Andrea Staack

Combined determination of plasma MMP2, MMP9, and TIMP1 improves the non-invasive detection of transitional cell carcinoma of the bladder

BackgroundMatrix metalloproteinases (MMPs) and their inhibitors (TIMPs) play a major role in the maintenance of extracellular matrix homeostasis and are involved in the process of tumour invasion and metastasis in several malignant tumour entities. The goal of this study is to evaluate the diagnostic value of various circulating MMPs and TIMPs in blood plasma for a non-invasive detection of transitional cell carcinoma of the bladder (TCC).MethodsIn this study the concentrations of MMP1, MMP2, MMP3, MMP9, their inhibitors TIMP1, TIMP2, and the MMP1/TIMP1-complex (MTC1) were quantified in blood plasma with the sandwich enzyme-linked immunosorbent assay (ELISA). Blood plasma samples were investigated from 68 patients (non-metastasized, n = 57 and metastasized, n = 11) with TCC of the bladder and from 79 healthy controls. The mROC program was used to calculate the best two- and three- marker combinations. The diagnostic values for all single markers and the marker combinations were estimated both by the overall diagnostic performance index area under the ROC curve (AUC) and the sensitivity and specificity at cutoff limits with the highest diagnostic accuracy and at the 90% and 95% limits of sensitivity and specificity, respectively.ResultsThe median MMP2 concentration was elevated in blood plasma in all patient groups with TCC in comparison to the controls (p < 0.001). The concentrations of TIMP1, TIMP2, and MTC1 in plasma probes were significantly lower from patients with non-metastasized TCC compared to the controls. MMP2 tested alone reached the highest sensitivity and specificity at 75%, respectively. The sensitivity and specificity increased when tested in combination with MMP9 and TIMP1 (97%, 94%, respectively). The combination of MMP9 and TIMP1 also showed an improved sensitivity (80%) and specificity (99%) than tested alone.ConclusionMMP2 is a statistically significant marker in blood plasma for bladder cancer detection with an increased diagnostic value in combination with MMP9 and TIMP1. This study showed that the highest sensitivities and specificities are not obtained by testing each marker alone. As shown by the best two-marker combination, which includes MMP9 and TIMP1, the optimized combination does not always include the best single markers.

Bladder tissue formation from cultured bladder urothelium

Tissue recombination is a powerful method to evaluate the paracrine‐signaling events that orchestrate the development of organs using the in vivo environment of a host rodent. Studies have reported the successful generation of primary cultures of rodent bladder urothelium, but none have reported their use to recapitulate bladder tissue with tissue recombination. We propose that primary cultured bladder urothelium, when recombined with inductive embryonic bladder mesenchyme, will form bladder tissue in a recombination model. Adult rat bladders were isolated and urothelium obtained. Sheets of bladder urothelium were re‐suspended in collagen and maintained in tissue culture. After expansion (>20 passages), the urothelium was recombined with embryonic day‐14 mouse bladder mesenchyme, then grafted beneath the renal capsule of immunocompromised mouse hosts. Grafts were harvested after 28 days. Control grafts were performed with bladder mesenchyme alone, cultured bladder urothelium alone, and collagen matrix alone. Final tissues were evaluated with staining and immunohistochemistry (H&E, Gomoris trichrome, broad‐spectrum uroplakin, and smooth muscle actin α and γ). Immunocytochemistry on cultured urothelium for broad‐spectrum keratin, vimentin, and broad‐spectrum uroplakin confirmed pure populations, void of mesenchymal contaminants. Staining of recombinant grafts demonstrated bladder tissue with mature urothelium and stromal differentiation. Control tissues were void of bladder tissue formation. We have successfully demonstrated that a chimeric bladder is formed from primary cultured bladder urothelium recombined with embryonic bladder mesenchyme. This is a powerful new tool for investigating the molecular mechanisms of bladder development and disease. Future applications may include the in vitro genetic manipulation of urothelium and examining those effects on growth and development in an in vivo environment. Developmental Dynamics 235:2795–2801, 2006.

Cathepsins B, H, and L activities in urine of patients with transitional cell carcinoma of the bladder

OBJECTIVES Cathepsin B, H, and L (CB, CH, CL) are lysosomal proteolytic enzymes that belong to the group of cysteine proteinases. The imbalance between proteinases and their inhibitors is believed to correlate with tumor progression and shortened patient survival. In transitional cell carcinoma (TCC) only limited data have been published. METHODS Using spectrofluorometric assays, catalytic activities of CB, CH, and CL in urine were measured to evaluate the potential diagnostic and prognostic value for patients with TCC of the bladder. Second morning urine was collected and used for measurements. CB, CH, and CL activities were determined for groups of patients with superficial disease (Ta-1, n = 43) and muscle-invasive tumors (T2, n = 18; or greater than T2, n = 9), as well as for different tumor grades (G1, n = 12; G2, n = 26; and G3, n = 31). For comparison, 14 urine samples from patients with bladder inflammation and 43 samples from a control group were also included. RESULTS Compared with the control group, patients with superficial Stage Ta-T1 disease and muscle-invasive Stage T2 or greater disease, as well as patients with G3 tumors, revealed significantly higher urinary CL activity. CB and CH did not show any tumor-related activity increase. CB was significantly lower in patients with nonrecurrent tumors. CONCLUSIONS These results suggest that elevated levels of CL in urine might be indicative of a cellular proteolytic imbalance in TCC of the bladder and may have a prognostic and/or diagnostic value.

Organ and Species Specificity in the Stimulation of Transitional Epithelial Cell Growth by Fibroblasts

Objectives: Culture of transitional epithelium for urinary tract reconstruction has been problematic due, in part, to the dependence of urothelial cells on a basal layer of bladder fibroblasts for growth. In vitro studies on the effect of bladder, ureter, and intestinal fibroblast cocultures and conditioned media upon urothelial cell growth were conducted to better characterize the dependence of urothelial cells of fibroblasts. Methods: Primary cultures of human and porcine bladder, ureter, and intestinal fibroblasts and bladder and ureter urothelial cells were established. The urothelial cells were incubated with fibroblasts in a coculture system and growth compared to that of individual fibroblast and urothelial cultures. Urothelium–specific medium was exposed to the fibroblast cultures for 6, 12, and 24 h. Urothelial cell growth in each of the fibroblast–conditioned media was evaluated. Results: Coculture of human urothelial cells with human bladder and ureter fibroblasts yielded increased cell growth when compared to the cells in individual culture. This improvement was greatest for the bladder fibroblasts in coculture with bladder epithelial cells. The media exposed to bladder and ureter fibroblasts for 24 h significantly increased bladder and ureter urothelial growth compared to fresh medium. Coculture with intestinal fibroblasts and exposure to intestinal fibroblast conditioned media did not significantly stimulate urothelial cell growth. Similarly, coculture and conditioned media studies of porcine urothelial cells with porcine bladder and ureter fibroblasts (but not intestinal fibroblasts) yielded increased cell growth when compared to the cells in individual culture, particularly with bladder fibroblasts. However, human urothelial cells were not stimulated by porcine bladder and ureter fibroblast conditioned media, nor were porcine urothelial cells stimulated by human bladder and ureter fibroblast conditioned media. Conclusions: The ability of urinary fibroblasts to stimulate urothelial cell proliferation resides in an unidentified soluble factor secreted into the medium, independent of the presence of the fibroblasts. This factor is relatively organ and species–specific.

Clinical value of vesical leukoplakia and evaluation of the neoplastic risk by mutation analyses of the tumor suppressor gene TP53.

Aim: Leukoplakia has been found to be precancerous in organs covered with squamous epithelium. The present study was conducted to determine whether leukoplakia described in the female bladder is also a premalignant lesion.

Applying translabial ultrasound to detect synthetic slings—You can do it too! A comparison of urology trainees to an attending radiologist

Translabial ultrasound (TUS) is a useful tool for identifying and assessing synthetic slings. This study evaluates the ability of urology trainees to learn basic pelvic anatomy and sling assessment on TUS.

Dextrose instillation as an alternative agent to observe ureteral efflux during pelvic reconstructive surgery

OBJECTIVE To evaluate the use, cost, postoperative urinary tract infection (UTI) rates, and complications of dextrose instillation during cystoscopy. METHODS The medical records of patients who underwent cystoscopy during pelvic reconstructive surgery between June 2016 and June 2017 were reviewed. Patients were divided into two groups: patients who had one ampule of dextrose 50% (D50) directly instilled and patients who did not have D50 instilled during cystoscopy. Preoperative demographics, UTI rates, and postoperative complications were compared. Pharmaceutical cost and availability were reported by the pharmacy at our institution. RESULTS Out of 63 patients identified, dextrose instillation was used in 20 patients and no dextrose was used in 43 patients. Each ampule of D50 cost

Translabial ultrasound evaluation of pelvic floor structures and mesh in the urology office and intraoperative setting

2.18 and there were no problems with supply shortage. As D50 was directly instilled into the bladder, there was immediate visualization of ureteral efflux at the time of surgery. Three patients (15%) in the dextrose group and 10 patients (23%) in the nondextrose group developed postoperative UTIs. There was no statistically significant difference in postoperative UTI rates between the two groups (p = 0.43) and there were no differences in postoperative complications. CONCLUSION Dextrose is a safe, cost-effective, readily available agent that provides instantaneous visualization of ureteral efflux without an increased risk of postoperative UTI.

Erosion of Polytetrafluoroethylene (Gore-Tex®) Sling Over 20 Years after Placement for Stress Urinary Incontinence

BACKGROUND Translabial ultrasound (TUS) can provide an inexpensive alternative imaging modality for evaluating pelvic floor structures and synthetic slings as mesh can be difficult to identify on pelvic exam or cystoscopy, patients may be unable to provide an accurate history of previous pelvic surgery, and cross-sectional imaging with computed tomography and magnetic resonance imaging can be inadequate for evaluating synthetic slings. OBJECTIVE To demonstrate the use of TUS in the evaluation of female pelvic floor structures and mesh. METHODS Translabial ultrasound can be used in the Urology clinic or intraoperative setting using a curvilinear transducer. Following identification of anatomic landmarks in the various planes of the pelvic floor, TUS can evaluate for pelvic floor disorders and the type and location of synthetic mesh material. Artifacts, such as air pockets in the vagina or rectum and the hypoechoic pubic symphysis, are also considered. RESULTS Real-time imaging allows for dynamic examination of pelvic organ prolapse and urethral hypermobility that can contribute to pelvic exam findings. Bladder ultrasound can help evaluate for lesions, calculi, and even mesh erosion. Translabial ultrasound can also be used to differentiate hyperechoic retropubic and transobturator slings by identifying the position of sling arms and the appearance of the sling at different planes. Evaluation with TUS can demonstrate sling disruption, folding, urethral impingement, and erosion into pelvic floor structures. This can be particularly useful in patients presenting with pain, recurrent infections, or voiding dysfunction in which problems with mesh may not be easily identified on pelvic exam or cystoscopy. This imaging modality can complement a patients history, aid in preoperative planning, and enable intraoperative identification of mesh slings. CONCLUSION Translabial ultrasound provides a quick, readily available, and easy-to-learn imaging modality for evaluating pelvic floor structures and mesh in the office or intraoperative setting.

Comparing the vaginal wall sling with autologous rectus fascia and polypropylene sling: Short term outcomes and patient satisfaction

As synthetic material has evolved to improve both the efficacy and biocompatibility of suburethral slings, soft polypropylene slings are currently the gold standard for treatment of stress urinary incontinence. However, reports of complications beyond 10 years are limited and patients can nevertheless present with erosion and other complications from other sling materials that have been used in the past. We present a case of synthetic sling erosion 21 years after placement of a polytetrafluoroethylene sling (Gore-tex).