Urine Of Interstitial Cystitis Patients Research Articles

Role of urinary cations in the etiology of interstitial cystitis: A multisite study.

To determine whether patients with interstitial cystitis have elevated levels of toxic urinary cations, to identify and quantify these cationic metabolites, and to assess their cytotoxicity. Isolation of cationic fraction was achieved by solid phase extraction using an Oasis MCX cartridge on urine specimens from interstitial cystitis patients and controls. C18 reverse phase high-performance liquid chromatography was used to profile cationic metabolites, and they were quantified by the area under the peaks and normalized to creatinine. Major cationic fraction peaks were identified by reverse phase high-performance liquid chromatography and liquid chromatography-mass spectrometry. HTB-4 urothelial cells were used to determine the cytotoxicity of cationic fraction and of individual metabolites. The reverse phase high-performance liquid chromatography analysis was carried out on cationic fraction metabolites isolated from urine samples of 70 interstitial cystitis patients and 34 controls. The mean for controls versus patients was 3.84 (standard error of the mean 0.20) versus 6.71 (0.37) mAU*min/µg creatinine, respectively (P=0.0001). The cationic fraction cytotoxicity normalized to creatinine for controls versus patients in mean percentage was -7.79% (standard error of the mean 3.32%) versus 20.03% (standard error of the mean 2.75%; P<0.0005). The major toxic cations were 1-methyladenosine, 1-methylguanine, N2 ,N2 -dimethylguanosine and L-tryptophan. These data confirm significant elevation of toxic cations in the urine of interstitial cystitis patients. These toxic cations likely represent a primary cause of interstitial cystitis, as they can injure the bladder mucus and initiate an epithelial leak.

1169 - Concentration of high mobility group box 1 proteins increases in the urine of interstitial cystitis patients in correlation with epithelial growth factor

1169 - Concentration of high mobility group box 1 proteins increases in the urine of interstitial cystitis patients in correlation with epithelial growth factor

MP72-01 URINE LEVELS OF MACROPHAGE MIGRATION INHIBITORY FACTOR ARE INCREASED IN PATIENTS WITH BLADDER INFLAMMATION

You have accessJournal of UrologyInfections/Inflammation/Cystic Disease of the Genitourinary Tract: Interstitial Cystitis1 Apr 2016MP72-01 URINE LEVELS OF MACROPHAGE MIGRATION INHIBITORY FACTOR ARE INCREASED IN PATIENTS WITH BLADDER INFLAMMATION Fei Ma, Dimotrios E. Kouzoukas, Magnus M. Fall, Robert Moldwin, David M. Preston, Deborah R. Erickson, Barbara Kahn, Katherine Meyer-Siegler, and Pedro L. Vera Fei MaFei Ma More articles by this author , Dimotrios E. KouzoukasDimotrios E. Kouzoukas More articles by this author , Magnus M. FallMagnus M. Fall More articles by this author , Robert MoldwinRobert Moldwin More articles by this author , David M. PrestonDavid M. Preston More articles by this author , Deborah R. EricksonDeborah R. Erickson More articles by this author , Barbara KahnBarbara Kahn More articles by this author , Katherine Meyer-SieglerKatherine Meyer-Siegler More articles by this author , and Pedro L. VeraPedro L. Vera More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2016.02.1614AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Macrophage migration inhibitory factor (MIF) is a pro-inflammatory cytokine, which appears to mediate bladder inflammation and pain in rodents (Vera et al, 2010; Kouzoukas et al, 2014). We hypothesized that urinary MIF levels would increase in patients with inflammatory diseases of the bladder, and therefore examined urine from patients with conditions including: interstitial cystitis (IC) with and without Hunner lesions, urinary tract infection (UTI), and radiation cystitis. We compared these samples to those of a control population. METHODS Urine samples were collected and studied in 3 male and 3 female groups: Specimens from males included those from patients with culture confirmed UTI, those with radiation cystitis (after undergoing radiation therapy) and a control population. Specimens from females include those from IC patients with and without Hunner lesions, and those from control subjects. Urine samples from patients with Interstitial Cystitis met NIDDK diagnostic criteria. All IC patients were symptomatic at the time of urine collection. Urinary MIF and Creatinine levels were assayed using commercially available kits (R&D; Enzo). RESULTS MIF levels were significantly increased in the urine of UTI patients (>4-fold; N=51) or radiation cystitis patients (10-fold; N=18) when compared to controls (N=117), either as total concentration or normalized to creatinine levels (Table 1). Similarly, MIF levels were significantly increased (5-fold; Table 2) in the urine of IC patients with Hunner lesions (N=47) when compared to female controls (N=102). Urine MIF levels in IC patients without Hunner lesions (N=63) were not different from controls. CONCLUSIONS Urinary MIF is increased in a variety of bladder inflammatory conditions including interstitial cystitis with Hunner lesions, UTI and radiation cystitis. Conversely, urine MIF levels from IC patients without Hunner lesions are similar to controls. These findings may represent differences in pathogenesis between these conditions and may serve as a marker to distinguish them from each other. MIF may be useful for diagnosis or prognosis of IC patients with Hunner lesions and may represent a novel therapeutic target for reducing bladder pain in IC and other bladder inflammatory disorders. © 2016FiguresReferencesRelatedDetails Volume 195Issue 4SApril 2016Page: e952 Advertisement Copyright & Permissions© 2016MetricsAuthor Information Fei Ma More articles by this author Dimotrios E. Kouzoukas More articles by this author Magnus M. Fall More articles by this author Robert Moldwin More articles by this author David M. Preston More articles by this author Deborah R. Erickson More articles by this author Barbara Kahn More articles by this author Katherine Meyer-Siegler More articles by this author Pedro L. Vera More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Interstitial cystitis antiproliferative factor (APF) as a cell-cycle modulator.

BackgroundInterstitial cystitis (IC) is a chronic bladder disorder of unknown etiology. Antiproliferative factor (APF), a peptide found in the urine of IC patients, has previously been shown to decrease incorporation of thymidine by normal bladder epithelial cells. This study was performed to determine the effect of APF on the cell cycle of bladder epithelial cells so as to better understand its antiproliferative activity.MethodsExplant cultures from normal bladder biopsy specimens were exposed to APF or mock control. DNA cytometry was performed using an automated image analysis system. Cell cycle phase fractions were calculated from the DNA frequency distributions and compared by two-way analysis of variance (ANOVA).ResultsAPF exposure produced statistically significant increases in the proportion of tetraploid and hypertetraploid cells compared to mock control preparations, suggesting a G2 and/or M phase cell cycle block and the production of polyploidy.ConclusionsAPF has a specific effect on cell cycle distributions. The presence of a peptide with this activity may contribute to the pathogenesis of interstitial cystitis through disruption of normal urothelial proliferation and repair processes.

NF-kappaB-dependent gene expression of proinflammatory cytokines in T24 cells: possible role in interstitial cystitis.

Our previous report of predominant activation of nuclear transcription factor NF-kappaB in the bladder urothelium of interstitial cystitis (IC) patients suggests a potential role for this nuclear factor in the pathogenesis of the disease. Although NF-kappaB has been implicated in the pathogenesis of several inflammatory diseases, the downstream mechanism(s) by which it can mediate its effects are still fragmentary. In this study, we examined the role of this nuclear factor on the induction of proinflammatory cytokine gene expression in human bladder carcinoma T24 cells and further examined their corresponding protein levels in the urine of IC patients. T24 cells transduced with a dominant-negative super-repressor IkappaB mutant (pAxCAmIkappaB-M) or wild-type adenoviral vectors in the presence or absence of rhTNF-alpha. Transduction efficiency and ability of pAxCAmIkappaB-M to inhibit NF-kappaB activation were monitored by in situ reporter beta-galactosidase and gel mobility shift assays, respectively. Expression profile analysis of proinflammatory cytokines was measured in cells and urine of IC patients using RT-PCR and ELISA, respectively. The activation of NF-kappaB by rhTNF-alpha was associated with 27, eight, ten and sevenfold increases in the TNF-alpha, IL-1beta, IL-6 and IL-8 transcripts, respectively. In contrast, abrogation of the TNF-alpha-induced cytokine gene expression by an adenovirus super-repressor IkappaB mutant vector demonstrate that these effects were NF-kappaB-dependent. Interestingly, the NF-kappaB-induced expression of these transcripts correlates with increased protein levels of NF-kappaB-regulated proinflammatory factors in the urine of IC patients in comparison to controls. That these factors are capable of activating NF-kappaB in urothelial cells suggests a pivotal role for this nuclear transcription factor in the pathophysiology of the disease, possibly by inducing aberrant immune and inflammatory responses within the bladder of IC patients.

INTRAVESICAL SODIUM HYALURONATE INHIBITS THE RAT URINARY MAST CELL MEDIATOR INCREASE TRIGGERED BY ACUTE IMMOBILIZATION STRESS

Mast cell activation and stress have been suggested as factors in the pathogenesis of interstitial cystitis, a painful disorder of the bladder that is diagnosed more frequently in women and characterized by increased urgency and frequency with absent infection. Intravesical sodium hyaluronate has been used to treat interstitial cystitis due to its possible replenishment of bladder glycosaminoglycans. We investigated the effect of sodium hyaluronate on the activation of bladder mast cell and release of proinflammatory mediators in the urine induced by acute immobilization stress in rats. Using anesthesia a catheter was inserted in the bladder of 170 gm. female Sprague-Dawley rats. After emptying post-void residual urine a solution of normal saline, 0.08% or 0.4% sodium hyaluronate was introduced for 30 minutes. Each rat was allowed to recover from anesthesia and stressed for 30 minutes by confining it in a clear acrylic plastic immobilizer, while urine was continuously collected. Urinary histamine, rat mast cell protease-I and interleukin (IL)-6 were then determined. At the end of the experiments each rat was sacrificed by CO2 asphyxiation, and the bladder was removed and fixed with 4% paraformaldehyde. Frozen sections were stained with acidified toluidine blue, and the mast cell number and degree of activation were determined by granule extrusion and reduced cellular staining. Mean bladder mast cell activation plus or minus standard deviation in 6 control rats was 30.4% +/- 3.7% but it increased to 76.2% +/- 6.1% in 6 stressed animals (p = 0.0001). Intravesical administration of 0.4% sodium hyaluronate for 30 minutes in 6 rats before stress reduced mean bladder mast cell activation by 69.7% to 23.1% +/- 6.1% compared with stressed controls (p = 0.0003). However, compared to itself before stress there was no significant difference, indicating complete inhibition in 6 rats. Intravesical 0.08% sodium hyaluronate had a weaker inhibitory effect in 6 rats, decreasing mean degranulation by 22.5% to 59.1% +/- 7.6% (p = 0.02). In 6 rats stress increased the total mean amount of urinary rat mast cell protease-I by 271% from 0.14 +/- 0.09 to 0.52 +/- 0.17 ng. (p = 0.008). Pretreatment with 0.4% sodium hyaluronate reduced mean rat mast cell protease-I 80.8% compared with stressed controls (p = 0.008) and prevented any increase in response to stress in the same group of 8 rats with a mean pre-stress and post-stress level of 0.09 +/- 0.04 and 0.1 +/- 0.04 ng., respectively (p = 0.8). Acute stress increased mean urinary histamine in 6 rats 40.2% from 137.3 +/- 29.7 before to 193.7 +/- 7.6 ng./ml. after stress (p = 0.004). Pretreatment with 0.4% sodium hyaluronate reduced mean histamine 7.1% compared with stressed controls but completely prevented any increase in the same group of 8 rats, in which it was 174.5 +/- 23.1 before and remained 179.4 +/- 9.9 ng./ml. after stress (p = 0.75). Acute stress in 7 rats also increased the mean amount of IL-6 released in the urine by 31.5% from 775.9 +/- 69.2 to 1,021.1 +/- 93.3 pg./ml. (p = 0.007). Pretreatment with 0.4% sodium hyaluronate in 9 rats reduced mean IL-6 17% compared with stressed controls but again prevented any increase from baseline, since the value was 898.6 +/- 299.3 before and 824.4 +/- 196.4 pg./ml. after stress (p = 0.03). Immobilization stress induces bladder mast cell activation and the secretion of proinflammatory mediators, which are inhibited by sodium hyaluronate. Intravesical sodium hyaluronate may be a useful therapeutic option for interstitial cystitis, especially in patients with bladder mastocytosis who have symptom exacerbation with stress.

AUGMENTED STRETCH ACTIVATED ADENOSINE TRIPHOSPHATE RELEASE FROM BLADDER UROEPITHELIAL CELLS IN PATIENTS WITH INTERSTITIAL CYSTITIS

Extracellular adenosine triphosphate (ATP) has been shown to mediate inflammation and nociception and, therefore, it may have a role in symptoms associated with interstitial cystitis. We theorized that the bladder uroepithelium releases ATP in response to stretch and, furthermore, this process is augmented in interstitial cystitis. We quantitated ATP using the luciferin-luciferase assay. Urinary ATP levels were compared in 35 patients with interstitial cystitis and in 33 normal controls after pH correction. Cultured interstitial cystitis and normal urothelial cells from the bladder biopsies of 5 patients each were stretched with the Flexcell 2000 machine (Flexcell International Corp., McKeesport, Pennsylvania) and supernatant ATP concentrations were measured. Mean urinary ATP plus or minus standard error of mean was significantly higher in patients with interstitial cystitis than in controls (L value 985 +/- 161 versus 377 +/- 27, p = 0.0007). Supernatant ATP released by stretched interstitial cystitis cells was stretch intensity dependent when comparing 0%, 10% and 20% elongation, and was also significantly higher in stretched interstitial cystitis than in stretched normal cells. Adenosine triphosphate was significantly elevated in the urine of individuals with interstitial cystitis and the stretch activated release of ATP was augmented in interstitial cystitis urothelium. Increased extracellular ATP may have a role in mechanosensory transduction and to our knowledge it represents a novel hypothesis.

BLADDER EPITHELIAL CELLS FROM PATIENTS WITH INTERSTITIAL CYSTITIS PRODUCE AN INHIBITOR OF HEPARIN-BINDING EPIDERMAL GROWTH FACTOR-LIKE GROWTH FACTOR PRODUCTION

The etiology of interstitial cystitis is unknown. We previously identified an interstitial cystitis urine factor, antiproliferative factor, that inhibits proliferation of bladder epithelial cells in vitro and complex changes in epithelial growth factor levels, including profound decreases in heparin-binding epidermal growth factor-like growth factor (HB-EGF). Bladder and renal pelvic catheterization of patients with interstitial cystitis indicated that the antiproliferative factor is made and/or activated in the distal ureter or bladder. Therefore, we determined whether bladder epithelial cells from interstitial cystitis cases produced the antiproliferative factor and whether purified antiproliferative factor could alter production of growth factors known to be abnormal in interstitial cystitis. Antiproliferative factor activity was determined by 3H-thymidine incorporation into primary bladder epithelial cells. The antiproliferative factor was purified by size fractionation followed by sequential chromatography involving ion exchange, hydrophobic interaction and high performance liquid chromatography. HB-EGF, epidermal growth factor, insulin-like growth factor and insulin-like growth factor binding protein 3 levels were determined by enzyme-linked immunosorbent assay. Bladder epithelial cells from patients with interstitial cystitis produced a single antiproliferative factor with the same purification profile as that purified from interstitial cystitis urine. Purified antiproliferative factor specifically inhibited HB-EGF production by bladder epithelial cells in vitro, and the effect of interstitial cystitis urine or purified antiproliferative factor on bladder cell proliferation was inhibited by recombinant human HB-EGF in a dose dependent manner. Similar to urine HB-EGF, serum HB-EGF was also significantly lower in interstitial cystitis cases than in controls. Bladder epithelial abnormalities in interstitial cystitis may be caused by a negative autocrine growth factor that inhibits cell proliferation by down-regulating HB-EGF production. Furthermore, decreased levels of urine and serum HB-EGF indicate that interstitial cystitis may be a urinary tract manifestation of a systemic disorder.

CYTO-INJURY FACTORS IN URINE: A POSSIBLE MECHANISM FOR THE DEVELOPMENT OF INTERSTITIAL CYSTITIS

In most of our patients with interstitial cystitis (IC), the disease is associated with an increased urothelial permeability whose cause has not been identified. We postulate that both normal urine and the urine of IC patients contains factors capable of injuring the mucosa and causing an increased permeability that would allow urine components to leak into the bladder muscle. To test this hypothesis, we examined fractions of normal urine for toxic effects on bladder smooth muscle and epithelial cells in vitro. In the same in vitro system, we measured the effects of Tamm-Horsfall protein (THP), a normal urinary glycoprotein that may be a scavenger of injurious agents capable of "detoxifying" normal metabolic products. Human urothelial cells (T24) and rabbit bladder smooth muscle cells were incubated overnight with various fractions prepared from healthy volunteers' urine. The urine fractions of molecular weights >100 Da were incubated overnight with either urothelial or smooth muscle target cells after no treatment or after heating to 56C, preincubation with THP, exposure to heparin, or elution from heparin. Cytotoxicity was determined for each group using a neutral red uptake assay. Urine fractions of molecular weight 500 to 1000 Da were cytotoxic to smooth muscle cells (39%) and urothelial cells (50%). Cytotoxicity levels for THP-treated fractions were significantly lower than those for untreated fractions in both urothelial cells (7% versus 89%, p <0. 001) and smooth muscle cells (8% versus 70%, p <0.01). Fractions exposed to heparin were less cytotoxic to smooth muscle cells (20%) than were untreated fractions (27%). Fractions eluted from heparin were also cytotoxic to urothelial cells (42%). Normal human urine contains heat labile, cationic components of low molecular weight that bind to heparin. These components, when separated from the bulk of the urinary wastes, are cytotoxic to urothelial cells as well as underlying smooth muscle cells, indicating their potential for causing bladder mucosal injury. The cytotoxic activity can be blocked by the presence of THP. This urinary cytoprotective activity of THP may play an important but unrecognized role in the development of IC.

ANTIPROLIFERATIVE ACTIVITY IS PRESENT IN BLADDER BUT NOT RENAL PELVIC URINE FROM INTERSTITIAL CYSTITIS PATIENTS

To determine whether an antiproliferative urine factor that we previously discovered to be specific for urine from interstitial cystitis (IC) patients originated in the lower urinary tract or a more proximal site. Sequential catheterized urine specimens were collected under sterile conditions from the bladder and renal pelvis of 20 IC patients and one control patient (with stress incontinence). Antiproliferative activity was determined by 3H-thymidine incorporation of primary normal adult bladder epithelial cells cultured with pH- and osmolality-corrected bladder or ureteral urine specimens; significant inhibition was defined as a change in 3H-thymidine incorporation greater than 2 standard deviations from the mean of control cells. Bladder urine specimens from 19 of 20 IC patients significantly inhibited 3H-thymidine incorporation as compared to cell medium alone (mean change for bladder specimens = -68.7+/-7.5%), while a renal pelvic specimen from only 1 of 20 IC patients inhibited proliferation significantly (mean change for renal pelvic specimens = 3.2+/-3.4%) (p<.001 by Fisher's exact test). The one inhibitory IC renal pelvic specimen inhibited by 31% while a bladder specimen obtained during the same procedure inhibited by 94%. In comparison, neither bladder nor renal pelvic urine from the control patient had inhibitory activity. The antiproliferative factor previously found in the urine of IC patients appears to be made and/or activated in the distal ureter or urinary bladder.

A hypothesis for the etiology of interstitial cystitis based upon inhibition of bladder epithelial repair

Interstitial cystitis (IC) is a chronic bladder disease characterized by distinct bladder mucosal abnormalities, for which the etiology is unknown. Although the epidemiology of this disorder is similar to that of bacterial cystitis, prospective studies using sensitive culture techniques and polymerase chain reaction assay for a variety of microorganisms have failed to identify a specific infectious etiology for IC. We have identified a low-molecular-weight peptide in the urine of IC patients that inhibits the proliferation of normal bladder epithelial cells in vitro. We therefore propose a model of IC, in which this peptide inhibits bladder epithelial regeneration following damage (such as that caused by bacterial cystitis). The chronically damaged epithelium is prone to colonization with various microorganisms, and the resulting exposure to these microorganisms, other urinary antigens, and/or damaged epithelial cells prompts the low-level inflammatory response commonly seen in this disorder.

CONCENTRATIONS OF SPECIFIC EPITHELIAL GROWTH FACTORS IN THE URINE OF INTERSTITIAL CYSTITIS PATIENTS AND CONTROLS

Purpose Interstitial cystitis (IC) is a chronic bladder disease for which the etiology is unknown. Because the bladder epithelium is often abnormal in IC, we determined whether the levels of specific urine growth factors postulated to be important for bladder epithelial proliferation are altered in IC. Materials and Methods ELISAs were used to determine levels of epidermal growth factor (EGF), insulin-like growth factor 1 (IGF1), insulin-like growth factor binding protein 3 (IGFBP3), and heparin binding epidermal growth factor-like growth factor (HB-EGF) in urine specimens from women with IC, asymptomatic women without bladder disease, and women with bacterial cystitis. Results Urine HB-EGF levels were specifically and significantly decreased in IC patients as compared to asymptomatic controls or patients with bacterial cystitis, whether expressed as concentration (amount per volume of urine) or the amount relative to urine creatinine in each specimen. In contrast, urine EGF, IGF1, and IGFBP3 levels were all significantly elevated in IC patients compared to asymptomatic controls. Further, the amounts of urine EGF and IGF1 were also elevated in IC patients as compared to patients with bacterial cystitis, and urine IGFBP3 levels were significantly elevated when expressed per milligram of urine creatinine. Conclusions These findings indicate that complex changes in the levels of urine epithelial cell growth factors (EGF, IGF1, and HB-EGF) and a growth factor binding protein (IGFBP3) are associated with IC. While EGF, IGF1, and IGFBP3 levels are either the same or increased in the urine of IC patients as compared to patients with bacterial cystitis or asymptomatic controls, HB-EGF levels are significantly decreased in the urine of IC patients. Understanding the reasons for these changes may lead to understanding the pathogenesis of this disorder.

Excretion of fluorescein in the urine of women with interstitial cystitis.

Altered bladder permeability may have a role in the pathogenesis of interstitial cystitis. Fluorescein, a fluorescent dye of molecular weight 325, has been used to assess membrane permeability. Orally ingested fluorescein normally is rapidly conjugated to glucuronate by the liver and excreted in the urine. To test its use as a marker of bladder permeability, we administered fluorescein orally to 6 patients with interstitial cystitis who satisfied National Institutes of Health, National Institute for Diabetes and Digestive and Kidney Diseases criteria and to 6 normal female control subjects. After emptying the bladder and collection of a baseline blood sample, fasted subjects ingested 20 mg. fluorescein and blood samples were collected 1, 2, 3, 4 and 24 hours later. Urine was collected during each of the first 4 hours, and then from 4 to 10, 10 to 16 and 16 to 24 hours. Urine volume was measured, and all plasma and urine samples were analyzed for fluorescein. Plasma fluorescein concentrations (ng./ml.) were significantly (p < 0.05) higher in interstitial cystitis patients than in control subjects at 1 and 2 hours after fluorescein ingestion. Urine fluorescein excretion (mg.) was significantly (p < 0.05) lower in interstitial cystitis patients than in control subjects at 4 to 10 hours after fluorescein ingestion, and for the entire 24 hours. The increased fluorescein concentration in the plasma and decreased excretion in the urine of interstitial cystitis patients suggest that fluorescein may be a useful marker of altered membrane permeability.

Excretion of Fluorescein in the Urine of Women With Interstitial Cystitis

Purpose Altered bladder permeability may have a role in the pathogenesis of interstitial cystitis. Fluorescein, a fluorescent dye of molecular weight 325, has been used to assess membrane permeability. Orally ingested fluorescein normally is rapidly conjugated to glucuronate by the liver and excreted in the urine. Materials and Methods To test its use as a marker of bladder permeability, we administered fluorescein orally to 6 patients with interstitial cystitis who satisfied National Institutes of Health, National Institute for Diabetes and Digestive and Kidney Diseases criteria and to 6 normal female control subjects. After emptying the bladder and collection of a baseline blood sample, fasted subjects ingested 20 mg. fluorescein and blood samples were collected 1, 2, 3, 4 and 24 hours later. Urine was collected during each of the first 4 hours, and then from 4 to 10, 10 to 16 and 16 to 24 hours. Urine volume was measured, and all plasma and urine samples were analyzed for fluorescein. Results Plasma fluorescein concentrations (ng./ml.) were significantly (p <0.05) higher in interstitial cystitis patients than in control subjects at 1 and 2 hours after fluorescein ingestion. Urine fluorescein excretion (mg.) was significantly (p <0.05) lower in interstitial cystitis patients than in control subjects at 4 to 10 hours after fluorescein ingestion, and for the entire 24 hours. Conclusions The increased fluorescein concentration in the plasma and decreased excretion in the urine of interstitial cystitis patients suggest that fluorescein may be a useful marker of altered membrane permeability.

Do infectious agents cause interstitial cystitis?

The possibility that infectious agents play a role in the etiology of interstitial cystitis (IC) has been investigated for a number of years. Early studies were directed toward attempts to cultivate bacteria and fungi on routine culture media and microscopic examinations of urine or bladder tissue specimens for the presence of microorganisms. In more recent years, this approach has been expanded to include sophisticated culture techniques to search for the presence of fastidious and unusual organisms that would not be detected by routine culture methods. Similarly, the presence of viruses has been sought by incubating specimens from interstitial cystitis patients in mammalian cell cultures to detect cytopathic effects. None of these approaches has provided convincing evidence that micro-organisms or viruses are associated with IC. The latest attempts to search for the presence of bacteria have made use of the polymerase chain reaction (PCR) to amplify bacterial 16S rRNA genes that would be present if bacteria were present in bladder tissue or urine of IC patients. This approach allows bacteria to be detected and even identified without culture. However, the results from the great majority of bladder biopsy samples analyzed by these molecular techniques have been negative. PCR strategies have also been used to search for the presence of certain viruses in IC specimens, again without success. At this time, the results from laboratory culture, light and electron microscopy, and various molecular strategies to detect micro-organisms and viruses in IC specimens all argue against an infectious etiology for IC.

Decreased sup 3 H-Thymidine Incorporation By Human Bladder Epithelial Cells Following Exposure To Urine From Interstitial Cystitis Patients

Purpose Interstitial cystitis (IC) is a chronic bladder disease of unknown etiology. We sought to determine whether a cytotoxin is present in the urine of IC patients that could cause the epithelial damage seen in this disease. Materials and Methods Evidence for a cytotoxin was sought using both a neutral red uptake viability assay in T24 bladder epithelial cells, and a 3 H-thymidine incorporation assay in primary normal adult bladder epithelial cells and FHS 738 Bl human fetal bladder cells. Results The neutral red assay in T24 cells indicated the presence of a cytotoxin in 2 of 9 IC patient urine specimens. However, the more sensitive assay of cell proliferation (sup 3 H-thymidine incorporation) in normal adult human bladder epithelial cells revealed antiproliferative activity in urine from 10 of 13 (77 percent) IC patients vs. 3 of 19 (16 percent) controls (two-way analysis of variance, p = .019). The antiproliferative activity of IC urine specimens was confirmed using FHS 738 Bl human fetal bladder cells. The antiproliferative urinary substance(s) appeared to be a low molecular weight (less than 10,000 Da), heat stable, trypsin-sensitive factor(s). Conclusions Because a denuded or damaged bladder epithelium is a central finding in IC, it is possible that the antiproliferative protein(s) contributes to the pathogenesis of this disease.

Activation of Bladder Mast Cells in Interstitial Cystitis: A Light and Electron Microscopic Study

Interstitial cystitis, a sterile bladder condition, is characterized by urinary frequency, urgency, burning and suprapubic pain. Increasing evidence indicates that interstitial cystitis is a heterogeneous syndrome that reflects an immune response to a variety of triggers. More than 50% of the patients have allergies, 30% have the irritable bowel syndrome and almost 20% suffer from migraine headaches. Increased numbers of mast cells have been reported in interstitial cystitis. Mast cell activation, which is critical if these cells were to be implicated in this syndrome, has been investigated by electron microscopy, which definitively shows mast cell secretion. Recently, methylhistamine, the major metabolite of histamine, and the specific mast cell marker, tryptase, were shown to be significantly elevated in urine of interstitial cystitis patients. Bladder biopsies from 53 patients were analyzed blindly for the number and degree of activation of mast cells using 4 different stains for light microscopy, as well as electron microscopy. Controls included 16 patients with incontinence and chronic bacterial cystitis. Mast cells in controls were less than 10/mm.2 and were all nearly intact. Surprisingly, mast cells from 11 cancer patients averaged 50/mm.2 but almost all were intact. In contrast, mast cells from 26 interstitial cystitis patients averaged 40/mm.2 and more than 90% were activated to various degrees. Therefore, bladder mast cell activation is a characteristic pathological finding in at least a subset of patients with interstitial cystitis.

A prospective study of microorganisms in urine and bladder biopsies from interstitial cystitis patients and controls

Objectives. Interstitial cystitis (IC) is a chronic inflammatory condition of the bladder of unknown etiology. We tested the hypothesis that a microorganism would be found at higher prevalence in urine or bladder tissue from women with IC than from control women. Methods. Urine and bladder tissue were obtained at cystoscopy from 11 IC patients and 7 control subjects. These specimens were cultured for a variety of fastidious and nonfastidious bacteria, mycobacteria, fungi, and viruses. In addition, special staining techniques were used to examine biopsy specimens and cytospun urine, and tissue sections and outgrowths of explanted bladder cells were examined by electron microscopy. Results. Cultures of urine from 6 of 11 IC patients grew five different bacteria ( Corynebacterium sp, Klebsiella pneumoniae, Lactobacillus sp, Streptococcus constellatus, and Streptococcus morbillorum), human cytomegalovirus, or Torulopsis glabrata; one of these organisms ( Lactobacillus sp) was found in urine from 2 patients. Although contamination by urethral organisms is possible, the prevalence of microorganisms in urine of IC patients (6 of 11) was significantly greater than in urine of control subjects (0 of 7) ( P <0.05). Acridine orange staining revealed rods with appropriate morphology in urine from 4 of the 5 IC patients who had positive bacterial cultures and yeastlike organisms in urine and bladder tissue specimens that grew Torulopsis. Additionally, rodlike organisms were seen in urine from 2 IC patients with negative bacterial cultures and cocci were seen in the urine of 1 control patient. Biopsy specimens from 2 IC patients grew Torulopsis sp or Lactobacillus sp, in agreement with the results of acridine orange staining and culture of urine from these patients; in contrast, specimens from 3 control subjects grew small numbers of Pseudomonas sp of Staphylococcus epidermidis, but no organisms were cultured from urine or seen in acridine orange-stained tissue smears. All other cultures and stains were negative. Conclusions. These data do not provide evidence that IC is associated with infection or colonization by a single microorganism. However, they do generate the hypothesis that the prevalence of microorganisms, especially bacteria at low concentrations, is greater in the urine of IC patients than of control subjects. If these results are confirmed by other controlled studies, the question of whether the presence of these oranisms is a cause or a result of IC should be addressed.