Extended-spectrum ß-lactamase in the rectal flora of patients undergoing transrectal prostate biopsy: a study of the prevalence in a major metropolitan hospital.

OBJECTIVES: To determine the prevalence of extended-spectrum ß-lactamase (ESBL) in patients undergoing transrectal prostate biopsy, to assess the incidence of postoperative sepsis, to correlate the development of sepsis with the presence of preoperative ESBL on rectal swabs, and to assess the adequacy of prophylactic antibiotic guidelines in the context of local ESBL prevalence. METHODS: Patients undergoing transrectal ultrasonography (TRUS)-guided biopsy at the Royal Melbourne Hospital between January 2012 and July 2016 had rectal swabs taken immediately prior to TRUS with specific cultures to identify the presence of ESBL. Patients were given a prophylactic antibiotic, 500 mg oral ciprofloxacin, 1 h before the TRUS procedure. Data were collected prospectively, with retrospective review of all readmitted patient files and audit data to ensure complete capture of events. RESULTS: A total of 387 TRUS-guided biopsy procedures were performed. Rectal swabs were correctly collected in 352 patients (91%). The median patient age was 65 years. In all, 25 (7%) ESBL-positive swabs were identified. Most ESBL were Escherichia coli. Half (50%) of ESBL were resistant to ciprofloxacin and all were sensitive to meropenem. A small increase in ESBL prevalence over time was not significant (R2 = 0.35). Four patients (1.1%) were readmitted with sepsis; ESBL Pseudomonas had previously grown in one patient, but sepsis was attributable to non-ESBL E. coli. In one of the readmitted patients ESBL E. coli was present, but this patient did not have ESBL preoperatively. There were no deaths or high-dependency/intensive care unit admissions. CONCLUSIONS: This study represents the largest Australian series to investigate ESBL prevalence, and reveals a rate lower than that of many other nations. Our sepsis rate is lower than many international series, perhaps because of our low ESBL rate and strict antibiotic prophylaxis. Preoperative swab results did not predict postoperative sepsis, and the process was therefore not useful for guiding antibiotic therapy. In this patient population, TRUS biopsy, with ciprofloxacin prophylaxis, remains a safe option for diagnostic prostate biopsy.

A urinary microRNA signature can predict the presence of bladder urothelial carcinoma in patients undergoing surveillance.

BACKGROUND: The objective of this study was to determine whether microRNA (miRNA) profiling of urine could identify the presence of urothelial carcinoma of the bladder (UCB) and to compare its performance characteristics to that of cystoscopy. METHODS: In the discovery cohort we screened 81 patients, which included 21 benign controls, 30 non-recurrers and 30 patients with active cancer (recurrers), using a panel of 12 miRNAs. Data analysis was performed using a machine learning approach of a Support Vector Machine classifier with a Student's t-test feature selection procedure. This was trained using a three-fold cross validation approach and performance was measured using the area under the receiver operator characteristic curve (AUC). The miRNA signature was validated in an independent cohort of a further 50 patients. RESULTS: The best predictor to distinguish patients with UCB from non-recurrers was achieved using a combination of six miRNAs (AUC=0.85). This validated in an independent cohort (AUC=0.74) and detected UCB with a high sensitivity (88%) and sufficient specificity (48%) with all significant cancers identified. The performance of the classifier was best in detecting clinically significant disease such as presence of T1 Stage disease (AUC=0.92) and high-volume disease (AUC=0.81). Cystoscopy rates in the validation cohort would have been reduced by 30%. CONCLUSIONS: Urinary profiling using this panel of miRNAs shows promise for detection of tumour recurrence in the surveillance of UCB. Such a panel may be useful in reducing the morbidity and costs associated with cystoscopic surveillance, and now merits prospective evaluation.

Video labelling robot-assisted radical prostatectomy and the role of artificial intelligence (AI): training a novice.

Video labelling is the assigning of meaningful information to raw videos. With the evolution of artificial intelligence and its intended incorporation into the operating room, video datasets can be invaluable tools for education and the training of intelligent surgical workflow systems through computer vision. However, the process of manual labelling of video datasets can prove costly and time-consuming for already busy practising surgeons. Twenty-five robot-assisted radical prostatectomy (RARP) procedures were recorded on Proximie, an augmented reality platform, anonymised and access given to a novice, who was trained to develop the knowledge and skills needed to accurately segment a full-length RARP procedure on a video labelling platform. A labelled video was subsequently randomly selected for assessment of accuracy by four practising urologists. Of the 25 videos allocated, 17 were deemed suitable for labelling, and 8 were excluded on the basis of procedure length and video quality. The labelled video selected for assessment was graded for accuracy of temporal labelling, with an average score of 93.1%, and a range of 85.6-100%. The self-training of a novice in the accurate segmentation of a surgical video to the standard of a practising urologist is feasible and practical for the RARP procedure. The assigning of temporal labels on a video labelling platform was also studied and proved feasible throughout the study period.

Management of local recurrences in the irradiated bladder: a systematic review.

OBJECTIVE: • To review and report the management and outcomes of patients with recurrence of bladder cancer (BC) who initially had complete response to bladder conservation therapy. METHODS: • A comprehensive review of all published literature reporting outcomes in bladder-sparing treatments for muscle-invasive BC (MIBC) was conducted in a systematic fashion by two independent authors across several databases including PubMed and Medline using the following keywords, alone or in combination: bladder cancer, radiotherapy, recurrence, outcome, cystoscopy, follow up, combined modality therapy, and organ preservation. • In all, 17 studies reporting prevalence and management of local recurrences were included in final analysis. RESULTS: • Complete response rates to CRT ranged from 56 to 100%. • Local recurrence rates ranged from 13 to 40%. Initial BC recurrences were equally likely to be non-muscle-invasive (NMI) or muscle invasive (MI). Average time to local recurrence from RT was 18-36 months but can extend to 10 years. • For first NMIBC recurrences, transurethral resection of bladder tumour (TURBT) with or without intravesical therapy was most commonly used with cystectomy and TURBT used equally for subsequent recurrences. Cystectomy was advocated most commonly for MIBC recurrences. • In most studies, the 5-year cancer-specific survival for patients with NMIBC recurrences was 50-70%, with that for MIBC recurrences being much less at 16-40%. Patients with recurrences have a lower probability of surviving with an intact bladder compared with those who do not have a recurrence. • Age, resection status, T-stage and presence of carcinoma in situ at time of RT were adversely associated with local control and overall survival. CONCLUSION: • In patients with MIBC managed with CRT, who have a complete response at initial cystoscopy, survival after NMIBC recurrence is comparable with those without any recurrence but with a smaller chance of surviving with an intact bladder. However, prognosis for patients with MIBC recurrences remains poor. • Cystoscopic follow-up to 10 years is mandated.

Nurse-led flexible cystoscopy in Australia: initial experience and early results.

OBJECTIVE: • To present our initial experience implementing a nurse-led flexible cystoscopy (NLFC) service in a Victorian tertiary hospital and our initial results from that service, as NLFC has developed over the past decade with reports suggesting that adequately trained nurses can undertake FC competently. PATIENTS AND METHODS: • We describe the implementation of a NLFC service including approval, funding, nurses' training, and protocols. • Outcomes of all patients having a NLFC or subsequent interventions were recorded prospectively and analysed retrospectively. • To gauge patients' response to NLFC, an anonymous feedback questionnaire was administered to 60 consecutive participating patients in the recovery unit. • The effect of NLFC on waiting times was determined from surgical scheduling records. RESULTS: • In all, 272 patients had 720 NLFC done over a 2-year period. In all, 150 (21%) FCs had a suspected bladder cancer recurrence and were referred for a rigid cystoscopy. Of those, 83 (58%) revealed a recurrence comprising of 14 (17%) high-grade lesions, 45 (54%) low-grade lesions and 24 (29%) were diathermied without a biopsy. In all, 41 (27%) had benign pathology on biopsy and 21 (14%) had normal rigid cystoscopy. • There were two significant adverse events. • There was a 65% reduction in the waiting list for surveillance FC after introduction of the service. • Of 60 patients who completed the feedback questionnaire, 95% reported that they were given enough information by the nurses, 92% had all their questions answered satisfactorily and 97% had enough confidence and trust in the nurse. In all, 90% had a positive perception of the service overall and 93% were happy to have a FC performed by a nurse rather than a doctor. CONCLUSIONS: • Results from our NLFC audit compare favourably with other published reports. NLFC is a safe and feasible option when established alongside strong departmental support, comprehensive nurses' training according to established guidelines, service supervision by a designated consultant and regular audits. • NLFC clinics can provide an efficient service and excellent continuity of care for patients with non-muscle-invasive bladder cancer.

Re-evaluating the biological significance of seminal vesicle invasion (SVI) in locally advanced prostate cancer.

OBJECTIVE: • To examine the impact of seminal vesicle invasion (SVI) in patients with locally advanced (pT3) prostate cancer on clinical outcome. • To explore the clinical association of SVI with metastatic disease. • To distinguish between the possibilities that either seminal vesicles possess their own biological significance and represent a privileged staging site for systemic tumour cell dissemination, or that their invasion is a surrogate marker for an aggressive large-volume poorly differentiated cancer. PATIENTS AND METHODS: • Patients with extraprostatic extension (EPE) and/or SVI were identified from a prospectively recorded and maintained prostate cancer database. • Patients were categorised according to the presence of SVI as determined by routine pathological assessment. Tumour volumes were measured routinely by computed planimetry at the time of histological assessment. • The impact of SVI on biochemical recurrence with a definition of a prostate-specific antigen (PSA) level of ≥0.2 ng/mL, as well as a clinically significant recurrence defined as failure with a PSA doubling time of <6 months, was determined by univariable and multivariable Cox regression analysis. RESULTS: • Of 249 patients with pT3 disease, 46 (18%) had SVI, 40 (87%) by direct extension and six (13%) metastatic. • Tumours with SVI had significantly greater tumour burden as determined by total tumour volume (7.2 vs 3.7 mL, P < 0.001), index tumour volume (6.8 vs. 3.4 mL, P < 0.001) and percentage tumour volume (21.8 vs 12.4 %, P= 0.001). • After controlling for tumour volume and Gleason score, the presence of SVI did not significantly predict for the development of a significant PSA recurrence. CONCLUSIONS: • Our results suggest that SVI is a surrogate marker of larger and more aggressive tumours with higher Gleason scores rather than a privileged site of tumour cell dissemination.

Superior Mesenteric Artery Injury During Robot-assisted Laparoscopic Nephrectomy: A Robotic Nightmare.

Major vascular injuries during robotic renal surgery are rare, but the close proximity of the superior mesenteric artery (SMA) to the left renal artery means that it is liable to iatrogenic injury with potentially catastrophic implications. In this review, we present a case of accidental SMA ligation during a robot-assisted laparoscopic nephrectomy for a 12-cm upper pole renal mass. Prompt recognition and early vascular surgical assistance with conversion to open surgery allowed a primary vascular anastomosis to be made. A computed tomography angiogram at 6 wk was normal. On review of the imaging, the left renal artery take-off was higher than the SMA, which represents an anatomical variant and may have contributed to the injury. The risk of accidental SMA ligation is highest in left-sided tumours and in larger medial tumours that lead to significant distortion of the anatomy. The anatomy of the renal artery can also vary greatly. Surgeons must be knowledgeable of common variations and meticulously review preoperative imaging for the number and course of renal vessels as well as the location of the SMA. In cases of significant bleeding, rapid conversion to open surgery and urgent vascular consultation are critical. PATIENT SUMMARY: In this article, we describe an accidental injury to a major blood vessel (the superior mesenteric artery) during a left robotic radical nephrectomy (kidney removal) for a tumour. We discuss the anatomical relationships of the blood vessels of the small bowel and kidneys, and how to anticipate, recognise, and manage such accidental injuries.

Streamlining the assessment of haematuria: 3-year outcomes of a dedicated haematuria clinic.

BACKGROUND: Urgent assessment of haematuria is critical to exclude malignancy. The objective of this study is to report the outcomes of the first 3 years of a dedicated haematuria clinic at the Royal Melbourne Hospital, a Victorian tertiary hospital. METHODS: All patients assessed in the haematuria clinic from April 2010 to April 2013 were included in the analysis. Outcomes were recorded prospectively and analysed retrospectively. RESULTS: A total of 643 patients were seen in the haematuria clinic with non-visible (170, 26%) and visible haematuria (463, 72%) during this time period, all within 28 days of referral being triaged. Sixty-five (10%) patients were diagnosed with urothelial carcinoma, 63 with lower tract disease and two with upper tract urothelial carcinoma and another five (1%) patients with other tumours. Thirty out of 63 (48%) of the bladder urothelial carcinomas were invasive or high-grade. Two hundred and sixty-seven (42%) patients were discharged from the clinic after a single point of contact. One hundred and fifty-three (24%) patients were referred for further definitive management of suspected pathology. Two hundred and twenty-three (34%) patients were referred to outpatients clinic for further investigations. Urothelial carcinoma was diagnosed more often in males, older patients and patients with visible haematuria. CONCLUSION: The Royal Melbourne Hospital haematuria clinic has served as an effective tool for rapid, streamlined assessment of patients presenting with haematuria. Follow-up of investigations by nurses and moving towards a 'one-stop' approach are helping to further decrease the number of patients requiring a second clinic visit.

Tracking the origins and drivers of subclonal metastatic expansion in prostate cancer.

Tumour heterogeneity in primary prostate cancer is a well-established phenomenon. However, how the subclonal diversity of tumours changes during metastasis and progression to lethality is poorly understood. Here we reveal the precise direction of metastatic spread across four lethal prostate cancer patients using whole-genome and ultra-deep targeted sequencing of longitudinally collected primary and metastatic tumours. We find one case of metastatic spread to the surgical bed causing local recurrence, and another case of cross-metastatic site seeding combining with dynamic remoulding of subclonal mixtures in response to therapy. By ultra-deep sequencing end-stage blood, we detect both metastatic and primary tumour clones, even years after removal of the prostate. Analysis of mutations associated with metastasis reveals an enrichment of TP53 mutations, and additional sequencing of metastases from 19 patients demonstrates that acquisition of TP53 mutations is linked with the expansion of subclones with metastatic potential which we can detect in the blood.

Gene-based urinary biomarkers for bladder cancer: an unfulfilled promise?

OBJECTIVE: Noninvasive biomarkers are used routinely in the clinical management of several cancers but bladder cancer detection and surveillance remains dependent on invasive procedures such as cystoscopy. No validated biomarker currently exists in routine clinical practice other than cytology. Gene-based testing has shown great promise for biomarker profiling and this review addresses the current state of biomarker research in bladder cancer. MATERIALS AND METHODS: A comprehensive review of all published literature on urinary biomarkers from 1970 - 2012 was conducted in PubMed. Keywords used alone or in combination were bladder cancer, diagnosis, surveillance, urinary biomarker, molecular biomarkers, methylation, gene expression, single nucleotide polymorphism and microRNA. The cited references of the manuscripts included in the review were also screened. RESULTS: We have reviewed various strategies currently used for gene-based biomarker profiling of bladder cancer. We have comprehensively summarized the performance of several biomarkers in the diagnosis and surveillance of bladder cancer. Finally we have identified biomarkers that have shown potential and now deserve the opportunity to be validated in the clinical setting. CONCLUSION: Several gene-based urinary biomarkers have demonstrated promise in initial studies, which now need to be rigorously validated in the clinical setting for them to be translated into clinically useful tests in diagnosis, surveillance or risk-stratification of bladder cancer.

Molecular biomarkers for predicting outcomes in urothelial carcinoma of the bladder.

Molecular biomarkers are used routinely in the clinical management of several tumours such as prostate, colon, ovarian and pancreatic cancer but management decisions in bladder cancer remain dependent on clinical and pathological criteria, which are limited in their ability to predict outcomes. Molecular markers are urgently needed in detection, surveillance and prognostication of bladder cancer as well as to predict treatment response to intravesical and systemic therapies. Advances in cancer genomics and platforms for biomarker profiling have led to a plethora of biomarkers, which must now be rigorously validated in the clinical setting. Pre-clinical and clinical studies exploring the role of emerging targeted therapies to risk stratify and reduce cancer progression are also needed.

Curated microRNAs in urine and blood fail to validate as predictive biomarkers for high-risk prostate cancer.

PURPOSE: The purpose of this study was to determine if microRNA profiling of urine and plasma at radical prostatectomy can distinguish potentially lethal from indolent prostate cancer. MATERIALS AND METHODS: A panel of microRNAs was profiled in the plasma of 70 patients and the urine of 33 patients collected prior to radical prostatectomy. Expression of microRNAs was correlated to the clinical endpoints at a follow-up time of 3.9 years to identify microRNAs that may predict clinical response after radical prostatectomy. A machine learning approach was applied to test the predictive ability of all microRNAs profiled in urine, plasma, and a combination of both, and global performance assessed using the area under the receiver operator characteristic curve (AUC). Validation of urinary expression of miRNAs was performed on a further independent cohort of 36 patients. RESULTS: The best predictor in plasma using eight miRs yielded only moderate predictive performance (AUC = 0.62). The best predictor of high-risk disease was achieved using miR-16, miR-21 and miR-222 measured in urine (AUC = 0.75). This combination of three microRNAs in urine was a better predictor of high-risk disease than any individual microRNA. Using a different methodology we found that this set of miRNAs was unable to predict high-volume, high-grade disease. CONCLUSIONS: Our initial findings suggested that plasma and urinary profiling of microRNAs at radical prostatectomy may allow prognostication of prostate cancer behaviour. However we found that the microRNA expression signature failed to validate in an independent cohort of patients using a different platform for PCR. This highlights the need for independent validation patient cohorts and suggests that urinary microRNA signatures at radical prostatectomy may not be a robust way to predict the course of clinical disease after definitive treatment for prostate cancer.

Percutaneous image-guided biopsy of prostate cancer metastases yields samples suitable for genomics and personalised oncology.

Personalised oncology through mutational profiling of cancers requires the procurement of fresh frozen tumour samples for genomics applications. While primary cancers are often surgically excised and therefore yield such tissue, metastases in the setting of a known cancer diagnosis are not routinely sampled prior to systemic therapy. Our study aimed to determine the suitability of extracted nucleic acids for genomics applications using distant metastatic prostate cancer samples obtained via percutaneous or surgical biopsy. Patients with metastatic prostate cancer were recruited for image-guided biopsy of metastases. Patients undergoing surgical procedures for the complications of metastases were also recruited. Tissue samples were flash frozen and cryosectioned for histological examination. DNA and RNA were simultaneously extracted and genomic DNA hybridised onto SNP arrays for genome-wide copy number analysis. 37 samples of metastatic tissue from seven patients with prostate cancer were obtained. Five of these underwent image-guided biopsies whilst two had therapeutic surgical procedures performed. 22 biopsy samples were obtained across the image-guided biopsy patients with 80 % of samples being successfully processed for downstream analysis. Nucleic acid yield from these samples were satisfactory for genomics applications. Copy number analysis revealed a median estimated tumour purity of 53 % and all samples showed chromosomal abnormalities suggestive of malignancy. The procurement of osseous metastatic prostate cancer from live patients, including the use of image-guided biopsy, is safe and feasible. Sufficient tissue can be obtained in a manner such that extracted nucleic acids are suitable for genomics research.

Circulating MicroRNAs as Biomarkers of Prostate Cancer: The State of Play.

MicroRNAs are key regulators of gene expression and play critical roles in both normal physiology and pathology. Recent research has demonstrated that these molecules are present in body fluids, such as serum, plasma, and urine, and can be readily measured using a variety of techniques. More importantly, emerging evidence suggests that circulating or urine miRNAs are useful indicators of disease. Here, we consider the potential utility of such miRNAs as noninvasive biomarkers of prostate cancer, a disease that would benefit substantially from novel diagnostic and prognostic tools. The studies aimed at identifying diagnostic, prognostic, and/or predictive miRNAs for prostate cancer are summarised and reviewed. Finally, practical considerations that will influence the translation of this recent research into clinical implementation are discussed.

Tumor vascularity in prostate cancer: an update on circulating endothelial cells and platelets as noninvasive biomarkers.

In order to individually tailor prostate cancer (PCa) treatment, clinicians need better tools to predict prognosis and treatment response. Given the relationship between angiogenesis and cancer progression, circulating endothelial cells (CECs) and their progenitors have logically been proposed as potential biomarkers. The utility of their baseline levels and kinetics has been investigated for years. However, owing to a lack of standardization and validation of CEC and circulating endothelial progenitors enumeration protocols, results have been inconsistent in prostate and other cancers. Similarly, platelets play a significant part in cancer progression, yet the role of platelet-related biomarkers in PCa is unclear. While there have been a number of theoretically interesting platelet-related markers proposed, limited research has been conducted in PCa patients. Currently, CECs and platelets do not have a clear role as biomarkers in routine PCa care. Given the theoretical merits of these cells, prospective trials are warranted.

Bladder cancer biorepositories in the "-omics" era: integrating quality tissue specimens with comprehensive clinical annotation.

Advances in genomic platforms have led to the need for well-characterized, high quality biospecimens for research. Bladder cancer, despite being a major epidemiological concern, has been under-represented in genomic programs due to unique challenges in collection of clinical informatics and tissues. Currently no targeted therapy exists for management of the disease. We report our experiences and lessons learnt in establishing an integrated model of bladder cancer that uses a dedicated bladder cancer team and relational databases. It streamlines both clinical activity and serial harvesting of biospecimens to obtain tissue that is consistently suitable for high-throughput genomic research. Fresh tissue, blood, and urine samples were prospectively collected and stored. RNA and DNA were extracted simultaneously, quality control was performed, and whole transcriptome sequencing also performed using the illumina series of platforms. Over a 15-month period, urine was banked for 209 patients, plasma and whole blood for 185 patients, and tissue for 71 patients. The collections included normal mucosa from patients with and without bladder cancer and cancer tissue across the entire histopathogical spectrum of bladder cancer from low-grade noninvasive cancers to metastatic lymph nodes. We used a relational database to link clinical information to tumor inventory and provide access to richly annotated specimens and matched clinical informatics. We found that tumor tissue was successfully banked more often in patients who had macroscopic papillary disease compared to those without. We also show that the median RNA integrity number (RIN) scores are significantly higher in patients whose tissues were banked using cold-cup biopsies compared to those banked using electrocautery. Transcriptome sequencing of all samples banked using cold-cup biopsies passed bioinformatics quality assessment and had a mean Q30 quality score well over the illumina quality control benchmark. Such an infrastructure will allow genomic profiling of bladder cancer to help us understand the "global picture" in bladder cancer pathogenesis.

Prostate tumour volume is an independent predictor of early biochemical recurrence in a high risk radical prostatectomy subgroup.

AIMS: To assess if accurately determined tumour volume variables could serve as independent predictors of early biochemical recurrence in high risk prostate cancer patients who underwent radical prostatectomy. METHODS: Tumour volume variables were calculated by digital planimetry in 269 prostatectomy specimens of patients with high risk prostate cancer. The associations to biochemical progression of tumour volume and clinicopathological variables, including age, pre-operative prostate specific antigen (PSA) levels, final Gleason score, pathological T stage, and surgical margins, were examined using univariate and multivariate Cox proportional hazards analyses. RESULTS: Median tumour volume was 3.7 ml [interquartile range (IQR) 2.1-6.1 mL] and median follow-up time was 12 months (IQR 6-24 months). Biochemical recurrence occurred in 64 men (24%) during this period, with a median time to recurrence of 7.5 months (IQR 3.0-15.5 months). On univariate analysis all of the tumour volume variables were strongly correlated with the clinicopathological variables, as well as biochemical recurrence (p < 0.001). On multivariate analysis, we found that tumour volume variables served as independent predictors of PSA progression whilst other routinely reported pathological variables did not. CONCLUSION: Accurately assessing tumour volume in the high risk setting may aid in identifying patients at greatest risk of developing early biochemical recurrence and most in need of adjuvant therapy.