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.

This study attempts to find a correlation between commuting modes in Washington DC and characteristics of the city and the people that they serve. It investigates why some census tracts have experienced increases in the commuting share of alternative transportation, such as public transit, walking, and bicycling, while others haven't. Findings demonstrate that demographic variables such as percent Hispanic and foreign born were the strongest predictors of change in commute mode share followed by distance to train station. Land use variables demonstrated weak correlations with variations in mode share due most likely to a lack of density gradient within the study area. The creation of variables to determine land use mix by census tract posed technical challenges as well. Recommendations include policy addressing rising demand for more diverse transportation systems be implemented and further research be conducted on creating more accurate land use variables to include in the model.

Background: Group B streptococcus (GBS) is a leading cause of invasive bacterial disease in neonates. The possibility of maternal immunization with GBS-vaccines is being explored. Vaccine candidates include serotype-specific polysaccharide-protein conjugates and GBS surface proteins, including pilus island proteins. In this project, we aimed to undertake capsular serotype identification, pilus island identification and genotypic characterization of GBS isolates associated with colonization in mothernewborn dyads and invasive disease in infants. Methods: Colonizing GBS isolates were identified by vaginal swabbing of mothers (n=541) during active labor and from skin of their newborns post-delivery (n=395). Invasive GBS isolates from infants (n=284) were identified through laboratory-based surveillance. GBS serotyping was done by latex agglutination. Serologically nontypeable isolates were typed by a serotype-specific PCR method. The pilus islands from 541 colonizing isolates and 284 invasive isolates were characterized by real-time PCR targeting the ancillary protein 1 and 2. We undertook sequence typing based on the three most heterogeneous genes (adhP, atr and glnA) of multilocus sequence typing (MLST) on GBS isolates identified in young-infants with invasive disease (n=283) and those associated with maternal (n=525) and newborn colonization at birth (n=369). A total of 121 colonizing and 131 invasive disease GBS isolates that were representative of 55 and 35 clusters respectively were analyzed by the remaining four MLST genes. The gbs2018 locus was characterized by DNA sequencing.

For centuries Aloe vera has been exploited for several verified and unverified medicinal uses such as wound healing, treatment of gastrointestinal ulcers and for its many biological effects including anti-microbial, laxative, anti-inflammatory and immunostimulatory activities. Studies have generally focused on its effects in vitro and in adults. When nursing mothers use Aloe vera extracts, their suckling infants are at risk of indirect exposure to Aloe vera via breast feeding or directly as dietary/health supplements. The gastrointestinal tract (GIT) of the neonate is sensitive to dietary manipulations during the suckling period with long lasting effects that can be irreversible. Thus babies may be at risk if administered Aloe vera extracts directly as dietary supplements or indirectly via breast milk. The main objectives of this study were to evaluate the effects of orally administered aqueous and alcohol extracts of Aloe vera on growth performance, the morphometry and morphology of the gastrointestinal tract and accessory organs, and liver function of suckling rats. Suckling Sprague-Dawley rats (77), males (n=38) and females (n=39) of 6 days old were randomly assigned to one of five treatment groups and given once daily by oral gavage a suspension of lyophilized crude alcohol or aqueous extracts of Aloe vera suspended in distilled water. Group I (control) was gavaged with distilled water (vehicle). Group II received a low dose of the aqueous extract (AqL) at 50mg. kg-1; Group III received a high dose of the aqueous extract (AqH) at 500mg. kg-1; Group IV received a low dose of the alcohol extract (AlcL) at 50mg. kg-1 whilst Group V received a high dose of the alcohol extract (AlcH) at 500mg. kg-1. The extracts and distilled water were 2 administered at a volume of 10ml.kg-1. The pups remained with their dams for the duration of the study and after 8 days on the treatments, the pups were humanely killed to harvest their tissues for measurements and physiological analysis. All data were expressed as mean ± SD and analyzed by one way ANOVA, the values were considered statistically significant when p < 0.05 and then a Bonferroni Post hoc test was applied. The suckling rats fed respectively with high doses of AlcH and AqH had a significantly higher body mass gain than the other groups (p < 0.05, one way ANOVA). Linear growth as measured by tibial length was significantly increased in the AqH group compared to the other groups. There was no significant difference in the mass and relative density of the tibia bones of the rats from the different treatment groups. The differences in growth could not be attributed to circulating concentrations of the somatotrophic hormone, Insulin-like growth factor-1 (IGF-1) which was not significantly different between the groups. The treatments did not result in any significant differences in lengths, and mass of the small and large intestine, however the caecum was significantly enlarged (hypertrophy of muscularis, submucosa and mucosa) in the rats that received the Aloe vera extracts. Although, there was no significant difference in the mass of the rats’ livers, the lipid and glycogen content were significantly higher (p < 0.001) for the AqH group compared to the other groups. Histologically, the hepatocytes showed enlarged nuclei, granular cytoplasm and dilated sinusoids for AqH and AlcH as compared to the control group. An indirect assessment of liver function by measurement of blood concentrations of alkaline phosphatase (ALP) and alanine amino transaminase (ALT) did not reveal a significant difference between the groups. The non fasting concentration of metabolic substrates (glucose and triglycerides) was also not significantly different between the groups. The pups given high doses of the extracts had a significantly greater (p < 0.05) thymus mass (hyperplastic) than the other groups. The short term administration of Aloe vera extracts has shown a growth promoting effect, enhanced hepatic storage of metabolic substrates and hypertrophy of the caecum and thymus of neonatal rats. These effects need to be explored further to enhance animal production and health.

A análise de sinais mioelétricos provenientes do tubo gastro-intestinal de animais de laboratório (ratos), conseguidos por meio de eletrodos cronicamente implantados, é peça-chave no entendimento das desordens associadas ao sistema digestivo. Esta análise enfrenta consideráveis dificuldades quando realizadas por métodos clássicos, em especial os baseados na transformada de Fourier. A interação de várias componentes mioelétricas torna muito complicado e trabalhoso o acompanhamento destes sinais ao longo do tubo digestivo e a obtenção de parâmetros típicos como a velocidade de propagação entre eletrodos. Estuda-se aqui uma alternativa mais nova e promissora: a transformada Wavelet. Utilizando esta ferramenta matemática, torna-se possível obter uma melhor resolução tempo-freqüencial dos sinais estudados, permitindo encontrar padrões referentes à propagação do sinal mesmo em leituras ruidosas e compostas de várias freqüências. Foram analisados 82 leituras de 9 animais normais do Laboratório de Investigação Médica da Faculdade de Medicina da USP, sendo possível determinar dois parâmetros: a velocidade de propagação média entre eletrodos (cerca de 1.2 cm/s) e as componentes principais da freqüência basal (0.63 e 0.65 Hz). / The analysis of myoelectric signals from the gastro-intestinal tube of laboratory animals (mice), recorded by chronically implanted electrodes, is a key stone in understanding the disorders associated to the digestive system. This analysis meets considerable difficulties when done by classical methods, specially those based in the Fourier transform. The many myoelectric components interactions makes the following of these signals along the digestive tract and the retrieval of typical parameters (such as the propagation velocity between electrodes) a very complicated and laborious task. Here is studied a newer and more promising alternative: the Wavelet transform. Using this mathematical tool, it becomes possible to obtain a better time-frequency resolution of the studied signals, allowing to find patterns related to the signal propagation even in noisy and multifrequencial readings. 82 readings from 9 normal animals belonging to the Medical Investigation Laboratory of the Medicine Faculty of University of São Paulo were analyzed, becoming feasible to determine two parameters: the mean propagation velocity between electrodes (about 1.2 cm/s) and the main components of the basal frequency (0.63 e 0.65 Hz).

O período embrionário se estende do 15º ao 45º dia de gestação na vaca, onde ocorre um rápido crescimento e diferenciação, durante o qual os principais órgãos, tecidos e sistemas são estabelecidos e as principais características são reconhecidas. Nos bovinos, a maioria dos órgãos e partes do corpo é formada entre a 2ª e a 6ª semana de gestação. Durante este período, o trato digestivo, os pulmões, o fígado e o pâncreas se desenvolvem do intestino primitivo, sendo estabelecidos os primórdios do sistema muscular, esquelético, nervoso e urogenital. Aproximadamente 25 a 45% dos embriões bovinos são perdidos até o processo final de implantação, no entanto, a literatura relacionada a sua organogênese é escassa, uma vez que crescem as inovações tecnológicas ligadas a reprodução bovina. Assim, julgamos oportuno e necessário um estudo sobre a biologia do desenvolvimento do aparelho digestório de embriões bovinos oriundos de monta natural, utilizando microscopia de luz e microscopia eletrônica de transmissão. Nossos resultados revelam que em embriões com idade gestacional estimada de 20 dias (CR 9,0 mm) ocorreu a formação do septo traqueoesofágico dividindo o intestino anterior no tubo laringotraqueal e esôfago, separando-se totalmente aos 28/29 dias de gestação. Ao redor do 60º de gestação (CR 70,0 mm) um estômago pluricavitário característico dos ruminantes é evidente e apresentam epitélio endodérmico em diferenciação, tecido conjuntivo mesodérmico e tecido muscular liso. Os núcleos das células epiteliais acompanham o formato da célula e a lâmina basal delimita área de epitélio e mesênquima, estando as células unidas umas às outras através dos desmossomos, que promovem sustentação celular. / The embryonary period extends itself from 15th to 45th day of gestation in cows, when a fast growing and differentiation occurs, during which the main organs, tissues and systems are established and the main characteristics are recognized. In bovines, most of the organs and parts of body are formed between 2th and 6th week of gestation. During this period, the digestive tract, the lungs, the liver and the pancreas are developed from the primitive intestine; being established the origins of the muscle, skeletal, nervous and urogenital systems. Approximately 25 to 45% of bovine embryos are lost until the final process of implantation; however, the literature related to their organogenesis is scant, since the technologic innovations related to bovine reproduction are growing. Thus, we think it is opportune and necessary a study about the biology of development of digestive tract of bovine embryos derived from natural coition, using optic microscopy and transmission electronic microscopy. Our results disclose that in embryos having a estimated gestational age of 20 days (CR 9,0 mm), it occurred a formation of tracheoesophagic septum, dividing the anterior intestine in laringotracheal tube and esophagus, separating itself totally on 28/29 days of gestation. At about 60th day of gestation (CR 70,0 mm), a multicavity stomach, characteristic of ruminants, is evident and presents endodermic epithelium on differentiation, mesodermic connective tissue and smooth muscle tissue. The epithelial cells nuclei go along the cell shape and the basal lamina delimits the area of epithelium and mesenchyme, being the cells bound each other through desmosoms, which promote the cellular sustentation.

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.  "