Evaluation of the metabolism of glycosaminoglycans in patients with interstitial cystis.

INTRODUCTION: Painful bladder syndrome/interstitial cystitis (PBS/IC) pathogenesis is not fully known, but evidence shows that glycosaminoglycans (GAG) of bladder urothelium can participate in its genesis. The loss of these compounds facilitates the contact of urine compounds with deeper portions of bladder wall triggering an inflammatory process. We investigated GAG in urine and tissue of PBS/IC and pure stress urinary incontinence (SUI) patients to better understand its metabolism. MATERIALS AND METHODS: Tissue and urine of 11 patients with PBS/IC according to NIDDK criteria were compared to 11 SUI patients. Tissue samples were analyzed by histological, immunohistochemistry and immunofluorescence methods. Statistical analysis were performed using t Student test and Anova, considering significant when p < 0.05. RESULTS: PBS/IC patients had lower concentration of GAG in urine when compared to SUI (respectively 0.45 ± 0.11 x 0.62 ± 0.13 mg/mg creatinine, p < 0.05). However, there was no reduction of the content of GAG in the urothelium of both groups. Immunofluorescence showed that PBS/IC patients had a stronger staining of TGF-beta, decorin (a proteoglycan of chondroitin/dermatan sulfate), fibronectin and hyaluronic acid. CONCLUSION: the results suggest that GAG may be related to the ongoing process of inflammation and remodeling of the dysfunctional urothelium that is present in the PBS/IC.

Evaluation of the metabolism of glycosaminoglycans in patients with interstitial cystis

Introduction: Painful bladder syndrome/interstitial cystitis (PBS/IC) pathogenesis is not fully known, but evidence shows that glycosaminoglycans (GAG) of bladder urothelium can participate in its genesis. The loss of these compounds facilitates the contact of urine compounds with deeper portions of bladder wall triggering an inflammatory process. We investigated GAG in urine and tissue of PBS/IC and pure stress urinary incontinence (SUI) patients to better understand its metabolism. Materials and Methods: Tissue and urine of 11 patients with PBS/IC according to NIDDK criteria were compared to 11 SUI patients. Tissue samples were analyzed by histological, immunohistochemistry and immunofluorescence methods. Statistical analysis were performed using t Student test and Anova, considering significant when p < 0.05. Results: PBS/IC patients had lower concentration of GAG in urine when compared to SUI (respectively 0.45 ± 0.11 x 0.62 ± 0.13 mg/mg creatinine, p < 0.05). However, there was no reduction of the content of GAG in the urothelium of both groups. Immunofluorescence showed that PBS/IC patients had a stronger staining of TGF-beta, decorin (a proteoglycan of chondroitin/dermatan sulfate), fibronectin and hyaluronic acid. Conclusion: the results suggest that GAG may be related to the ongoing process of inflammation and remodeling of the dysfunctional urothelium that is present in the PBS/IC. .

Testing for urinary hyaluronate improves detection and grading of transitional cell carcinoma.

OBJECTIVE: The purpose of this study is to establish a method for the diagnosis and grading of transitional cell carcinoma (TCC), which is responsible for 90% of bladder tumors, using a recently developed ultrasensitive assay for the measurement of hyaluronan (HA). MATERIALS AND METHODS: Urine samples were collected prior to surgery (cystoscopy, transurethral resection for bladder cancer (TURBT), and cystectomy) in 350 patients. After the procedure, pathologic examination revealed that 160 patients had TCC. HA was measured directly in the urine by a noncompetitive enzyme-linked immunosorbent assay (ELISA)-like fluorometric assay. Using the receiver operator characteristic curve (ROC), t-test, Dunn test, Kruskal-Wallis test, and Mann-Whitney test, we evaluated the differences between groups (those with TCC vs. those without TCC). RESULTS: By analyzing the ROC curve, we chose a urinary HA cutoff value of 13.0 µg/l for indicating risk of TCC. Using the value this of 13.0 µg/l, we found that this test had an overall sensitivity of 82.3% and an overall specificity of 81.2%. The positive predictive value of this assay was 78.9%, the negative predictive negative value was 84.2%, and the predictive accuracy was 81.7%. Logistic regression analysis revealed that every 1 µg/l increase in HA increased a patient's likelihood of having TCC by 3.9%. The sensitivity of this test to detect superficial tumors was 76.6%, whereas its sensitivity for detecting invasive tumors was 94.6%. The urinary HA excretion of patients with TCC, classified according to the TNM staging system and the World Health Organization (WHO) grading system, were compared, and a significant difference was observed between the HA levels of patients with superficial tumors compared with invasive tumors (P = 0.005) as well as between patients with low- vs. high-grade carcinomas (P < 0.001). Patients with urinary HA levels >35 µg/l had a 4.63 times increased risk of having an aggressive, invasive, high grade tumor (P = 0.005). CONCLUSIONS: Our results support the postulate that urinary HA may be used as a tumor marker to aid in the diagnosis and grading of TCC. Additionally, more invasive tumors produce and release more HA in urine than superficial tumors, thus higher HA levels indicate more aggressive disease.

Urinary glycosaminoglycans excretion and the effect of dimethyl sulfoxide in an experimental model of non-bacterial cystitis.

PURPOSE: We reproduced a non-bacterial experimental model to assess bladder inflammation and urinary glycosaminoglycans (GAG) excretion and examined the effect of dimethyl sulfoxide (DMSO). MATERIALS AND METHODS: Female rats were instilled with either protamine sulfate (PS groups) or sterile saline (control groups). At different days after the procedure, 24 h urine and bladder samples were obtained. Urinary levels of hyaluronic acid (HA) and sulfated glycosaminoglycans (S-GAG) were determined. Also to evaluate the effect of DMSO animals were instilled with either 50% DMSO or saline 6 hours after PS instillation. To evaluate the effect of DMSO in healthy bladders, rats were instilled with 50% DMSO and controls with saline. RESULTS: In the PS groups, bladder inflammation was observed, with polymorphonuclear cells during the first days and lymphomononuclear in the last days. HA and S-GAG had 2 peaks of urinary excretion, at the 1st and 7th day after PS injection. DMSO significantly reduced bladder inflammation. In contrast, in healthy bladders, DMSO produced mild inflammation and an increase in urinary HA levels after 1 and 7 days and an increase of S-GAG level in 7 days. Animals instilled with PS and treated with DMSO had significantly reduced levels of urinary HA only at the 1st day. Urinary S-GAG/Cr levels were similar in all groups. CONCLUSIONS: Increased urinary levels of GAG were associated with bladder inflammation in a PS-induced cystitis model. DMSO significantly reduced the inflammatory process after urothelial injury. Conversely, this drug provoked mild inflammation in normal mucosa. DMSO treatment was shown to influence urinary HA excretion.

Urinary glycosaminoglycans excretion and the effect of dimethyl sulfoxide in an experimental model of non-bacterial cystitis

PURPOSE: We reproduced a non-bacterial experimental model to assess bladder inflammation and urinary glycosaminoglycans (GAG) excretion and examined the effect of dimethyl sulfoxide (DMSO). MATERIALS AND METHODS: Female rats were instilled with either protamine sulfate (PS groups) or sterile saline (control groups). At different days after the procedure, 24 h urine and bladder samples were obtained. Urinary levels of hyaluronic acid (HA) and sulfated glycosaminoglycans (S-GAG) were determined. Also to evaluate the effect of DMSO animals were instilled with either 50 percent DMSO or saline 6 hours after PS instillation. To evaluate the effect of DMSO in healthy bladders, rats were instilled with 50 percent DMSO and controls with saline. RESULTS: In the PS groups, bladder inflammation was observed, with polymorphonuclear cells during the first days and lymphomononuclear in the last days. HA and S-GAG had 2 peaks of urinary excretion, at the 1st and 7th day after PS injection. DMSO significantly reduced bladder inflammation. In contrast, in healthy bladders, DMSO produced mild inflammation and an increase in urinary HA levels after 1 and 7 days and an increase of S-GAG level in 7 days. Animals instilled with PS and treated with DMSO had significantly reduced levels of urinary HA only at the 1st day. Urinary S-GAG/Cr levels were similar in all groups. CONCLUSIONS: Increased urinary levels of GAG were associated with bladder inflammation in a PS-induced cystitis model. DMSO significantly reduced the inflammatory process after urothelial injury. Conversely, this drug provoked mild inflammation in normal mucosa. DMSO treatment was shown to influence urinary HA excretion.

Urinary hyaluronan as a marker for the presence of residual transitional cell carcinoma of the urinary bladder.

OBJECTIVE: The purpose of this report is to evaluate the value of urinary hyaluronan (HA) as a maker of residual transitional cell carcinoma (TCC). PATIENTS AND METHODS: Urine samples were collected from 83 patients hospitalized for transurethral resection (TUR). Patient ages ranged from 36 to 86 years. Samples were taken both before and after surgery. HA analysis was performed using an "ELISA-like" fluorometric assay. RESULTS: Patients were divided into two groups: a control group whose previous diagnosis was negative for tumors (n=22) and another with positive diagnosis for tumors (n=61) which was further sub-divided into with and without residual tumor. After the second procedure 47 individuals did not display residual tumor, whereas 14 (23%) did. The average HA in the control group was 8.3 microg/L pre- and 7.1 post-operatively, hence, no change occurred (p=0.471). In the group with TCC patients, the HA dropped from 885.5 microg/L to 215.3 microg/L with residual tumors and from 234.3 microg/L to 11.2 microg/L for those without residual tumor. Using a cut-off value of 20 microg/L, the sensitivity to detect residual tumor is 92.9% and specificity is 83%. CONCLUSION: HA in addition to being one of the best markers for the initial evaluation of bladder carcinoma can be used to determine the presence of a residual tumor. This is associated with poor prognosis.

Glycosaminoglycan structure and content differ according to the origins of human tumors.

The glycosaminoglycans of the tumor mass and from the urine of patients with a nephroblastoma of embryonic origin (Wilms' tumor) and hypernephroma were analyzed. The urine of patients with Wilms' tumors prior to treatment, and two patients with metastasis contained high levels of hyaluronic acid (2-5 mg/l of urine) when compared to patients after surgery or chemotherapy where the content of hyaluronic acid was less than 0.1 mg/l. Urine of patients with hypernephroma and normal individuals contained even smaller amounts of hyaluronic acid. Normal kidneys contain mainly dermatan sulfate and heparan sulfate, while the hypernephroma and Wilms' tumor contain substantial amounts of chondroitin sulfate. The amount of glycosaminoglycans isolated from Wilms' tumor and hypernephroma were 10 times and 3 times, respectively, greater than normal kidneys. The amounts of hyaluronic acid in Wilms' tumor varied from 56 to 73% whereas normal kidneys contained about 13%. Chondroitin sulfate was also increased in Wilms' tumor and hypernephroma. It corresponded to 11% and 42%, respectively, of the total glycosaminoglycans. These and other findings indicate that the glycosaminoglycans of Wilms' tumors resemble those present during embryonic development of normal tissues whereas those in hypernephroma are typical of other carcinomas of different origins.

Glycosaminoglycan structure and content differ according to the origins of human tumors

The glycosaminoglycans of the tumor mass and from the urine of patients with a nephroblastoma of embryonic origin (Wilms' tumor) and hypernephroma were analyzed. The urine of patients with Wilms/ tumors prior to treatment, and two patients with metastasis contained high levels of hyaluronic acid (2-5 mg/l of urine) when compared to patients after surgery or chemotherapy where the content of hyaluronic acid was less than 0.1 mg/l. Urine of patients with hypernephroma and normal individuals contained even smaller amounts of hyaluronic acid. Normal kidneys contain mainly dermatan sulfate and heparan sulfate, while the hypernephroma and Wilms' tumor contain substantial amounts of chondroitin sulfate. The amount of glycosaminoglycans isolated from Wilms' tumor and hypernephroma were 10 times and 3 times, respectively, greater than normal kidneys. The amonunts of hyaluronic acid in Wilms' tumor varied from 56 to 73 per cent whereas normal kidneys contained about 13 per cent. Chondroitin sulfate was also increased in Wilms' tumor and hypernephroma. It corresponded to 11 per cent and 42 per cent, respectively, of the total glycosaminoglycans. These and other findings indicate that the glycosaminoglycans of Wilms' tumors resemble those present during embryonic development of normal tissues whereas those in hypernephroma are typical of other carinomas of different origins