Background: High recurrence rates for non-muscle-invasive bladder cancer (NMIBC) remain a clinical challenge. Recommended post-operative treatments are underutilized, highlighting the need for alternative strategies. Given the variability in bladder cancer prognosis, personalized treatment approaches are highly relevant. In this study, we evaluated post-operative two-hour continuous sterile water bladder irrigation (CSWBI) regarding recurrence and safety, as a potential addition to the treatment arsenal for bladder cancer. Method: In 2018, two-hour CSWBI was implemented as routine treatment after all transurethral resection procedures of the bladder (TURB), at the urology department of Sundsvall Hospital. All patients who underwent TURBs four years prior (control group) and four years after the implementation of CSWBI (intervention group) were analyzed. Primary NMIBC were included, MIBC and CIS were excluded. Data were collected retrospectively from patient records, including baseline characteristics, adverse events, and recurrence rates within 12 months follow-up. Statistical analyses included Chi-squared test, Wilcoxon rank-sum test, univariate and multivariate logistic regression analyses, Kaplan-Meier curves and log-rank test. Results: A total of 168 patients were included (control group n = 90, irrigation group n = 78). Median age was 73 years, 23% were female, 77% were male, and 74% were active or previous smokers. The recurrence rate within twelve months for the intervention group vs. the control group was: 27% vs. 21% (p = 0.4) respectively. CSWBI had no statistically significant impact on recurrence (OR 1.25, 95% CI 0.58-2.68, p = 0.6). Adverse effects were limited and equal between groups. Conclusions: Post-operative two-hour CSWBI did not significantly reduce NMIBC recurrence within twelve months in this cohort.

Background: Comorbidity alterations during neoadjuvant chemotherapy (NAC) in patients with muscle invasive bladder cancer (MIBC) are not well studied. Comorbidity-estimates are mainly based on calculations at diagnosis. It is unknown if comorbidities alter in this patient group. This study aimed to investigate if comorbidity alterations during treatment need to be considered, for scientific evaluations. Methods: A retrospective study of comorbidity alterations on MIBC patients undergoing NAC using age adjusted Charlson Comorbidity Index (CACI). We identified 684 patients between 2007-2022 from four Swedish cystectomy centers, a total of 320 were enrolled in the study. Inclusion criteria were nondisseminated MIBC, cT2-4aN0M0, NAC-treatment or NAC-eligibility. Medical files were reviewed and CACI were assessed and graded at three checkpoints during treatment: At diagnosis, post-NAC and pre-surgery. The cohort was divided into three subgroups, NAC complete, NAC incomplete and NAC-eligible. Data was retrospectively analyzed in SPSS Statistics 29.0 using descriptive statistics, one-way analysis of variance (ANOVA) and t-test. Results: The total cohort (n=320) had an increase in CACI, 0.11 CACI points, between diagnosis and prior to surgery [95% confidence interval (CI): 0.06-0.15, P<0.001]. The NAC-incomplete group had the highest increase in CACI, 0.21 CACI points from diagnosis to pre-surgery (95% CI: 0.03-0.39, P=0.01). The NACcomplete group also had an increase in CACI, 0.11 CACI points from diagnosis to pre-surgery (95% CI: 0.06-0.15, P<0.001). The NAC-eligible group had no significant CACI-alterations. The most common events causing an increase in CACI was suspected peptic ulcer (85.2%). Conclusions: There is a statistically significant increase of comorbidity in the total cohort as well as for two of the three subgroups, the NAC complete group and the NAC incomplete group. However, the increases are subtle and of limited clinical significance. This suggests that regardless of adverse events, complications, or progression, comorbidities only slightly change over time in the patient group. All together, answering the primary question if a comorbidity measurement at any moment on the timeline is sufficient.

Single particle profiling (SPP) is a unique methodology to study nanoscale bioparticles such as liposomes, lipid nanoparticles, extracellular vesicles, and lipoproteins in a single particle and high throughput manner. The initial version requires the single photon counting modules for data acquisition, which limits its adoptability. Here, we present imaging-based SPP (iSPP) that can be performed by imaging a spot over time in the common imaging mode with confocal detectors. We also provide user-friendly software with a graphical user interface to analyze such data and give quantitative insights on the content and properties of nanoscale bioparticles. We use iSPP to decipher lipid-protein interactions, membrane modifications by drugs, and the heterogeneity of extracellular vesicles isolated from cell lines and human urine. This easily applicable modality of the single particle profiler will facilitate nanoscale bioparticle research in laboratories with access to any confocal microscope.

Urinary Bladder Cancer (UBC) ranks among the most prevalent cancers worldwide, has a high recurrence rate and unpredictable treatment responses. Thus, biomarkers are urgently needed. Extracellular vesicles (EVs) are released from both cancer- and immune cells and provide a snapshot of the originating cell. They are abundant in urine and are therefore candidate biomarkers for UBC.Isolated urinary EVs from 39 UBC patients were compared with EVs from healthy controls, prostate cancer patients and whole urine. Samples were from bladder urine at time of both transurethral resection of the bladder tumour (TURB) and cystectomy, as well as urine taken from the ureter at cystectomy. EVs were isolated by tangential flow filtration and differential ultracentrifugation and their protein composition was detected by Proximity Extension Assay (PEA; Olink, immuno-oncology panel).In UBC patients, the proteomic signature of bladder urine EVs differed from ureter urine EVs from the same individuals, and from bladder urine derived EVs of both healthy and prostate cancer controls. Pairwise comparison was performed with matched whole urine revealing proteins solely detected in isolated vesicles. Additionally, a distinct signature was identified in bladder urine EVs correlating with muscle invasiveness, and a trained classifier could predict UBC with 92% accuracy. Some differentially expressed proteins, HO-1 and MMP7, were analysed by bead- based flow cytometry, where HO-1 was detected on the EV surface.Taken together, these results strengthen the rationale of using EVs as non-invasive biomarkers and prognostic tools for UBC.

OBJECTIVE:The aim of this study was to evaluate the early experiences of prostate artery embolization (PAE) in patients with benign prostatic hyperplasia (BPH). MATERIAL AND METHODS:This retrospective study included all patients treated for BPH who were referred to the radiology department for PAE in Västmanland between 2018 and 2021. Data were collected on patient demographics, International Prostate Symptom Score (IPSS), prostate-specific antigen level, and peri- and post-procedure outcomes. Clinical success was defined as a decrease in the IPSS by ≤3 points or catheter freedom. RESULTS:A total of 96 patients (median age 74 years) underwent PAE. Before PAE, 65% of the patients were treated with α-blockers and 83% with 5α-reductase inhibitors. Clinical success was achieved in 60 patients (63%). Among catheterized patients, 33 (53%) achieved catheter freedom. In the non-catheterized group, 27 (79%) experienced clinical success. No significant differences in outcomes were observed based on the presence of a median lobe. CONCLUSIONS:PAE appears to be a viable treatment option for BPH and may provide substantial symptom relief and catheter freedom for most patients. These findings suggest that PAE may be an effective alternative to more invasive procedures. Further research is needed to refine the patient selection criteria.

Background:The 5-year survival rate in muscle-invasive bladder cancer (MIBC) is approximately 50%, cross over computed tomography (CT) stage in chemo-naive patients. Studies indicate lower survival rates in females when compared to males. The theories that explain the sex disparity are hormonal factors and delayed diagnosis for females. New investigations suggest that neoadjuvant chemotherapy (NAC) might be a possible method for bridging the gender survival gap. The aim of this study was to investigate whether complete treatment with NAC (≥3 cycles) prior to cystectomy reduces the gender gap in survival rates for MIBC and improves the surrogate marker of downstaging. Methods:A multicenter retrospective cohort from five Swedish urological centers, from 1st January 2005 to 17th July 2023 based on NAC-eligible patients divided in NAC-receiving and non-NAC-receiving subgroups and further divided by sex (males and females). Survival was analyzed based on the Kaplan-Meier method, using log-rank test and adjusted analyses were made with the Cox regression model. Outcome measurements were overall survival (OS), disease-free survival (DFS), and downstaging. Results:In the analysis of the total cohort (n=412), we could not detect any statistically significant differences in OS between NAC and non-NAC, nor between sexes, in the unadjusted analysis. In the adjusted analysis, we did not observe any significant differences in OS between sexes, either in total or within the NAC subgroups. Further analyzing the NAC group, we could see a significant increased downstaging rate in the NAC group compared to the non-NAC group (P<0.001) which indicates an increased survival in those receiving NAC treatment. There was no relationship between sexes and downstaging (P=0.72). Neither could we see any significant difference in downstaging between males and females in the NAC/non-NAC subgroups (P=0.41 and P=0.92, respectively). Conclusions:NAC-eligible female and male MIBC patients who underwent radical cystectomy (RC) after at least three cycles of NAC, demonstrated similar OS and DFS. NAC seems to obliterate survival differences between genders in MIBC patients.

Background: A previously published study at Norrland University Hospital, Ume & aring;, Sweden, found that in 29.5% of patients with urinary bladder cancer (UBC) who underwent cystectomy, incorrect cT-stage (clinical T-stage) was registered in the Swedish National Register of Urinary Bladder Cancer (SNRUBC). Tumor in bladder diverticulum (TIBD) and tumor-associated hydronephrosis (TAH) were common causes for misclassification. Our aim was to further investigate cT-staging, as well as pathoanatomical markers, in the SNRUBC, compared to detailed data from medical records in a larger, retrospective multicenter cohort. Our secondary objective was to describe the frequency of pathoanatomical markers in pathology reports (PAD) after transurethral resection of the bladder (TURb): variant histology (VH), concomitant carcinomain situ(CIS), lymphovascular invasion (LVI) and perineural invasion (PNI). Methods: Medical records of 630 patients planned for radical cystectomy in the years 2009-2022 in the Northern Healthcare Region, Region of G & auml;vleborg and Region of V & auml;stmanland were reviewed. Factors impacting risk of misclassification of cT-staging were identified through logistic regression. In TURb pathology reports, all comments on pathoanatomical markers were identified. For each pathoanatomical marker, respectively, comments were then registered as positive or negative. The absence of a comment on a marker was registered as "not commented". Results: A total discrepancy rate of 36.5% was found between validated cT-staging and the SNRUBC, of which 13.3% were upstaged from 2 to >= T2. The results are presented as odds ratios (ORs) with corresponding 95% confidence intervals (CIs). Registrations with discrepancy were significantly associated with TIBD (OR: 10.28, 95% CI: 5.20-20.34), TAH (OR: 9.60, 95% CI: 6.12-15.10) and year of cystectomy 2009-2011 (OR: 3.38, 95% CI: 2.13-5.36). Incorrect CIS registration: 134 (35.8%); incorrect histology registration: 98 (25.6%). Total frequencies of recorded pathoanatomical markers in TURb-reports were for VH =23.8%, concomitant CIS =36.9%, LVI =30.4%, PNI =2.3%. Conclusions: The SNRUBC has a significant prevalence of misclassification of cT-staging with a large proportion due to TAH and TIBD. Misclassification of VH and CIS is also common. Improved guidelines could increase consistency. Total rates of recorded pathoanatomical markers in TURb-reports are low.

Objective: To assess the safety and diagnostic accuracy of renal tumour biopsy (RTB) in patients with small renal masses (SRM) and to assess if RTB prevents overtreatment in patients with benign SRM. Material and methods: In a retrospective, single-centre study from Västmanland, Sweden, 195 adult patients (69 women and 126 men) with SRM ≤ 4 cm who had undergone RTB during 2010–2023 were included. The median age was 70 years (range 23–89). The sensitivity, specificity and predictive values of RTB were calculated using the final diagnosis as the reference standard. Treatment outcomes were recorded for a median 42-month follow-up. Complications following the biopsies were assessed according to the Clavien–Dindo system. Results: The overall sensitivity of RTB was 95% (95% confidence interval [CI] 90% – 98%) and specificity was 100% (95% CI 95% – 100%). The positive predictive value was 100% and negative predictive value was 92%. The rate of agreement between RTB and the final diagnosis measured using kappa statistics was 0.92. Of the 195 patients, 62 underwent surgery and 48 were treated with ablation. The concordance rate between the RTB histology and final histology after surgery was 89%. Treatment was withheld in 67 of 195 patients with a benign or inconclusive RTB. No patients developed renal cell carcinoma or metastasis during follow-up. Complications occurred in two patients that were classified with Clavien–Dindo grades I and IV. Conclusions: Percutaneous renal tumour biopsy appears to be a safe diagnostic method that provides accurate histopathological information about small renal masses and reduces overtreatment of benign SRM.

B cells are multifaceted immune cells responding robustly during immune surveillance against tumor antigens by presentation to T cells and switched immunoglobulin production. However, B cells are unstudied in prostate cancer (PCa). We used flow cytometry to analyze B-cell subpopulations in peripheral blood and lymph nodes from intermediate–high risk PCa patients. B-cell subpopulations were related to clinicopathological factors. B-cell-receptor single-cell sequencing and VDJ analysis identified clonal B-cell expansion in blood and lymph nodes. Pathological staging was pT2 in 16%, pT3a in 48%, and pT3b in 36%. Lymph node metastases occurred in 5/25 patients (20%). Compared to healthy donors, the peripheral blood CD19+ B-cell compartment was significantly decreased in PCa patients and dominated by naïve B cells. The nodal B-cell compartment had significantly increased fractions of CD19+ B cells and switched memory B cells. Plasmablasts were observed in tumor-draining sentinel lymph nodes (SNs). VDJ analysis revealed clonal expansion in lymph nodes. Thus, activated B cells are increased in SNs from PCa patients. The increased fraction of switched memory cells and plasmablasts together with the presence of clonally expanded B cells indicate tumor-specific T-cell-dependent responses from B cells, supporting an important role for B cells in the protection against tumors.

Supplemental figure 1 illustrates gating strategy appliead to FACS data. Supplemental figure 2 shows gene expression correlation to Treg frequency overlaid on KEGG bladder cancer pathway. Supplemental figure 3 shows expression of functional Treg markers in FOXP3+ and FOXP3- cells from PBMCs and TILs. Supplemental figure 4 compares the phenotype of TIL Tregs from MIBC and non-MIBC tumors. Supplemental figure 5 illustrates post-sort purity sorted populations. Supplemental figure 6 illustrates the correlation of Treg frequency at tumor sublocations to tumor stage and survival. Supplemental figure 7 shows expression level of different MMPs in UBC tumor tissue.

ObjectivesAndrogen deprivation therapy (ADT) has long been a cornerstone in treatment of advanced prostate cancer (PCa), and is known to improve the results of radiotherapy (RT) for high-risk disease. The purpose of our study was to use a multiplexed immunohistochemical (mIHC) approach to investigate the infiltration of immune cells in PCa tissue after eight weeks of ADT and/or RT with 10 Gy.MethodsFrom a cohort of 48 patients divided into two treatment arms, we obtained biopsies before and after treatment and used a mIHC method with multispectral imaging to analyze the infiltration of immune cells in tumor stroma and tumor epithelium, focusing on areas with high infiltration.ResultsTumor stroma showed a significantly higher infiltration of immune cells compared to tumor epithelium. The most prominent immune cells were CD20(+) B-lymphocytes, followed by CD68(+) macrophages, CD8(+) cytotoxic T-cells, FOXP3(+) regulatory T-cells (Tregs), and T-bet(+) Th1-cells. Neoadjuvant ADT followed by RT significantly increased the infiltration of all five immune cells. Numbers of Th1-cells and Tregs significantly increased after single treatment with ADT or RT. In addition, ADT alone increased the number of cytotoxic T-cells and RT increased the number of B-cells.ConclusionsNeoadjuvant ADT in combination with RT results in a higher inflammatory response compared to RT or ADT alone. The mIHC method may be a useful tool for investigating infiltrating immune cells in PCa biopsies to understand how immunotherapeutic approaches can be combined with current PCa therapies.

Purpose: Previous studies have indicated that patients with muscle-invasive bladder cancer with non-O blood types have an increased risk of experiencing thromboembolic events (TEEs). This is finding is in relation to neoadjuvant-chemotherapy (NAC)-naïve patients. Aim: to establish the risk of TEEs and any association with blood types among NAC patients as well as NAC-naïve patients. Methods: Cystectomized patients at four centres treated from 2009 to 2018 (n = 244) were analysed. The quantities of patients corresponding to each blood group were as follows: A—108 (44%); O—99 (41%); B—30 (12%); and AB—7 (3%). NAC patients (n = 167) and NAC-naïve NAC-eligible patients (n = 77) were assessed. In total, 54 women (22%) and 190 men (78%), with a median age of 69 years, were included in the study. The occurrence of any type of TEE from six months pre-cystectomy to 12–24 months after was analysed using logistic regression adjusted for NAC and confounders. Results: Sixty-six TEEs were detected in 21% of the patients (n = 52). Pulmonary embolus (n = 33) and deep venous thrombosis (n = 11) were the most common forms. No significant differences between blood types were found in the analysis, although B blood type had a nearly significant increased crude risk compared with O blood type, for which there was an OR of 2.48 (95% CI 0.98–6.36). Adjustment for NAC and covariates weakened the OR, which plummeted to 1.98 (95% CI 0.71–5.51). Conclusions: No significant associations were found between blood types and TEE occurrences in this cohort including both NAC and NAC-naïve NAC-eligible patients.

Table S1 shows patient characteristics. Table S2 summarizes the antibodies used. Table S3 shows primer sequences used.

Background: A high pre-treatment De Ritis ratio, the aspartate transaminase/alanine aminotransferase ratio, has been suggested to be of prognostic value for mortality in muscle-invasive bladder cancer (MIBC). Our purpose was to evaluate if a high ratio was associated with mortality and downstaging. Methods: A total of 347 Swedish patients with clinically staged T2-T4aN0M0, with administered neoadjuvant chemotherapy (NAC) or eligible for NAC and undergoing radical cystectomy (RC) 2009–2021, were retrospectively evaluated with a low ratio < 1.3 vs. high ratio > 1.3, by Log Rank test, Cox regression and Mann–Whitney U-test (MWU), SPSS 27. Results: Patients with a high ratio had a decrease of up to 3 years in disease-free survival (DFS), cancer-specific survival (CSS) and overall survival (OS) (p = 0.009, p = 0.004 and p = 0.009) and 5 years in CSS and OS (p = 0.019 and p = 0.046). A high ratio was associated with increased risk of mortality, highest in DFS (HR, 1.909; 95% CI, 1.265–2.880; p = 0.002). No significant relationship between downstaging and a high ratio existed (p = 0.564 MWU). Conclusion: A high pre-treatment De Ritis ratio is on a population level, associated with increased mortality post-RC in endpoints DFS, CSS and OS. Associations decrease over time and require further investigations to determine how strong the associations are as meaningful prognostic markers for long-term mortality in MIBC. The ratio is not suitable for downstaging-prediction.