Role of Gut Microbiome in Neoadjuvant Chemotherapy Response in Urothelial Carcinoma: A Multi-institutional Prospective Cohort Evaluation.

Neoadjuvant chemotherapy (NAC) is linked with clinical advantages in urothelial carcinoma for patients with muscle-invasive bladder cancer (MIBC). Despite comprehensive research into the influence of tumor mutation expression profiles and clinicopathologic factors on chemotherapy response, the role of the gut microbiome (GM) in bladder cancer chemotherapy response remains poorly understood. This study examines the variance in the GM of patients with bladder cancer compared with healthy adults, and investigates GM compositional differences between patients who respond to chemotherapy versus those who exhibit residual disease.Our study reveals distinct clustering, effectively separating the bladder cancer and healthy cohorts. However, no significant differences were observed between chemotherapy responders and nonresponders within community subgroups. Machine learning models based on responder status outperformed clinical variables in predicting complete response (AUC 0.88 vs. AUC 0.50), although no single microbial species emerged as a fully reliable biomarker.The evaluation of short chain fatty acid (SCFA) concentration in blood and stool revealed no correlation with responder status. Still, SCFA analysis showed a higher abundance of Akkermansia (rs = 0.51, P = 0.017) and Clostridia (rs = 0.52, P = 0.018), which correlated with increased levels of detectable fecal isobutyric acid. Higher levels of fecal Lactobacillus (rs = 0.49, P = 0.02) and Enterobacteriaceae (rs = 0.52, P < 0.03) correlated with increased fecal propionic acid.In conclusion, our study constitutes the first large-scale, multicenter assessment of GM composition, suggesting the potential for a complex microbial signature to predict patients more likely to respond to NAC based on multiple taxa. SIGNIFICANCE: Our study highlights results that link the composition of the GM to the efficacy of NAC in MIBC. We discovered that patients with higher levels of Bacteroides experienced a worse response to NAC. This microbial signature shows promise as a superior predictor of treatment response over traditional clinical variables. Although preliminary, our findings advocate for larger, more detailed studies to validate these associations.

Candida nivariensis: Identification strategy in mycological laboratories.

Candida nivariensis is a cryptic fungal species classified within the Candida glabrata complex. It was described for the first time in 2005 by the means of DNA sequencing. We report a rare case of C. nivariensis deep-seated infection occurring in a 77-year-old man hospitalized for cysto-prostatectomy. Phenotypic testing based on the direct examination and the macroscopic features of the in vitro culture initially suggested C. glabrata species, while MALDI-TOF mass spectrometry enables correct identification. The isolate was found resistant to fluconazole, like in almost 20% of the reported cases. Herein, we present our practical strategy to reliably characterize this rare cryptic species. To date, MALDI-TOF mass spectrometry-based analysis showed very good results for such a purpose.

Concomitant Antibiotic Use and Survival in Urothelial Carcinoma Treated with Atezolizumab.

Antibiotic effects on the gut microbiota may negatively impact survival with immune checkpoint inhibitors (ICIs). However, there is minimal evidence regarding whether antibiotic impacts are specific to ICIs or impacts in urothelial carcinoma (UC). In a post hoc analysis of IMvigor210 (single-arm atezolizumab) and IMvigor211 (phase III randomised trial of atezolizumab vs chemotherapy), the association between antibiotic use and overall survival (OS) and progression-free survival (PFS) was assessed via Cox proportional hazard analysis. Antibiotic use was defined as any antibiotic administration between 30 d prior to and 30 d after treatment initiation. Antibiotic use was associated with worse OS (n = 847, hazard ratio or HR [95% confidence interval {CI}] = 1.44 [1.19-1.73]) and PFS (1.24 [1.05-1.46]) with atezolizumab, but not chemotherapy (n = 415, 1.15 [0.91-1.46] and 1.09 [0.88-1.36], respectively). In the randomised cohort of IMvigor211, the OS treatment effect (HR [95% CI]) of atezolizumab versus chemotherapy was 0.95 (95% CI 0.71-1.25) for antibiotic users, compared with 0.73 (0.60-0.88) for nonusers (p[interaction] = 0.1). Similar associations were noted in the PD-L1 IC2/3 population. In conclusion, antibiotic use was associated with worse survival outcomes in UC patients treated with atezolizumab. The study does not justify a change in antibiotic selection for infections; however antibiotic overuse occurs in cancer care and this needs to be evaluated for ICIs. PATIENT SUMMARY: In this report from clinical trials IMvigor210 and IMvigor211, it was demonstrated that antibiotic use is consistently associated with worse survival in patients with urothelial carcinoma treated with atezolizumab. No antibiotic association was observed in patients treated with chemotherapy, suggesting that antibiotics may specifically reduce the effectiveness of cancer immunotherapies. Future research will continue to explore the effect of antibiotics on other immune checkpoint inhibitors and confirm whether immune checkpoint inhibitors remain the treatment of choice in cancer patients requiring antibiotics.

Sex-specific Alterations in the Urinary and Tissue Microbiome in Therapy-naïve Urothelial Bladder Cancer Patients.

Comprehensive characterization of the urinary and urothelium-bound microbiomes in bladder cancer (BCa) and healthy state is essential to understand how these local microbiomes may play a role in BCa tumorigenesis and response to therapy, as well as to explain sex-based differences in BCa pathobiology. Performing 16 s rDNA microbiome analysis on 166 samples (urine and paired bladder tissues) from therapy-naïve BCa patients undergoing radical cystectomy and healthy controls, we defined (1) sex-specific microbiome differences in the urine and bladder tissue, and (2) representativeness of the tissue microenvironment by the voided urinary microbiome. The genus Klebsiella was more common in the urine of female BCa patients versus healthy controls, while no clinically relevant bacteria were found differently enriched in men. In tissues, the genus Burkholderia was more abundant in the neoplastic versus the non-neoplastic tissue in both sexes, suggesting a potential role in BCa pathobiology. Lastly, we found that the urinary microbiome shares >80% of the bacterial families present in the paired bladder tissue, making the urinary microbiome a fair proxy of the tissue bacterial environment. PATIENT SUMMARY: We identified specific bacteria present in the urine and tissues of male and female bladder cancer patients. These novel data represent a first step toward understanding the influence of the bladder microbiome on the development of bladder cancer and on the response to intravesical and systemic therapies.

The Urinary Microbiome: Implications in Bladder Cancer Pathogenesis and Therapeutics.

Recent investigation has proven that the bladder is not sterile. However, the implications of this finding in the pathophysiology and management of urothelial cell carcinoma have not been fully described. In this review, we summarize the literature relating to the urinary and gastrointestinal microbiomes in the context of urothelial cell carcinoma. The bladder microbiome may relate to urothelial cell carcinoma pathogenesis/progression, act as a noninvasive and modifiable urinary biomarker and have implications in treatment using immunotherapy agents such as intravesical bacillus Calmette-Guerin. Investigators should continue to optimize techniques to characterize this intriguing new area of human health.

Effect of Preoperative Bacteriuria and Pyuria on Intravesical Recurrence in Patients with Upper Tract Urothelial Carcinoma Undergoing Radical Nephroureterectomy.

BACKGROUND/AIM: We investigated the effect of bacteriuria and pyuria on intravesical recurrence (IVR) in patients with upper tract urothelial carcinoma (UTUC) undergoing radical nephroureterectomy (RNU). PATIENTS AND METHODS: Preoperative bacteriuria and pyuria were defined as urine containing ≥5 bacteria/high-power field (HPF) and >5 white blood cells/HPF, respectively. Their associations with IVR were evaluated in 97 patients with UTUC undergoing RNU. RESULTS: Preoperative bacteriuria [n=15 (15%)] was significantly associated with preoperative pyuria [n=42 (43%), p<0.001]. During follow-up (median of 19 months), 45 (46%) patients developed IVR (median IVR-free survival=38 months). On multivariate analysis, preoperative bacteriuria was an independent predictor for reduced risk of IVR (hazard ratio=0.23, p=0.010). The 2-year IVR-free survival of patients with preoperative bacteriuria and pyuria was significantly longer than that of patients without preoperative bacteriuria (83% vs. 54%, p=0.028) and pyuria (69% vs. 50%, p=0.024), respectively. CONCLUSION: Bacteriuria and pyuria may reduce the risk of IVR in patients with UTUC undergoing RNU.

Bacterial urinary tract infections associated with transitional cell carcinoma in dogs.

BACKGROUND: Urinary tract infections (UTI) are believed to be common in dogs with transitional cell carcinoma (TCC), but incidence and contributing factors have not been reported. OBJECTIVES: To determine the frequency and bacterial agents associated with UTI in dogs with TCC and define contributing factors. ANIMALS: Eighty-five dogs with a history of urogenital TCC undergoing treatment with chemotherapy that had at least 1 urine culture performed. METHODS: Medical records and culture results were retrospectively reviewed and ultrasound images were reviewed when available. Clinical factors were evaluated statistically for association with positive culture. RESULTS: Fifty-five percent (47/85) of dogs had at least 1 positive culture during the course of treatment. Female dogs (80%, 40/50) were more likely than male dogs (29%, 10/35) to have at least 1 positive culture. Ultrasound examination determined that female dogs were more likely to have urethral (74%, 31/42) or trigonal tumor involvement (71%, 30/42) compared to male dogs (32%, 9/28 and 43%, 12/28, respectively). The most commonly isolated organisms were Staphylococcus spp. (23.9%, 29/121) and Escherichia coli (19.8%, 24/121). Dogs with urethral involvement of TCC were significantly more likely to have at least 1 positive culture than dogs without urethral involvement (75%, 30/40 versus 30%, 9/30). CONCLUSIONS: Urinary tract infection is common in dogs with TCC highlighting the importance of regular monitoring for bacterial cystitis in dogs with TCC. In addition, clinical factors such as tumor location and sex may be predictive of positive culture and can help clinicians assess the risk of UTI.

The microbiome of the urinary tract--a role beyond infection.

Urologists rarely need to consider bacteria beyond their role in infectious disease. However, emerging evidence shows that the microorganisms inhabiting many sites of the body, including the urinary tract--which has long been assumed sterile in healthy individuals--might have a role in maintaining urinary health. Studies of the urinary microbiota have identified remarkable differences between healthy populations and those with urologic diseases. Microorganisms at sites distal to the kidney, bladder and urethra are likely to have a profound effect on urologic health, both positive and negative, owing to their metabolic output and other contributions. Connections between the gut microbiota and renal stone formation have already been discovered. In addition, bacteria are also used in the prevention of bladder cancer recurrence. In the future, urologists will need to consider possible influences of the microbiome in diagnosis and treatment of certain urological conditions. New insights might provide an opportunity to predict the risk of developing certain urological diseases and could enable the development of innovative therapeutic strategies.

La hibridación in situ fluorescente en la predicción de recurrencia del carcinoma urotelial / Fluorescent in situ Hybridization as a Predictor of Relapse in Urotelial Carcinoma

Objetivo: Evaluar la utilidad del estudio de alteraciones cromosómicas mediante hibridación in situ fluorescente en una serie de pacientes diagnosticados de carcinoma urotelial con un seguimiento mínimo de 24 meses, y analizar su posible efecto anticipador. Material y métodos: La serie global incluye 338 muestras procedentes de 98 pacientes con 84 episodios de carcinoma urotelial. Para evaluar la capacidad de predicción del test se estudió un subgrupo de 24 pacientes que presentaron como mínimo una recurrencia a lo largo del seguimiento (serie de recurrencia). Se consideraron 3 categorías (episodio coherente positivo, episodio coherente negativo y episodio no coherente) en función de la relación entre los resultados de la hibridación in situ fluorescente del estudio concomitante al nuevo episodio y las muestras precedentes. Resultados: La hibridación in situ fluorescente presentó mayor sensibilidad independientemente del grado, valor predictivo negativo y exactitud, mientras que la especificidad y el valor predictivo positivo fueron superiores para la citología convencional. En la serie de recurrencia 19/29 episodios resultaron coherentes, 11/19 fueron coherentes positivos, todos con carcinoma urotelial de alto grado, y 8/19 coherentes negativos, la mayoría de bajo grado. Conclusiones: La hibridación in situ fluorescente muestra una alta sensibilidad en un seguimiento de 24 meses, y es capaz de predecir recurrencias, especialmente en casos de alto grado. Nuestros datos demuestran también la existencia de carcinomas uroteliales sin alteraciones cromosómicas detectables con la metodología actualmente disponible. Los resultados apoyan un seguimiento multidisciplinar con la utilización combinada de la hibridación in situ fluorescente, citología y cistoscopia (AU)

Objective: To assess the value of the study of chromosomal alterations by fluorescent in situ hybridization in a series of patients diagnosed of urothelial carcinoma and a minimum follow up of twenty four months, as well as evaluate its putative predictive potential. Material and methods: The overall series includes 338 samples from 98 patients with 84 episodes of urothelial carcinoma. A subgroup of 24 patients who had at least one recurrence during the follow up was used to evaluate the predictive potential of the test. Three categories were considered (positive coherent episode, negative coherent episode, and incoherent episode) depending on the relationship between the fluorescent in situ hybridization result in the concomitant study of the new episode and those of the preceding samples. Results: Fluorescent in situ hybridization showed higher sensitivity regardless of grade, negative predictive value and accuracy, while specificity and positive predictive value were superior with conventional cytology. In the recurrence, series 19/29 episodes were coherent, 11/19 were positive coherent with urothelial carcinoma all high grade and 8/19 negative coherent, most low grade. Conclusions: Fluorescent in situ hybridization test shows good sensitivity during a follow up of twenty four months and is able to predict recurrence, especially in cases of high grade. Our data demonstrate the existence of urothelial carcinomas without detectable chromosomal abnormalities by currently available methodology. The results support a multidisciplinary follow up combining fluorescent in situ hybridization, cytology and cystoscopy (AU)

Clostridium septicum can cause distant myonecrosis in patients with ovarian cancer.

A case report of lethal distant myonecrosis with gas gangrene is presented. Blood cultures and tissue biopsies revealed Clostridium septicum. The 55-year-old female patient presented with recurrent ovarian cancer of transitional cell type, initially diagnosed as FIGO IIb in January 2011, with hepatic metastasis and invasion of the ceacal wall. She underwent several operations, including partial bowel and liver resection in September 2011. Second-line therapy with topotecan three weekly was started in October 2011 while the patient was still in the hospital. During this chemotherapy, the patient revealed symptoms of severe pain and erythema of the skin. Within hours she died of sceptic shock after a debridement. The diagnosis was gas gangrene due to Clostridium septicum. Because it is a rare and severe disease and the time slot in which therapeutic measures can be taken is narrow, we discuss clinical symptoms and therapeutic options.

Mycotic aneurysm of the aorta as a complication of Bacillus Calmette-Guérin instillation.

Bacillus Calmette-Guérin (BCG) is a live attenuated strain of Mycobacterium bovis that has been widely used for the treatment of superficial transitional cell carcinoma of the bladder. We describe a rare case of supra-renal mycotic aortic aneurysm secondary to BCG instillation in a 75-year-old male. Patients presenting with systemic symptoms post-instillation, possibly with an aneurysm, should raise suspicion of BCG dissemination, which requires early instigation of anti-mycobacterial drugs.

[Study on the inducible expression of toll-like receptors in human bladder cell line].

OBJECTIVE: To investigate and compare the expression of Toll-like receptors (TLRs) in human bladder cells under the stimulus of Escherichia coli (E. coli) versus that under the stimulus of Bacillus Calmette-Guérin (BCG). METHODS: Human bladder cancer cell line T24 was cultured for 1 hour under the stimuli of various doses (bacillus-cell ratio) of E. coli and BCG respectively. The levels of expression of TLR-2 and TLR-4 mRNA in T24 cells were assessed by RT-PCR method. RESULTS: By comparison with control, there was no difference observed on the expression of either TLR-2 or TLR-4 in T24 cells under the stimulus of E. coli. The expression level of either TLR-2 or TLR-4 was increased under the stimulus of BCG in a dose-dependent manner. The effects reached the highest level when the dose of BCG was 10 bacilli per cell. CONCLUSION: There exist different expression patterns of TLRs in bladder transitional cells under the different stimuli of E. coli and BCG respectively.

Bacillus Calmette-Guerin initiates intracellular signaling in a transitional carcinoma cell line by cross-linking alpha 5 beta 1 integrin.

PURPOSE: The adherence of bacillus Calmette-Guerin (BCG) to the surface of transitional carcinoma tumor cells initiates nuclear factor (NF)-kappa B signal transduction pathways that modulate the expression of proteins important in the antitumor response to BCG. We tested the hypothesis that BCG initiates NF-kappa B signaling as a consequence of cross-linking alpha 5 beta 1 integrin receptors present on the tumor cell surface. MATERIALS AND METHODS: The effect of alpha 5 beta 1 antibody mediated cross-linking on interleukin (IL)-6 mRNA expression, IL-6 promoter activation and activation of a specific NF-kappa B reporter construct was determined. A series of reporter constructs containing nonfunctional mutations in the AP-1, NF-IL-6 and NF-kappa B sites were used to determine the relative importance of these response elements in alpha 5 beta 1 cross-linking mediated activation of the IL-6 promoter. A final series of experiments assessed the role of alpha 5 beta 1 receptor occupancy by fibronectin (FN) in initiating antibody or BCG mediated signaling. RESULTS: Anti alpha 5 and anti beta 1 mediated cross-linking of alpha 5 beta 1 integrin initiated NF-kappa B signaling, IL-6 promoter activation and IL-6 mRNA expression. Deletion mutants demonstrated that alpha 5 beta 1 cross-link initiated, IL-6 promoter transactivation required intact NF-kappa B and AP-1 response elements. Receptor occupancy by FN was required for BCG but not for antibody initiated signaling. CONCLUSIONS: Cross-linking the alpha 5 beta 1 receptor present on the surface of human transitional carcinoma cells lines initiates signal transduction in a manner identical to that observed for BCG. We propose a model in which multiple FN binding sites present on BCG interact with alpha 5 beta 1 receptor bound FN molecules to cross-link alpha 5 beta 1 receptors and initiate intracellular signaling.

[Investigation of hydrophobicity of Proteus vulgaris strains and ability of Proteus vulgaris and Proteus penneri strains to penetrate bladder membrane HCV T-29 cells ]. / Badania hydrofobowosci szczepów Proteus vulgaris oraz zdolnosci szczepów Proteus vulgaris i Proteus penneri do penetracji komórek nablonka pecherza moczowego HCV T-29.

Proteus bacilli play a particularly important role in urinary tract infections (UTI). Fimbriae and adherence ability and hemolysins production (HpmA, HlyA) are one of the factors of pathogenicity of these bacteria. In this paper we describe the invasion of HCV T-29 transitional bladder urothelial cells carcinoma strains of P. penneri, as well as P. vulgaris strains belonging to different serogroups. The cytotoxic effect was observed at 8 hour of incubation of the tested cells with P. vulgaris O21 and the same effect (complete lysis) at 6 hours by P. vulgaris O4 (this strain manifests maximal activity in the production of HlyA hemolysin). P. penneri strains, produce different types of fimbriae, expressed similar bacterial invasiveness. The hydrophobic properties of 25 P. vulgaris strains were also tested and only 3 strains occur to have hydrophobic cell surface.

Absence of Helicobacter pylori in oral mucosal lesions.

A prospective study was conducted in 26 patients with benign oral ulcers and oral carcinoma and 26 age and sex matched controls to determine the prevalence of Helicobacter pylori in oral mucosal biopsies. Oral mucosal biopsies were subjected to rapid urease test, campylobacter like organism (CLO) test, histopathological examination and bacteriological culture for demonstration of H pylori. Urease test was positive for H pylori in 3 (11.4%) out of 26 cases and CLO test in 2 (14%) out of 14 cases. On histology, H pylori was identified in 4 (15.38%) out of 26 cases. The spiral organism was universally absent in culture of both patients and controls. These pilot data negate the casual association of H pylori in oral mucosal lesions.

The persistence of bacille Calmette-Guérin in the bladder after intravesical treatment for bladder cancer.

OBJECTIVE: To determine the incidence of bacille Calmette-Guérin (BCG) bacilli persisting in the urinary tract of patients treated previously with intravesical BCG for carcinoma in situ or multiple Ta.T1 transitional cell carcinoma. PATIENTS AND METHODS: One-hundred and twenty-five patients were treated at the Freeman Hospital, Newcastle upon Tyne, UK over an 8-year period, 90 of whom submitted early morning urine samples for culture for acid-fast bacilli at varying intervals following BCG treatment. The records of all patients were reviewed to determine the incidence of caseating granulomata containing acid-fast bacilli together with the incidence of toxicity and the outcome of treatment. RESULTS: Five patients were found to have persisting acid-fast mycobacteria in their urine or bladder up to 16.5 months after completing intravesical instillations of BCG. In one patient this probably accounted for bladder symptoms that required palliative cystectomy. In four patients the 'infection' was not severe. Two patients were treated with antituberculous chemotherapy without complication. Three years after intravesical BCG therapy 36 of 69 patients (52%) had remained tumour free. CONCLUSION: BCG organisms can persist in the urinary tract for at least 16.5 months after the completion of intravesical BCG instillation therapy.

Intravesical gene therapy: in vivo gene transfer using recombinant vaccinia virus vectors.

Intratumoral gene transfer may be a significant tool in active immunotherapy. The ability to insert functional genes into a tumor in vitro and in vivo using recombinant vaccinia vectors was examined in the murine bladder tumor model. Vaccinia recombinants expressing the influenza hemagglutinin or nucleoprotein antigens infected/transfected murine (MB-49 and MBT-2) and human (T24) bladder tumor cell lines in vitro. Systemic vaccinia immunity was induced with as few as 10 plaque-forming units of recombinant vaccinia instilled intravesically, and the encoded protein was expressed in vivo in tumor and urothelium. However, preimmunity to vaccinia did not inhibit intravesical tumor transfection. Thus, recombinant vaccinia virus is effective in introducing foreign antigens locally into tumor in vivo, supporting its use in clinical immunotherapy.

High-risk human papillomavirus infections and overexpression of p53 protein as prognostic indicators in transitional cell carcinoma of the urinary bladder.

Ninety Japanese patients with transitional cell carcinoma of the urinary bladder were investigated for tumor incorporation of DNA for high-risk human papillomavirus (HPV) types 16, 18, and 33 by in situ hybridization with biotinylated DNA probes. In addition, immunohistochemical analysis of p53 protein expression was performed with an antibody to p53 protein. Twenty-eight tumors were positive for HPV DNA, and multiple HPV infection was detected in 17 cases. Positive nuclear staining of cancer cells by the antibody to p53 protein was detected in 32 cases. DNA for HPV 16, 18, and/or 33 and the overexpression of p53 protein were simultaneously observed in 6 tumors by using a mirror section method. The overexpression of p53 protein was frequently detected in invasive and nonpapillary tumors (P < 0.05) and in high grade tumors (P < 0.05). In contrast, HPV infection was more common in noninvasive and papillary tumors (P < 0.01). The patients with tumors positive for HPV DNA and/or p53 antibody had a significantly worse survival rate (P < 0.05). These results suggest that HPV infection or overexpression of p53 protein may be related to tumor behavior and may indicate a relatively poor prognosis in patients with transitional cell carcinoma.