Bladder Epithelium Research Articles

Effects of chronic stress on bladder morphology of rats and impact of comfort food diet as an ameliorating agent

Objectives To investigate the effects of chronic stress on bladder morphology and the impact of food preference (standard or comfort foods) on the bladder of stressed rats. Methods In total, 32 Wistar male rats (3 months old) were divided into four groups: control (C), stressed (S), control + comfort food (C + CF), and stressed + comfort food (S + CF). Groups C and C + CF were maintained under normal conditions, while groups S and S + CF were subjected to chronic stress by the restraint method. Groups C and S received standard rat chow, while groups C + CF and S + CF received comfort food (Froot Loops®) and standard chow. The stress stimuli were induced daily for 2 h over 8 weeks. After 8 weeks, all animals were killed, and the bladders were removed and used for histomorphometric analysis. Results Body mass was similar among the groups. Stress did not promote differences regarding food intake, but animals receiving comfort food showed higher calories intake (in kcal/Kg) than animals receiving only standard chow. The C + CF and S + CF groups preferred comfort food over the standard chow; this preference was higher in the S + CF than in the C + CF group. The surface density of smooth muscle was reduced in stressed animals, while connective tissue and elastic system fiber content were increased in stressed groups. Further, epithelial height was increased in rats submitted to chronic stress. The surface density of elastic system fibers was decreased by the consumption of comfort food. Conclusions Chronic stress induces morphological modifications on the bladder wall and epithelium. These modifications may be related to lower urinary tract symptoms. Additionally, chronic stress caused a higher preference for comfort food intake which did not ameliorate or aggravate the stress-induced bladder alterations.

Regeneration of Functional Bladder Using Cell-seeded Amnion and P(LA/CL) Scaffolds.

Long-term bladder regeneration has not been successful instead of augmentation with gastrointestinal segments, as is commonly performed for bladder reconstruction. To evaluate whether or not cell-seeded bioabsorbable materials regenerate half-resected bladder in a rabbit model. Female Japanese white rabbits were divided two groups: cell-seeded material (CSM) group and Control (n = 6 each). Control rabbits underwent resection of half the bladder. CSM rabbits were sutured with cell-seeded amniotic membrane and P(LA/CL) material after bladder resection. After 6, 12, and 18 months, rabbits underwent X-ray and cystometry, and bladder tissues after 18 months were subjected to functional and histological analyses. X-ray confirmed the peristaltic movements of the reconstructed bladders in the CSM group. On cystometry, the mean maximum bladder volume, maximum bladder pressure, and 25 mL bladder volume compliance in the CSM group were significantly greater than in the Control group at 6, 12, and 18 months. In addition, organ bath studies showed good contraction under electrical stimulation with increasing stimulation frequency in the CSM group, while, the Control group showed weak contraction on both tests in the central marginal zone. Furthermore, the rates of neovascularization, urothelial and smooth muscle formation, and neurofilamentation in the CSM group were significantly greater than in the Control group. Oral mucosal cell-seeded amniotic membrane and stomach smooth muscle cell-seeded P(LA/CL) scaffold with omentum after abdominal implantation regenerated functional bladder with satisfactory epithelium and smooth muscle without scarring more than 1 year. Impact Statement Regeneration of functional bladder without using gastrointestinal segments has been a huge challenge to urological reconstruction. Various materials, such as nonbioabsorbable materials and biomaterials have been attempted to reconstruct bladder in animal models. However, the long-term results more than a year failed due to the low biocompatibility, high risks, and difficulty creating the materials. In this study, we revealed long-term bladder regeneration using cell-seeded amniotic membrane and P(LA/CL) material in a rabbit model. The new method of bladder reconstruction seems able to regenerate functional bladder with satisfactory bladder epithelium and bladder smooth muscle function without scarring for more than 1 year successfully.

Ketamine-Induced Cystitis: A Comprehensive Review of the Urologic Effects of This Psychoactive Drug

Ketamine is a common medical anesthetic and analgesic but is becoming more widely used as a recreational drug. Significant side effects on the urinary tract are associated with frequent recreational ketamine use most notably ketamine-induced cystitis (KIC). Regular ketamine consumption has been shown to increase the risk of cystitis symptoms by 3- to 4-fold, and cessation of ketamine use is usually associated with improvement of symptoms. Common KIC-related problems are urinary pain and discomfort, bladder epithelial barrier damage, reduced bladder storage and increased pressure, ureter stenosis, and kidney failure, all of which significantly impact patients' quality of life. Furthermore, it becomes a vicious cycle when KIC patients attempt to manage their urinary pain with increased ketamine use. The precise pathophysiology of KIC is still unknown but several theories exist, most of which highlight the inflammatory signaling pathways leading to bladder epithelium damage due to presence of ketamine in the urine. Empirical treatment options for KIC are available and consist of ketamine cessation, noninvasive therapies, and surgery, and should be decided upon based on the time course and severity of the disease. Of note, cessation of use is strongly recommended for all KIC patients, and should be supplemented with motivational interviews and psychological and social support. It is crucial for clinicians to be familiar with KIC diagnosis and treatment, and to be prepared to have informed discussions with ketamine-using patients about the potential health consequences of ketamine.

A new catheter-integrated drug-delivery system for controlled intravesical mitomycin C release

Intravesical treatment of bladder cancer is preferred over systemic administration. However, the efficacy of intravesical instillations is challenged by the periodic voiding that flushes out the instilled drug and ultimately reduces drug exposure to the bladder epithelium. Here, we demonstrate a new catheter-integrated drug-delivery concept that utilizes a silicone-based interpenetrating polymer network (IPN) as material for the catheter balloon, to facilitate continuous release of the bladder cancer adjuvant, Mitomycin C, from a balloon-reservoir to the urinary bladder. Long-term release properties and anti-carcinoma cell efficacy of released drug was investigated in vitro. Short-term release experiments were performed in live pigs to evaluate the IPN prototype catheter in a physiological relevant environment in vivo. Sustained zero-order release of Mitomycin C was achieved for 12 days in vitro without refilling the balloon. Mitomycin C was released from the IPN-balloons into the urinary bladder of live pigs in concentrations adequate to inhibit carcinoma cell growth. The IPN catheter represents a new drug-delivery concept for prolonged Mitomycin C delivery to the urinary bladder.

Computational modeling of stretch induced calcium signaling at the apical membrane domain in umbrella cells

The urinary bladder epithelium comprises a specialised population of superficially placed cells called the umbrella cells. The apical membrane domain of umbrella cells has several intriguing morphological properties and is the site for various signaling activities. A key function of umbrella cells is to sense mechanical stimuli as the bladder stretches in response to filling. More specifically, the mechanotransduction of stretch into subcellular signals is brought about by the activation of Piezo1 channels that mediate calcium into the cell interior. The incoming calcium is critical to several aspects of umbrella cell signaling, including regulation of exocytosis, ATP release and downstream purinergic signaling. We report here a computational framework that models stretch-induced mechanotransduction via Piezo1 channels and the resulting calcium signaling in umbrella cells factoring in morphological details of subcellular compartment volumes. Our results show the following: (i) activation of Piezo1 conductance in response to stretch; (ii) development of varying Piezo1 mediated [Ca2+] profiles in subcellular compartments, namely, the apical sub-plasma membrane space, cytosol and mitochondria. The varying calcium amplitudes and temporal profiles in the subcellular compartments indicate highly specialised roles for stretch-mediated calciumin umbrella cells, including its potential effect on the energetics of mitochondria and the regulation of exocytosis.

EP-157 Pyocystis: A silent complication of a supravesical urinary diversion

Abstract Aim Pyocystis is an important yet overlooked rare complication of a non-functioning urinary bladder. It is due to collection, liquefaction and decomposition of shed bladder epithelium, resulting in accumulation of pus in the bladder causing sepsis. We describe a case of Pyocystis in a patient with bilateral nephrostomies who presented with urosepsis multiple times due to this complication. Case Report A 78-year-old man, known case of Ca Prostate wit bilateral nephrostomies warranted multiple subsequent hospital admissions in one month due to urosepsis presenting with fever, urethral discharge, suprapubic pain and very high inflammatory markers. He underwent a CT Abdomen & Pelvis which revealed markedly distended bladder despite nephrostomies in place. Upon catheterization, no urine output was observed which raised concerns. An attempt at bladder washout displayed a large amount of thick foul-smelling pus which was aspirated. This was sent for culture and patient was commenced on appropriate IV antibiotics. After a thorough bladder wash out under flexible cystoscopy, patient was eventually discharged on a long-term catheter with regular bladder washouts in community as required. Patient was successfully managed on conservative treatment. Conclusion Pyocystis can sometimes be a missed complication of urinary diversion or a non-functioning bladder that can present silently under sepsis and should therefore be considered as a differential diagnosis of sepsis in patient with supravesical urinary diversion. We wanted to highlight this condition which if left undiagnosed will progress to severe sepsis and possible death. Hence prompt intervention and aggressive management is vital. Treatment for this complication requires catheterization, bladder irrigation and antibiotics.

The Significance of Expression Pattern of Dab1 and Reelin in the Postnatal Bladder of Dab1 -/- (Yotari) Mice

Background. The aim of our study is to provide an insight into the genetic expression landscape of Dab1 and Reelin, which are important in human genitourinary tract development and could help elucidate the critical stages of the onset of bladder anomalies.Methods. Yotari, heterozygote and wild type mice were sacrificed on the 4th, 11th and 14th postnatal day. Morphological parameters were analyzed using immunohistochemistry on mice bladder samples.Results. Significant expression of Dab1 was observed in the bladder epithelium during the 4th postnatal day and in the wt -wild type, but also in the yot -/- mouse. Reelin expression was significant in bladder epithelium during the 4th postnatal day in the wild-type and mutant mouse type. A significant expression of Dab1 positive cells in lamina propria was observed on the 4th postnatal day in yot -/- and in wt. The percentage of Reelin positive cells in lamina propria was significant on the 4th postnatal day in mutated and wild-type mice.Conclusions. An increase in the expression of Dab1 and Reelin proteins in the bladder of a yotari mouse may indicate bladder damage, due to congenital renal abnormalities and caused by a mutation in the Dab1 gene.Key words: Dab1; Reelin, CAKUT, bladder development, yotari mice

Urinary Tract Infections Caused by Uropathogenic Escherichia coli Strains-New Strategies for an Old Pathogen.

Urinary tract infections (UTIs) are among the most common infections worldwide. Uropathogenic Escherichia coli (UPECs) are the main causative agent of UTIs. UPECs initially colonize the human host adhering to the bladder epithelium. Adhesion is followed by the bacterial invasion of urothelial epithelial cells where they can replicate to form compact aggregates of intracellular bacteria with biofilm-like properties. UPEC strains may persist within epithelial urothelial cells, thus acting as quiescent intracellular bacterial reservoirs (QIRs). It has been proposed that host cell invasion may facilitate both the establishment and persistence of UPECs within the human urinary tract. UPEC strains express a variety of virulence factors including fimbrial and afimbrial adhesins, invasins, iron-acquisition systems, and toxins, which cooperate to the establishment of long lasting infections. An increasing resistance rate relative to the antibiotics recommended by current guidelines for the treatment of UTIs and an increasing number of multidrug resistant UPEC isolates were observed. In order to ameliorate the cure rate and improve the outcomes of patients, appropriate therapy founded on new strategies, as alternative to antibiotics, needs to be explored. Here, we take a snapshot of the current knowledge of coordinated efforts to develop innovative anti-infective strategies to control the diffusion of UPECs.

Molecular Mechanisms and Key Processes in Interstitial, Hemorrhagic and Radiation Cystitis.

Simple SummaryPathologies of the bladder are called cystitis. They cause discomfort for the patient. Due to persistent pain, bleeding, urinary incontinence, and uncontrolled urination, the chronic forms cause considerable degradation to patient quality of life. Currently, there is no curative treatment for the most severe forms. This is both an economic and a societal problem. Although the different forms of cystitis have different causes, they share common mechanisms. We propose to describe in detail the key processes and the associated mechanisms involved in abacterial cystitis.Cystitis is a bladder disease with a high rate of prevalence in the world population. This report focuses on Interstitial Cystitis (IC), Hemorrhagic Cystitis (HC) and Chronic Radiation Cystitis. These pathologies have different etiologies, but they share common symptoms, for instance, pain, bleeding, and a contracted bladder. Overall, treatments are quite similar for abacterial cystitis, and include bladder epithelium protective or anti-inflammatory agents, alleviating pain and reducing bleeding. This review summarizes the mechanisms that the pathologies have in common, for instance, bladder dysfunction and inflammation. Conversely, some mechanisms have been described as present in only one pathology, such as neural regulation. Based on these specificities, we propose identifying a mechanism that could be common to all the above-mentioned pathologies.

Indirect immunofluorescence on rat bladder epithelium inpatients with pemphigus vulgaris with an extended follow-up.

BackgroundIndirect immunofluorescence (IIF) on rat bladder epithelium (RBE) has been widely used to detect anti‐plakin antibodies present in paraneoplastic pemphigus (PNP). However, anti‐plakin antibodies have also been found in a group of patients with pemphigus vulgaris (PV).ObjectivesTo assess the reactivity rate of PV sera in IIF using RBE as substrate and the diagnostic usefulness of the aforementioned test.MethodsPatients diagnosed with PV presenting to Razi Hospital, Tehran, Iran, were recruited. The patients' demographics, disease severity, and response to the initial treatment were recorded. Sera were collected and tested by IIF on RBE and by desmoglein 3/1 (Dsg 3/1) enzyme‐linked immunosorbent assay. Patients were followed up closely for a mean of 53.9 months for any evidence of malignancy.ResultsForty‐six patients were enroled (mean age of 42.9 years old, 31 females). Nine sera (19.6%) showed reactivity in IIF on RBE. Mean serum anti‐Dsg levels did not differ significantly among the two groups with positive and negative IIF results. Negative anti‐Dsg3 was related to a higher positive rate in IIF on RBE. There was no significant correlation between the reactivity of IIF on RBE and patients' demographic, clinical, or serological characteristics.ConclusionsIIF on RBE is a sensitive test for detecting antibodies against plakins. However, it has a relatively high false‐positive rate in PV, probably due to the epitope spreading phenomenon. This test should be suggested when there is a clinical or immunohistopathological suspicion of PNP and should be interpreted with caution.

Antibiofilm and Antimicrobial Activities of Chloroindoles Against Uropathogenic Escherichia coli.

Uropathogenic Escherichia coli (UPEC) is a nosocomial pathogen associated with urinary tract infections and expresses several virulence factors that cause recurring infections and cystitis of the bladder, which can lead to pyelonephritis. UPEC uses different types of extracellular appendages like fimbriae and pili that aid colonization and adherence to bladder epithelium and can form persistent biofilm-like bacterial communities that aid its survival after the deployment of host immune responses. We investigated the antibiofilm, antimicrobial, and antivirulence properties of three indole derivatives namely, 4-chloroindole, 5-chloroindole, and 5-chloro 2-methyl indole. All the three chloroindoles had MICs of 75 μg/ml and inhibited biofilm formation by an average of 67% at 20 μg/ml. In addition, they inhibited swarming and swimming motilities, which are essential for dissemination from bacterial communities and colonization, reduced cell surface hydrophobicity, and inhibited indole production and curli formation. Gene expression analysis showed all three chloroindoles significantly downregulated the expressions of virulence genes associated with adhesion, stress regulation, and toxin production. A 3D-QSAR analysis revealed substitutions at the fourth and fifth positions of the indole moiety favored antimicrobial activity. Furthermore, these chloroindoles potently inhibited biofilm formation in other nosocomial pathogens and polymicrobial consortia.

Abstract 6032: Establishment of a bladder cancer model using organoids derived from bladder epithelium in genetically engineered mice

Abstract Despite recent advances in molecular profiling of bladder cancer, molecular mechanisms for urothelial carcinogenesis have not fully understood. It is partly because the lack of faithful disease models that accurately recapitulate human bladder cancer. In our previous study, we have analyzed mutation landscape of chemical-induced mouse bladder cancer and human bladder cancer by exome-sequence and found that some components of histone methyltransferase complex like Kmt2c, Kmt2d, Kdm6a were highly prevalent as common mutations and associated with Trp53. Additionally, the downregulation of tumor suppressor gene PTEN is known in human muscle invasive bladder cancer (MIBC). We also investigated cell of origin of chemical-induced mouse bladder cancer using lineage tracing method and found that Krt5 expressing cells with Trp53 alteration efficiently give a rise of high-grade MIBC with squamous differentiation. Based on these results, we have tried to make MIBC mouse model using genetically engineered mouse in which Cas9 and target gene mutation express specifically in Krt5 expressing cells by Cre-LoxP system, combined with CRISPR/Cas9 gene editing technique and 3D organoid culture system. Eight-week-old mice harboring Krt5CreERT2/+; LSL-Cas9; LSL-Trp53+/R172H were used in this study. After introducing gene recombination by administration of tamoxifen, we generated organoids from bladder epithelial cells. For further gene editing, we infected adeno-associated virus with sgPten and sgMll3 to the organoids, which successfully edited the target genes. There seemed to be no significant differences in growth of the organoids in culture between the wild-type and gene-edited organoids. However, gene-edited organoids showed tumorigenesis in vivo when inoculated in subcutaneous space, renal subcapsular space or bladder submucosa of athymic mice, whereas wild-type organoids did not show tumorigenesis in vivo. Histologically, the organoid-derived tumors showed morphological similarity to human urothelial carcinoma with squamous differentiation and intra-tumoral heterogeneity. Furthermore, tumor derived organoid has a potential of tumor development in immune-competent mice. These tumors also maintain squamous phenotype and tumor heterogeneity. These findings suggest that the technique is a promising approach toward the establishment of a useful model of human bladder cancer for the elucidation of bladder carcinogenesis including driver genes. In addition, we think our syngeneic mouse model may be useful tool for further research of tumor immunity and immune checkpoint inhibitor drugs. Citation Format: Akihiro Hamada, Yuki Kita, Hideaki Takada, Kenji Nakamura, Toru Sakatani, Takeshi Sano, Takayuki Goto, Atsuro Sawada, Shusuke Akamatsu, Takashi Kobayashi. Establishment of a bladder cancer model using organoids derived from bladder epithelium in genetically engineered mice [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 6032.

Development and biological evaluation of protective effect of kidney targeted N-acetylated chitosan nanoparticles containing thymoquinone for the treatment of DNA damage in cyclophosphamide-induced haemorrhagic cystitis

Thymoquinone (TQ), the most prominent constituent of Nigella sativa seeds, essential oil, is reported to possess an organ protective effect via Nrf2 expression and activation of Phase-II antioxidant enzymes. Haemorrhagic cystitis is the sudden onset of haematuria combined with bladder pain and irritable bladder symptoms are the known toxic effects of cyclophosphamide (CYP) chemotherapy. The objective of the present study was to investigate and compare the protective effect of thymoquinone (TQ) and thymoquinone nanoparticles (TQ-NP) in the kidney against CYP-induced haemorrhagic cystitis. Primarily, TQ-NP was fabricated by synthesis of N-acetylated chitosan and nanoparticle preparation by the ionic gelation technique. They were characterized by particle size, polydispersive index (PDI), zeta potential, entrapment efficiency (EE), SEM, and dynamic scattering calorimetry (DSC). Moreover, fluorescein isothiocyanate (FITC) labeled NPs were prepared for biodistribution studies. The protective mechanisms of TQ-NP included its anti-inflammatory activity, inhibitory effects on cytokine levels, and protection against the DNA damage in the bladder epithelium. The cystitis was induced in rats by orally administering 200 mg/kg of CYP. The dose-dependent protective effect of the TQ-NP was determined by intravenously administering 1, 2, and 5 mg/kg of the TQ-NP to CYP-treated rats. The present study revealed that the TQ-NP prepared by ionic gelation method provides kidney targeted delivery of TQ as compared to TQ solution. The mean particle size, PDI, and %EE of TQ-NP were 272.6 nm, 0.216, 70.81 ± 0.12% respectively. The zeta potential of thymoquinone-loaded nanoparticles was found to be −20.7 mV and − 22.6 mV respectively before and after lyophilization. SEM study also confirmed the small size and spherical shape. Pharmacokinetic studies revealed the improvement in half-life and prolonged action of the TQ-NP as compared to the TQ solution. Also, TQ-NP administration showed more protection against the characteristic histological alterations in the bladder in comparison to TQ solution. The present study indicates that TQ-NP exerts potent anti-oxidant, DNA protective and cytokine inhibitory activity at considerably lower concentrations as compared to plain TQ solution. The nano formulation of TQ using N-acetylated chitosan provides effective kidney targeted delivery of TQ, which in turn improves its retention and protective efficacy against CYP-induced haemorrhagic cystitis.

Urinary calculi in larvae of striped trumpeter, Latris lineata.

Urinary calculi are observed in some cultured marine fish larvae and may negatively impact larval health and survival. This study assessed urinary calculi in striped trumpeter Latris lineata larvae from hatching to 28 days post-hatching (dph). The prevalence of urinary calculi was variable over time and ranged from 15% to 50%, whereas the average size of calculi increased with larval age. Urinary calculi were semi-translucent, light cream to white colour with irregular morphology and a uniform internal structure. The calculi resulted in pressure atrophy causing a distended epithelium of the urinary bladder of fish with calculi. The calculi were predominantly formed of calcium hydrogen phosphate (CaHPO4 ). Routine assessment of the prevalence of urinary calculi in marine fish larvae is recommended, along with other parameters, to monitor larval quality and inform hatchery management actions.

Suprapubic-assisted transurethral excision of eroded transvaginal cerclage suture.

The aim was to demonstrate our minimally invasive technique for excision of eroded transvaginal cervical cerclage suture through the bladder mucosa using a suprapubic-assisted transurethral approach. Transvaginal cervical cerclage is a common treatment for cervical insufficiency in pregnancy. Complications such as erosion are rare, as the duration of treatment is typically several months, with cerclage placement in the second trimester and complete removal prior to the onset of labor. Retained suture can lead to erosion through the vaginal epithelium and into other organs, as seen in our case. Our technique offers a minimally invasive approach to the excision of eroded transvaginal cervical cerclage suture through the bladder mucosa. A narrated, stepwise video demonstration for removal of eroded cervical cerclage through bladder epithelium with suprapubic-assisted transurethral technique in a single patient was carried out. Key strategies for a successful outcome include: use of a Carter-Thomason device for suprapubic assistance in lieu of suprapubic trocar or suprapubic incision, use of rigid biopsy forceps for improved traction on the eroded suture, performing a methylene blue test for evaluation of vesicovaginal fistula after excision procedure. At her 2-week postoperative evaluation, the patient reported resolution of all symptoms. The Carter-Thomason incision was well healed, and postoperative urinalysis was negative for hematuria. A suprapubic-assisted transurethral approach can be used as a minimally invasive technique for excision of eroded transvaginal cervical cerclage suture through the bladder mucosa.

Establishment of an experimental model of normal dog bladder organoid using a three-dimensional culture method

Dog bladder cancer (BC) is mostly muscle-invasive (MI) with poor prognosis, and its pathogenesis is close to human MIBC. Three-dimensional (3D) organoid culture ensures novel knowledge on cancer diseases including BC. Recently, we have established dog BC organoids (BCO) using their urine samples. BCO recapitulated the epithelial structures, characteristics, and drug sensitivity of BC-diseased dogs. However, organoids from dog normal bladder epithelium are not established yet. Therefore, the present study aimed to establish dog normal bladder organoids (NBO) for further understanding the pathogenesis of dog BC and human MIBC. The established NBO underwent various analyzes including cell marker expressions, histopathological structures, cancer-related gene expression patterns, and drug sensitivity. NBO could be produced non-invasively with a continuous culturing and recapitulated the structures and characteristics of the dog's normal bladder mucosal tissues. Different drug sensitivities were observed in each NBO. The analysis of RNA sequencing revealed that several novel genes were changed in NBO compared with BCO. NBO showed a higher expression of p53 and E-cadherin, but a lower expression of MDM2 and Twist1 compared with BCO. These results suggest that NBO could be a promising experimental 3D model for studying the developmental mechanisms of dog BC and human MIBC.

The C5a/C5aR1 Axis Contributes to the Pathogenesis of Acute Cystitis Through Enhancement of Adhesion and Colonization of Uropathogenic E. coli.

Our previous work using a murine model of pyelonephritis demonstrated that the C5a/C5aR1 axis plays a pathogenic role in acute kidney infection. In this study, we report that the C5a/C5aR1 axis also plays a pathogenic role in acute bladder infection. C5aR1-deficient mice had reduced bladder bacterial load and attenuated bladder tissue injury, which is associated with reduced expression of terminal α-mannosyl residues (Man) (a potential ligand for type 1 fimbriae of E. coli) at the luminal surface of the bladder epithelium and reduced early bacterial colonization of the bladder. In vitro, C5a stimulation enhanced mannose expression in and facilitated bacterial adhesion/colonization to human bladder epithelial cells. C5a stimulation also upregulated the activation of ERK1/2 and NF-κB signaling and gene expression of proinflammatory cytokines (i.e., Il6, Il1b, Cxcl1, Ccl2) in the epithelial cells, which could drive pro-inflammatory responses leading to tissue injury. Administration of the C5aR1 antagonist effectively reduced bladder bacterial load and tissue injury. Thus, our findings demonstrate a previously unknown pathogenic role for the C5a/C5aR1 axis in bladder infection and suggest that the C5a/C5aR1 axis-mediated upregulation of Man expression, enhancement of bacterial adhesion/colonization, and excessive inflammatory responses contribute to acute bladder infection. These findings improve our understanding of the pathogenesis of bladder infection with therapeutic implications for UTI.

Pharmacologic Activation of STING in the Bladder Induces Potent Antitumor Immunity in Non-Muscle Invasive Murine Bladder Cancer.

Stimulator of interferon genes (STING) is an innate immune receptor activated by natural or synthetic agonists to elicit antitumoral immune response via type I IFNs and other inflammatory cytokines. Bacillus Calmette-Guerin (BCG) is the standard of care as intravesical therapy for patients with high-risk non-muscle invasive bladder cancer (NMIBC). There are limited options available for patients with NMIBC who developed BCG unresponsiveness. In this study, we characterized in vitro and in vivo antitumor effects of E7766, a macrocyle-bridged STING agonist, via intravesical instillation in two syngeneic orthotopic murine NMIBC tumor models resistant to therapeutic doses of BCG and anti-PD-1 agents. E7766 bound to recombinant STING protein with a Kd value of 40 nmol/L and induced IFNβ expression in primary human peripheral blood mononuclear cells harboring any of seven major STING genotypes with EC50 values of 0.15 to 0.79 μmol/L. Intravesical E7766 was efficacious in both NMIBC models with induction of effective immunologic memory in the treated animals. Pharmacologic activation of the STING pathway in the bladder resulted in IFN pathway activation, infiltration of T cells and natural killer (NK) cells, dendritic cell activation, and antigen presentation in bladder epithelium, leading to the antitumor activity and immunity. In addition, measurements of the pharmacodynamic markers, Ifnβ1 and CXCL10, in bladder, urine, and plasma, and of STING pathway intactness in cancer cells, supported this mode of action. Taken together, our studies reveal an antitumor immune effect of pharmacologic activation of the STING pathway in bladder epithelium and thus provide a rationale for subsequent clinical studies in patients with NMIBC.

Factor VIII as a potential player in cancer pathophysiology

BackgroundTrousseau sign was the first demonstration of a close relationship between cancer and thrombosis. Currently, venous thromboembolism (VTE) is five to six times more likely to occur in cancer patients, whereas there is a greater risk of cancer diagnoses following thromboses. In considering novel players, factor VIII (FVIII), an essential coagulation cofactor with emerging extracoagulative functions, has been identified as an independent VTE risk factor in cancer; however, the basis of this increase is unknown. ObjectiveTo investigate the possible direct expression and secretion of FVIII by cancer cells. MethodsBladder cancer, with a high VTE risk, and normal bladder tissue and epithelium, were used to investigate FVIII. Factor VIII protein and secretion were examined in bladder cancer cell lines. Expanding to other cancers, the Cancer Cell line Encyclopedia database was used to analyze FVIII, tissue factor, FV, FVII, FIX, FX, and von Willebrand factor (VWF) mRNA in 811 cell lines subdivided according to origin. Factor VIII protein synthesis, secretion, and bioactivity were investigated in a profile of cancer cell lines of differing origins. Results and conclusionsAlthough expressed in the normal bladder epithelium, FVIII mRNA and protein were higher in matched bladder neoplasms, with synthesis and secretion of bioactive FVIII evident in bladder cancer cells. This can be extended to other cancer cell lines, with a pattern reflecting the tumor origin, and that is independent of VWF and other relevant players in the coagulation cascade. Here, evidence is provided of a possible independent role for FVIII in cancer‐related pathophysiology.

Sphaeromyxa azevedoi n. sp. (Myxozoa: Sphaeromyxidae) infecting the gall bladder of Gobioides grahamae (Perciformes: Gobiidae) in the Amazon region.

Sphaeromyxa azevedoi n. sp. is described from the gall bladder of the goby Gobioides grahamae (Gobiidae) captured on the Paracauari River in Salvaterra, on Marajó Island, northern Brazil. A total of 50 G. grahamae specimens were analysed, and 15 (30%) were parasitised by the plasmodia and myxospore of Sphaeromyxa azevedoi n. sp. Large plasmodia were observed floating in the bile. These plasmodia were flat, rounded, oval or elongated, and of varying sizes. The mature myxospores, found singly or in pairs, were 27.1 ± 2.7 (20.5-30.1) μmLength and 3.8 ± 0.2 (3.5-4.4) μmWidth in the valvular view. The myxospore has two polar capsules of equal size, 8.1 ± 0.6 (7.4-9.4) μm in length and 2.9 ± 0.2 (2.3-3.3) μm in width. A polar tubule was observed in each capsule, arranged perpendicularly to the principal axis, with three or four coils. The histological analysis showed that the plasmodia and myxospore are located in the lumen of the gall bladder, arranged in pairs, and the epithelium of the gall bladder presented multifocal necrosis. The SSU rDNA of Sphaeromyxa azevedoi n. sp. clusters in the 'balbianii' group of the Sphaeromyxa clade. The morphological characteristics and molecular phylogeny of Sphaeromyxa azevedoi n. sp. support its classification as a new species of the genus Sphaeromyxa, which represents an important advancement in the understanding of the diversity of the myxozoan parasite fauna of Brazilian fishes, especially considering that the new species may be detrimental to the host, a commercially important Brazilian fish species.