Thesis (D.P.H.)--University of Michigan, 1994.

Published Article / The 2012 residents of Huis Welgemoed, a residence for female students at Central University of Technology, Free State Bloemfontein Campus have reported a high prevalence of infections of the vagina and the urinary tract. They assume that this problem is associated with poor hygienic conditions in the toilets of their residence. However, this assumption may not be entirely true, as other factors may also contribute to their problem. Previous studies have shown that there is an evident relation of vaginitis and urinary tract infections (UTI) among students and the practices of personal hygiene or the level of toilet sanitation in their residences. Especially in facilities where the students have complained about the hygiene state of their residence toilets as unsatisfactory. This preliminary study has shown that the residence conforms to the standards set out by the S.A. requirements for toilets in student housing of 1 toilet per 6 students and that the cleaning materials and methods used by staff are adequate. Interestingly, the authors have found that a lack of knowledge around UTI's and VI's regarding basic prevention strategies is most probably the cause of the high incidence of these two types of infections.

Lactobacillus is a genus of lactic acid bacteria frequently used as healthpromoting probiotics. Using probiotics to treat or prevent infections is a novel experimental approach with vast impact on future therapy. Lactobacillus rhamnosus GR-1 is a probiotic investigated for its ability to reduce urogenital disease including urinary tract infections caused by pathogenic Escherichia coli. L. rhamnosus GR-1 has been shown to modulate immunity, thought to influence its probiotic effect. In this thesis, the aim was to study immunomodulation by L. rhamnosus GR-1 and other lactobacilli, with emphasis on elicited immune responses such as nuclear factor-kappaB (NF-κB) activation and cytokine release from human urothelial cells. Viable, heat-killed, and isolated released products from L. rhamnosus GR-1 augmented NF-κB activation in E. coli-challenged urothelial cells. Blocking of lipopolysaccharide binding to toll-like receptor 4 completely quelled this augmentation. Size-fractionation, urothelial cell challenge, and two-dimensional gel electrophoresis of L. rhamnosus GR-1 released products presented several candidate proteins with NF-κB modulatory actions including chaperonin GroEL, elongation factur Tu, and a protein from the NLP/P60 protein family. While tumor necrosis factor was correspondingly augmented by L. rhamnosus GR-1, the release of two other cytokines, interleukin (IL)-6 and CXCL8, was reduced. Similar effects were observed in macrophage-like cells stimulated with L. rhamnosus GR-1. Many immunomodulatory effects of lactobacilli are believed to be species and strain dependent. Therefore, twelve Lactobacillus strains were used to screen for their effects on CXCL8 release from urothelial cells. A majority of these strains were able to influence CXCL8 release from the cells. Phylogenetic analysis revealed close evolutionary linkage between lactobacilli with similar actions on CXCL8. Increased knowledge on probiotic bacterial products and the mechanism(s) of action could lead to improved future treatments for infections.

Master of Science / Department of Clinical Sciences / Greg Grauer / A urinary tract infection (UTI) is defined as a temporary or permanent breach in host defense mechanisms that allows microbes to adhere, multiply, and persist within the urinary tract. Development of a UTI is multi-factorial with bacterial number and virulence and the health status of the patient (normal urogenital tract anatomy and physiology and systemic immunocompetence) playing important roles in determining the outcome. A UTI can involve a single site, such as the renal pelvis, ureter, bladder, urethra, prostate or vagina, or can include multiple sites. Infection of any portion of the urinary tract may increase the likelihood of infection in other locations. Diagnosis of a UTI incorporates findings from the history, physical examination, complete urinalysis, and urine culture. Proper classification and localization of the UTI are important when formulating a treatment regime as well as evaluating treatment success and failure. Most UTI can be successfully managed with appropriate antibiotic treatment; however, bacterial resistance and compromised host defense mechanisms can result in persistent or recurrent infections. In patients with recurrent UTI, identification of underlying predisposing conditions will often improve treatment success. In patients where underlying causes cannot be identified or treated, therapies designed to prevent recurrent UTI may be employed. Proanthrocyanidins found in cranberry juice inhibit E. coli attachment to human uroepithelial cells, impairing bacterial adherence and colonization. These characteristics have encouraged widespread usage of cranberry extract as a prevention strategy for woman predisposed to urinary tract infections. E. coli is a common cause of canine urinary tract infection. Current treatment emphasizes eradication of established infection rather than infection prevention, but increased antibiotic resistance necessitates strategies to prevent infection. We hypothesized that purified cranberry extract (CE) inhibits bacterial adhesion to canine uroepithelial cells. The results of our study show that CE supplementation can reduce adhesion of uropathogenic E. coli to canine uroepithelium and suggests one mechanism by which CE might improve urinary tract health.

by Phillip Y.P. Jen. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1994. / Includes bibliographical references (leaves 97-110). / Abstract --- p.iii / Acknowledgements --- p.x / Chapter 1. --- Review of literature --- p.1 / Chapter 2. --- Materials and Methods --- p.8 / Chapter 2.1 --- Collection and preparation of tissues --- p.8 / Chapter 2.2 --- Immunofluorescence --- p.9 / Chapter 3. --- Results --- p.15 / Chapter 3.1 --- Urinary bladder --- p.15 / Chapter 3.1.1 --- Bladder detrusor muscle --- p.15 / Chapter 3.1.2 --- Intramural ureters and superficial trigone --- p.17 / Chapter 3.1.3 --- Bladder mucosa --- p.19 / Chapter 3.1.4 --- The bladder neck --- p.20 / Chapter 3.2 --- Vas deferens and seminal vesicle --- p.22 / Chapter 3.2.1 --- The smooth muscle coat --- p.30 / Chapter 3.2.2 --- The mucosa --- p.24 / Chapter 3.3 --- Prostate --- p.26 / Chapter 3.4 --- Urethra --- p.30 / Chapter 3.4.1 --- Rhabdosphincter --- p.31 / Chapter 3.4.2 --- Smooth muscle coat and lamina propria --- p.32 / Chapter 3.5 --- Autonomic ganglia and paraganglia --- p.34 / Chapter 4. --- Discussion --- p.70 / Chapter 4.1 --- Urinary bladder --- p.70 / Chapter 4.2 --- Vas deferens & seminal vesicle --- p.81 / Chapter 4.3 --- Prostate --- p.84 / Chapter 4.4 --- Autonomic ganglia --- p.87 / Chapter 5. --- Suggestions for further study --- p.93 / Chapter 6. --- References --- p.101

Extra-intestinal pathogenic Escherichia coli (ExPEC) are a significant cause of urinary tract infections and bacteraemia in the UK and around the world. These E. coli primarily belong to phylogenetic groups B2 and D, with the clones ST131, ST127, ST95, ST73 and ST69 responsible for the majority of these infections. In the UK, studies of ExPEC have focused on isolates from the North of England, ST131 strains and ExPEC that possess extended-spectrum beta-lactamase (ESBL) enzymes. Therefore, very little is understood about the UK ExPEC population as a whole, the breadth of virulence factors contributing to these infections and the differences between urinary and bloodstream-derived ExPEC. In this study ST131 was more frequently detected in bloodstream isolates and ST95 was most prevalent in urinary isolates. Comparative virulence of the major clones in the Galleria mellonella infection model revealed ST131 isolates to effect the highest mortality, although serogroup O6, which is linked with ST73, was also associated with high mortality, potentially explaining the success of ST73-O6 in bacteraemia. Analysis of virulence factors identified pap, afa/dra and kpsMTII as important determinants in isolates causing urosepsis and those of ST131, while fyuA and fimH were distinctly lacking, demonstrating their role as colonisation factors rather than virulence factors. Although these findings are important, with appropriate treatment of urinary tract infections they can become redundant, as ExPEC would be eradicated before causing a severe infection such as bacteraemia or urosepsis. In urinary isolates, resistance to trimethoprim approached 50% and ampicillin resistance was >70%, while ST131 isolates as a whole demonstrated ciprofloxacin and trimethoprim resistance >50%. Together these indicate that empirical UTI guidelines need to be revisited, to prevent recurrence of infection and ascension to the kidneys and bloodstream. In addition, data from this study can be used to develop a point-of-care test to detect ST131, to guide appropriate treatment, without the delay associated with referring a urine specimen for microbiological investigation.

"American cranberry (Vacciniumm macrocarpon) has been long known for its benefits in maintaining urinary tract health. Clinical trials have shown that drinking cranberry juice can prevent urinary tract infections (UTIs) in various subpopulations that are prone to UTIs, especially women, but the mechanisms by which cranberry acts against uropathogenic bacteria are still unclear. Studies showed that when exposed to cranberry juice or A- PACs, a group of tannins that are unique to cranberry, the adhesion activity and biofilm formation of uropathogenic bacteria were reduced. However, the metabolism of cranberry juice has not be elucidated, therefore further study is needed to find out whether the anti-bacterial components in cranberry could survive the digestive system and reach the urinary tract, and how the components or metabolites remaining in urine act against uropathogenic bacteria. We used atomic force microscopy (AFM) to study the surface adhesion force of uropathogenic E. coli incubated with urine samples that were collected from volunteers after drinking 16 oz. of cranberry juice cocktail (CJC) or water. The urine samples were collected at 0, 2, 4, 6, and 8 hours after CJC or water consumption. When incubated with post-water urine, the adhesion forces of pathogenic bacteria that have fimbriae (E. coli B37, B73, B78, BF1023, CFT 073, and J96) did not change; whereas the adhesion forces of these strains decreased over the 8 hour period after CJC consumption. The control strain that does not have frimbriae, E. coli HB101, showed low adhesion force when incubated with post-water and post-CJC urine. In a human red blood cell agglutination (HRBC) assay, the attachment of pathogenic E. coli to red blood cells was significantly lower after exposed to post-CJC urine, compared to those exposed to post-water urine. These results indicate the anti-bacteria components or metabolites of CJC stay active in urine, and these compounds prevent adhesion of E. coli by reducing fimbriae-mediated adhesion. We also examined the effects of drinking CJC on biofilm formation of uropathogenic bacteria. Female volunteers were given 16 oz. of CJC or placebo, and their urine was collected at 0, 2, 8, 24, and 48 hours after consumption. Bacteria (E. coli B37, CFT073, BF1023, HB101, and S. aureus ATCC43866) were cultured in a mixture of urine and growth media in 96 well microtiters. The biofilm formed was quantified by staining the biofilm dissolved in a solvent with crystal violet and measuring the absorbance at 600 nm. The results showed that biofilm formation was reduced within 24 hours after CJC consumption, and it started to increase after 48 hours, possibly due to the washout of CJC in the system. These studies suggest that CJC can be an effective preventive measure for UTIs as it inhibits adhesion and biofilm formation of uropathogenic bacteria."

"Regular ingestion of American cranberry (Vaccinium macrocarpon) has been traditionally utilized for its health benefits against urinary tract infections. The proanthocyanidins (PACs), in particular, the unique A-type double linkages of PACs present in cranberry, have been identified as the active components. However, A-type PACs and any other active agents have not yet been detected or identified in urine. Additional experiments are required to investigate the inhibitory effects and persistence of cranberry metabolites present in urine collected following CJC consumption, and to determine how these compounds act against uropathogenic Escherichia coli for the prevention of urinary tract infections. Two separate bioassays (a biofilm formation assay and a bacterial cell viability assay) were used to determine the in vitro effect of cranberry juice cocktail (CJC) oral consumption on bacterial anti-adhesion activity in a double-blind, placebo-controlled pilot clinical trial. A single dose of 16 oz. of CJC or a placebo beverage was given to ten healthy women, ages ranging from 18 to 27, and urine samples were collected in the following 48 hours. A washout period of seven days was allowed. Bacteria (Escherichia coli B37, CFT073, BF1023, HB101, and Staphylococcus aureus ATCC43866) were cultured in the urine samples, supplemented with media, and the amount of biofilm formed was measured using a crystal violet absorbance assay in a 96-well plate. In the urine of volunteers who had consumed CJC, biofilm formation was inhibited within 24 hours after CJC consumption, and started to increase after 48 hours by 49-67%. S. aureus showed the least biofilm formation after incubation with post-CJC urine. The results indicated that drinking CJC can be an effective preventive measure for bacterial adhesion and biofilm formation in healthy women. The anti-biofilm activity peaks between 24 and 48 hours after drinking CJC. The viability assay showed that the colony count after culturing in urine collected following consumption of CJC or placebo were not significantly different, implying that CJC works as an inhibitor by blocking bacterial adhesion instead of killing the bacteria or restraining its growth. Another randomized, placebo-controlled, double-blind, crossover study was conducted to further investigate the molecular-scale effect of cranberry juice metabolites on two P-fimbriated E. coli strains: B37 and CFT 073, as assessed by atomic force microscopy (AFM). Three female subjects were asked to consume 8 oz. CJC or water. The washout period was 7 days. The urine samples were collected at 2, 4 and 6 hours post-ingestion of CJC or water. Urine collected before consumption of CJC was used as a control. For this control urine, the average adhesion force between E. coli and uroepithelial cells was 13.09 ± 11.60 nN for CFT073 and 10.30 ± 5.50 nN for B37. For post-CJC urine treatment, the adhesion forces decreased to 2.94 ± 1.82 nN at 2 hours after consumption then increased slightly to 5.51 ± 2.78 nN at 6 hours after ingestion for CFT073, while they decreased to 4.77 ± 3.33 nN after consuming for 2 hours and seemed to be stable in the next 4 hours following consumption (5.52 ± 4.04 nN after drinking for 4 hours; 5.05 ± 4.42 nN after drinking for 6 hours) for B37. The adhesion forces in post-water consumption urine were similar to those of the background for E. coli B37; meanwhile a downward trend for the adhesion forces in post-water consumption urine compared to the background was observed for E. coli CFT073. However, these adhesion forces in post-water consumption urine were still higher than those measured after CJC consumption at the same time intervals. The mean differences between the cranberry and placebo groups were statistically different according to the two way ANOVA procedure followed by Holm-Sidak test. Our results suggest a significant inhibitory interaction between the daily consumption of 8 oz. cranberry juice and bacterial adhesive activity. These results help form the mechanistic understanding of how cranberry products can be used to prevent bacterial attachment to host tissue, and may lead to new therapeutic strategies to prevent the rising problem of bacteria antibiotic resistance.  "

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniv., 2004. / Härtill 3 uppsatser.

Thesis (Ph. D.)--University of Sydney, 2004. / Title from title screen (viewed 8 May 2008). A reference list of published articles is provided in Appendices. Includes copies of seven published papers, co-authored by Fanrong Kong. Submitted in fulfilment of the requirements for the degree of Doctor of Philosophy to the Dept. of Medicine. Includes bibliographical references. Also available in print form.