Improved mycobacterial protein production using a Mycobacterium smegmatis groEL1ΔC expression strain

<p>Abstract</p> <p>Background</p> <p>The non-pathogenic bacterium <it>Mycobacterium smegmatis </it>is widely used as a near-native expression host for the purification of <it>Mycobacterium tuberculosis </it>proteins. Unfortunately, the Hsp60 chaperone GroEL1, which is relatively highly expressed, is often co-purified with polyhistidine-tagged recombinant proteins as a major contaminant when using this expression system. This is likely due to a histidine-rich C-terminus in GroEL1.</p> <p>Results</p> <p>In order to improve purification efficiency and yield of polyhistidine-tagged mycobacterial target proteins, we created a mutant version of GroEL1 by removing the coding sequence for the histidine-rich C-terminus, termed GroEL1ΔC. GroEL1ΔC, which is a functional protein, is no longer able to bind nickel affinity beads. Using a selection of challenging test proteins, we show that GroEL1ΔC is no longer present in protein samples purified from the <it>groEL1ΔC </it>expression strain and demonstrate the feasibility and advantages of purifying and characterising proteins produced using this strain.</p> <p>Conclusions</p> <p>This novel <it>Mycobacterium smegmatis </it>expression strain allows efficient expression and purification of mycobacterial proteins while concomitantly removing the troublesome contaminant GroEL1 and consequently increasing the speed and efficiency of protein purification.</p

How COVID-19 has changed the unselected medical take: an observational study

INTRODUCTION: COVID-19 has had a profound effect on the NHS. Little information has been published as to how the unselected medical take has been affected. METHODS: We retrospectively reviewed patients who were referred to general medicine during March 2020. We compared clinical outcomes of patients with and without COVID-19. RESULTS: 814 patients were included, comprising 777 unique patients. On average, 26 patients were admitted per day. 38% of admitted patients were suspected of COVID-19, with greater numbers of COVID-19 patients in the second half compared to the first half of the month (p<0.001). Logistic regression analyses showed suspected COVID-19 was an independent predictor for inpatient mortality (odds ratio [OR] = 6.09, p<0.001) and 30-day mortality (OR = 4.66, p<0.001). CONCLUSIONS: COVID-19 patients had worse clinical outcomes and increased healthcare use compared to non-COVID-19 patients. Our study highlights some of the challenges in healthcare provision faced during this pandemic

GLP-1 Receptor Agonists and SGLT2 Inhibitors for the Treatment of Type 2 Diabetes: New Insights and Opportunities for Cardiovascular Protection

The risk of cardiovascular disease (CVD) (myocardial infarction, stroke, peripheral vascular disease) is twice in type 2 diabetes (T2D) patients compared to non-diabetic subjects. Furthermore, cardiovascular disease (CV) is the leading cause of death in patients with T2D.In the last years several clinical intervention studies with new anti-hyperglycaemic drugs have been published, and they have shown a positive effect on the reduction of mortality and cardiovascular risk in T2D patients. In particular, these studies evaluated sodium/glucose-2 cotransporter inhibitors (SGLT2i) and Glucagon-like peptide-1 receptor agonists (GLP-1RA).In secondary prevention, it was clearly demonstrated that SGLT2i and GLP-1RA drugs reduce CV events and mortality, and new guidelines consider now these drugs as first choice (after metformin) in the treatment of T2D; there are also some signs that they may be effective also in primary prevention of CVD. However, the mechanisms involved in cardiovascular protection are not yet fully understood, but they appear to be both "glycaemic" and "extra-glycaemic".In this review, we will examine the fundamental results of the clinical trials on SGLT2i and GLP-1RA, their clinical relevance in term of treatment of T2D, and we will discuss the mechanisms that may explain how these drugs exert their cardiovascular protective effects

Proceedings of the 6th International Conference on Modeling and Simulation in Civil Engineering

This conference proceedings contains articles on the various research ideas of the academic community and technical researchers presented at the 6th International Conference on Modeling and Simulation in Civil Engineering (ICMSC 2022). ICMSC 2022 was organized by the Department of Civil Engineering, TKM College of Engineering, Kollam, Kerala, India on December 1-3, 2022. The main aim of this conference is to bring together leading academicians, researchers, technocrats, practitioners, and students to exchange and share their experiences and research outputs on all aspects of Civil Engineering, especially related to the modeling and simulation in Civil Engineering.  Conference Title: 6th International Conference on Modeling and Simulation in Civil EngineeringConference Acronym:  ICMSC 2022Conference Date: 1-3 December 2022Conference Location: IndiaConference Organizer: Department of Civil Engineering, TKM College of Engineering, Kollam, Kerala, Indi