Female urinary microbiota.

PURPOSE OF REVIEW: The newly discovered female urinary microbiota has the potential to deepen our understanding of urinary tract health and disease, including common lower urinary tract conditions such as urinary incontinence and urinary tract infection. The spectrum of painful bladder disorders and other less common conditions also may benefit from additional research that includes consideration of the resident bacterial community of the female bladder. The present review provides a clinical context for the rapidly emerging research regarding the female urinary microbiota and its relationships with urinary tract conditions of interest. RECENT FINDINGS: Studies using culture-independent techniques confirm prior reports of bacteria that reside in the female urinary bladder. These resident communities, the female urinary microbiota, possess characteristics that differ between women affected by urgency urinary incontinence and matched, unaffected controls. Enhanced urine culture techniques permit cultivation of organisms, including uropathogens, missed by standard urine culture, but detected by culture-independent sequencing techniques. SUMMARY: New technology is available. Clinical laboratories can modify traditional standard urine culture methods to enhance detection of uropathogens. However, given the existence of the female urinary microbiota, the simple presence of bacteria in the lower urinary tract should not be taken as evidence of infection.

Hedgehog/Wnt feedback supports regenerative proliferation of epithelial stem cells in bladder.

Epithelial integrity in metazoan organs is maintained through the regulated proliferation and differentiation of organ-specific stem and progenitor cells. Although the epithelia of organs such as the intestine regenerate constantly and thus remain continuously proliferative, other organs, such as the mammalian urinary bladder, shift from near-quiescence to a highly proliferative state in response to epithelial injury. The cellular and molecular mechanisms underlying this injury-induced mode of regenerative response are poorly defined. Here we show in mice that the proliferative response to bacterial infection or chemical injury within the bladder is regulated by signal feedback between basal cells of the urothelium and the stromal cells that underlie them. We demonstrate that these basal cells include stem cells capable of regenerating all cell types within the urothelium, and are marked by expression of the secreted protein signal Sonic hedgehog (Shh). On injury, Shh expression in these basal cells increases and elicits increased stromal expression of Wnt protein signals, which in turn stimulate the proliferation of both urothelial and stromal cells. The heightened activity of this signal feedback circuit and the associated increase in cell proliferation appear to be required for restoration of urothelial function and, in the case of bacterial injury, may help clear and prevent further spread of infection. Our findings provide a conceptual framework for injury-induced epithelial regeneration in endodermal organs, and may provide a basis for understanding the roles of signalling pathways in cancer growth and metastasis.

Conformational inactivation induces immunogenicity of the receptor-binding pocket of a bacterial adhesin.

Inhibiting antibodies targeting receptor-binding pockets in proteins is a major focus in the development of vaccines and in antibody-based therapeutic strategies. Here, by using a common mannose-specific fimbrial adhesin of Escherichia coli, FimH, we demonstrate that locking the adhesin in a low-binding conformation induces the production of binding pocket-specific, adhesion-inhibiting antibodies. A di-sulfide bridge was introduced into the conformationally dynamic FimH lectin domain, away from the mannose-binding pocket but rendering it defective with regard to mannose binding. Unlike the native, functionally active lectin domain, the functionally defective domain was potent in inducing inhibitory monoclonal antibodies that blocked FimH-mediated bacterial adhesion to epithelial cells and urinary bladder infection in mice. Inhibition of adhesion involved direct competition between the antibodies and mannose for the binding pocket. Binding pocket-specific inhibitory antibodies also were abundant in polyclonal immune serum raised against the functionally defective lectin domain. The monoclonal antibodies elicited against the binding-defective protein bound to the high-affinity conformation of the adhesin more avidly than to the low-affinity form. However, both soluble mannose and blood plasma more strongly inhibited antibody recognition of the high-affinity FimH conformation than the low-affinity form. We propose that in the functionally active conformation the binding-pocket epitopes are shielded from targeted antibody development by ligand masking and that strong immunogenicity of the binding pocket is unblocked when the adhesive domain is in the nonbinding conformation.

[A Case Diagnosed as Iatrogenic Vesical Tuberculosis 4 Years after Intravesical Immunotherapy Using Bacillus Calmette-Guérin (BCG) for the Treatment of Bladder Carcinoma].

We report a case of iatrogenic vesical tuberculosis diagnosed 4 years after intravesical immunotherapy using Bacillus Calmette- Guérin (BCG) for the treatment of bladder carcinoma. A 72-year-old man underwent a transurethral resection (TUR) of multiple noninvasive urothelial carcinomas and intravesical BCG infusion (40 mg/week) for 7 weeks to prevent the recurrence of bladder carcinoma. BCG infusion therapy was terminated because of the appearance of Reiter's syndrome, including arthritis of the left toe joint, conjunctivitis and non-gonococcal urethritis as complications. The patient suffered from repeated cystitis, bladder atrophy and urethral stenosis. The cystitis improved with the administration of antibiotics (Levofloxacin) but persisted without a complete cure. Four years later, a cystoscopy revealed mucosal erosion and a white coating. An acid-fast bacteria examination of a urine sample using bacteria incubation and DNA PCR revealed the presence of Mycobacterium bovis. Finally, anti-tuberculosis therapy (INH＋REP＋EB) was initiated after the patient was diagnosed as having iatrogenic bladder tuberculosis resulting from BCG immunotherapy. The tuberculosis bacteria subsequently disappeared from the urine samples, and the gross appearance of the bladder mucosa improved. Bladder carcinoma has not recurred to date. Intravesical BCG infusion therapy has a good anti-tumor effect and can help prevent tumor recurrence after TUR therapy in case of noninvasive bladder carcinoma. However, there is a risk of severe complications arising from the BCG infusion. In the present case, an adequate bacteria examination was not performed, even though antibiotics were repeatedly administered for cystitis. In particular, the patient was not tested for the presence of acid-fast bacteria for 4 years after the intravesical BCG infusion therapy. Furthermore, among patients who received anti-bacteria therapy for repeated cystitis after BCG infusion, a bacteria examination including bacteria incubation, was not ordered in 19 out of 30 cases treated at our hospital over the past 5 years. In conclusion, bacteria examination, including tests for acid-fast bacteria, should be immediately performed when repeated and/or persistent cystitis occurs after BCG infusion therapy.

[REASONS OF DELAYED DIAGNOSIS OF BLADDER TUBERCULOSIS].

UNLABELLED: The fourth, terminal, stage of bladder tuberculosis (BT) manifests itself in irreversible changes and requires surgical treatment. OBJECTIVE: To identify the reasons for delayed diagnosis of this urogenital tuberculosis complication. Medical history of 26 urogenital tuberculosis patients with a complicated form of stage 4 BT, referred to the Novosibirsk TB Research Institute for reconstructive surgery were analysed. In 22 patients, bladder volume ranged from 55 to 100 ml, 4 patients previously underwent cystostomy due to extremely small bladder volume. Average duration of BT hidden in the guise of "urogenital infection" was 6.2 years. Patients were treated with norfloxacin (a total of 104 courses), ciprofloxacin (86 courses), amikacin (43 courses), nitroxoline (27 courses), third generation cephalosporins (32 courses), lomefloxacin (17 courses), levofloxacin (11 courses), Amoxicillin clavulanate (4 courses), ampicillin (2 courses). It was demonstrated that all cases of BT stage 4 were iatrogenic. Irreversible debilitating complications occurred due to suboptimal therapy, primarily due to administration of amikacin and fluoroquinolones for urogenital infections, which was tuberculosis in disguise. Absence of M. tuberculosis growth does not exclude tuberculosis; pathological specimens must be further examined at least by PCR. Interventional material must be mandatory examined histologically and stained by Ziehl-Neelsen method to identify M. tuberculosis. Effective and not masking tuberculosis, optimal therapy for urogenital infections includes fosfomycin, furazidin (nitrofurantoin), gentamicin, III generation cephalosporins (in outpatient settings dispersible form of efixime should be preferable).

A rare case of histopathological bladder necrosis associated with Actinobaculum schaalii: the incremental value of an accurate microbiological diagnosis using 16S rDNA sequencing.

We describe here a rare case of bladder wall necrosis associated with Actinobaculum schaalii in a 72-year-old patient with non-muscle-invasive bladder cancer (NMIBC). A. schaalii microbiological diagnosis requires high index of suspicion and accurate identification methods such as 16S rDNA sequencing or MALDI-TOF Mass spectrometry.

Host-pathogen interactions mediating pain of urinary tract infection.

BACKGROUND: Pelvic pain is a major component of the morbidity associated with urinary tract infection (UTI), yet the molecular mechanisms underlying UTI-induced pain remain unknown. UTI pain mechanisms probably contrast with the clinical condition of asymptomatic bacteriuria (ASB), characterized by significant bacterial loads without lack symptoms. METHODS: A murine UTI model was used to compare pelvic pain behavior elicited by infection with uropathogenic Escherichia coli strain NU14 and ASB strain 83972. RESULTS: NU14-infected mice exhibited pelvic pain, whereas mice infected with 83972 did not exhibit pain, similar to patients infected with 83972. NU14-induced pain was not dependent on mast cells, not correlated with bacterial colonization or urinary neutrophils. UTI pain was not influenced by expression of type 1 pili, the bacterial adhesive appendages that induce urothelial apoptosis. However, purified NU14 lipopolysaccharide (LPS) induced Toll-like receptor 4 (TLR4)-dependent pain, whereas 83972 LPS induced no pain. Indeed, 83972 LPS attenuated the pain of NU14 infection, suggesting therapeutic potential. CONCLUSIONS: These data suggest a novel mechanism of infection-associated pain that is dependent on TLR4 yet independent of inflammation. Clinically, these findings also provide the rational for probiotic therapies that would minimize the symptoms of infection without reliance on empirical therapies that contribute to antimicrobial resistance.

FimH adhesin of type 1 fimbriae is a potent inducer of innate antimicrobial responses which requires TLR4 and type 1 interferon signalling.

Components of bacteria have been shown to induce innate antiviral immunity via Toll-like receptors (TLRs). We have recently shown that FimH, the adhesin portion of type 1 fimbria, can induce the innate immune system via TLR4. Here we report that FimH induces potent in vitro and in vivo innate antimicrobial responses. FimH induced an innate antiviral state in murine macrophage and primary MEFs which was correlated with IFN-beta production. Moreover, FimH induced the innate antiviral responses in cells from wild type, but not from MyD88(-/-), Trif(-/-), IFN-alpha/betaR(-/-) or IRF3(-/-) mice. Vaginal delivery of FimH, but not LPS, completely protected wild type, but not MyD88(-/-), IFN-alpha/betaR(-/-), IRF3(-/-) or TLR4(-/-) mice from subsequent genital HSV-2 challenge. The FimH-induced innate antiviral immunity correlated with the production of IFN-beta, but not IFN-alpha or IFN-gamma. To examine whether FimH plays a role in innate immune induction in the context of a natural infection, the innate immune responses to wild type uropathogenic E. coli (UPEC) and a FimH null mutant were examined in the urinary tract of C57Bl/6 (B6) mice and TLR4-deficient mice. While UPEC expressing FimH induced a robust polymorphonuclear response in B6, but not TLR4(-/-) mice, mutant bacteria lacking FimH did not. In addition, the presence of TLR4 was essential for innate control of and protection against UPEC. Our results demonstrate that FimH is a potent inducer of innate antimicrobial responses and signals differently, from that of LPS, via TLR4 at mucosal surfaces. Our studies suggest that FimH can potentially be used as an innate microbicide against mucosal pathogens.

The immune response to infection in the bladder.

The bladder is continuously protected by passive defences such as a mucus layer, antimicrobial peptides and secretory immunoglobulins; however, these defences are occasionally overcome by invading bacteria that can induce a strong host inflammatory response in the bladder. The urothelium and resident immune cells produce additional defence molecules, cytokines and chemokines, which recruit inflammatory cells to the infected tissue. Resident and recruited immune cells act together to eradicate bacteria from the bladder and to develop lasting immune memory against infection. However, urinary tract infection (UTI) is commonly recurrent, suggesting that the induction of a memory response in the bladder is inadequate to prevent reinfection. Additionally, infection seems to induce long-lasting changes in the urothelium, which can render the tissue more susceptible to future infection. The innate immune response is well-studied in the field of UTI, but considerably less is known about how adaptive immunity develops and how repair mechanisms restore bladder homeostasis following infection. Furthermore, data demonstrate that sex-based differences in immunity affect resolution and infection can lead to tissue remodelling in the bladder following resolution of UTI. To combat the rise in antimicrobial resistance, innovative therapeutic approaches to bladder infection are currently in development. Improving our understanding of how the bladder responds to infection will support the development of improved treatments for UTI, particularly for those at risk of recurrent infection.

Ovarian and vesical actinomycosis: a case report and literature review.

We present a patient with a tubo-ovarian abscess pathologically confirmed to be actinomycosis in a 44-year-old woman with an intrauterine device (IUD). An ultrasound showed that the IUD was imposed on an apparently degenerated myoma. A pelvic MRI was performed to differentiate the uterine findings from a sarcoma. The MRI showed a heterogeneous pelvic mass and a bladder mass suggesting chronic inflammation caused by an organism such as actinomycosis. An exploratory laparotomy was performed, which revealed a right tubo-ovarian mass with abscess formation as well as a bladder mass. A subtotal hysterectomy, right salpingoophorectomy, partial cystectomy, and appendectomy were performed in addition to drainage of the abscess. Histopathological examination revealed a tubo-ovarian abscess and a bladder mass with colonies of actinomycoses.

[Leucoplakia vesicae in females: diagnosis and treatment].

Sixty patients suffering from leucoplakia vesicae (LV) were examined using cystoscopy with biopsy of the urinary bladder wall, blood enzyme immunoassay for detection of antibodies to agents of sexually transmitted infections (STI), uroflowmetry, culture analysis of cervical canal and mucosa samples for STI. As shown by a pathomorphological examination of the vesical mucosa biopsy specimens, long-term persistence of pathogenic (chlamydia, trichomonades) and opportunistic (mycoplasma, ureaplasma, fungi) flora underlies development of LV. Morphogenesis of LV is characterized by hyperplastic reactions of urothelium and its metaplasy in laminated squamous keratosic epithelium, often with para- and dyskeratosis, developing in the presence of inflammatory reactions in the lamina in the presence of persisting infection. In LV, specific infection agents are often found in the urogenital tract. The spectrum of these agents is identical for samples from the cervical canal and vesical mucosa from leucoplakia foci. Vesical mucosa is most frequently contaminated with Mycoplasma hominis (57.2%), Candida albicans (51.4%), Ureaplasma urealiticum (37.1%) and Trichomonas vaginalis (22.9%). Associations of the infection agents are detected in 70% of LV patients. Persistent dysuria is a basic clinical symptom of leucoplakia. The following therapeutic measures should be taken: transurethral coagulation of the vesical mucosa, intravesical therapy, immunocorrection, antibacterial treatment by standard schemes or according to the isolated flora sensitivity.

An update on lower urinary tract tuberculosis.

Tuberculosis of the genitourinary tract presents with atypical manifestations. Only 20% to 30% of patients with genitourinary tuberculosis have a history of pulmonary infection. Tuberculosis often affects the lower genitourinary system rather than the kidney. Tuberculosis of the lower genitourinary tract most commonly affects the epididymis and the testis, followed by bladder, ureter, prostate, and penis. Use of bacillus Calmette-Guérin therapy for bladder cancer can cause symptomatic tubercular infections of the lower genitourinary tract. Tuberculosis of the lower genitourinary tract can present with irritative voiding symptoms, hematuria, epididymo-orchitis, prostatitis, and fistulas. Tuberculosis of the seminal vesicles, vas, fallopian tubes, and the uterus can cause infertility. Urinalysis may demonstrate sterile pyuria, hematuria, or albuminuria. Identification of acid-fast bacilli in culture or tissue or by polymerase chain reaction studies is diagnostic. Medical treatment may not result in resolution of symptoms. Surgical intervention and reconstruction of the urinary tract are frequently indicated.

Anterior spinal cord syndrome as a rare complication of acute bacterial meningitis in an adult.

Acute bacterial meningitis is not an uncommon central nervous system infection. In severe cases, it can be associated with various neurological or systemic complications. However, acute spinal cord dysfunction rarely occurs. We report a case of bacterial meningitis complicated with spinal cord infarction despite adequate treatment with antibiotics and corticosteroid therapy. He had residual paraplegia and was fully dependent in the activity of daily living.

[Efficacy of instillagel against methicillin-resistant strains of Staphylococcus aureus in urinary bladder catheterization].

Infections caused by methicillin-resistant Staphylococcus aureus (MRSA) present a global problem. Catheterization of the bladder is a risk factor of MRSA infection. Bacteriological tests showed that a lubricating agent for catheterization installagel has a special formulation killing MRSA for 5 minutes. Thereby, instillagel is recommended for catheterization in risk groups attending clinics and old people's homes.

[Bladder fungus ball by Candida tropicalis: a pediatric case report]. / Fungoma vesical por Candida tropicalis: un caso clínico pediátrico.

Vesical fungus ball is a mobile, oval and echogenic mass as a result of accumulation of long and wide numerous hyphae. Fungal urinary tract infection incidence has increased notoriously and there are isolated yeast in 7 to 8% of urine cultures. Different species of Candida are cause of urinary tract infection. Epidemiologically, the first isolated pathogen is Candida albicans, followed by Candida tropicalis. Bladder poll has been documented as the most important risk factor for candiduria in critical patients into intensive care.

Aqueous extract from Orthosiphon stamineus leaves prevents bladder and kidney infection in mice.

BACKGROUND: Extracts from the leaves of Orthosiphon stamineus are used in phytotherapy for treatment of uncomplicated urinary tract infections. PURPOSES: Evaluation of an aqueous extract against infection with uropathogenic Escherichia coli in vivo; investigation of underlying microbiological mechanisms. STUDY DESIGN: In vivo studies in mice and in vitro investigations on cytotoxicity, antiadhesive potential, influence on bacterial gene expression and quorum sensing. METHODS: Extract OWE was prepared by hot water extraction. For in vivo studies BALB/c mice were used in an UPEC infection model. The effect of OWE on bacterial load in bladder/kidney tissue was monitored in pre- and posttreatment. Cytotoxicity of OWE against different UPEC strains, T24 bladder/A498 kidney cells, gene expression analysis, monitoring of phenotypic motility and quorum sensing was investigated by standard methods of microbiology. RESULTS: OWE was quantified (UHPLC) according to the content of rosmarinic acid, cichoric acid, caffeic acid. Three- and 5-day treatment of animals with OWE (750mg/kg) after transurethral infection with UPEC CFT073 reduced the bacterial load in bladder and kidney, similar to norfloxacin. Four- and 7-day pretreatment of mice prior to the infection with UPEC NU14 reduced bacterial bladder colonization. In vitro investigations indicated that OWE (≤2mg/ml) has no cytotoxic or proliferation-inhibiting activity against different UPEC strains as well as against T24 bladder and A498 kidney cells. OWE exerts a dose dependent antiadhesive activity against UPEC strains NU14 and UTI89. OWE reduced gene expression of fimH, but evoked increase of the expression of motility/fitness gene fliC. Increase of bacterial motility on gene level was confirmed by a changed bacterial phenotype by an increased bacterial motility in soft agar assay. OWE inhibited in a concentration-dependent manner bacterial quorum sensing. CONCLUSION: OWE is assessed as a strong antiadhesive plant extract for which the traditional use in phytotherapy for UTI might be justified.

Does the Urinary Microbiome Play a Role in Urgency Urinary Incontinence and Its Severity?

OBJECTIVES: Traditionally, the urinary tract has been thought to be sterile in the absence of a clinically identifiable infection. However, recent evidence suggests that the urinary tract harbors a variety of bacterial species, known collectively as the urinary microbiome, even when clinical cultures are negative. Whether these bacteria promote urinary health or contribute to urinary tract disease remains unknown. Emerging evidence indicates that a shift in the urinary microbiome may play an important role in urgency urinary incontinence (UUI). The goal of this prospective pilot study was to determine how the urinary microbiome is different between women with and without UUI. We also sought to identify if characteristics of the urinary microbiome are associated with UUI severity. METHODS: We collected urine from clinically well-characterized women with UUI (n = 10) and normal bladder function (n = 10) using a transurethral catheter to avoid bacterial contamination from external tissue. To characterize the resident microbial community, we amplified the bacterial 16S rRNA gene by PCR and performed sequencing using Illumina MiSeq. Sequences were processed using the workflow package QIIME. We identified bacteria that had differential relative abundance between UUI and controls using DESeq2 to fit generalized linear models based on the negative binomial distribution. We also identified relationships between the diversity of the urinary microbiome and severity of UUI symptoms with Pearson's correlation coefficient. RESULTS: We successfully extracted and sequenced bacterial DNA from 95% of the urine samples and identified that there is a polymicrobial community in the female bladder in both healthy controls and women with UUI. We found the relative abundance of 14 bacteria significantly differed between control and UUI samples. Furthermore, we established that an increase in UUI symptom severity is associated with a decrease in microbial diversity in women with UUI. CONCLUSIONS: Our study provides further characterization of the urinary microbiome in both healthy controls and extensively phenotyped women with UUI. Our results also suggest that the urinary microbiome may play an important role in the pathophysiology of UUI and that the loss of microbial diversity may be associated with clinical severity.