Expression of Cell-Adhesion Molecules in E. coli: A High Throughput Screening to Identify Paracellular Modulators.

Cell-adhesion molecules (CAMs) are responsible for cell-cell, cell-extracellular matrix, and cell-pathogen interactions. Claudins (CLDNs), occludin (OCLN), and junctional adhesion molecules (JAMs) are CAMs' components of the tight junction (TJ), the single protein structure tasked with safeguarding the paracellular space. The TJ is responsible for controlling paracellular permeability according to size and charge. Currently, there are no therapeutic solutions to modulate the TJ. Here, we describe the expression of CLDN proteins in the outer membrane of E. coli and report its consequences. When the expression is induced, the unicellular behavior of E. coli is replaced with multicellular aggregations that can be quantified using Flow Cytometry (FC). Our method, called iCLASP (inspection of cell-adhesion molecules aggregation through FC protocols), allows high-throughput screening (HTS) of small-molecules for interactions with CAMs. Here, we focused on using iCLASP to identify paracellular modulators for CLDN2. Furthermore, we validated those compounds in the mammalian cell line A549 as a proof-of-concept for the iCLASP method.

Will our junior doctors be ready for the next major incident? A questionnaire audit on major incident awareness across three NHS Trusts in Wales.

AIM: The aim was to assess junior doctors' understanding of their role in the Major Incident Contingency Plan at their hospital, and to evaluate the effectiveness of a teaching intervention on increasing awareness. BACKGROUND: In this audit, 'junior doctor' refers to the first 2 years of work after qualifying from medical school. Once a major incident is confirmed, junior doctors should go to their ward, contact the senior nurse in charge and compile a list of the patients who could safely be discharged from the hospital. This action is standard across NHS Trusts in Wales. METHOD: A questionnaire was given to 89 junior doctors across three NHS Trusts in Wales. It involved general aspects of a major incident, as well as ascertaining perceptions of their role as junior doctors. They then received formal teaching by Emergency Planning Faculty. Following this, a repeat questionnaire was completed. RESULTS: 91% felt they did not know what would be expected of them during a major incident; 47% would initially go to the Emergency Department; 27% were unclear where to go; 31% were unaware who to contact on hearing of a major incident; and 16% would telephone switchboard. Junior doctors believe their primary role would be triage (16%); clerking in the emergency department (36%); clerking in Medical Assessment Unit/Surgical Assessment Unit (17%); and practical work (15%). Only 3% would first go to their ward; 12% believe their primary role would involve patients on the ward; and only 1% would list patients for discharge. 90% of completed questionnaires included a request for teaching. Following teaching, 97% knew who to call, where to go and what to do during a major incident. CONCLUSION: Junior doctors' awareness of major incidents within Wales was near absent prior to teaching. This teaching is vital to help ensure smooth running of the hospital on the day.