published_or_final_version / Pharmacology / Master / Master of Philosophy

This thesis uses the methods of sychometrics and clinimetrics to develop and evaluate a Pediatric Ulcerative Colitis Activity Index (PUCAI). The initial phases of item generation and reduction were performed previously. This thesis comprises five main studies. Study one: the weighting and formatting of an initial draft PUCAI using a cohort of 157 children with ulcerative colitis, enrolled prospectively in five pediatric IBD centers. Study two: the validation of the final draft on a separate prospective cohort of 48 children undergoing complete colonoscopy. The PUCAI was highly correlated with physician global assessment (PGA) (r=0.91), Mayo score (r=0.95) and colonoscopic appearance (r=0.77). The PUCAI was able to differentiate the different categories of disease activity, and cutoff points were defined. Study three: Assessment of the responsiveness of the PUCAI. The index demonstrated excellent responsiveness on 75 children seen twice during the study period (effect size=1.9, standardized response mean=2.2, responsiveness statistics=2.6, correlation with PGA of change=0.84, and area under the ROC curve=0.97 95%CI 0.93- 0.99). Study four was aimed at evaluating the predictive validity of the PUCAI, on a retrospective cohort of 99 children with severe ulcerative colitis admitted for intravenous corticosteroid therapy. The PUCAI, calculated on the third and fifth day of therapy was highly predictive of therapy failure at discharge and one year post discharge (area under the ROC curve 0.84 (95%CI 0.76-0.92). Study five: a methodological study evaluating the preferred way to determine the minimal clinically important difference (MCID) of health-related outcome measures. This study was conducted using the PUCAI and three other well established instruments. It was concluded that the MCID should be determined primarily by the anchor-based approach using the ROC curve method on the entire cohort, supplemented by calculating the minimal detectable difference beyond statistical error using the standard error of measurement. Small, moderate and large MCID values could be presented based on the degree of expected relevant change. Together, these studies have contributed to the rigorous development and thorough evaluation of a novel, non-invasive tool for assessing disease activity in pediatric ulcerative colitis clinical studies and practice.

This thesis uses the methods of sychometrics and clinimetrics to develop and evaluate a Pediatric Ulcerative Colitis Activity Index (PUCAI). The initial phases of item generation and reduction were performed previously. This thesis comprises five main studies. Study one: the weighting and formatting of an initial draft PUCAI using a cohort of 157 children with ulcerative colitis, enrolled prospectively in five pediatric IBD centers. Study two: the validation of the final draft on a separate prospective cohort of 48 children undergoing complete colonoscopy. The PUCAI was highly correlated with physician global assessment (PGA) (r=0.91), Mayo score (r=0.95) and colonoscopic appearance (r=0.77). The PUCAI was able to differentiate the different categories of disease activity, and cutoff points were defined. Study three: Assessment of the responsiveness of the PUCAI. The index demonstrated excellent responsiveness on 75 children seen twice during the study period (effect size=1.9, standardized response mean=2.2, responsiveness statistics=2.6, correlation with PGA of change=0.84, and area under the ROC curve=0.97 95%CI 0.93- 0.99). Study four was aimed at evaluating the predictive validity of the PUCAI, on a retrospective cohort of 99 children with severe ulcerative colitis admitted for intravenous corticosteroid therapy. The PUCAI, calculated on the third and fifth day of therapy was highly predictive of therapy failure at discharge and one year post discharge (area under the ROC curve 0.84 (95%CI 0.76-0.92). Study five: a methodological study evaluating the preferred way to determine the minimal clinically important difference (MCID) of health-related outcome measures. This study was conducted using the PUCAI and three other well established instruments. It was concluded that the MCID should be determined primarily by the anchor-based approach using the ROC curve method on the entire cohort, supplemented by calculating the minimal detectable difference beyond statistical error using the standard error of measurement. Small, moderate and large MCID values could be presented based on the degree of expected relevant change. Together, these studies have contributed to the rigorous development and thorough evaluation of a novel, non-invasive tool for assessing disease activity in pediatric ulcerative colitis clinical studies and practice.

This thesis explored the effects of cooked whole asparagus and its purified bioactive, rutin, on colitis symptoms and disease progression in mice using a chemically-induced model of colitis. This model mimics active colitis and recovery states of ulcerative colitis. C57BL/6 mice were fed a basal diet supplemented with 2% asparagus or 0.025% rutin for 3 weeks. Colitis was induced by 2% dextran sodium sulfate in drinking water for 7 days. Asparagus diet was determined to contain higher antioxidant capacities than rutin diet through antioxidant assays. During active colitis, consumption of asparagus alleviated some clinical symptoms (stool consistency, stool blood, and spleen hypertrophy) of colitis. In recovery, asparagus-fed mice were improving in terms of regenerating crypts, surface epithelial, and goblet cells, potentially due to its rutin content. Overall, these findings advocate that asparagus can be therapeutic in treating symptoms during active colitis and recovery phases of ulcerative colitis.

A series of investigations examining the importance of genetic factors in the development of the inflammatory bowel diseases (IBD) namely Crohn’s disease (CD), Ulcerative Colitis (UC) and Indeterminate Colitis (IC) has been undertaken in Scottish children. This has been performed by collection of clinical details and DNA from children with IBD, then analysing the contribution of various candidate genes to both disease susceptibility and disease phenotype. In order to carry out these studies the presenting features of a large cohort of children from across Scotland with IBD diagnosed at less than 16 years were collected, both by examination of hospital case records and by patient interview and questionnaire. For each patient a detailed analysis was made of disease phenotype at presentation including detailed examination of disease location, disease behaviour and growth parameters. A repository of clinical material (DNA, plasma and lymphocytes) was collected from children to accompany the detailed clinical parameters allowing genotype-phenotype analysis at a later stage. Additionally, DNA was also collected from parents where possible to facilitate family based association analysis of candidate genes by transmission disequilibrium testing. A previous DNA repository of healthy Scottish controls had been collected previously and the data generated was available for use in this study. The phenotypic data was collected using an established phenotypic classification (the Vienna classification) used in adult studies as well as a personally devised paediatric phenotypic classification designed for use in this thesis. Firstly, the contribution of the three common mutations within the NOD2/CARD15 gene (R702W, G908R and Leu1007finsC) was analysed in 247 children with IBD. The Leu1007finsC variant was associated with Crohn’s disease by case-control (p = 0.01) and TDT analysis (p = 0.006). Genotype phenotype analysis demonstrated NOD2/CARD15 variants were strongly associated with several markers of disease severity in CD most notably with an increased need for surgery on multifactorial analysis. Then to examine the further contribution of other mutations within the whole NOD2/CARD15 gene, the 12 exons of the gene were sequenced in 24 paediatric CD patients, to identify any additional SNPs that may have conferred an increased susceptibility to CD. Two mutations (V955I, M863V) identified in xii sequencing were genotyped in a large patient cohort, but were not found to confer increased disease susceptibility. Next, the contribution of IBD5 locus was analysed in 299 children with IBD studying 5 SNPs, including mutations in the proposed candidate genes OCTN 1 and 2. Allele frequencies of OCTN1/2 variants were significantly higher in IBD/CD cases (p<0.04). The homozygous mutant OCTN1/2 haplotype was increased in IBD and UC patients (p = 0.02 for both) compared to healthy controls. OCTN1/2 variants however were not independent of the background IBD5 risk haplotype in conferring disease susceptibility. Genotype- phenotype analysis demonstrated association of the risk haplotype with both lower weight and body mass index centiles at diagnosis as analysed by multifactorial analysis. The contribution of the 113 G/A mutation within the discs, large homolog 5 (DLG5) gene was examined in 296 children with IBD. TDT analysis demonstrated a significant association with IBD (p<0.05). Genotype-phenotype analysis demonstrated associations with higher social class, male sex and taller children. Finally, the Anti-Saccharomyces cerevisiae antibodies (ASCA) status of 301 IBD patients was determined. CD patients had a higher prevalence of ASCA antibodies compared to UC patients and healthy controls (p<0.001 for both). A positive ASCA antibody was more common in CD patients with markers of more severe disease and on multifactorial analysis in patients with CD involvement of the oral cavity (p = 0.001). In summary, the candidate genes examined thus far in children with IBD in Scotland have demonstrated a relatively minor contribution to disease susceptibility but have been demonstrated to be associated with specific disease phenotypes in patients with Crohn’s disease. The use of a novel paediatric phenotypic classification in this thesis has allowed description of these novel genotype-phenotype associations.

Bifidobacterium infantis (BI) and other probiotics are non-pathogenic living organisms that have recently gained attention for their possible therapeutic implications on the health of the digestive tract. The mechanisms by which probiotics exert their effects are largely unknown. This study explored the protective and regulatory effect of oral BI on the enteric nervous system (ENS) in the TNBS-induced colitis rats. Electrical field stimulation and chemical stimulation by serotonin (5-HT) were used to elicit changes in the short-circuit current (Isc) response of the colonic rat tissue. BI-fed colitis rats expressed trends of higher secretomotor activity and revealed signs of decreased macroscopic inflammatory damage when compared to sham-fed colitis rats, suggesting a protective and preventative role of oral BI. These findings may provide additional insights for understanding the prophylactic and therapeutic value of specific probiotics in intestinal inflammatory disorders, offering the possibility of a noninvasive alternative to toxic and immune-compromising drugs. / Access to thesis permanently restricted to Ball State community only / Department of Physiology and Health Science

The overall aims of this thesis were to investigate the incidence of inflammatory bowel disease in the Uppsala Region of Sweden, to study the clinical course and the impact of the disease with regards to complications. Patients in Uppsala County were included in the study from the 1st of January 2005 and patients in Falun, Eskilstuna and Åland counties from the 1st of January 2007. The study was closed for all centres on the 31st of December 2009. Mean population in the study region was 305,381 in 2005–2006 and 642,117 in 2007–2009. The mean incidence for ulcerative colitis (UC) during the time period 2005-2009 was 20.0 /100,000/year (95% CI: 16.1-23.9) and for Crohn’s disease (CD) it was 9.9/100,000/year (95% CI: 7.1-12.6). The combined incidence for UC or CD in the area was thus 29.9/100,000/year (95% CI: 25.1-34.7). Half of the UC patients relapsed during the first year. Risk factors for relapse were female gender and young age. Colectomy during the first year was uncommon (2.5%). CD patients with complicated disease had longer symptom duration before diagnosis and less often diarrhoea and blood in stools compared to patients with non-complicated disease. The risk for surgery during the first year was 12%. The prevalence of anaemia at the time of diagnosis was 30% and after one year 18%. Anaemia was more common among newly diagnosed patients with CD compared with UC. 13% of the UC patients developed an acute severe episode. During the first 90 days 22% of these patients were subjected to colectomy. There was a significant difference between University and County hospitals in colectomy frequency (7.5% vs. 41%). The cumulative prevalence of treatment complications was 12% at the hospital with low colectomy rate versus 41% at the hospitals with high colectomy rate. In conclusion, the incidence of UC and CD in Sweden was high compared to international studies. Colectomy frequency for UC during the first year was low. Patients with complicated CD at the time of diagnosis had longer symptom duration and less alarming symptoms compared to uncomplicated disease. Anaemia was a common trait among patients with newly diagnosed IBD and more effort is needed to treat anaemia in these patients. Severe UC can be treated safely with prolonged medical therapy instead of colectomy.

Diss. (sammanfattning) Stockholm : Karol. inst., 2003. / Härtill 5 uppsatser.

Thesis (M.S.)--Virginia Polytechnic Institute and State University, 1993. / Vita. Abstract. Includes bibliographical references (leaves 72-74). Also available via the Internet.

Orientador: Josias Rodrigues / Resumo: Retocolite Ulcerativa (RU) é um tipo de patologia que acomete o cólon intestinal, se apresentando na forma de lesões superficiais de gravidade variável. Não possui causa definida, mas sabe-se que é influenciada por fatores genéticos e ambientais, na qual, esse último, inclui um desequilíbrio na composição de espécies da microbiota intestinal. Escherichia coli (E. coli), uma das bactérias que se encontra aumentada nesses pacientes, tem sido foco de estudos de caracterização, com o objetivo de esclarecer sua participação na etiologia ou complicação dos sintomas da doença. Esse trabalho adotou essa abordagem para a caracterização de uma coleção de E. coli isoladas de portadores de RU atendidos no Hospital das Clínicas da Universidade Estadual Paulista Júlio de Mesquita Filho (HC/UNESP) de Botucatu, com base em sua capacidade de produção de biofilme, sorotipagem e filotipagem. Juntamente a esses testes, foi realizada uma revisão bibliográfica sobre a possível relação da E. coli com a RU. O objeto de estudo dos testes foi uma coleção de E. coli composta por 68 isolados bacterianos de 34 portadores de RU e 44 de 22 indivíduos controle (CO). A tipagem bacteriana teve como foco genes que identificam os sorogrupos O25 e O83 e determinação de filogrupos da coleção de referência de E. coli (EcoR – A, B1, B2 e D). Os resultados obtidos foram: 1) predomínio de E. coli dos filogrupos B2 e A nos grupos CO (54,5% x 26,5%, p=0,01) e de portadores de RU (32,4% x 9,1%, p=0,04) respectivamente, ... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Ulcerative colitis (UC) is a type of pathology that affects the intestinal colon, presenting as superficial lesions of different severity. It has no defined cause, but it is known to be influenced by genetic and environmental factors, which includes an imbalance in the composition of species of the intestinal microbiota. Escherichia coli (E. coli), one of the bacteria that is increased in these patients, has been the focus of characterization studies, to clarify its participation in the etiology or complication of the disease’s symptoms. Following a line of research already consolidated in our laboratory, this work adopted this approach for the characterization of a collection of E. coli isolated from UC patients treated at the HC / UNESP of Botucatu, based on its biofilm production capacity, serotyping and filotyping. Also, a literature review was performed on the possible relationship between E. coli and UC. The study’s object of these tests was a collection of E. coli composed of 68 bacterial isolates from 34 UC carriers and 44 from 22 control individuals (CO). Bacterial typing focused on genes that identify the O25 and O83 serogroups and determination of phylogroups from the E. coli reference collection (EcoR - A, B1, B2 and D). The results obtained were: 1) Predominance of E. coli of the phylogenetic groups B2 and A in the CO groups (54.5% x 26.5%, p = 0.01) and in the UC group (32.4% x 9, 1, p = 0.04), respectively. 2) In the UC group, 8.8% and 11.8% of the individuals ... (Complete abstract click electronic access below) / Mestre