Background The relationship between intestinal epithelial integrity and the development of intestinal disease is of increasing interest. A reduction in mucosal integrity has been associated with ulcerative colitis, Crohn’s disease and potentially could have links with colorectal cancer development. The Ussing chamber system can be utilised as a valuable tool for measuring gut integrity. Here we describe step-by-step methodology required to measure intestinal permeability of both mouse and human colonic tissue samples ex vivo, using the latest equipment and software. This system can be modified to accommodate other tissues. Methods An Ussing chamber was constructed and adapted to support both mouse and human tissue to measure intestinal permeability, using paracellular flux and electrical measurements. Two mouse models of intestinal inflammation (dextran sodium sulphate treatment and T regulatory cell depletion using C57BL/6-FoxP3 DTR mice) were used to validate the system along with human colonic biopsy samples. Results Distinct regional differences in permeability were consistently identified within mouse and healthy human colon. In particular, mice showed increased permeability in the mid colonic region. In humans the left colon is more permeable than the right. Furthermore, inflammatory conditions induced chemically or due to autoimmunity reduced intestinal integrity, validating the use of the system. Conclusions The Ussing chamber has been used for many years to measure barrier function. However, a clear and informative methods paper describing the setup of modern equipment and step-by-step procedure to measure mouse and human intestinal permeability isn’t available. The Ussing chamber system methodology we describe provides such detail to guide investigation of gut integrity. Electronic supplementary material The online version of this article (10.1186/s12876-019-1002-4) contains supplementary material, which is available to authorized users.
Background: Assessment of lymph node reactivity pattern is an important indicator of the host response status and prognosis of oral squamous cell carcinoma (OSCC).
We report a 57-year-old man who developed subacute small bowel obstruction after running a marathon. A fibrofatty band was identified restricting the terminal ileum upon subsequent imaging. Surgical division of the band resulted in complete resolution of the patient’s symptoms. Fibrofatty bands are embryonic remnants of the vitellointestinal duct and have not previously been reported to cause small bowel obstruction at the terminal ileum. We discuss the origin of remnant fibrofatty bands and physiological impact of running a marathon upon the gastrointestinal tract that would have contributed to development of subacute small bowel obstruction.
Azathioprine (AZA) and 6-mercaptopurine (6-MP) are the most widely used immunosuppressive therapies in inflammatory bowel disease. Pretreatment measurement of thiopurine methyltransferase (TPMT) activity is recommended and although conventional practice is to use a dose of 2 mg/kg AZA (1 mg/kg 6-MP), higher doses of 2.5 mg/kg AZA or more may be required in some patients, particularly if TPMT activity is high. Dose raising is limited by toxicity, and a robust monitoring system is mandatory. Patients with side effects to AZA may tolerate 6-MP but pancreatitis is a contraindication to switching. Metabolite monitoring is not widely available but may be useful, particularly if non-compliance is possible or where metabolite shunting to 6-methylmercaptopurine is suspected, on the basis of non-response or toxicity. It may allow dose optimisation before switching to alternative immunosuppressants. The drug appears safe in pregnancy and breast feeding. Long term duration of therapy is a balance between benefits in relation to the underlying disease extent, activity and aggressiveness, and the risk of neoplasia, particularly lymphoma.
involved in mechano-transduction. PDLIM5 mRNA expression was confirmed by qPCR, and PDLIM5 protein expression was demonstrated by WB and ICC in both LX-2 cells and primary HSCs. Stimulation of LX-2 cells with TGFb (2 ng/ml) for 24 hrs significantly increased expression of enigma proteins. siRNA knock down of PDLIM5 reduced the expression of fibrotic genes including ACTA2, CTGF, and COL1; and was accompanied by increased cytoplasmic localization and phosphorylation (inactivation) of YAP1. Conclusion In brief, our work defined a new mechanism for activation and nuclear translocation of YAP1 in HSCs via the enigma family protein PDLIM5. Understanding hippo independent mechanisms of YAP1 activation in HSCs may reveal novel targets for urgently needed anti-fibrotics.
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