Surface-modified nanoparticles enhance transurothelial penetration and delivery of survivin siRNA in treating bladder cancer.

Penetration of the bladder permeability barrier (BPB) is a major challenge when treating bladder diseases via intravesical delivery. To increase transurothelial migration and tissue and tumor cell uptake, poly(lactic-co-glycolic acid; PLGA) nanoparticles (NP) were modified by addition of a low molecular weight (2.5 or 20 kDa) positively charged mucoadhesive polysaccharide, chitosan, to the NP surface. In designing these NPs, we balanced the adhesive properties of chitosan with the release and bioactivity of the siRNA. Chitosan-functionalized NPs demonstrated increased binding to and uptake in intravesically instilled mouse bladders and human ureter at 10 times the level of unmodified NPs. Furthermore, we extended the bioactivity of survivin siRNA in vitro for up to 9 days and demonstrated a decrease in proliferation when using chitosan-modified NPs relative to unmodified NPs. In addition, treatment of xenograft tumors with chitosan-modified NPs that encapsulate survivin siRNA (NP-siSUR-CH2.5) resulted in a 65% reduction in tumor volume and a 75% decrease in survivin expression relative to tumors treated with blank chitosan NPs (NP-Bk-CH2.5). Our low molecular weight chitosan delivery system has the capacity to transport large amounts of siRNA across the urothelium and/or to the tumor site, thus increasing therapeutic response.

Nanoparticles for urothelium penetration and delivery of the histone deacetylase inhibitor belinostat for treatment of bladder cancer.

Nearly 40% of patients with non-invasive bladder cancer will progress to invasive disease despite locally-directed therapy. Overcoming the bladder permeability barrier (BPB) is a challenge for intravesical drug delivery. Using the fluorophore coumarin (C6), we synthesized C6-loaded poly(lactide-co-glycolide) (PLGA) nanoparticles (NPs), which were surface modified with a novel cell penetrating polymer, poly(guanidinium oxanorbornene) (PGON). Addition of PGON to the NP surface improved tissue penetration by 10-fold in intravesically-treated mouse bladder and ex vivo human ureter. In addition, NP-C6-PGON significantly enhanced intracellular uptake of NPs compared to NPs without PGON. To examine biological activity, we synthesized NPs that were loaded with the histone deacetylase (HDAC) inhibitor belinostat (NP-Bel-PGON). NP-Bel-PGON exhibited a significantly lower IC50 in cultured bladder cancer cells, and sustained hyperacetylation, when compared to unencapsulated belinostat. Xenograft tumors treated with NP-Bel-PGON showed a 70% reduction in volume, and a 2.5-fold higher intratumoral acetyl-H4, when compared to tumors treated with unloaded NP-PGON. FROM THE CLINICAL EDITOR: These authors demonstrate that PLGA nanoparticles with PGON surface functionalization result in greatly enhanced cell penetrating capabilities, and present convincing data from a mouse model of bladder cancer for increased chemotherapy efficacy.

A phase 2 cancer chemoprevention biomarker trial of isoflavone G-2535 (genistein) in presurgical bladder cancer patients.

The soy compound genistein has been observed preclinically to inhibit bladder cancer growth with one potential mechanism being the inhibition of epidermal growth factor receptor phosphorylation (p-EGFR). A phase 2 randomized, placebo-controlled trial investigated whether daily, oral genistein (300 or 600 mg/d as the purified soy extract G-2535) for 14 to 21 days before surgery alters molecular pathways in bladder epithelial tissue in 59 subjects diagnosed with urothelial bladder cancer (median age, 71 years). G-2535 treatment was well tolerated; observed toxicities were primarily mild to moderate gastrointestinal or metabolic and usually not attributed to study drug. Genistein was detected in plasma and urine of subjects receiving G-2535 at concentrations greater than placebo subjects' but were not dose-dependent. Reduction in bladder cancer tissue p-EGFR staining between the placebo arm and the combined genistein arms was significant at the protocol-specified significance level of 0.10 (P = 0.07). This difference was most prominent when comparing the 300-mg group with placebo (P = 0.015), but there was no significant reduction in p-EGFR staining between the 600-mg group and placebo. No difference in normal bladder epithelium p-EGFR staining was observed between treatment groups. No significant differences in tumor tissue staining between treatment groups were observed for COX-2, Ki-67, activated caspase-3, Akt, p-Akt, mitogen-activated protein kinase (MAPK), or p-MAPK. No significant differences in urinary survivin or BLCA-4 levels between treatment groups were observed. Genistein displayed a possible bimodal effect (more effective at the lower dose) on bladder cancer tissue EGFR phosphorylation that should be evaluated further, possibly in combination with other agents.

Effect of mitomycin C on concentrations of vascular endothelial growth factor and its receptors in bladder cancer cells and in bladders of rats intravesically instilled with mitomycin C.

OBJECTIVES: • To examine, using in vitro and in vivo models, the largely unexamined effect of mitomycin C (MMC), an effective intravesical treatment for superficial bladder cancer and carcinoma in situ, on expression of vascular endothelial growth factor (VEGF) and vascular endothelial growth factor receptor-2 (VEGFR-2), which mediates many of the angiogenic properties of VEGF. • To measure, as a positive control, concentrations of the inhibitor of apoptosis, survivin, as an assessment of MMC effectiveness. • To measure MMC-induced changes in proliferation in the presence and absence of VEGF-A small interfering RNA (siRNA). MATERIALS AND METHODS: • After treatment with increasing MMC concentrations (5-200 µg/mL), we measured proliferation, as well as VEGF, survivin, VEGF receptor-1 (VEGFR-1) and VEGFR-2 concentrations in RT-4 and T-24 bladder cancer cells. • The effect of pre-treatment of VEGF siRNA and survivin siRNA on MMC-induced decreases in proliferation was measured. • Urinary VEGF concentrations and bladder and kidney concentrations of VEGF-A, VEGFR-1, VEGFR-2 and interleukin-6 (IL-6) mRNA were measured in rats intravesically instilled with saline or MMC (200 µg/mL). RESULTS: • Although MMC treatment inhibited cell proliferation and decreased survivin mRNA expression in T-24 and RT4 cells, MMC (12-50 µg/mL) increased VEGF-A mRNA and VEGFR-2 mRNA and protein expression. • Pre-treatment with VEGF-A siRNA or survivin siRNA before MMC treatment reduced proliferation more than MMC alone. • MMC-induced reductions in proliferation were reduced additively by pre-treatment with survivin siRNA, but were potentiated by pre-treatment with VEGF-A siRNA. • VEGFR-2 mRNA and protein concentrations and urinary VEGF concentrations were increased in bladders of rats instilled with MMC. CONCLUSIONS: • Intravesically instilled MMC increases urinary VEGF and bladder VEGFR-2 protein and mRNA in rats. • MMC increases VEGF mRNA and VEGFR-2 protein and mRNA concentrations in bladder cancer cells. Therefore, we speculate that MMC could increase the angiogenic potential of both cancer and normal cells. • In cancer cells this effect is largest at lower MMC concentrations. • Combining MMC with agents that reduce EGF concentrations could be of value in treatment of transitional cell carcinoma of the bladder (TCC).

Urine markers do not predict biopsy findings or presence of bladder ulcers in interstitial cystitis/painful bladder syndrome.

PURPOSE: We tested for associations between urine markers, bladder biopsy features and bladder ulcers in interstitial cystitis/painful bladder syndrome. MATERIALS AND METHODS: Subjects were 72 patients with interstitial cystitis/painful bladder syndrome undergoing bladder distention and biopsy. Urine was collected before the procedure. Urine marker levels were correlated with biopsy and cystoscopic findings. Patients with no previous interstitial cystitis/painful bladder syndrome treatments (47) were analyzed separately from previously treated patients (25). RESULTS: For untreated patients urine interleukin-6 and cyclic guanosine monophosphate were associated with urothelial epidermal growth factor receptor staining (for interleukin-6 r = 0.29; 95% CI 0.07, 0.51; p = 0.01 and for cyclic guanosine monophosphate r = 0.34; 95% CI 0.13, 0.55; p = 0.002). Urine interleukin-8 was negatively associated with urothelial heparin-binding epidermal growth factor-like growth factor staining (r = -0.34; 95% CI -0.55, -0.12; p = 0.002) and positively associated with lamina propria mast cell count (r = 0.29; 95% CI 0.06, 0.52; p = 0.01). The latter association also was seen in treated patients (r = 0.46; 95% CI 0.20, 0.73; p <0.001). None of the urine markers was significantly different for ulcer vs nonulcer groups. All of the patients with ulcer had extensive inflammation on bladder biopsy including severe mononuclear cell infiltration, moderate or strong interleukin-6 staining in the urothelium and lamina propria, and leukocyte common antigen staining in more than 10% of the lamina propria. However, these features also were seen in 24% to 76% of the patients without ulcer. CONCLUSIONS: Overall urine markers did not associate robustly with biopsy findings. The strongest association was a positive association between urine interleukin-8 levels and bladder mast cell count. Patients with ulcer consistently had bladder inflammation but the cystoscopic finding of ulcers was not a sensitive indicator of inflammation on bladder biopsy.

Downregulation of survivin is associated with reductions in TNF receptors' mRNA and protein and alterations in nuclear factor kappa B signaling in urothelial cancer cells.

Because survivin is selectively expressed in and associated with an unfavorable prognosis in transitional cell carcinoma of the bladder (TCC), we treated T-24 cells with survivin siRNA. Survivin siRNA treatment caused a profound decrease of survivin protein that was associated with decreased cell growth, a specific G2/M arrest and increased cytochrome c release. Microarray analysis of apoptosis genes showed that levels of 14/114 gene products were decreased after 72 hours treatment with survivin siRNA, including survivin, three TNF receptors, Akt, c-Abl, caspases and their related genes and Bcl-2 and NF-kappaB signaling related genes. TNFR1, pro-caspase-2 and Akt protein levels were decreased after survivin siRNA treatment for 48 and 72 hours. Downregulation of survivin causes changes in mitosis and apoptosis, which may be related to changes in TNF receptors and NF-kappaB signaling.

Changes in urine markers and symptoms after bladder distention for interstitial cystitis.

PURPOSE: We evaluated changes in urine markers and symptom scores after bladder distention in patients with interstitial cystitis. MATERIALS AND METHODS: Study subjects were 33 new patients who had undergone no prior interstitial cystitis treatment. Urine specimens were taken before and 1 month after bladder distention. University of Wisconsin symptom scores were done the same day as the urine specimen collection. Urine marker levels and symptom scores before and after distention were compared. Changes in markers were tested for associations with changes in symptom scores and other markers. Markers and specific symptoms before distention were tested for their association with post-distention symptom improvement. RESULTS: After distention the median total University of Wisconsin score decreased significantly (28.5 before, 20 after, p <0.001). A total of 12 patients (36%) had at least 30% improvement in University of Wisconsin score, and 8 patients (24%) had at least 50% improvement. No pre-distention markers or symptoms predicted which patients would have a good response. There were 2 urine markers that improved significantly after distention: anti-proliferative factor activity (median -96% before, -17% after, p <0.001) and heparin-binding epidermal growth factor-like growth factor levels (median 0.34 ng/mg creatinine before, 4.1 after, p <0.001). None of the changes in urine markers associated with changes in symptom scores. CONCLUSIONS: The median symptom score for newly diagnosed patients with interstitial cystitis decreased after distention, but only a minority of patients had at least 30% symptom improvement. Bladder distention altered urine anti-proliferative factor activity and heparin-binding epidermal growth factor-like growth factor levels toward normal, but the mechanism of symptom relief after distention is still unknown.

Pacemaker activity in the upper urinary tract.

Ureteral peristaltic activity begins with the origin of electrical activity at pacemaker sites. These sites are located in the proximal portion of the urinary collecting system. The 'atypical' smooth muscle cells at these sites fire 'pacemaker' potentials at a frequency higher than the 'driven' action potentials recorded from typical smooth muscle cells. In contrast to typical smooth muscle cells, these atypical pacemaker cells have less than 40% of their cellular area occupied by contractile filaments and demonstrate a sparse immunoreactivity for alpha-smooth muscle actin. Expression of c-Kit, a tyrosine kinase receptor, correlates with the onset of organized ureteral peristalsis in the embryo. Capsaicin-sensitive sensory afferents and the endogenous release of tachykinins and prostaglandins are involved in the maintenance of normal ureteral peristalsis.

A role for Akt in the rapid regulation of inflammatory and apoptotic pathways in mouse bladder.

Akt is linked to both inflammatory and neoplastic pathways. Akt activation is dependent on the phosphatidylinositol-3 kinase (PI3K) signaling pathways. Upon phosphorylation by PI3K, Akt can phosphorylate nuclear factor kappa B (NF-kappaB) and members of the forkhead family of transcription factors, which includes AFX. Our goal is to examine the effect of Escherichia coli lipopolysaccharide (LPS) on early cellular signaling in inflammatory (NF-kappaB) and apoptotic pathways (AFX) in a mouse-bladder model and in T-24 urothelial cancer cells. Female C57BL/6 mice were given an intraperitoneal (IP) injection of LPS or LPS free water and sacrificed 0-120 minutes later. Bladders were harvested, and immunohistochemistry (IHC) and/or immunoblotting performed using antibodies to PI3K, inhibitor kappa B-alpha (IkappaB-alpha), and total and phosphorylated Akt, NF-kappaB and AFX. Levels of IkappaB-alpha and total and phosphorylated Akt and NF-kappaB were determined in T-24 cells treated with LPS for 0-120 minutes. Bladders and T-24 cells were treated with PI3K inhibitors in some experiments. Protein amounts in different samples were normalized to immunoreactive actin. Phosphorylated and non-phosphorylated species of Akt, NF-kappaB, and AFX were localized to the urothelium. IP LPS injection rapidly (within 30 minutes) increased Akt phosphorylation. IP LPS injection decreased IkappaB-alpha levels, and increased NF-kappaB and AFX phosphorylation. Wortmannin effectively blocked phosphorylation of Akt in LPS-treated mice, and also reduced phosphorylation of AFX and, to a lesser extent, NF-kappaB. After treatment with LPS, Akt and NF-kappaB phosphorylation was rapidly increased in T-24 cells. Akt phosphorylation, and to a lesser extent NF-kappaB phosphorylation, were blocked by LY-294,002. LPS/PI3K/Akt is a cellular signaling pathway which rapidly activates downstream pathways of inflammation and neoplasia in bladder urothelium.

Regulation of cyclic nucleotides in the urinary tract.

Cyclic nucleotide levels are controlled through their synthesis from nucleotide triphosphates by cyclases and their degradation to 5'-monophosphates by phosphodiesterases (PDEs). Components controlling cyclic AMP-induced relaxation in the urinary tract include receptors, inhibitory and stimulatory G-proteins, isoforms of adenylyl cyclase and PDEs. The responsiveness of PDEs to a variety of physiological challenges is related to the presence of multiple families of isoenzymes with specific localization within tissues and within cells. At least 11 families of PDEs encode more than 50 PDE proteins produced in mammalian cells. In the urinary tract, characterization of PDE isoforms has lagged behind other systems and much of the literature was published prior to identification of PDE7, 8, 9, 10, 11. Specific PDE inhibitors regulate smooth muscle function in the bladder, urethra, prostate and ureter. The pharmacological potential of these inhibitors may include treatment of urge incontinence and the low compliance bladder, and treatment of prostate cancer. G-proteins also regulate cyclic AMP production. Changes in specific G- protein isoforms with aging, most prominently Gialpha2, cause decreased relaxation response in the aging bladder. As we have seen here with aging and certainly in other disease processes, levels of the components of adenylyl cyclase/phosphodiesterase/protein kinase can change and thus affect the relaxation response. By exploitation of differences in PDE expression in disease, such as the overexpression of PDEs in cancer, treatment options may present themselves.

Do the National Institute of Diabetes and Digestive and Kidney Diseases cystoscopic criteria associate with other clinical and objective features of interstitial cystitis?

PURPOSE: We compared urine markers, bladder biopsy findings and clinical features of patients with symptoms of interstitial cystitis (IC) who did or did not meet the cystoscopic criteria defined by the National Institute of Diabetes and Digestive and Kidney Diseases. MATERIALS AND METHODS: Urine markers and symptom questionnaires were measured before and 1 month after bladder distention for IC. Bladder biopsies were taken at the time of distention. At distention patients were defined as meeting or not meeting the National Institute of Diabetes and Digestive and Kidney Diseases established cystoscopic criteria for IC. The 2 patient groups were compared. RESULTS: Urine marker levels were similar for the 2 groups, including antiproliferative factor, epidermal growth factor, heparin binding epidermal growth factor-like growth factor, cyclic guanosine monophosphate, interleukins 6 and 8, and methylhistamine. Bladder biopsy features were similar for the 2 groups. Bladder capacity with the patients under anesthesia was higher in those who did not meet the criteria (median 750 vs 1,000 ml, p = 0.005). Median age at symptom onset was 26 years in both groups. On the University of Wisconsin symptom scale patients who met the criteria had higher scores for daytime frequency (p = 0.002) and nocturia (p = 0.01). Symptom characteristics and symptom response after bladder distention were similar for the 2 groups. CONCLUSIONS: Patients who met the cystoscopic criteria had worse daytime frequency and nocturia, and lower bladder capacity under anesthesia. However, the 2 groups had similar urine markers and bladder biopsy findings. The cystoscopic criteria do not appear to identify a distinct pathophysiological subset of patients with IC symptoms.

Rapid up-regulation of endothelial nitric-oxide synthase in a mouse model of Escherichia coli lipopolysaccharide-induced bladder inflammation.

Increases in the signaling molecule nitric oxide (NO) during inflammation may be linked not only to inducible nitric-oxide synthase (iNOS) but also to endothelial (e)NOS. Escherichia coli lipopolysaccharide (LPS) induces an inflammatory response in the bladder and rapidly increases phosphorylation of Akt/protein kinase B (Akt), a key enzyme regulating proliferation, apoptosis, and inflammation. Activated Akt phosphorylates human eNOS at serine 1177 and subsequently increases NOS activity. Because Akt and eNOS are both localized in the bladder urothelium, phosphorylation of eNOS by Akt provides an attractive mechanism for rapid increases in urinary NO production. Female mice were intraperitoneally injected with LPS (25 mg/kg) or pyrogen-free water (control). Four hours before LPS injection, some mice were injected with wortmannin, which inhibits Akt phosphorylation. Levels of urinary cyclic GMP, a downstream product of NO, increase 75% within 1 h after intraperitoneal injection of LPS, and this increase is blocked by wortmannin. Bladder eNOS and phosphorylated eNOS protein increase 94 and 151%, respectively, 1 h after LPS treatment, whereas iNOS was not detected. Wortmannin decreases eNOS phosphorylation by 60%. Furthermore, bladder Ca(2+)-dependent NOS activity (eNOS, neuronal NOS) is increased 79 +/- 20% 1 h after LPS treatment, whereas there is no increase in Ca(2+)-independent (iNOS) activity (n = 4). Increases in urinary cyclic GMP, NOS activity, and eNOS protein and phosphorylation 1 h after induction of inflammation with LPS, indicate that eNOS plays a role in the early response to bladder inflammation.

Urinary interleukin-8 levels are elevated in subjects with transitional cell carcinoma.

OBJECTIVES: Interleukin (IL)-8 is one of several angiogenic factors produced in bladder cancer cell lines. Although many studies have indicated that IL-8 is increased in infectious/inflammatory processes, elevated levels of IL-8 in urine also may be indicative of active transitional cell carcinoma (TCC). METHODS: Urinary IL-8 levels were measured with an enzyme-linked immunosorbent assay in subjects with TCC (n = 51), prostate cancer (n = 17), or a history of successfully treated TCC (n = 23) and in healthy subjects (n = 49). In addition, urinary IL-8 levels were measured in 21 subjects before, during, and 1 month after intravesical therapy with bacille Calmette-Guérin or mitomycin C. RESULTS: The median urinary IL-8 levels were greater in subjects with TCC (64 pg/mL urine) than in healthy subjects (6 pg/mL urine), subjects with prostate cancer (0.5 pg/mL urine), or subjects with a history of successfully treated TCC (5.0 pg/mL urine). Urinary IL-8 levels were greater in subjects with invasive (high-stage) TCC than in those with low-stage TCC. Furthermore, the postintravesical instillation levels of urinary IL-8 levels were greater in patients whose TCC recurred compared with those whose TCC was in remission. CONCLUSIONS: IL-8 levels were greater in subjects with TCC compared with those with successfully treated TCC. IL-8 levels increased with TCC stage, indicating that they are greater in more invasive (ie, angiogenic) tumors. A reduction in urinary IL-8 levels after treatment with bacille Calmette-Guérin or mitomycin C may reflect a decrease in bladder cancer cells and/or in other cells that produce IL-8.

Effect of intravesical treatment of transitional cell carcinoma with bacillus Calmette-Guerin and mitomycin C on urinary survivin levels and outcome.

PURPOSE: Urine survivin is a predictive/prognostic molecular marker that detects transitional cell carcinoma (TCC) with high specificity and sensitivity. The presence of urine survivin in patients with TCC who receive intravesical instillation of bacillus Calmette-Guerin or mitomycin C may predict recurrence. MATERIALS AND METHODS: Urine from 25 subjects receiving 27 intravesical treatments of bacillus Calmette-Guerin or mitomycin C for TCC were collected prior to, during and after treatment. Urinary survivin levels were compared with outcome, as assessed by cytology and cystoscopy with or without biopsy 1 month and up to 12 months after the completion of treatment. RESULTS: Pretreatment survivin levels were higher in subjects in whom TCC recurred following treatment compared with those who achieved remission. Survivin levels increased several-fold during treatment with the highest survivin levels measured in subjects with recurrence. Median posttreatment values of survivin were zero in those who achieved remission and 1.0 ng/ml urine in subjects in whom TCC recurred. CONCLUSIONS: The presence of urinary survivin 1 month after the completion of treatment predicts TCC recurrence with 100% sensitivity and 78% specificity. Specificity to predict TCC recurrence increases to 92% after 1 year. No TCC recurred for 1 year in 12 of the 14 subjects with a posttreatment survivin level of 0.1 ng or less per ml urine. Three of the 4 subjects who were survivin positive but in remission 1 month after the completion of treatment had recurrent TCC within 1 year. Subjects who have urinary survivin after the completion of intravesical instillation have a high likelihood of TCC recurrence.

Prostaglandin E2 production and cyclooxygenase-2 induction in human urinary tract infections and bladder cancer.

PURPOSE: Increased prostaglandin production correlates positively with cancer risk and cyclooxygenase (COX)-2, the inducible rate limiting enzyme for prostaglandin synthesis, is elevated in bladder cancer cases. Urinary prostaglandin levels and COX-2 expression in urine particulates may increase in urogenital cancer, including bladder cancer, and with infectious and inflammatory processes, including urinary tract infections and that resulting from bacillus Calmette-Guerin (BCG) treatment for bladder cancer. MATERIALS AND METHODS: Urinary prostaglandin E2 levels were measured in patients with a urinary tract infection before and after treatment, urogenital cancer (including bladder cancer), bladder cancer in remission and bladder cancer with BCG treatment. COX-1 and COX-2 mRNA and protein were assessed in the human ureter, in normal human bladder muscle and urothelium, and in urine particulates from patients with urinary tract infections, bladder cancer and bladder cancer with BCG treatment. RESULTS: COX-1 protein, and mRNA and COX-2 mRNA were expressed in the ureter, and bladder muscle and urothelium. Urinary prostaglandin E2 levels and COX-2 protein expression in urine particulates were elevated in patients with urinary tract infections and with bladder cancer compared with age matched controls. Successful treatment for urinary tract infections and bladder cancer lowers urinary prostaglandin E2 levels. Urinary prostaglandin E2 and COX-2 protein levels are elevated during BCG treatment. CONCLUSIONS: Increased urinary prostaglandin E2 production and COX-2 protein expression correlate with urogenital cancer, urinary tract infections and inflammatory processes, such as those induced by BCG. Patients in whom urinary tract infection was treated with antibiotics or in whom bladder cancer is in remission have reduced urinary prostaglandin E2 compared with those who have active disease.

A comparison of multiple urine markers for interstitial cystitis.

PURPOSE: We measured several urine markers in 24-hour specimens from patients with interstitial cystitis and healthy controls. For each marker we determined whether the urine level was significantly different in interstitial cystitis and control cases, and whether the marker level correlated with the symptom score. MATERIALS AND METHODS: Study participants included 36 female patients with interstitial cystitis and 36 age matched female volunteers. Multiple urine aliquots were obtained to measure the various markers. RESULTS: Certain markers were significantly increased in interstitial cystitis, including anti-proliferative factor, epidermal growth factor, insulin-like growth factor (IGF) binding protein-3 and interleukin (IL)-6. Markers significantly decreased in interstitial cystitis were heparin-binding epidermal growth factor-like growth factor, cyclic guanosine monophosphate and methylhistamine. Other markers were not significantly different in the interstitial cystitis and control groups, including total glycosaminoglycans, epitectin, hyaluronic acid, IL-8, IL-1 and nitrates plus nitrites. IGF-1 was undetectable in 24-hour urine samples but spot voided samples from the same interstitial cystitis population had IGF-1 levels similar to previously reported levels. The only significant association of marker with symptom score was a positive correlation of IL-6 with nocturia. For all markers the conclusions were the same whether the marker was normalized to creatinine or to 24 hours. CONCLUSIONS: This study confirmed several previously reported urine alterations in interstitial cystitis, including increased anti-proliferative factor, epidermal growth factor, IGF binding protein-3 and IL-6, and decreased heparin-binding epidermal growth factor-like growth factor and cyclic guanosine monophosphate. Of all markers studied anti-proliferative factor had the least overlap in the interstitial cystitis and control groups, and so it is the most likely candidate to become a diagnostic test.

Bladder cancer detection with urinary survivin, an inhibitor of apoptosis.

The current "gold standard" for the diagnosis of bladder cancer is cystoscopy and urine cytology. Cystoscopy, a naked eye assessment of the bladder, is invasive, uncomfortable and costly while cytology has high specificity but low sensitivity (40-60%) particularly for low-grade lesions. Therefore, there is a need for a molecular tumor marker assay that is simple to perform and sensitive, particularly for low-grade lesions. By looking to the pathophysiology of bladder cancer, we identified survivin, an inhibitor of apoptosis that is not generally expressed in fully differentiated adult tissue and is highly expressed in bladder cancer. Survivin is detected in whole urine of patients with TCC using a simple antibody based test. The sensitivity of survivin testing for new or recurrent bladder cancer is 100% while the specificity for other neoplastic and non-neoplastic genitourinary disease is 95%. The high sensitivity of this simple, noninvasive test is well suited to bladder cancer, a disease with high rates of recurrence.