Effect of α-Hemolysin Producing <i>E. coli</i> in Two Different Mouse Strains in a DSS Model of Inflammatory Bowel Disease

Background: Phylogroup B2 Escherichia coli have been associated with ulcerative colitis (UC). In this study, we aimed to compare colonization with the UC-associated E. coli p19A in different mice strains, to investigate the role of alpha hemolysin in a UC mouse model. Methods: In this study, Sigirr &minus;/&minus; and C57BL/6 mice were chosen, and UC was induced by adding dextran sulfate sodium (DSS) to the drinking water. The mice were pre-treated with ciprofloxacin. p19A expressing luminescence and GFP, alpha-hemolysin knock out p19A-&Delta;hlyI II, and non-pathogenic lab E. coli DH10B were cultured in LB broth, and orally gavaged into the mice. Colonization with p19A WT was visualized using an in vivo imaging system. Results: p19A WT colonized the colon, ileum, Peyer&rsquo;s patches, liver, and spleen of infected C57BL/6 and Sigirr &minus;/&minus; mice. A total of 99% of the p19A WT infected C57BL/6 mice and 29% of the p19A WT infected Sigirr &minus;/&minus; mice survived to the 4th post infection day. Conclusion: UC-associated E. coli p19A WT colonized the intestines of DSS-treated mice and caused extra-intestinal infection. Hemolysin is an important factor in this pathogenesis, since isogenic hemolysin mutants did not cause the same inflammation

Comparison of the diagnostic performance of CT colonography interpreted by radiologists and radiographers

OBJECTIVE: To compare computed tomographic colonography (CTC) performance of four trained radiographers with the CTC performance of two experienced radiologists. METHODS: Four radiographers and two radiologists interpreted 87 cases with 40 polyps ≥6 mm. Sensitivity, specificity, and positive predictive value (PPV) were assessed on a per-patient basis. On a per-polyp basis, sensitivity was calculated according to the respective size categories (polyps ≥6 mm as well as polyps ≥10 mm). RESULTS: Overall per-patient sensitivity for polyps ≥6 mm was 76.2 % (95 % CI 61.4–91.0) and 76.2 % (95 % CI 61.7–90.6), for the radiographers and radiologists, respectively. Overall per-patient specificity for polyps ≥6 mm were 81.4 % (95 % CI 73.7–89.2) and 81.1 % (95 % CI 73.8–88.3) for the radiographers and the radiologists, respectively. For the radiographers, overall per-polyp sensitivity was 60.3 % (95 % CI 50.3–70.3) and 60.7 % (95 % CI 42.2–79.2) for polyps ≥6 mm and ≥10 mm, respectively. For the radiologists, overall per polyp sensitivity was 59.2 % (95 % CI 46.4–72.0) and 69.0 % (95 % CI 48.1–89.6) for polyps ≥6 mm and ≥10 mm, respectively. CONCLUSION: Radiographers with training in CT colonographic evaluation achieved sensitivity and specificity in polyp detection comparable with that of experienced radiologists. MAIN MESSAGES: • The diagnostic accuracy of trained radiographers was comparable to that of experienced radiologists. • The use of radiographers in reading CTC examinations is acceptable, however radiologists would still be necessary for the evaluation of extracolonic findings. • Skilled non-radiologists may play a vital role as a second reader of intraluminal findings or by performing quality control of examinations before patient dismissal

MR imaging of scaphoid fractures. Fat-saturated T2-weighted and Short tau inversion recovery images.

Objective: Traumatic injuries of os scaphoideum are serious, and might lead to two main grades of consequences (i.e. osteoarthrosis or avascular necrosis), if a fracture remains undiagnosed. Bone bruise may be the only pathological sign of pain which can last for week or month. Articles describe the importance of early MRI and hereby predict bone bruise with the help of fat suppression sequence; however, only a limited selection articles compares various fat suppression techniques. The purpose of this prospective study was to compare the short tau inversion recovery (STIR) and T2 fat saturation (FAT SAT) sequences, sectional directed along the scaphoid bone axis. In relation to background fat intensity suppression, this study sought the sequence that best evaluated posttraumatic bone marrow edema (bone bruise) on scaphoid injury musculoskeletal magnetic resonance imaging (MRI, 1.5 T extremity scanner). Materials and methods: Two hundred and fifty-one patients with relevant trauma and positive clinical test for scaphoid bone fractures, exceeding no more than 14 days, underwent MRI examinations. A fast STIR and T2 FAT SAT fast spin echo sequence (FSE) were obtained using a comparable parameter setting (scan time ca. 3 minutes). Three experienced readers (one radiographer and two radiologists) carried out the evaluation blinded to each other’s, based on a quantitative assessment of size (area) and image quality (image contrast, IC and contrast-to-noise ratio, CNR). The study period lasted March 2014-April 2015. Sixty patients met the inclusion criteria and were enrolled. This prospective study was ethically approved by the institutional review board. Results: There were no significant difference between the bone bruise areas (P=0.45, P=0.44 and P=0.83) or CNR (P=0.31, P=0.38 and P=0.17). However, image contrast showed significant difference in favour of T2 FAT SAT in all three readers’ reports (P<0.05, P<0.05 and P<0.05). Conclusions: The two sequences appear almost identical. An interchangeable usage of the two sequences was found being acceptable for the diagnosis if the protocol is composed appropriately (1.5T). However, the T2 FAT SAT provided a higher image contrast by specific settings (e.g. short TI = 125 ms) compared to STIR

Secretion of alpha-hemolysin by<i> Escherichia coil</i> disrupts tight junctions in ulcerative colitis patients

OBJECTIVES: The potential of Escherichia coli (E. coli) isolated from inflammatory bowel disease (IBD) patients to damage the integrity of the intestinal epithelium was investigated. METHODS: E. coli strains isolated from patients with ulcerative colitis (UC) and healthy controls were tested for virulence capacity by molecular techniques and cytotoxic assays and transepithelial electric resistance (TER). E. coliisolate p19A was selected, and deletion mutants were created for alpha-hemolysin (α-hemolysin) (hly) clusters and cytotoxic necrotizing factor type 1 (cnf1). ProbioticE. coliNissle and pathogenicE. coliLF82 were used as controls. RESULTS: E. colistrains from patients with active UC completely disrupted epithelial cell tight junctions shortly after inoculation. These strains belong to phylogenetic group B2 and are all α-hemolysin positive. In contrast, probioticE. coliNissle, pathogenicE. coliLF82, fourE. colifrom patients with inactive UC and threeE. colistrains from healthy controls did not disrupt tight junctions.E. colip19A WT as well ascnf1, and single loci ofhlymutants from cluster I and II were all able to damage Caco-2 (Heterogeneous human epithelial colorectal adenocarcinoma) cell tight junctions. However, this phenotype was lost in a mutant with knockout (Δ) of bothhlyloci (P<0.001). CONCLUSIONS: UC-associated E. coliproducing α-hemolysin can cause rapid loss of tight junction integrity in differentiated Caco-2 cell monolayers. This effect was abolished in a mutant unable to express α-hemolysin. These results suggest that high Hly expression may be a mechanism by which specific strains of E. colipathobionts can contribute to epithelial barrier dysfunction and pathophysiology of disease in IBD