No abstract

Cell-free microRNAs have been reported as biomarkers for several diseases. For testicular germ cell tumors (GCT), circulating microRNAs 371a-3p and 372-3p in serum and plasma have been proposed as biomarkers for diagnostic and disease monitoring purposes. The most widely used method for quantification of specific microRNAs in serum and plasma is reverse transcriptase real-time quantitative PCR (RT-qPCR) by the comparative Ct-method. In this method one or several reference genes or reference microRNAs are needed in order to normalize and calculate the relative microRNA levels across samples. One of the pitfalls in analysis of microRNAs from serum and plasma is the release of microRNAs from blood cells during hemolysis. This is an important issue because varying degrees of hemolysis are not uncommon in routine blood sampling. Thus, hemolysis must be taken into consideration when working with circulating microRNAs from blood. miR-93-5p, miR-30b-5p, and miR-20a-5p have been reported as reference microRNA in analysis of the miR-371a-373 cluster. We here show how these three microRNAs are influenced by hemolysis. We also propose a new reference microRNA, miR-191-5p, which is relatively stable in serum samples with mild hemolysis. In addition, we show how hemolysis can have effect on the reported microRNA levels in patient samples when these reference microRNAs are used in samples with varying levels of hemolysis.

No superior results were found at the hospital with the routine use of an SGW. It may be questioned if the SGW still should be considered mandatory.

Background and purpose: Ureteral stricture is a rare, but serious complication following ureteroscopy (URS) for stones. The aim of this study was to investigate how many patients ended up with a ureteral stricture after URS at our hospital and how these were treated. We also wanted to identify potential risk factors for post endoscopic stricture formation. Materials and methods:A retrospective evaluation of 1001 URS for stone treatment at the day-case surgery unit between 2013 -2018 was performed. Data on pretreatment status, the surgical procedure and follow-up were recorded. Exact χ 2 and independent-samples ttests were used comparing data among those who developed strictures and those who did not. Multiple logistic regression was performed analyzing risk factors for stricture formation. Results:In total, 1001 URS were performed in 725 patients, 289 women and 436 men. Of these, 995 cases were eligible for analysis. At follow-up with computed tomography after 3 months, 28 (3.0%) strictures were identified. Of these, 20 received endourological treatment with balloon dilatation of which 15 (75%) were successful. Definitive treatment in the 13 patients with failed or unattempted endourological treatment included nephrectomy, reconstructive surgery, permanent nephrostomy or observation with no further treatment.In multiple regression analysis, use of access sheath (UAS) (OR 4.6, p = 0.011), ureteral perforation (OR 11.8, p < 0.0001) and surgical time > 60 minutes (OR 5.7, p < 0.005) were found to be risk factors for stricture formation. Conclusion:Ureteral stricture is a rare complication of URS. Balloon dilatation should be the first line of treatment. Use of UAS, perforation and excessive operating time were found to be risk factors for post endoscopic ureteral stricture formation. Special attention to these risk factors should be given to reduce the incidence of ureteral strictures.

MicroRNA-371a-3p (miR371) has been suggested as a sensitive biomarker in testicular germ cell cancer (TGCC). We aimed to compare miR371 with the classical biomarkers α-fetoprotein (AFP) and β-human chorionic gonadotropin (hCGβ). Overall, 180 patients were prospectively enrolled in the study, with serum samples collected before and after orchiectomy. We compared the use of digital droplet PCR (RT-ddPCR) with the quantitative PCR used by others for detection of miR371. The novel RT-ddPCR protocol showed high performance in detection of miR371 in serum samples. In the study cohort, miR371 was measured using RT-ddPCR. MiR371 detected CS1 of the seminoma and the non-seminoma sub-types with a sensitivity of 87% and 89%, respectively. The total sensitivity was 89%. After orchiectomy, miR371 levels declined in 154 of 159 TGCC cases. The ratio of miR371 pre- and post-orchiectomy was 20.5 in CS1 compared to 6.5 in systemic disease. AFP and hCGβ had sensitivities of 52% and 51% in the non-seminomas. MiR371 is a sensitive marker that performs better than the classical markers in all sub-types and clinical stages. Especially for the seminomas CS1, the high sensitivity of miR371 in detecting TGCC cells may have clinical implications.