9 research outputs found
A case controlled study examining the bladder microbiome in women with Overactive Bladder (OAB) and healthy controls
Objective: To characterise the microbiome in healthy women with no bladder symptoms and to compare this to the bladder microbiome in patients with overactive bladder syndrome (OAB).Study design: MSU specimens from 63 women with OAB were compared to urine from 35 controls. Urine was centrifuged and the resulting sediment pellet was re-suspended in supernatant and plated under aerobic conditions for 48 h and anaerobic conditions for 7 days. Each morphologically distinct colony was purity plated. Bacterial colonies were lysed and polymerase chain reaction undertaken to amplify the 16 s ribosomal RNA gene. This DNA was purified and sequenced allowing identification of bacterial genera.Results: The mean number of different bacterial genera was 5.0 in both controls and OAB patients (p = 0.99). The uropathogenic bacteria Proteus (P = 0.01) was more commonly isolated from women with OAB. The genus lactobacillus was present less commonly in urine from OAB patients when compared to urine taken from controls (p = 0.02). Overall the most commonly grown bacteria were staphylococcus (grown in 59% of samples), streptococccus (51%), corynebacterium (37%) and lactobacillus (28%). A total of 95 different genera were identified from the urine samples.Conclusion: The female human bladder has a diverse microbiome with stastistically significant differences between bacterial species present in OAB patients and controls
Age, Menopausal Status and the Bladder Microbiome
Objectives: The bladder is not sterile but contains a healthy community of microbes termed the microbiome. Alterations in the bladder microbiome have been demonstrated in disease states such as the overactive bladder. The microbiome in other anatomical niches is known to alter with age eg the vagina. The objective of this study was to identify if the bladder microbiome in healthy women varies with age and menopausal status.
Study design: Urine from 79 healthy women attending secondary care gynaecological clinics with no urinary symptoms provided clean catch mid-stream urine specimens. Urine was centrifuged and the resultant pellet was re-suspended and inoculated onto chocolate agar plates and cultured under either aerobic or anaerobic conditions. Morphologically different colonies were purity plated and 16s rRNA gene sequencing was performed. A microbe genomic basic local alignment search tool (BLAST) was used to identify the genus of the bacteria.
Results: There was no significant correlation between the age of a woman and the number of different genera identified (r=-0.034, p=0.79). There were few significant differences in the frequency with which the majority of organisms were found in pre and post-menopausal women. The exceptions however were lactobacillus, which was more common in pre-menopausal women (31 vs 3 p=0.002) and Mobiluncus, which was more common in post-menopausal women (0 vs 3 p=0.02).
Conclusions: There was no significant correlation between patient age and diversity of the bladder microbiome but large numbers of different organisms were identified. Significant differences were however observed for Lactobacillus which is more common in pre-menopausal women and Mobiluncus which is more common in in post-menopausal women
Altered urothelial ATP signaling in a major subset of human overactive bladder patients with pyuria
Overactive Bladder (OAB) is an idiopathic condition, characterized by urgency, urinary frequency, and urgency incontinence, in the absence of routinely traceable urinary infection. We have described microscopic pyuria (?10 wbc/?l) in patients suffering from the worst symptoms. It is established that inflammation is associated with increased ATP release from epithelial cells, and extracellular ATP originating from the urothelium following increased hydrostatic pressure is a mediator of bladder sensation. Here, using bladder biopsy samples, we have investigated urothelial ATP signaling in OAB patients with microscopic pyuria. Basal, but not stretch-evoked, release of ATP was significantly greater from the urothelium of OAB patients with pyuria than from non-OAB patients or OAB patients without pyuria (<10 wbc/?l). Basal ATP release from the urothelium of OAB patients with pyuria was inhibited by the P2 receptor antagonist suramin and abolished by the hemichannel blocker carbenoxolone, which differed from stretch-activated ATP release. Altered P2 receptor expression was evident in the urothelium from pyuric OAB patients. Furthermore, intracellular bacteria were visualized in shed urothelial cells from ?80% of OAB patients with pyuria. These data suggest that increased ATP release from the urothelium, involving bacterial colonization, may play a role in the heightened symptoms associated with pyuric OAB patients
A long-term cohort study of surgery for recurrent prolapse comparing mesh augmented anterior repairs to anterior colporrhaphy
Cystodistension versus cystoscopy in patients with refractory detrusor overactivity: a randomized controlled trial
Altered urothelial ATP signaling in a major subset of human overactive bladder patients with pyuria
Overactive Bladder (OAB) is an idiopathic condition, characterized by urgency, urinary frequency, and urgency incontinence, in the absence of routinely traceable urinary infection. We have described microscopic pyuria (≥10 wbc/μl) in patients suffering from the worst symptoms. It is established that inflammation is associated with increased ATP release from epithelial cells, and extracellular ATP originating from the urothelium following increased hydrostatic pressure is a mediator of bladder sensation. Here, using bladder biopsy samples, we have investigated urothelial ATP signaling in OAB patients with microscopic pyuria. Basal, but not stretch-evoked, release of ATP was significantly greater from the urothelium of OAB patients with pyuria than from non-OAB patients or OAB patients without pyuria (<10 wbc/μl). Basal ATP release from the urothelium of OAB patients with pyuria was inhibited by the P2 receptor antagonist suramin and abolished by the hemichannel blocker carbenoxolone, which differed from stretch-activated ATP release. Altered P2 receptor expression was evident in the urothelium from pyuric OAB patients. Furthermore, intracellular bacteria were visualized in shed urothelial cells from ∼80% of OAB patients with pyuria. These data suggest that increased ATP release from the urothelium, involving bacterial colonization, may play a role in the heightened symptoms associated with pyuric OAB patients
Mg2+ facilitates leader peptide translation to induce riboswitch-mediated transcription termination
The 5′-UTR of the Salmonella Mg2+ transporter mgtA contains a magnesium sensitive riboswitch encompassing the ORF mgtL. MGTL translation is regulated by Mg2+-dependent opening of a ribosome-binding site and causes premature termination of mgtA transcription in cis
Modulation of the Regulatory Activity of Bacterial Two-component Systems by SlyA*
Activation of the transcriptional regulator SlyA by the PhoP/PhoQ
two-component system controls intracellular expression of numerous factors
influencing Salmonella virulence. By dissecting the SlyA regulon
using stable isotope labeling with amino acids in cell culture analysis, we
found that SlyA enhances overall transcription of PhoP-activated loci. This
amplification of cellular responses to Mg2+ occurs when SlyA binds
to the phoPQ promoter thereby activating phoP autoregulation
via a positive feedback mechanism. SlyA footprints a DNA region located one
helical turn upstream of the PhoP box, which overlaps the H-NS-binding motif
required for signal-dependent phoP repression in high Mg2+
conditions. Therefore, binding of SlyA likely antagonizes H-NS and facilitates
the interaction of PhoP to its own promoter, subsequently activating the
phoPQ operon. Establishment of this regulatory circuit allows SlyA to
exert its effect on the PhoP/PhoQ system specifically in Salmonella,
which may confer an additional transcriptional regulation. Thus, our results
provide a molecular mechanism that determines SlyA-dependent activation of
PhoP-regulated genes in modulating Salmonella virulence. Evidence
from this study also suggests a function of SlyA as a mediator in signal
transduction from the PhoP/PhoQ system to other bacterial two-component
systems in Salmonella