Two-Year Preclinical Evaluation of Long-Term Absorbable Poly-4-hydroxybutyrate Scaffold for Surgical Correction of Pelvic Organ Prolapse.

INTRODUCTION AND HYPOTHESIS: Fully absorbable implants may be an alternative to permanent meshes in the correction pf pelvic organ prolapse (POP) as they may reduce adverse events by promoting tissue regeneration and collagen metabolism. This study was aimed at evaluating the long-term host and biomechanical response to a fully absorbable poly-4-hydroxybutyrate (P4HB) scaffold in comparison with polypropylene (PP) mesh. METHODS: Poly-4-hydroxybutyrate scaffold (n = 16) and PP mesh (n = 16) were surgically implanted in the posterior vaginal wall of parous female Dohne Merino sheep. Vaginal explants were evaluated in terms of gross necropsy, host response (immune response, collagen deposition, tissue regeneration), biomechanics, and degradation of P4HB at 12 and 24 months post-implantation. RESULTS: Gross necropsy revealed no infection or fluid collection using P4HB or PP. At 12 months, exposures were observed with both P4HB (3 out of 8) and PP (4 out of 8), whereas at 24 months, exposures were observed only with PP (4 out of 8). The tensile stiffness of the P4HB explants was maintained over time despite complete absorption of P4HB. The collagen amount of the vaginal tissue after P4HB implantation increased over time and was significantly higher than PP at 24 months. P4HB scaffolds exhibited significantly lower myofibroblast differentiation than PP meshes at 24 months. CONCLUSIONS: The P4HB scaffold allowed for gradual load transfer to the vaginal wall and resulted in mechanically self-sufficient tissue. P4HB scaffold had a more favorable host response than PP mesh, with higher collagen content, lower myofibroblastic differentiation, and no exposures at 24 months. P4HB scaffolds have potential as an alternative to permanent implants in treating POP.

Introducing an lsoprenaline Eluting Guidewire: Report on its Design and the Results of the Dose-Determining Pilot Study.

Introduction: Retrograde intrarenal surgery (RIRS) is associated with complications, many of which are related to the intrarenal pressure (IRP). We aim to describe the design of a novel isoprenaline-eluting guidewire ("IsoWire") and present the results from the first in vitro release studies and the first animal studies showing its effect on IRP. Materials and Methods: The IsoWire comprises a Nitinol core surrounded by a stainless-steel wire wound into a tight coil. The grooves created by this coil provided a reservoir for adding a hydrogel coating into which isoprenaline, a beta-agonist, was loaded. Animal studies were performed using a porcine model. For the control, IRP, heart rate (HR), and mean arterial pressure (MAP) were measured continuously for 6 minutes with a standard guidewire in place. For the experiment, the standard hydrophilic guidewire was removed, the IsoWire was inserted into the renal pelvis, and the same parameters were measured. Results: In vitro analysis of the isoprenaline release profile showed that most (63.9 ± 5.9%) of the loaded drug mass was released in the 1st minute, and almost all of the drug was released in the first 4 minutes exponentially. Porcine studies showed a 25.1% reduction in IRP in the IsoWire that released 10 µg in the 1st minute; however, there was a marked increase in HR. The average percentage reduction in IRP was 8.95% and 21.3% in the IsoWire that released 5 and 7.5 µg of isoprenaline, respectively, with no changes in HR or MAP. Conclusions: The IsoWire, which releases 5 and 7.5 µg of isoprenaline in the 1st minute, appears to be safe and effective in reducing the IRP. Further studies are needed to establish whether the isoprenaline-induced ureteral relaxation will render easier insertion of a ureteral access sheath, reduce IRP during sheathless RIRS, or even promote the practice of sheathless RIRS.

Detection of Cancer-Associated Gene Mutations in Urinary Cell-Free DNA among Prostate Cancer Patients in South Africa.

Prostate cancer (PCa) is the most common cause of cancer death among African men. The presence of tumor-specific variations in cell-free DNA (cfDNA), such as mutations, microsatellite instability, and DNA methylation, has been explored as a source of biomarkers for cancer diagnosis. In this study, we investigated the diagnostic role of cfDNA among South African PCa patients. We performed whole exome sequencing (WES) of urinary cfDNA. We identified a novel panel of 31 significantly deregulated somatic mutated genes between PCa and benign prostatic hyperplasia (BPH). Additionally, we performed whole-genome sequencing (WGS) on matching PCa and normal prostate tissue in an independent PCa cohort from South Africa. Our results suggest that the mutations are of germline origin as they were also found in the normal prostate tissue. In conclusion, our study contributes to the knowledge of cfDNA as a biomarker for diagnosing PCa in the South African population.

Potential of miRNAs in Plasma Extracellular Vesicle for the Stratification of Prostate Cancer in a South African Population.

Prostate cancer (PCa) is the most common cause of cancer death among African men. The analysis of microRNAs (miRNAs) in plasma extracellular vesicles (EVs) can be utilized as a non-invasive tool for the diagnosis of PCa. In this study, we used small RNA sequencing to profile miRNAs cargo in plasma EVs from South African PCa patients. We evaluated the differential expression of miRNAs between low and high Gleason scores in the plasma EVs of South African patients and in the prostatic tissue from data available in the Cancer Genome Atlas (TCGA) Data Portal. We identified 7 miRNAs differently expressed in both EVs and prostatic tissues. We evaluated their expression using qPCR in a larger cohort of 10 patients with benign prostatic hyperplasia (BPH) and 24 patients with PCa. Here, we reported that the ratio between two of these miRNAs (i.e., miR-194-5p/miR-16-5p) showed a higher concentration in PCa compared to BPH and in metastatic PCa compared to localized PCa. We explored for the first time the profiling of miRNAs cargo in plasma EVs as a tool for the identification of putative markers in the South African population. Our finding indicated the ratio miR-194-5p/miR-16-5p as a non-invasive marker for the evaluation of PCa aggressiveness in this population.

A Pilot Study of a Novel Syphon Ureteral Access Sheath Shows Potential to Reduce Renal Pressures and Improve Irrigant Flow.

OBJECTIVE: To describe a novel syphon ureteral access sheath (UAS) intended for use during flexible uretero-renoscopy (fURS). We aimed to report on a pilot study as well as intrarenal pressures (IRP) and irrigant flow volumes compared to traditional UAS. METHODS: Patients undergoing routine fURS for single, <2cm intrarenal nephrolithiasis were identified, and written informed consent was obtained. Irrigation via the fURS was instilled through the novel 11/13 Fr UAS without (a proxy for a traditional UAS) and with the novel syphon box attached. Measured minute irrigant flow volume, steady state and bolus IRP were compared. RESULTS: Ten participants (6 males and 4 females) were treated with the syphon UAS. All procedures were completed safely without intraoperative complications. The mean baseline IRP with and without the syphon was 18 vs 29 mmHG (P < .001, SD 4.0 vs 4.8). The mean minute irrigant flow volume with and without the syphon was 31 vs 21 ml (P < .001, SD 6.4 vs 3.3). The mean peak IRP following a 10 ml bolus with and without the syphon was 71 vs 104 mmHg (P = .03, SD 74 vs 59). CONCLUSION: The described novel UAS is different from traditional devices by incorporating a syphon mechanism. This pilot trial demonstrates that the novel syphon UAS may hold clinical potential to reduce IRP and increase irrigant flow compared to traditional UAS. Firm conclusions about efficacy and safety require assessment of the device in a larger clinical trial.

Validation of an ovine vesicovaginal fistula model.

INTRODUCTION AND HYPOTHESIS: A representative, large animal model of vesicovaginal fistulas is needed for the training of surgeons and for the development of new surgical techniques and materials for obstetric fistula repair. METHODS: The safety, feasibility, and reproducibility of vesicovaginal fistula creation were studied in 4 adult female sheep. A 1-cm fistula was created between the vagina and the bladder through a transvaginal approach. The defect was allowed to heal for 8 weeks and the animals were then euthanized. The primary outcome was the fistula patency. Secondary outcomes were fistula size, urogenital dimensions, urodynamic evaluation, histology (inflammation, vascularization, collagen deposition) and biomechanical characteristics of the fistula edge (stress at break, maximum elongation, and stiffness). RESULTS: The transvaginal creation of a vesicovaginal fistula was safe. All animals survived the surgical procedure and follow-up period, without complications. Three of the four animals demonstrated a patent vesicovaginal fistula after 8 weeks. Baseline data are provided of the urogenital dimensions and the urodynamic, histological, and biomechanical characteristics of the model. CONCLUSIONS: The ewe is a safe, feasible, and reproducible model for vesicovaginal fistulas. The model can help to study new techniques and materials to boost surgical innovation for vesicovaginal fistula repair.

Novel syphon ureteric access sheath has the potential to improve renal pressures and irrigant flow.

OBJECTIVE: To describe a novel syphoning ureteric access sheath (UAS) intended for use during flexible ureterorenoscopy (URS). We aimed to assess if in vitro it could reduce intrarenal pressure (IRP) and increase irrigant flow compared to traditional UASs. MATERIALS AND METHODS: A validated phantom kidney with fibre optic pressure sensing capabilities was used to assess the IRP. Standardised 80 cmH2 O irrigation via a ureterorenoscope was instilled through traditional UASs (11/13 and 12/14 F) and compared to the novel 11/13-F syphoning UAS. The measured minute volume, calculated hourly flow volume, and steady state IRP were compared. RESULTS: The traditional 11/13 and 12/14-F UASs had statistically poorer irrigant flow than the novel syphoning UAS, at 19.3 vs 29.3 mL/min (P < 0.001) and 22.7 vs 29.3 mL/min (P = 0.002), respectively. The steady state IRP was 20 mmHg for the traditional 11/13 F and 13 mmHg for the 12/14 F compared to 0 mmHg for the novel UAS. CONCLUSION: The described novel UAS is different from traditional devices by incorporating a syphon mechanism. Our in vitro assessment demonstrates that the novel UAS holds clinical potential to reduce IRP while allowing a significant increase in irrigant flow compared to larger diameter traditional UASs.

Inflammatory metabolic profile of South African patients with prostate cancer.

BACKGROUND: Men with African ancestry are more likely to develop aggressive prostate cancer (PCa) and to die from this disease. The study of PCa in the South African population represents an opportunity for biomedical research due to the high prevalence of aggressive PCa. While inflammation is known to play a significant role in PCa progression, its association with tumor stage in populations of African descent has not been explored in detail. Identification of new metabolic biomarkers of inflammation may improve diagnosis of patients with aggressive PCa. METHODS: Plasma samples were profiled from 41 South African men with PCa using nuclear magnetic resonance (NMR) spectroscopy. A total of 41 features, including metabolites, lipid classes, total protein, and the inflammatory NMR markers, GlycA, and GlycB, were quantified from each NMR spectrum. The Bruker's B.I.-LISA protocols were used to characterize 114 parameters related to the lipoproteins. The unsupervised KODAMA method was used to stratify the patients of our cohort based on their metabolic profile. RESULTS: We found that the plasma of patients with very high risk, aggressive PCa and high level of C-reactive protein have a peculiar metabolic phenotype (metabotype) characterized by extremely high levels of GlycA and GlycB. The inflammatory processes linked to the higher level of GlycA and GlycB are characterized by a deep change of the plasma metabolome that may be used to improve the stratification of patients with PCa. We also identified a not previously known relationship between high values of VLDL and low level of GlycB in a different metabotype of patients characterized by lower-risk PCa. CONCLUSIONS: For the first time, a portrait of the metabolic changes in African men with PCa has been delineated indicating a strong association between inflammation and metabolic profiles. Our findings indicate how the metabolic profile could be used to identify those patients with high level of inflammation, characterized by aggressive PCa and short life expectancy. Integrating a metabolomic analysis as a tool for patient stratification could be important for opening the door to the development of new therapies. Further investigations are needed to understand the prevalence of an inflammatory metabotype in patients with aggressive PCa.

Optically Tracked Needle for Ultrasound-Guided Percutaneous Nephrolithotomy Puncture: A Preliminary Report.

Background and Aim: Precise needle puncture of the renal collecting system is an essential step for effective percutaneous nephrolithotomy (PCNL). The use of ultrasound for puncture is receiving increased attention. Ultrasound has recognized limitations related to poor observation of the needle tip. We aimed to assess whether an affordable open-source computerized needle navigation training system, using optically tracked ultrasonography, could improve performance of simulated PCNL puncture by urologic trainees, compared with conventional free hand manual sonographic puncture. Materials and Methods: This study describes a PCNL navigation system that can be recreated with any standard ultrasound machine using relatively inexpensive components. The system allows the needle tip to be precisely appreciated in the ultrasound image, its trajectory planned, and the appreciation of needle tip to target calix proximity sound effect. Eight participants (six trainees and two qualified urologists) assessed the PCNL training model. Alternating starting with freehand (control) and tracked needle (experimental) punctures were performed on a phantom kidney. Total procedure and the number of reinsertions required were recorded. Results: The mean time for freehand puncture was 89 seconds (range 13-173), whereas that of the optically tracked needle was 36 seconds (range 12-72). Thus, puncture time was significantly reduced by an average of 53 seconds (p = 0.045) in the experimental arm. The mean number of needle reinsertions was 3.3 with freehand compared with 1.3 in the optically tracked puncture (p = 0.005). The mean square root error of the optical tracking system was 1.8 mm (four calibrations averaged). Conclusion: This study demonstrates that affordable hardware and open-source software can be used to construct an optically tracked ultrasound navigation system for PCNL training. Statistically significant reduced puncture time and number of passes required for effective puncture are demonstrated. We feel that computerized needle tracking during PCNL puncture deserves further evaluation in a training and, potentially, in a clinical setting.

Management of urological injury at the time of urogynaecology surgery.

The presentation and management of bladder, ureteric and urethral injuries during and following urogynaecology surgery are discussed. Applied anatomy is reviewed, and the surgical management of injuries diagnosed intra- and post-operatively is discussed.

Novel potential serological prostate cancer biomarkers using CT100+ cancer antigen microarray platform in a multi-cultural South African cohort.

There is a growing need for high throughput diagnostic tools for early diagnosis and treatment monitoring of prostate cancer (PCa) in Africa. The role of cancer-testis antigens (CTAs) in PCa in men of African descent is poorly researched. Hence, we aimed to elucidate the role of 123 Tumour Associated Antigens (TAAs) using antigen microarray platform in blood samples (N = 67) from a South African PCa, Benign prostatic hyperplasia (BPH) and disease control (DC) cohort. Linear (fold-over-cutoff) and differential expression quantitation of autoantibody signal intensities were performed. Molecular signatures of candidate PCa antigen biomarkers were identified and analyzed for ethnic group variation. Potential cancer diagnostic and immunotherapeutic inferences were drawn. We identified a total of 41 potential diagnostic/therapeutic antigen biomarkers for PCa. By linear quantitation, four antigens, GAGE1, ROPN1, SPANXA1 and PRKCZ were found to have higher autoantibody titres in PCa serum as compared with BPH where MAGEB1 and PRKCZ were highly expressed. Also, p53 S15A and p53 S46A were found highly expressed in the disease control group. Statistical analysis by differential expression revealed twenty-four antigens as upregulated in PCa samples, while 11 were downregulated in comparison to BPH and DC (FDR = 0.01). FGFR2, COL6A1and CALM1 were verifiable biomarkers of PCa analysis using urinary shotgun proteomics. Functional pathway annotation of identified biomarkers revealed similar enrichment both at genomic and proteomic level and ethnic variations were observed. Cancer antigen arrays are emerging useful in potential diagnostic and immunotherapeutic antigen biomarker discovery.

In silico verification and parallel reaction monitoring prevalidation of potential prostate cancer biomarkers.

PURPOSE: Targeted proteomics of potential biomarkers is often challenging. Hence, we developed an intermediate workflow to streamline potential urinary biomarkers of prostate cancer (PCa). MATERIALS & METHODS: Using previously discovered potential PCa biomarkers; we selected proteotypic peptides for targeted validation. Preliminary in silico immunohistochemical and single reaction monitoring (SRM) verification was performed. Successful PTPs were then prevalidated using parallel reaction monitoring (PRM) and reconfirmed in 15 publicly available databases. RESULTS: Stringency-based targetable potential biomarkers were shortlisted following in silico screening. PRM reveals top 12 potential biomarkers including the top ranking seven in silico verification-based biomarkers. Database reconfirmation showed differential expression between PCa and benign/normal prostatic urine samples. CONCLUSION: The pragmatic penultimate screening step, described herein, would immensely improve targeted proteomics validation of potential disease biomarkers.

Discovery of novel candidate urinary protein biomarkers for prostate cancer in a multiethnic cohort of South African patients via label-free mass spectrometry.

PURPOSE: Improvement in diagnostic accuracy of prostate cancer (PCa) progression using MS-based methods to analyze biomarkers in our African, Caucasian, and Mixed Ancestry patients can advance early detection and treatment monitoring. EXPERIMENTAL DESIGN: MS-based proteomic analysis of pooled (N = 36) and individual samples (N = 45) of PCa, benign prostatic hyperplasia, normal healthy controls, and patients with other uropathies was used to identify differences in proteomics profile. Samples were analyzed for potential biomarkers and proteome coverage in African, Caucasian, and Mixed Ancestry PCa patients. RESULTS: A total of 1102 and 5595 protein groups and nonredundant peptides, respectively, were identified in the pooling experiments (FDR = 0.01). Twenty potential biomarkers in PCa were identified and fold differences ± 2SD were observed in 17 proteins using intensity-based absolute quantification. Analysis of 45 individual samples yielded 1545 and 9991 protein groups and nonredundant peptides, respectively. Seventy-three (73) proteins groups, including existing putative PCa biomarkers, were found to be potential biomarkers of PCa by label-free quantification and demonstrated ethnic trends within our PCa cohort. CONCLUSION AND CLINICAL RELEVANCE: Urinary proteomics is a promising route to PCa biomarker discovery and may serve as source of ethnic-related biomarkers of PCa.

Intravesical condylomata accuminata in HIV positive patient.

A 48-year-old HIV positive woman presented with urgency, frequency, recurrent cystitis and episodic macroscopic hematuria. Cystoscopy revealed papillary lesions involving most of the bladder. Histology of bladder biopsies revealed human papilloma virus (HPV) associated condyloma acuminata. We discuss the treatment of this rare lesion and review the literature.