Urogynecology Section of the Polish Society of Gynecologists and Obstetricians Guideline on the use of urodynamic testing in gynecological practice

Objectives: The aim was to present an interdisciplinary Guideline of the Urogynecology Section of the Polish Society of Gynecologists and Obstetricians (PSGO) for the use of urodynamics (UDS) in the diagnostic process of patients with lower urinary tract symptoms (LUTS) based on the available literature, expert knowledge, and everyday practice. Material and methods: A review of the literature concerning the use of UDS in women, including current international guidelines and earlier recommendations of the PSGO Urogynecology Section, was conducted. Results: Urodynamic testing allows to make the urodynamic diagnosis which, nevertheless, remains to be the preliminary diagnosis. Medical history, physical examination, and detailed analysis of the previous test results (laboratory, imaging, endoscopic) need to be taken into consideration before making the final diagnosis. Urodynamic testing before surgical treatment of SUI is allowable, but the decision remains at the discretion of the physician. Urodynamic testing is not necessary before primary surgical treatment of uncomplicated SUI, but it has been demonstrated to optimize the therapeutic methods in complicated SUI. The significance of UDS in the diagnostic process of patients with overactive bladder symptoms, voiding dysfunction, and bladder outlet obstruction was discussed. Conclusions: Urodynamic testing is a vital element of the urogynecological diagnostic process. The scope of UDS should reflect the individual needs and symptoms of each patient and be based on the current guidelines, expert knowledge and experience of the physician, indications, and eligibility, as well as additional test results of the affected patients. Due to formal and legal requirements, PSGO, in this Guideline, wishes to emphasize the need for an individualized approach to both, test performance and result interpretation

Open partial nephrectomy for entirely intraparenchymal tumors: a matched case-control study of oncologic outcome and complication rate

ABSTRACT Purpose To compare the oncologic and clinical outcomes for open partial nephrectomy (OPN) performed in patients with entirely intraparenchymal tumors versus case-matched controls, with exophytic lesions. Material and methods Patients having undergone OPN between 2007 and 2012 were investigated. Exclusion criteria included patients with a benign tumor, advanced malignancy, malignancies other than renal cell carcinoma, end-stage renal failure, or 3 or more co-existing chronic diseases. Individuals with tumors that were invisible at the renal surface were identified, and then matched with 2 controls chosen for tumor size, pathology, age, follow-up period, and presence of a solitary kidney. Oncological status, perioperative, and postoperative data were collected and compared between groups. Results 17 individuals with entirely endophytic RCC tumors and available oncologic status were identified. For five patients, only one suitable control could be identified, bringing the control group number to 29. All tumors were clear cell carcinomas staged at pT1a. Median tumor size was 25mm for endophytic lesions, and 27mm for exophytic masses (P=0.32). The operative period was extended by 20 minutes for intrarenal tumors (P=0.03), with one case of a positive surgical margin in each group (P=0.7). There were no significant differences in perioperative or postoperative complications. Median follow-up was 47 and 43 months for patients with endophytic and exophytic tumors respectively. Disease recurrence was recorded in one patient after endophytic tumor resection, and in four controls (P=0.4). Conclusions OPN shows equivalent safety and efficacy for both intrarenal RCC tumors and exophytic tumors of the same size and type

Immunohistochemical differentiation between muscularis mucosae and muscularis propria for improving the staging of bladder cancer in patients undergoing transurethral resection of bladder tumours

Microscopic differentiation between muscularis mucosae (MM) and muscularis propria (MP) of the bladder in the material obtained during transurethral resection (TUR) remains difficult. The study was aimed at determination of the usefulness of immunohistochemical staining in this context. Forty-seven TUR specimens were stained with 5 mouse anti-human antibodies: anti-desmin, anti-filamin, anti-type IV collagen, anti-smoothelin, and anti-vimentin. Slides were assessed under light microscopy and the intensity of the immune reaction within MM and MP was evaluated on a four-level visual scale as follows: negative (0) and weakly (1), moderately (2), or strongly (3) positive. MM was identified in 27 patients (57.4%). The modal values of reaction intensity in MM and MP was 0 and 2 for desmin (p > 0.05), 2 and 2 for filamin (p = 0.01), 2 and 2 for type IV collagen (p > 0.05), 1 and 2 for smoothelin (p = 0.03), and 2 and 0 for vimentin (p = 0.02), respectively. Identical intensity within MM and MP was observed in 7.1%, 28.6%, 20%, 30.1%, 5.6%, respectively. Immunohistochemistry can help differentiate between MM and MP in TUR specimens. As of yet, no single marker can reliably differentiate between MM and MP; however, a combination of anti-filamin, anti-smoothelin, and anti-vimentin antibodies may be reasonable for diagnostic purposes

In vivo imaging system for explants analysis—A new approach for assessment of cell transplantation effects in large animal models

Introduction: Despite spectacular progress in cellular transplantology, there are still many concerns about the fate of transplanted cells. More preclinical studies are needed, especially on large animal models, to bridge the translational gap between basic research and the clinic. Herein, we propose a novel approach in analysis of cell transplantation effects in large animals explants using in vivo imaging system (IVIS®) or similar equipment. Material and methods: In the in vitro experiment cells labeled with fluorescent membrane dyes: DID (far red) or PKH26 (orange) were visualized with IVIS®. The correlation between the fluorescence signal and cell number with or without addition of minced muscle tissue was calculated. In the ex vivo study urethras obtained from goats after intraurethral cells (n = 9) or PBS (n = 4) injections were divided into 0.5 cm cross-slices and analyzed by using IVIS®. Automatic algorithm followed or not by manual setup was used to separate specific dye signal from tissue autofluorescence. The results were verified by systematic microscopic analysis of standard 10 μm specimens prepared from slices before and after immunohistochemical staining. Comparison of obtained data was performed using diagnostic test function. Results: Fluorescence signal strength in IVIS® was directly proportional to the number of cells regardless of the dye used and detectable for minimum 0.25x106 of cells. DID-derived signal was much less affected by the background signal in comparison to PKH26 in in vitro test. Using the IVIS® to scan explants in defined arrangement resulted in precise localization of DID but not PKH26 positive spots. Microscopic analysis of histological specimens confirmed the specificity (89%) and sensitivity (80%) of IVIS® assessment relative to DID dye. The procedure enabled successful immunohistochemical staining of specimens derived from analyzed slices. Conclusions: The IVIS® system under appropriate conditions of visualization and analysis can be used as a method for ex vivo evaluation of cell transplantation effects. Presented protocol allows for evaluation of cell delivery precision rate, enables semi-quantitative assessment of signal, preselects material for further analysis without interfering with the tissue properties. Far red dyes are appropriate fluorophores to cell labeling for this application

Intraurethral co-transplantation of bone marrow mesenchymal stem cells and muscle-derived cells improves the urethral closure

Abstract Background Cell therapy constitutes an attractive alternative to treat stress urinary incontinence. Although promising results have been demonstrated in this field, the procedure requires further optimization. The most commonly proposed cell types for intraurethral injections are muscle derived cells (MDCs) and mesenchymal stem/stromal cell (MSCs). The aim of this study was to evaluate the effects of MDC-MSC co-transplantation into the urethra. Methods Autologous transplantation of labeled MDCs, bone marrow MSCs or co-transplantation of MDC-MSC were performed in aged multiparous female goats (n = 6 in each group). The mean number of cells injected per animal was 29.6 × 106(± 4.3 × 106). PBS-injected animals constituted the control group (n = 5). Each animal underwent urethral pressure profile (UPP) measurements before and after the injection procedure. The maximal urethral closure pressure (MUCP) and functional area (FA) of UPPs were calculated. The urethras were collected at the 28th or the 84th day after transplantation. The marker fluorochrome (DID) was visualized and quantified using in vivo imaging system in whole explants. Myogenic differentiation of the graft was immunohistochemically evaluated. Results The grafted cells were identified in all urethras collected at day 28 regardless of injected cell type. At this time point the strongest DID-derived signal (normalized to the number of injected cells) was noted in the co-transplanted group. There was a distinct decline in signal intensity between day 28 and day 84 in all types of transplantation. Both MSCs and MDCs contributed to striated muscle formation if transplanted directly to the external urethral sphincter. In the MSC group those events were rare. If cells were injected into the submucosal region they remained undifferentiated usually packed in clearly distinguishable depots. The mean increase in MUCP after transplantation in comparison to the pre-transplantation state in the MDC, MSC and MDC-MSC groups was 12.3% (± 11.2%, not significant (ns)), 8.2% (± 9.6%, ns) and 24.1% (± 3.1%, p = 0.02), respectively. The mean increase in FA after transplantation in the MDC, MSC and MDC-MSC groups amounted to 17.8% (± 15.4%, ns), 15.2% (± 12.9%, ns) and 17.8% (± 2.5%, p = 0.04), respectively. Conclusions The results suggest that MDC-MSC co-transplantation provides a greater chance of improvement in urethral closure than transplantation of each population alone

The presence of specific DID fluorescence in urethras from study group after IVIS® spectral unmixing algorithm application.

<p>The presence of specific DID fluorescence in urethras from study group after IVIS® spectral unmixing algorithm application.</p

Preparation of posthumously collected urethra for analysis in <i>ex vivo</i> experiment.

<p>A) Photograph of freshly isolated urethra with bladder and a method of length and diameter measuring; B) Slices of whole urethra for analysis with IVIS®; C) Schematic arrangement of tissue slices for IVIS® analysis. caudal–top, dorsal–upwards; D) Photograph of the microscopic preparation with drawn model "map" after fluorescence microscopy analysis. Lines shows the outline of the preparation (black), urethra light (red), location of spots showing specific fluorescence (blue).</p

The Mutual Interactions between Mesenchymal Stem Cells and Myoblasts in an Autologous Co-Culture Model

<div><p>Both myoblasts and mesenchymal stem cells (MSC) take part in the muscle tissue regeneration and have been used as experimental cellular therapy in muscular disorders treatment. It is possible that co-transplantation approach could improve the efficacy of this treatment. However, the relations between those two cell types are not clearly defined. The aim of this study was to determine the reciprocal interactions between myoblasts and MSC <i>in vitro</i> in terms of the features important for the muscle regeneration process. Primary caprine muscle-derived cells (MDC) and bone marrow-derived MSC were analysed in autologous settings. We found that MSC contribute to myotubes formation by fusion with MDC when co-cultured directly, but do not acquire myogenic phenotype if exposed to MDC-derived soluble factors only. Experiments with exposure to hydrogen peroxide showed that MSC are significantly more resistant to oxidative stress than MDC, but a direct co-culture with MSC does not diminish the cytotoxic effect of H₂O₂ on MDC. Cell migration assay demonstrated that MSC possess significantly greater migration ability than MDC which is further enhanced by MDC-derived soluble factors, whereas the opposite effect was not found. MSC-derived soluble factors significantly enhanced the proliferation of MDC, whereas MDC inhibited the division rate of MSC. To conclude, presented results suggest that myogenic precursors and MSC support each other during muscle regeneration and therefore myoblasts-MSC co-transplantation could be an attractive approach in the treatment of muscular disorders.</p></div