Predictive factors for successful testicular biopsy.

INTRODUCTION: Infertility, the inability to conceive, constitutes a major problem in modern societies. It affects 10 to 15 percent of couples in the United States. Evaluation of infertile men is usually complex and often demands a testicular biopsy. MATERIALS AND METHODS: We reviewed all azoospermic men submitted to testicular biopsy, in our center, during infertility investigation between January 2015 and December 2021. RESULTS: A total of 117 patients with a mean age of 36.5 was considered. Biopsy was positive, as defined by the presence of viable spermatozoids by microscopy, in 48.7% of patients (n = 57). Patients were divided in two separate groups based on positive (PB) or negative biopsy (NB) and compared. PB-group had normal serum total testosterone levels and higher than NB-group (3.7 ng/mL vs. 2.85 ng/mL, p = 0.021), and normal serum FSH levels and lower than NB-group (6.0 mIU/mL vs. 16.0 mIU/mL, p < 0.001). The groups were similar concerning serum LH levels (3.9 mIU/mL vs. 6.3 mIU/mL, p = 0.343. CONCLUSIONS: Predicting outcomes of testicular biopsy is a difficult task. Our study found that men with normal testicular volume, normal levels of testosterone and FSH and those with type 1 diabetes mellitus had a higher probability of positive testicular biopsy.

Surveillance as a safe and effective option for treatment of stage I seminoma.

Introdubction: Stage I seminoma has a very good prognosis, yet approximately 15% have subclinical metastatic disease and will relapse after orchidectomy alone. Several management approaches have been investigated. We aimed to evaluate the clinical outcomes of real-world patients with stage I seminoma, analysing prognostic factors influencing treatment choice and oncological outcomes. METHODS: Retrospective, single institution study, with 55 patients diagnosed with clinical stage I seminoma between 2007 and 2020. Selected patients were analysed regarding three management approaches - surveillance, adjuvant radiotherapy and adjuvant carboplatin AUC7. Overall survival and progression-free survival outcomes were analysed. Predictors of treatment choice were determined, and predictors of recurrence were analysed in patients on active surveillance. RESULTS: The median follow-up time was 91 months (13-165). Overall survival at 10 years was 98.2%. Stage I seminoma patients had a 1-, 3- and 10-year progression free survival of 98%, 94% and 89%, respectively. Three-year progression free survival was 92.0% for those on active surveillance (IC95%, 91.5-92.5%), 95.2% for carboplatin (IC95%, 94.8-95.6%) and 100% for those on adjuvant radiotherapy (p > 0.05). All relapses on active surveillance protocols occurred during the first 24 months. Overall, 43% of patients who underwent adjuvant treatment reported adverse effects of therapy, with higher incidence on radiotherapy group (63%). CONCLUSIONS: Stage I seminoma have excellent prognosis, high cure rates, and low treatment-associated morbidity. Active surveillance is a safe modality when applied to selected patients. Adjuvant radiotherapy and adjuvant chemotherapy with carboplatin show similar results, with fewer adverse effects on chemotherapy arm.

An overview of the latest in state-of-the-art murine models for prostate cancer.

INTRODUCTION: Prostate cancer (PCa) is a complex, heterogenous and multifocal disease, which is debilitating for patients and often fatal - due to bone metastasis and castration-resistant cancer. The use of murine models that mimic human disease has been crucial in the development of innovative therapies and for better understanding the mechanisms associated with initiation and progression of PCa. AREAS COVERED: This review presents a critical analysis of murine models for the study of PCa, highlighting their strengths, weaknesses and applications. EXPERT OPINION: In animal models, disease may not occur exactly as it does in humans, and sometimes the levels of efficacy that certain treatments obtain in animal models cannot be translated into clinical practice. To choose the most appropriate animal model for each research work, it is crucial to understand the anatomical and physiological differences between the mouse and the human prostate, while it is also important to identify biological similarities and differences between murine and human prostate tumors. Although significant progress has already been made, thanks to many years of research and study, the number of new challenges and obstacles to overcome mean there is a long and difficult road still to travel.

Acute Urinary Retention After Kidney Transplant: Effect on Graft Function, Predictive Factors, and Treatment.

BACKGROUND: Benign prostatic hyperplasia (BPH) is common in older adults. Although BPH may be asymptomatic in patients with chronic kidney disease (CKD) with low diuresis, the condition may become troublesome when diuresis resumes after transplantation. This study evaluated the effect that developing acute urinary retention (AUR) in first 4 months after kidney transplantation (KT) can have on graft function at 6 months. The study identified predictive factors and analyzed treatment of AUR in these patients. METHODS: This study retrospectively included 303 men who received KT. Independent samples Student t test was used to compare glomerular filtration rates (GFRs) at 6 months. Logistic regression was applied to identify predictors of AUR. RESULTS: The study found that 14 patients developed AUR within the first 4 months after KT. This group had lower GFR at 6 months post-KT. Nine patients required transurethral resection of the prostate, and 2 of these patients developed acute graft pyelonephritis following resection. Residual diuresis and recipient age were predictive factors. Recipient age >55 years was a risk factor. Medical therapy of BPH before transplantation was a protective factor. CONCLUSIONS: Developing AUR in the first 4 months after KT was associated with lower graft GFR at 6 months, and transurethral resection of the prostate was required in 64% of these patients, with good results. Medical therapy for BPH before the transplant was associated with a lower risk of AUR. Older patients and patients with pretransplant low urine output had a higher risk of AUR. These patients should be closely monitored in the posttransplant period for the presence of obstructive uropathy.

Early morning kidney transplantation: Perioperative complications.

INTRODUCTION: To reduce cold ischemia time (CIT), many kidney transplants are performed in the early morning. Conducting complex surgeries in the early morning may influence the surgeon's technical capacity and rate of surgical complications (SC). AIM: Evaluate the influence of surgery start hour (SSH) regarding duration of surgery (DS), immediate diuresis (ID), SC and acute rejection (AR); evaluate the influence of CIT regarding SC, ID, and AR. METHODS: 2855 cadaveric transplants performed between June 1980 and March 2018 were retrospectively evaluated. Regarding SSH, two groups were created: Group M (00: 00h-05.59h, n = 253) and Group D (06: 00h - 23: 59h, n = 2602). Analyzing the impact of SSH on DS, ID, SC and AR. Evaluate the relationship between CIT (< 18h, 18-30h and > 30h) on ID, SC and AR utilizing univariate and multivariate statistical analysis with SPSS. RESULTS AND CONCLUSION: Groups M and D were comparable in all evaluated demographic variables (p > 0.05), except cold ischemia time (Group M with higher CIT, p < 0.001). Regarding univariate analysis, Surgery start hour did not influence DS (p = 0.344), and SC (p = 0.264), but related with higher ID (p = 0.028) and AR (p = 0.018). CIT related with immediate diuresis (p = 0.020) and acute rejection (p < 0.001) but did not relate with complications (p = 0.734). Regarding multivariate analysis, SSH only influenced immediate diuresis (p = 0.026) and did not influenced acute rejection (p = 0.055). CIT influenced immediate diuresis (p = 0.019) and acute rejection (p < 0.001). Surgery start hour influences Immediate diuresis. With this study, we conclude that the priority must be a short cold ischemia time.

Caveolin-1 Modulation Increases Efficacy of a Galacto-Conjugated Phthalocyanine in Bladder Cancer Cells Resistant to Photodynamic Therapy.

Photodynamic therapy (PDT) has demonstrated encouraging anticancer therapeutic results, but the current clinically approved photosensitizers (PSs) are not ideal in the treatment of bladder cancer. Conventional PSs have low selectivity to the bladder tumor tissue and induce toxicity or bystander effects on nontumor urothelium. Previous studies demonstrated that the use of galactose-photosensitizer (PS) conjugates is a more selective method of delivering PDT-mediated toxicity due to their ability to recognize carbohydrate-binding domains overexpressed in bladder tumors. Using patient-derived bladder tumor specimens cultured ex vivo and bladder cancer cell lines with different PDT sensitivity, we find that a galactose-phthalocyanine (PcGal16) accumulates in bladder tumors expressing galactose-binding proteins and internalizes through an endocytic process. The endocytosis mechanism is cell line-dependent. In HT-1376 bladder cancer lines resistant to PDT, depletion of caveolin-1-the main structural protein of caveolae structures-increased the amount of sugar-binding proteins, i.e. GLUT1, at the cell membrane resulting in an improved PcGal16 uptake and PDT efficacy. These data show the potential of ex vivo cultures of bladder cancer, that ideally could mimic the original microenvironment, in screening galacto-PDT agents. Additionally, our studies demonstrate that PDT efficacy in bladder cancer depends on the endocytic mechanisms that regulate PS accumulation and internalization in cancer cells.

Transplantectomy in the First 3 Months After Renal Transplantation: Experience of a Reference Center.

INTRODUCTION/OBJECTIVE: Transplantation is the treatment of choice in end-stage renal disease. However, there are complications that require transplantectomy. The objective of this study was to evaluate predictive factors for transplantectomy in the first 3 months after renal transplantation. MATERIAL AND METHODS: This retrospective study included 770 kidney transplants performed between June 2011 and June 2017. Logistic regression was applied to study the relationship between independent variables and the occurrence of transplantectomy. RESULTS: Analyzing variables of the recipients, it was verified that age over 65 years; body mass index; dialysis time; history of previous transplant and comorbidities such as obesity, overweight, hypertension, diabetes mellitus, dyslipidemia, peripheral arterial disease; or history of a thrombotic episode were not predictive factors. It was found that the use of expanded criteria donors, their age, or cause of death were not predictive factors. The use of a right renal graft or grafts with multiple arteries; the duration of surgery; the performance of surgery at dawn; the need for transfusion; the cold ischemia time; and hemodynamic parameters at reperfusion (central venous pressure, systolic or diastolic blood pressure) were not predictive factors. The recipient age at transplantation (p = .014; B=-0.059; Exp(B)=0.943 [0.899-0.988]) and reoperation in the first 10 days after transplantation (p < .001; B= -2.574; Exp(B)=0.076 [0.028-0.210]) were predictive factors. CONCLUSION: Reoperation in the first 10 days after transplantation decreased the risk of transplantectomy in the first 3 months. The lower the age of the recipient, the greater the risk of transplantectomy.

Impact of Positive Surgical Margins After Partial Nephrectomy.

BACKGROUND: The impact of positive surgical margins (PSMs) after partial nephrectomy (PN) is controversial. OBJECTIVE: To evaluate the risk factors for a PSM and its impact on overall survival. DESIGN SETTING AND PARTICIPANTS: This is a retrospective study of 388 patients were submitted to PN between November 2005 and December 2016 in a single centre. Two groups were created: PSM and negative surgical margin (NSM) after PN. A p value of <0.05 was considered significant. OUTCOME MEASUREMENTS AND STATISTICAL ANALYSIS: Relationships with outcome were assessed using univariable and multivariable tests and log-rank analysis. RESULTS AND LIMITATIONS: The PSM rate was 3.8% (N = 16). The mean age at the time of surgery (PSM group: 64.1 ± 11.3 vs NSM group: 61.8 ± 12.8 yr, p = 0.5) and the mean radiological tumour size (4.0 ± 1.5 vs 3.4 ± 1.8 cm, p = 0.2) were similar. Lesion location (p = 0.3), surgical approach (p = 0.4), warm ischaemia time (p = 0.9), and surgery time (p = 0.06) had no association with PSM. However, higher surgeon experience was associated with a lower PSM incidence (2.6% if ≥30 PNs vs 9.6% if <30 PNs; p = 0.02). Higher operative blood loss (p = 0.02), higher-risk tumours (p = 0.03), and larger pathological size (p = 0.05) were associated with an increase in PSM. In the PSM group, recurrence rate (18.7% vs 4.2%, p = 0.007) and secondary total nephrectomy rate (25% vs 4.4%, p < 0.001) were higher. However, overall survival was similar. Multivariate analysis revealed that high-risk tumour (p = 0.05) and low experience (p = 0.03) could predict a PSM. Limitations include retrospective design and reduced follow-up time. CONCLUSIONS: PSMs were mainly associated with high-risk pathological tumour (p = 0.05) and low-volume surgeon experience. Recurrence rate and need for total nephrectomy were higher in that group, but no impact on survival was noticed. PATIENT SUMMARY: The impact of positive surgical margins (PSMs) after partial nephrectomy is a matter of debate. In this study, we found that PSMs were mainly associated with aggressive disease and low surgeon experience.

Surgical Complications in Kidney Transplantation: An Overview of a Portuguese Reference Center.

INTRODUCTION: Kidney transplantation (KT) is a surgery performed worldwide and has some complications. The objective of this study is to evaluate our surgical complications, comparing the outcomes with those KTs without surgical complications. PATIENTS AND METHODS: An observational cross-sectional study of all surgical complications among 3102 kidney transplants performed between June 1980 and April 2018. RESULTS: Of 3102 kidney transplantations, 490 (15.8%) had the following complications: surgical complications (n = 527); urinary (n = 184; 5.9%); vascular (n = 140; 4.5%); wound-related (n = 78; 2.5%); lymphocele (n = 56; 1.8%); and others (n = 69; 2.2%). The most common complications were ureteral obstruction (n = 85; 2.7%) and urinary fistula (n = 72; 2.3%). The immunosuppression regimen did not influence the surgical complications rate. Surgical complications mainly occurred in male (71.4% vs 66.7%) and heavier (67.6 ± 13.9 vs 65.9 ± 13.5 kg) recipients (P < .05). The hospitalization time was also different (26.3 ± 30.6 vs 15.0 ± 38.8 days, P < .05). Serum creatinine values were different until the second year. After that, the renal function was approximately the same. Nearly 26.1% of complicated kidney transplants had delayed graft function (vs 14.8%, P < .001). Only 23.9% of complicated kidney transplants needed transplant nephrectomy (vs 6.2%, P < .001). The survival of kidneys with surgical complications was lower (64.2 ± 4.5 vs 94.09 ± 2.6 months, P < .001). DISCUSSION/CONCLUSION: Kidney transplant surgical complications occur over time, especially urinary and vascular complications, remaining a problem that leads to prolonged hospitalization and decreased graft survival.

Transumbilical laparoendoscopic single-site adrenalectomy: A feasible and safe alternative to standard laparoscopy.

OBJECTIVES: Standard multi-port laparoscopic adrenalectomy (LA) is considered the gold standard for benign adrenal tumors. Single-site LA has been proposed as a feasible and safe alternative because of lower invasiveness, improved cosmetics, less pain and shorter hospital stay. The objective was to evaluate and compare results of single-site transumbilical laparoendoscopic adrenalectomy with standard LA for adrenal tumors. MATERIALS AND METHODS: One hundred consecutive adrenalectomies from 93 patients, performed between March 2009 and June 2017, were laparoscopically excised: 59 by standard multi-port LA (group 1) and 41 by transumbilical laparoendoscopic single-site adrenalectomy (group 2). Data gathered included demographics, comorbidities, preoperative imaging, tumor characteristics, perioperative data, surgical complications, pathology and follow-up. IBM SPSS Statistics 23 software was used and p value < 0.05 was considered significant. RESULTS: Patients of group 2 were younger (48.7 ± 13.9 versus 59.7 ± 15.1 years; p < 0.001) and had fewer comorbidities (p < 0.05). Mean tumor diameter in group 2 was lower than those of group 1 (27.52 ± 14.3 versus 47.9 ± 30.6 mm; p < 0.001). Tumor laterality did not influence the choice of technique nor the surgical morbidity. All procedures were successfully completed, although one standard LA needed conversion to open surgery. Mean operative time, hemorrhagic losses, postoperative opioid analgesic requirement and hospital stay were not statistically different between groups. Most patients in group 2 (31 patients, 85.4%) did not require drainage, compared to 14 (25.4%) patients of group 1 (p < 0.001). Patients who underwent single-site LA resumed normal diet earlier (1.0 ± 0.2 versus 1.6 ± 0.7 days; p < 0.001). There were no reoperations and no perioperative mortality. Overall mean follow-up time was 94.9 ± 3.1 months, not statiscally different between groups (p = 0.7). CONCLUSIONS: Our results revealed that transumbilical approach for laparoendoscopic single-site adrenalectomy for adrenal tumors is a feasible and safe alternative to standard laparoscopic adrenalectomy.

Supportive Care Network: Evaluation of Its Impact on the Performance of a Urology Department.

INTRODUCTION: Hospitals are dealing with patients who may have clinical discharge but cannot return to their home due to non-medical issues. MATERIAL AND METHODS: Cross-sectional analysis of all the cases referred to the Integrated Care Network during the year 2016. Evaluation of waiting times, typology, reason for referral and clinical parameters. IBM SPSS 24.0 software was used for all statisticalanalyses. RESULTS: In the evaluated period, 2294 patients were discharged from our department. Of these, 55 were referred to Integrated Care Network. The mean length of hospitalization of the patients referred to the network was 20.6 ± 11.4 days, and the mean overall length of hospital stay in the period analyzed was 4.8 ± 0.9 days. The mean time between hospitalization and referral for continuing care was 10.7 ± 7.2 days. The time between referral and discharge of the hospital was 10.0 ± 8.7 days. Thirty-nine (70.9%) patients were hospitalized for oncological diseases. The most common referral was to Palliative Care units (n = 16; 29.1%). Patients referred to Palliative Care units showed the largest waiting times between the referral for the network and the hospital discharge, 12.2 ± 10.51 days. We observed 289 hospitalization days with patients who had no need of specialized urological care. DISCUSSION: In order to reduce time between referral to the network and hospital discharge, there is a need for enhanced cooperation and coordination among doctors, nurses and social workers. CONCLUSION: Early identification by physicians and nurses of patients who will require care after discharge will provide a better response from social workers and increased hospital performance.

Introdução: Os hospitais deparam-se cada vez mais com doentes que, tendo alta clínica, não têm condições de ordem não clínica para regressar imediatamente ao domicílio. Material e Métodos: Estudo transversal dos casos referenciados para a Rede Nacional de Cuidados Continuados Integrados durante o ano de 2016 no nosso Serviço de Urologia. Foram avaliados os tempos de espera, tipologia, motivo de referenciação e os parâmetros clínicos. Análise estatística realizada com recurso ao software IBM SPSS 24.0. Resultados: No período analisado, 2294 pacientes tiveram alta hospitalar no nosso serviço. Destes, 55 foram referenciados para a Rede Nacional de Cuidados Continuados Integrados. O tempo médio de internamento dos pacientes referenciados foi de 20,6 ± 11,4 dias enquanto o tempo médio global de internamento foi de 4,8 ± 0,9 dias. O tempo médio entre o internamento e a referenciação para a Rede Nacional de Cuidados Continuados Integrados foi de 10,7 ± 7,2 dias. O tempo entre a referenciação e a alta hospitalar foi de 10,0 ± 8,7 dias. Trinta e nove (70,9%) pacientes foram internados por patologias oncológicas. A referenciação mais frequente foi para unidades de cuidados paliativos (n = 16; 29,1%). Os pacientes referenciados para cuidados paliativos foram os que apresentaram os maiores tempos de espera entre a referenciação e a alta hospitalar efetiva, 12,2 ± 10,51 dias. Foram despendidos 289 dias de hospitalização com pacientes que não precisavam de cuidados urológicos especializados. Discussão: Para que o tempo entre a referenciação para a Rede Nacional de Cuidados Continuados Integrados e a alta hospitalar sejam diminuídos, é necessário que haja uma otimização da cooperação e coordenação entre médicos, enfermeiros e assistentes sociais. Conclusão: A identificação precoce dos doentes que necessitarão de apoio após a alta clínica permitirá uma resposta mais atempada por parte dos assistentes sociais e uma consequente melhoria do desempenho dos serviços hospitalares e satisfação dos doentes.

Prognostic value of subclassification (pT2 stage) of pathologically organ-confined prostate cancer: Confirmation of the changes introduced in the 8th edition of the American Joint Committee on Cancer (AJCC) staging system.

OBJECTIVE: The last edition of the AJCC staging system eliminated the pT2 subclassification of prostate cancer (PCa). Our objective was to evaluate the association of pT2 subclassification with the oncological results of patients with PCa who underwent radical prostatectomy (RP). MATERIAL AND METHODS: We evaluated 367 patients who underwent RP between 2009 and 2016, with pT2 disease in the final pathological evaluation. We assessed differences in rates of biochemical recurrence (BCR), metastasis and mortality between T2 substages (pT2a/b vs pT2c). RESULTS: Fifty-three (14.4%) patients presented pT2a/b disease and 314 (85.6%) pT2c disease. The mean follow-up time was 4.9 ± 2.6 years. Grade group scores (p = 0.1) and prostate specific antigen (PSA) (p = 0.2) did not differed between pT2 substages. The rate of BCR in pT2a/b and pT2c patients was 11.3% and 18.2%, respectively (p = 0.2). Five (9.4%) patients with pT2a/b and 45 (14.3%) with pT2c substage underwent salvage radiotherapy (p = 0.3). The rate of positive surgical margins did not differ between groups (p = 0.2). Seven (2.2%) patients with pT2c had lymph nodes or distant metastases. The overall survival was 92.5% and 93.6% in pT2a/b and pT2c, respectively (p = 0.2). CONCLUSION: Our results are in accordance with the changes introduced in the 8th edition of the AJCC staging system in which the pT2 subclassification was eliminated.

Diabetes mellitus and prostate cancer metabolism: Is there a relationship?

OBJECTIVE: Our aim was to evaluate the effects of glucose levels and diabetes mellitus in prostate cancer (PCa) biology. MATERIALS AND METHODS: Two PCa cell lines (LNCap and PC3) were cultured in RPMI medium with different glucose concentrations [5mM (LG) and 25mM (HG)]. Expressions of androgen receptor, Her2/neu and glucose transporters (GLUT1, 3, 5 and 12) were evaluated by flow cytometry. Proliferation rate was assessed by colorimetric assay MTT and cellular characterization was performed by haematoxylin and eosin staining. Additionally, we performed a cross sectional analysis of 704 patients undergoing radical prostatectomy who were divided into two groups (diabetic and non-diabetic). An analysis of clinical and histological data seeking to identify the differences on tumor aggressiveness between the two groups was performed. RESULTS: In LNCaP cell line, when the glucose concentration in the medium increased, there was an increased in AR expression. Regarding expression of Her2/neu receptor, medium's glucose concentration significantly changed the expression of this receptor in both PC3 and LNCaP cell lines. Growth rate was higher on the HG medium for both cell lines. The clinical study of patients undergoing radical prostatectomy revealed no relationship between the presence of diabetes and the development of more aggressive tumours. Diabetic patients had significantly higher prostatic volumes, however, no significant difference was found between the relapse risk classification or the ISUP classification between the two groups. CONCLUSIONS: Our results showed that medium glucose concentration could influence prostate cancer cells growing but not the aggressiveness.

Intracaval misplacement of a double-J ureteral stent.

Double-J ureteral stent (DJUS) is an important therapeutic tool in endourology. There are well-known frequent complications associated with DJUS placement such as distal and proximal migration within the urinary tract. However, perforation and stent misplacement are uncommon but serious complications of this technique. We present a case of a 63-year-old man who had a misplacement of a DJUS into the inferior vena cava during an elective procedure of ureteral catheterisation. The stent placement was performed under fluoroscopic control and it seemed well positioned. Actually, the DJUS was misplaced in the inferior vena cava after drilling at the level of the crossing of the ureter with the ipsilateral iliac vessels. Diagnosis was incidentally made 3 months after the placement of the stent in a renal CT scan. The patient was always asymptomatic. We performed an endoscopic removal of the ureteral stent, which took place without complications.

Natural killer cell-based adoptive immunotherapy eradicates and drives differentiation of chemoresistant bladder cancer stem-like cells.

BACKGROUND: High-grade non-muscle invasive bladder cancer (NMIBC) has a high risk of recurrence and progression to muscle-invasive forms, which seems to be largely related to the presence of tumorigenic stem-like cell populations that are refractory to conventional therapies. Here, we evaluated the therapeutic potential of Natural Killer (NK) cell-based adoptive immunotherapy against chemoresistant bladder cancer stem-like cells (CSCs) in a pre-clinical relevant model, using NK cells from healthy donors and NMIBC patients. METHODS: Cytokine-activated NK cells from healthy donors and from high-grade NMIBC patients were phenotypically characterized and assayed in vitro against stem-like and bulk differentiated bladder cancer cells. Stem-like cells were isolated from two bladder cancer cell lines using the sphere-forming assay. The in vivo therapeutic efficacy was evaluated in mice bearing a CSC-induced orthotopic bladder cancer. Animals were treated by intravesical instillation of interleukin-activated NK cells. Tumor response was evaluated longitudinally by non-invasive bioluminescence imaging. RESULTS: NK cells from healthy donors upon activation with IL-2 and IL-15 kills indiscriminately both stem-like and differentiated tumor cells via stress ligand recognition. In addition to cell killing, NK cells shifted CSCs towards a more differentiated phenotype, rendering them more susceptible to cisplatin, highlighting the benefits of a possible combined therapy. On the contrary, NK cells from NMIBC patients displayed a low density on NK cytotoxicity receptors, adhesion molecules and a more immature phenotype, losing their ability to kill and drive differentiation of CSCs. The local administration, via the transurethral route, of activated NK cells from healthy donors provides an efficient tumor infiltration and a subsequent robust tumoricidal activity against bladder cancer with high selective cytolytic activity against CSCs, leading to a dramatic reduction in tumor burden from 80 % to complete remission. CONCLUSION: Although pre-clinical, our results strongly suggest that an immunotherapeutic strategy using allogeneic activated NK cells from healthy donors is effective and should be exploited as a complementary therapeutic strategy in high-risk NMIBC patients to prevent tumor recurrence and progression.

Functional and molecular characterization of cancer stem-like cells in bladder cancer: a potential signature for muscle-invasive tumors.

Striking evidence associates cancer stem cells (CSCs) to the high recurrence rates and poor survival of patients with muscle-invasive bladder cancer (BC). However, the prognostic implication of those cells in risk stratification is not firmly established, mainly due to the functional and phenotypic heterogeneity of CSCs populations, as well as, to the conflicting data regarding their identification based on a single specific marker. This emphasizes the need to exploit putative CSC-related molecular markers with potential prognostic significance in BC patients.This study aimed to isolate and characterize bladder CSCs making use of different functional and molecular approaches. The data obtained provide strong evidence that muscle-invasive BC is enriched with a heterogeneous stem-like population characterized by enhanced chemoresistance and tumor initiating properties, able to recapitulate the heterogeneity of the original tumor. Additionally, a logistic regression analysis identified a 2-gene stem-like signature (SOX2 and ALDH2) that allows a 93% accurate discrimination between non-muscle-invasive and invasive tumors.Our findings suggest that a stemness-related gene signature, combined with a cluster of markers to more narrowly refine the CSC phenotype, could better identify BC patients that would benefit from a more aggressive therapeutic intervention targeting CSCs population.

Omega-3 fatty acids inhibit tumor growth in a rat model of bladder cancer.

Omega-3 (ω-3) fatty acids have been tested on prevention and treatment of several cancer types, but the efficacy on "in vivo" bladder cancer has not been analyzed yet. This study aimed at evaluating the chemopreventive efficacy of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) mixture in an animal model of bladder cancer. Forty-four male Wistar rats were divided into 4 groups during a 20-week protocol: control; carcinogen-N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN); ω-3 (DHA + EPA); and ω-3 + BBN. BBN and ω-3 were given during the initial 8 weeks. At week 20 blood and bladder were collected and checked for the presence of urothelium lesions and tumors, markers of inflammation, proliferation, and redox status. Incidence of bladder carcinoma was, control (0%), ω-3 (0%), BBN (65%), and ω-3 + BBN (62.5%). The ω-3 + BBN group had no infiltrative tumors or carcinoma in situ, and tumor volume was significantly reduced compared to the BBN (0.9 ± 0.1 mm(3) versus 112.5 ± 6.4 mm(3)). Also, it showed a reduced MDA/TAS ratio and BBN-induced serum CRP, TGF-ß1, and CD31 were prevented. In conclusion, omega-3 fatty acids inhibit the development of premalignant and malignant lesions in a rat model of bladder cancer, which might be due to anti-inflammatory, antioxidant, anti-proliferative, and anti-angiogenic properties.

Chemopreventive efficacy of Atorvastatin against nitrosamine-induced rat bladder cancer: antioxidant, anti-proliferative and anti-inflammatory properties.

To investigate the anti-carcinogenic effects of Atorvastatin (Atorva) on a rat bladder carcinogenesis model with N-butyl-N-(4-hydroxibutil)nitrosamine (BBN), four male Wistar rat groups were studied: (1) CONTROL: vehicle; (2) Atorva: 3 mg/kg bw/day; (3) Carcinogen: BBN (0.05%); (4) Preventive Atorva: 3 mg/kg bw/day Atorva + BBN. A two phase protocol was used, in which the drug and the carcinogen were given between week 1 and 8 and tumor development or chemoprevention were expressed between week 9 and 20, when the bladders were collected for macroscopic, histological and immunohistochemical (p53, ki67, CD31) evaluation. Serum was assessed for markers of inflammation, proliferation and redox status. The incidence of bladder carcinoma was: control 0/8 (0%); Atorva 0/8 (0%); BBN 13/20 (65%) and Atorva + BBN 1/8 (12.5%). The number and volume of tumors were significantly lower in the Atorva + BBN group, with a marked reduction in hyperplasia, dysplasia and carcinoma in situ lesions. An anti-proliferative, anti-inflammatory and antioxidant profile was also observed in the preventive Atorva group. p53 and ki67 immunostaining were significantly increased in the BBN-treated rats, which was prevented in the Atorva + BBN group. No differences were found for CD31 expression. In conclusion, Atorvastatin had a clear inhibitory effect on bladder cancer development, probably due to its antioxidant, anti-proliferative and anti-inflammatory properties.

Inhibition of bladder tumor growth by chitooligosaccharides in an experimental carcinogenesis model.

Urinary bladder cancer is one of the most common cancers worldwide, with the highest incidence in industrialized countries. Patients with cancer commonly use unconventional and complementary therapy including nutraceuticals. In this study we evaluated the efficacy of chitooligosaccharides (in orange juice) in rat bladder cancer chemoprevention and as therapeutic agent, on a rat model of urinary bladder carcinogenesis induced with N-butyl-N-(4-hydroxybutyl) nitrosamine. Results indicate that chitooligosaccharides may have a preventive effect on bladder cancer development and a curative effect upon established bladder tumors, dependent on the concentration ingested 500 mg/kg b.w., every three days, showed capacity to inhibit and prevent the proliferation of bladder cancer; however, this was associated with secondary effects such as hypercholesterolemia and hypertriglyceridemia. The use of lower doses (50 and 250 mg/kg b.w.) showed only therapeutic effects. It is further suggested that this antitumor effect might be due to its expected anti-inflammatory action, as well as by mechanisms not directly dependent of COX-2 inhibition, such as cellular proliferation control and improvement in antioxidant profile.

Inhibition of bladder tumour growth by sirolimus in an experimental carcinogenesis model.

OBJECTIVE: To investigate the anticarcinogenic effects of sirolimus 2 mg/kg/day on a rat model of urinary bladder carcinogenesis induced with N-butyl-N(4-hydroxybutyl)nitrosamine (BBN). MATERIALS AND METHODS: Thirty-six male Wistar rats were divided into four groups: 1, a control group (eight), given tap water only; 2, a sirolimus control group (eight), given 2 mg/kg/day; 3, a carcinogen (BBN) group (12) exposed to 0.05% BBN; 4, a treatment group (sirolimus/BBN; eight) given 2 mg/kg/day + 0.05% BBN. In the tumour-induction phase, from week 1 to week 8, rats from groups 3 and 4 received BBN ad libitum in drinking water. In the treatment phase, from week 8 to week 20, rats from groups 2 and 4 received sirolimus 2 mg/kg/day by an oesophageal cannula. At week 20 the rats were killed humanely, and the number and size of tumours recorded. The bladders were collected for histological, immunohistochemical and gene expression evaluation. Blood was collected for the determination of several serum proliferative and inflammatory markers. Lipid peroxidation, through serum malondialdehyde (MDA) content, and total antioxidant status (TAS) were also evaluated. RESULTS: Sirolimus caused a marked inhibition of bladder tumour growth. When compared with group 3, group 4 had a reduced proportion of rats with tumour (three of eight vs eight of 12), and significantly fewer tumours per rat, with a mean (sd) of 1.00 (0.0) vs 1.88 (0.35), and tumour volume per tumour, of 0.30 (0.11) vs 66.1 (48.9) mm³, with less aggressive histological changes, i.e. a marked reduction in hyperplasia (four of eight vs 12/12), high-grade dysplasia (four of eight vs 11/12) and urothelial tumour. Rats in group 4 had no infiltrative bladder cancers and had a lower incidence of high-grade tumours than rats in group 3. The rats in group 4 had decreased serum levels of transforming growth factor-ß1, higher levels of tumour necrosis factor-α, and higher levels of serum TAS and a better serum MDA/TAS ratio, a marker of more favourable redox status. Furthermore, the down-regulation of bladder caspase 3 gene expression and the increased Ki67 immunostaining in group 3 were significantly attenuated in group 4. CONCLUSIONS: Sirolimus given as an oral agent, 2 mg/kg/day, significantly inhibited rat bladder carcinogenesis. Sirolimus reduced the number and volume of tumours and induced a less aggressive histological behaviour. This might be due to antiproliferative and antioxidant properties, as well as to the restoration of apoptotic pathways.