is a cardinal symptom of visceral inflammatory pain and has been demonstrated after turpentine-induced urinary bladder inflammation in the rat. The authors examined the effect of the endocannabinoids anandamide and palmitoylethanolamide on the referred hyperalgesia associated with this model. METHODS: After measurement of baseline limb withdrawal latencies to a noxious heat stimulus, the bladders of 50 female Wistar rats were inflamed by intravesical administration of 0.5 ml 50% turpentine. Ten or 25 mg/kg of anandamide or palmitoylethanolamide or vehicle were administered immediately before introduction of turpentine. Antagonists to both the cannabinoid CB1 and CB2 receptors were coadministered with the higher dose of endocannabinoids. Latencies were recorded 2, 4, 6, 8, and 24 h after removal of turpentine. The difference between forelimb and hind limb withdrawal latencies was plotted against time, and areas under these curves were compared. RESULTS: Inflammation of the urinary bladder was associated with a relative thermal hyperalgesia referred to the hind limb. Anandamide and palmitoylethanolamide attenuated this referred hyperalgesia at doses of 10 and 25 mg/kg. The CB1 receptor antagonist SR141716A reduced the antihyperalgesic effect of anandamide, but the CB2 antagonist SR144528 did not. Coadministration of SR141716A with palmitoylethanolamide did not affect the antihyperalgesic effect but was reduced by SR144528. CONCLUSIONS: Anandamide (via CB1 receptors) and palmitoylethanolamide (putatively via CB2 receptors) attenuated a referred hyperalgesia in a dose-dependent fashion. CB1 and CB2 receptors are strategically situated to influence the nerve growth factor-driven referred hyperalgesia associated with inflammation of the urinary bladder. These data implicate cannabinoids as a novel treatment for vesical pain.

Adjuvants, Immunologic↗
PubMed1983

The topical use of misonidazole in bladder cancer.

Penetration studies of MIS after intravesical administration showed adequate concentrations with a gradient across the tumor. After instillation of 2.5 g in 50 ml water the concentration in the deep parts of the tumor amounted to about 100 microgram per ml. This corresponds to a SER for hypoxic cell of the order of 1.7. A more uniform tissue distribution of the drug was noted 3.5 hours after an oral dose of 3 g/meter square. The concentration in the deep parts of the tumor and perivesical tissue was of the order of 100 micrograms/g. The concentration in these regions are relevant to preoperative irradiation which aims at sterilizing the deep infiltrating margins. The intravesical use with or without oral augmentation is suitable for use in association with concentrated preoperative radiotherapy regimens. The topical use of MIS in such regimens markedly reduces the risk of neurotoxicity. The tissue concentration resulting from the two routes proved to be additive. The higher concentration in lymph nodes after the oral route the greater concentration and prolonged contact after combined administration may have therapeutic merits.

Administration, Oral↗
PubMed1978

Dimethyl sulfoxide in treatment of inflammatory genitourinary disorders.

Intravesical dimethyl sulfoxide (DMSO) has been used in the treatment of 213 patients with various inflammatory conditions involving the lower genitourinary tract, including intractable interstitial cystitis, radiation cystitis, chronic prostatitis, and chronic female trigonitis. Significant symptomatic relief has been achieved in the majority of patients so treated, and no systemic or local toxicity has been noted. Some patients failed to respond entirely, and others relapsed after DMSO treatment periods of several months, ultimately coming to augmentation cystoplasty or urinary diversion. However, because of its simplicity and ease of administration, intravesical DMSO therapy is recommended in all noninfectious or non-neoplastic inflammatory conditions presenting initially with severe symptoms, or that have failed to respond to conventional therapy.

Adult↗
PubMed1987

The practical use of tumor marker determination in bladder washing specimens. Assessing the urothelium of patients with superficial bladder cancer.

Bladder washing specimens from 81 patients with recurrent multifocal superficial bladder cancer were evaluated for DNA profile by flow cytometry and cell-surface blood group (ABH) antigen reactivity by a modified specific erythrocyte adherence test. The study was conducted in a prospective, blind, nonrandomized fashion. Fifteen patients were treated with transurethral resection (TUR) alone and 66 with TUR and intravesical administration of bacillus Calmette-Guerin weekly for 6 weeks (TUR + BCG). Among the patients treated with TUR only, there was a notably greater rate of tumor recurrence and progression in patients with unfavorable tumor markers (aneuploidy and ABH-negative reactivity) than in those with favorable markers (diploidy and ABH-positive reactivity). The difference was less striking in patients treated with BCG, which reduced the frequency of recurrence or progression at 30 months from 87% to 44% and from 60% to 23%, respectively. This favorable effect of BCG was virtually confined to patients with initially favorable markers and to those whose initially unfavorable markers became favorable during BCG administration. Aneuploidy and negative ABH are phenotypic expressions of undifferentiation that can forecast the potential of the urothelium to form new tumors and predict invasion. To an extent, these markers are independent of the grade and stage of the disease. BCG can induce prognostically favorable conversion of the markers expression. Lack of such conversion indicates lack of response to BCG and should be regarded as evidence of persistent disease even if conventional methods do not reveal it. Therefore, sequential determination of markers is useful in monitoring patients with superficial bladder cancer treated with intravesical BCG.

Adult↗
PubMed2002

Vanilloid receptor 1 expression in the rat urinary tract.

Previous findings have shown that the capsaicin sensitivity of sensory fibres is due to the expression of a specific membrane protein, the vanilloid receptor type 1 (VR1). In the present work we studied the distribution, morphology and the neurochemical content of nerve fibres expressing this receptor in the rat urinary tract. Immunolabelling was performed against the VR1 and the positive fibres were examined by light and electron microscopy. Colocalisation of VR1 and substance P or calcitonin gene-related peptide immunoreactivities, and isolectin B4 binding, was evaluated under the confocal microscope. In addition, the effect of intravesical administration of resiniferatoxin, an ultra-potent vanilloid receptor agonist, in the receptor expression in the bladder was also studied. Numerous VR1-immunoreactive fibres were found in the mucosa and muscular layer of the entire urinary tract except the kidney. In the bladder, most fibres were also substance P- or calcitonin gene-related peptide-immunoreactive but did not bind isolectin B4. Under the electron microscope VR1 immunoreactivity was confined to unmyelinated axons and varicosities containing small clear and large dense-core synaptic vesicles. They occurred beneath or among epithelial cells or closely apposed to smooth muscle cells. Intravesical resiniferatoxin decreased VR1 immunoreactivity transiently. These data indicate that primary sensory fibres expressing VR1 are extremely abundant in the rat urinary tract and that, in contrast to the skin, they belong almost exclusively to the peptide-containing sub-population of primary afferents. As capsaicin-sensitive bladder afferents are involved in nociception and reflex micturition control, the numerous free terminal nerve endings expressing VR1 in the mucosa seem more adequate to accomplish the former function. However, the close apposition between VR1-expressing fibres and smooth muscle cells suggests that they may also encode the tonus of the muscular layer.

Animals↗
PubMed1988

Inhibition of mouse bladder tumor proliferation by alpha difluoromethylornithine and interferon in vitro and in vivo.

Previous studies have reported that alpha difluoromethylornithine (DFMO), an enzyme-activated, irreversible inhibitor of ornithine decarboxylase (ODC), has anti-tumor activity in several tumor systems. Recently, investigations have revealed that combinations of DFMO and Interferon (IFN) are synergistic in inhibiting tumor cell growth. We tested the effects of DFMO and IFN alpha, beta alone and in combination on the growth of mouse bladder tumor (MBT-2) cells both in vitro and in vivo. MBT-2 cells were incubated for 72 hours in 96 well microtiter plates with DFMO, IFN alpha, beta and combinations of both agents and percentage inhibition was calculated. In vivo studies utilized the intravesical implantation method as well as subcutaneous implantation. DFMO was administered as a 1% solution in the drinking water. IFN alpha, beta was given bi-weekly by intravesical administration. DFMO effectively inhibited MBT-2 growth in vitro. The ID50 was 0.08 mM and peak inhibitory activity was reached at concentrations of 0.16 mM and remained constant with concentrations of up to 10 mM. IFN alpha, beta also inhibited the in vitro proliferation of MBT-2 with maximum inhibition (46%) at 2,000 U/ml. Combinations of DFMO and IFN alpha, beta showed increased anti-proliferative activity. The degree of enhancement varied with synergism, additivity, or sub-additivity at varying drug concentrations. In vivo, DFMO significantly retarded the growth of tumors implanted subcutaneously (p less than .05) and significantly delayed the outgrowth of tumors implanted intravesically (p less than .01). IFN alpha, beta alone was ineffective in vivo and produced no additive effect in vivo when used in combination with DFMO. Results of our investigation show that DFMO inhibits proliferation of MBT-2 cells in vitro and exhibits a similar effect in vivo against subcutaneous and intravesical tumor implants. IFN alpha, beta alone demonstrated anti-proliferative activity in vitro but did not affect MBT-2 growth in vivo. Although the combination of DFMO and IFN alpha, beta exhibited enhanced activity in vitro, no enhancement was observed with combination therapy in vivo.

Animals↗
PubMed2004

Mechanisms involved in new therapies for overactive bladder.

During the last few years, vanilloid substances and botulinum-A toxin were extensively investigated as new therapies for overactive bladder. Intravesical administration of capsaicin or resiniferatoxin--2 members of the vanilloid family--has been shown to increase bladder capacity and decrease urge incontinence in patients with neurogenic, as well as nonneurogenic, forms of detrusor overactivity. In addition, vanilloids have been shown also to reduce bladder pain in patients with hypersensitive disorders. Vanilloids are exogenous ligands of vanilloid receptor type 1 (VR1), an ion channel present in the membrane of type C primary afferent nerve fibers. This receptor, which plays a key role in pain perception and control of the micturition reflex, may be upregulated by nerve growth factor (NGF), a neurotrophic molecule detected in high concentrations in overactive detrusor tissue. Vanilloids, by reducing uptake of NGF through sensory neurons, may counteract VR1 upregulation. Intravesical injections of botulinum-A toxin, a neurotoxin produced by Clostridium botulinum, were shown to increase bladder capacity and to decrease urge incontinence episodes in patients with neurogenic detrusor overactivity. Botulinum-A toxin impedes the release of acetylcholine from cholinergic nerve endings at the neuromuscular junction, leading to paralysis of the detrusor smooth muscle.

Animals↗
PubMed2004

Bladder C-fiber desensitization induces a long-lasting improvement of BPH-associated storage LUTS: a pilot study.

OBJECTIVES: To evaluate the effect of bladder C-fiber desensitization on BPH-associated storage LUTS. METHODS: Twelve patients with predominant BPH-associated storage LUTS gave written informed consent to enter the study. IPSS and QoL scores, micturition chart, uroflowmetry, cystometry and post-void residual (PVR) were obtained at baseline and at 1, 3 and 6 months after a single intravesical administration of 50 nM resiniferatoxin (RTX) solution. RESULTS: Mean IPSS score decreased from 20.1 +/- 6.0 to 10.5 +/- 4.4 at 1 month (p = 0.0001), to 10.3 +/- 4.4 at 3 months (p = 0.00001) and to 9.8 +/- 5.7 at 6 months (p < 0.00001). Mean QoL score decreased from 4.5 +/- 1 to 2.5 +/- 1 at 1 month (p = 0.0001), to 2.3 +/- 1 at 3 months (p = 0.0004) and to 2.6 +/- 1.2 at 6 months (p = 0.0003). Mean urinary frequency decreased from 15.2 +/- 8.5 to 10.8 +/- 7.3 at 1 month (p = 0.0002), to 10.2 +/- 4.9 at 3 months (p = 0.002) and to 11.7 +/- 9.2 at 6 months (p = 0.005). Urge incontinence, which was present in six cases, disappeared in 4 patients and decreased to less than half in the other two. Mean first desire to void and maximal cystometric capacity increased significantly after RTX whereas uroflowmetry and PVR were not altered. CONCLUSIONS: Intravesical desensitization of bladder C fibres with intravesical RTX might be useful in the treatment of patients with predominant BPH associated storage LUTS.

Aged↗
PubMed1992

Morphological aspects of the interaction of Bacillus Calmette-Guérin with urothelial bladder cells in vivo and in vitro: relevance for antitumor activity?

Intravesical administration of Bacillus Calmette-Guérin (BCG) has been shown to be effective in the treatment of patients with superficial bladder cancer. For a better understanding of the mechanism of this antitumor activity, scanning and transmission electron microscope (SEM, TEM) studies were carried out to investigate morphological aspects of the interaction of BCG with the bladder wall in vivo and in vitro. Adherence of BCG to the bladder wall in vivo was studied 1 and 24 h after single or multiple (6x) BCG instillations in intact and in electrocauterized guinea pig bladders. Despite extensive search with SEM for its presence, virtually no BCG was found on the intact urothelium, and BCG was only occasionally observed in the coagulation lesions. SEM and TEM studies revealed adherence and phagocytosis of BCG by the T24 human bladder carcinoma cell line in vitro. Time sequence studies on the phagocytosis and fate of BCG showed that T24 cells are capable of progressively degrading the mycobacteria in phagolysosomes. However, BCG did not alter MHC class II antigen expression on T24 cells in vitro. In contrast, 54 urine sediments and bladder washings of 11 bladder cancer patients, taken prior to or after several intravesical BCG instillations, failed to demonstrate urothelial (tumor) cells showing evidence of BCG phagocytosis (682 cells screened by TEM), while BCG was phagocytized avidly by leukocytes. These data suggest that a direct interaction of BCG with urothelial bladder cells in vivo can be called in question.

Animals↗
PubMed2004

Cytokine-modified Mycobacterium smegmatis as a novel anticancer immunotherapy.

Intravesical administration of live M. bovis BCG organisms for carcinoma in situ of the urinary bladder is the most successful immunotherapy for solid malignancy. Nevertheless BCG-therapy is associated with significant toxicity and is ineffective in 30-40% of cases. Recently it has been proposed that cytokine-modified mycobacteria may give greater efficacy. As any immunotherapy involving administration of live BCG organisms (wild-type or recombinant) is likely to have associated toxicity (notably in the immunocompromised), we examined the anti-tumour potential of the closely related nonpathogenic organism, Mycobacterium smegmatis, and a TNFalpha gene-modified recombinant M. smegmatis. When wild-type M. smegmatis were delivered to immunocompetent C57Bl/6 mice bearing the transplantable MB49 bladder tumour, efficacy comparable to live BCG was observed with 10-20% long-term survival. However, this effect was lost in both Nude and Beige mice, lacking functional T and NK cells, respectively. Recombinant M. smegmatis secreting TNFalpha, however, gave a 70% durable tumour-free survival. Lymphocytes from draining lymph-nodes and spleens of these mice exhibited pronounced IFNgamma production to mycobacterial-antigen and tumour-lysate, indicating a bias towards cell-mediated immunity. This was further supported by histopathological examination of the tumour site, which revealed significantly increased numbers of CD3+ lymphocytes in animals receiving the recombinant vaccine, but not in those receiving wild-type bacteria. Importantly, tumour rejection following M. smegmatis/TNFalpha was independent of T lymphocytes, as athymic Nude mice efficiently eradicated MB49 tumours. In contrast, the therapeutic efficacy of M. smegmatis/TNFalpha was reduced in animals deficient in NK cytolytic function, suggesting a role for NK-cells in initial tumour destruction. Furthermore the absence of NK-function in Beige mice did not prevent the establishment of tumour-protective memory. No toxicity was observed with wild-type or recombinant M. smegmatis in immunocompetent, T-deficient or NK-deficient models. We demonstrate for the first time that recombinant mycobacteria expressing mammalian cytokines have markedly increased anti-tumour properties. The lack of toxicity suggests that M. smegmatis is a "safe" vehicle for use in immunocompromised patients.

Animals↗
PubMed1992

Epidermal growth factor and its receptor: markers of--and targets for--chemoprevention of bladder cancer.

Epidermal growth factor (EGF) is excreted in urine in high concentrations in a biologically active form. Several lines of evidence indicate that EGF plays a role in transitional cell carcinoma (TCC) development and growth. These include: (1) EGF in the normal urine of rats promotes chemically initiated TCC; (2) EGF in normal human urine stimulates the clonal growth of human TCC cells in vitro; (3) EGF stimulates the in vitro growth of human TCC cells, but not normal human urothelial cells; (4) the density and distribution of the EGF receptor (EGF-R) on human urothelial tissues permits significant access of premalignant, dysplastic, and malignant cells to EGF; and (5) the concentration of EGF in the voided urine of patients with TCC is reduced, implying that EGF may be "extracted" from urine by the greater number of EGF-Rs in patients with urothelial malignancy. Abnormal expression of the urothelial EGF-R and/or altered excretion of EGF may well precede overt evidence of TCC and thus may serve as markers of risk or exposure. Similarly, reversion of EGF-R expression or the return of excreted EGF to normal levels may provide a marker of response for preventive and therapeutic strategies. Interference with the EGF/EGF-R interaction through dietary or pharmacological manipulations of the urine, or via targeting strategies employing intravesical administration of conjugated toxins or isotopes is already being employed in experimental and clinical studies. These approaches offer promising new tools in the detection, monitoring, prevention, and management of early stage bladder cancer.

Antineoplastic Agents↗
PubMed1983

Development and clinical application of an AFC regimen (adriamycin, futraful, cytosine arabinoside) for superficial bladder tumors.

Our AFC regimen was originally developed to treat superficial bladder tumors. It consists of a daily intravesical administration of 20 mg Adriamycin (ADM) and 200 mg Cytosine arabinoside (CA) dissolved in 20 ml of sterilized distilled water, simultaneously with a daily rectal suppository of 750 mg tegafur (Futraful) for a total of 20 applications. Our study demonstrated the following characteristics of the AFC modality: (1) It is simple; (2) it is completed in 3 weeks, which is shorter than conventional bladder instillation therapy; (3) the effective rate is reasonable (70%); (4) no local side-effects were observed because the concentrations of ADM and CA were low; (5) interruption of therapy was unnecessary, because whatever general side-effects were caused, they disappeared within several days of completion of treatment.

Adult↗
PubMed1990

Presence of interleukin-2 in urine of superficial bladder cancer patients after intravesical treatment with bacillus Calmette-Guérin.

Urine samples were obtained from patients with superficial bladder cancer after immunotherapy with bacillus Calmette-Guérin (BCG). The patients were repeatedly (once a week for 6 consecutive weeks) treated with intravesical administration of approximately 5 X 10(8) culturable particles of BCG. Some patients received more than six BCG instillations. The urine samples were investigated for the presence of interleukin-2 (IL-2) in an in vitro bioassay using a murine cytotoxic T cell line (CTTL-16) that shows IL-2-dependent growth. Preliminary experiments indicated the presence of inhibitory factors in the urine. This inhibitory activity was abolished after 24 h dialysis. In a neutralization assay with both polyvalent and monoclonal anti-(human IL-2) antibody it was demonstrated that there was indeed IL-2 in the urine samples. In 8 of 11 patients the presence of IL-2 in the urine was demonstrated. The IL-2 production was directly related to the BCG administration as samples obtained just before the BCG instillation were always negative. In IL-2-positive samples a maximum level of IL-2 was observed between 2 h and 6 h after the BCG instillation. In urine samples obtained 24 h after the BCG IL-2 was not detected. In most patients the urine became positive after the third or fourth PCG instillation.

BCG Vaccine↗
PubMed2006

[Interstitial cystitis. An important differential diagnosis].

Interstitial cystitis (IC) is a chronic, debilitating inflammatory disease of the urinary bladder, characterized by chronic pelvic pain, urgency/frequency symptoms, and decrease of the functional bladder capacity. Both the etiology and pathogenesis of the condition that predominantly affects women are still not fully understood. Thus, a causal therapeutic approach still does not exist. IC remains a strict diagnosis of exclusion; distinct histopathological findings within the bladder wall are frequently found, although they are not pathognomonic. The diagnostic criteria elaborated by the NIH are helpful in clinical research; their dogmatic utilization will however lead to an underdiagnosis of IC in 60% of the patients. IC-specific symptom questionnaires and validated symptom scales help the physician to assess the therapeutic response during the follow-up of the patients.

Administration, Intravesical↗
PubMed2001

Production and characterisation of a C595 antibody-99mTc conjugate for immunoscintigraphy of bladder cancer.

Current radiological techniques for staging bladder cancer are inaccurate, especially in the identification of pelvic lymph node metastases. Immunoscintigraphy has the potential to offer improved staging for bladder cancer. The aim of this study was to label the anti-MUC1 monoclonal antibody C595 with 99mtechnetium (Tc), the most widely used diagnostic radionuclide, and assess the potential of the resultant conjugate for intravenous immunoscintigraphy of bladder cancer. A direct, reduction-mediated technique was used to label the antibody. The resultant conjugate was shown to be highly immunoreactive, stable and bound specifically to MUC1. The ability of the conjugate to bind to bladder tumours was demonstrated in an ex vivo model where the mean tumour:normal urothelial uptake was 5.7:1 and by intravesical administration in patients with bladder cancer where the mean tumour:normal urothelial uptake was 20.4:1. The ability of the conjugate to localise MUC1-expressing tumours was demonstrated in a nude mouse xenograft model. A conjugate of 99mTc-C595 has been produced and characterised, and it may be suitable for intravenous immunoscintigraphy, a potential novel staging tool for bladder cancer.

Animals↗
PubMed1998

Targeted diagnosis of bladder and ureteral carcinoma using radiolabelled BDI-1.

The aim of this study is to investigate the possibility of radioimmunoimaging (RII) by radiolabelled anti-bladder carcinoma monoclonal antibody BDI-1 applied to diagnosis of bladder cancer and ureteral cancer. BDI-1 was labelled with 131I and 99mTc. The immunoreactivity, pharmacokinetics and biodistribution in mice were studied. RII was performed in 46 patients. The results showed that 131I, 99mTc-BDI-1 have satisfactory immunoreactivity and excellent tumor-locating properties. The blood clearance half-life T1/2alpha and T1/2beta were 35 h in the first phase and 151 h in the second phase, respectively. Thirty-nine patients were studied by an intravesical administration method; the sensitivity was 90.5%. Seven patients were studied by an intravenous administration method. The RII results of three cases with primary or recurrent bladder cancer and three cases with ureteral cancer were confirmed histologically. RII was negative in one patient with suspected lung metastasis that was shown on radiography. The investigation revealed that RII can be used as an auxiliary method for the detection of bladder cancer and may be valuable for the diagnosis of ureteral cancer.

Aged↗