Ligilactobacillus ubinensis sp. nov., a novel species isolated from the wild ferment of a durian fruit (Durio zibethinus).

A Gram-stain-positive, rod-shaped, non-spore-forming, catalase-negative, urease-negative, homofermentative and facultatively anaerobic strain, named WILCCON 0076T, was isolated from a wild ferment of pieces of a 'Kampung' durian fruit collected on the island of Ubin (Pulau Ubin), Singapore. The durian had fallen to the ground from a durian tree (Durio zibethinus), on which a group of long-tailed macaques had been observed picking and eating the fruits. Comparative analyses of 16S rRNA gene sequences indicated that WILCCON 0076T potentially represented a novel species within the genus Ligilactobacillus, with the most closely related type strain being Ligilactobacillus agilis DSM 20509T (16S rRNA gene sequence similarity of 97.2 %). Average nucleotide identity and digital DNA-DNA hybridization prediction values were only 86.0% and 18.9 %, respectively. On the basis of the results of a polyphasic approach that included phylogenomic, chemotaxonomic and morphological analyses, we propose a novel species with the name Ligilactobacillus ubinensis sp. nov. (type strain WILCCON 0076T=DSM 114293T=LMG 32698T).

Genome-wide high-throughput signal peptide screening via plasmid pUC256E improves protease secretion in Lactiplantibacillus plantarum and Pediococcus acidilactici.

BACKGROUND: Proteases catalyze the hydrolysis of peptide bonds of proteins, thereby improving dietary protein digestibility, nutrient availability, as well as flavor and texture of fermented food and feed products. The lactobacilli Lactiplantibacillus plantarum (formerly Lactobacillus plantarum) and Pediococcus acidilactici are widely used in food and feed fermentations due to their broad metabolic capabilities and safe use. However, extracellular protease activity in these two species is low. Here, we optimized protease expression and secretion in L. plantarum and P. acidilactici via a genetic engineering strategy. RESULTS: To this end, we first developed a versatile and stable plasmid, pUC256E, which can propagate in both L. plantarum and P. acidilactici. We then confirmed expression and secretion of protease PepG1 as a functional enzyme in both strains with the aid of the previously described L. plantarum-derived signal peptide LP_0373. To further increase secretion of PepG1, we carried out a genome-wide experimental screening of signal peptide functionality. A total of 155 predicted signal peptides originating from L. plantarum and 110 predicted signal peptides from P. acidilactici were expressed and screened for extracellular proteolytic activity in the two different strains, respectively. We identified 12 L. plantarum signal peptides and eight P. acidilactici signal peptides that resulted in improved yield of secreted PepG1. No significant correlation was found between signal peptide sequence properties and its performance with PepG1. CONCLUSION: The vector developed here provides a powerful tool for rapid experimental screening of signal peptides in both L. plantarum and P. acidilactici. Moreover, the set of novel signal peptides identified was widely distributed across strains of the same species and even across some closely related species. This indicates their potential applicability also for the secretion of other proteins of interest in other L. plantarum or P. acidilactici host strains. Our findings demonstrate that screening a library of homologous signal peptides is an attractive strategy to identify the optimal signal peptide for the target protein, resulting in improved protein export.

The impact of a virtual cardiology outpatient clinic in the COVID-19 era.

BACKGROUND: Restrictions as a result of the COVID-19 pandemic have demanded an innovative approach to provide appropriate patient review. We have been running virtual cardiology clinics as per Health Service Executive guidance. AIMS: Our study aims to determine how virtual clinics change practice vs traditional clinics. METHODS: A retrospective cohort analysis was conducted on patients attending cardiology clinics in our hospital from 6 January to 13 March 2020 ('traditional clinic', n = 1644), compared with clinics during the COVID-19 outbreak, from 16 March to 22 April 2020 ('virtual clinic', n = 691), with the same medical staff. RESULTS: There was no difference in age (61 vs 60), case mix or new vs return appointments in virtual vs traditional clinics. There were similar rates of clinic participation, 71.8% vs 74.2%. A lower proportion of investigations (e.g. imaging) were booked in virtual (38.5%) vs traditional (55.7%) clinics, p < 0.00001. Management changes (e.g. medication changes) were less frequent in virtual (19.9%) vs traditional (38.5%) clinics, p < 0.00001. However, the discharge rate was higher in virtual (28.8%) vs traditional (19.5%) clinics, p = 0.00003. CONCLUSION: This study highlights that virtual clinic consultations are associated with fewer investigations, fewer management changes, and increased discharge rates compared with traditional consultations. These practice changes would reduce costs and hospital outpatient congestion by avoiding unnecessary hospital reviews. Nonetheless, it is unknown whether patients requiring face-to-face consultations could be missed as a result of this virtual approach. Longitudinal studies are required to assess clinical outcomes as a result of these practice changes and whether patient satisfaction is altered.

Real World Prescribing Strategies for Patients With an Indication for Oral Anticoagulation Undergoing Percutaneous Coronary Intervention.

BACKGROUND: Limited data are available regarding real-world prescribing in patients with an indication for oral anticoagulation (OAT) who undergo percutaneous coronary intervention (PCI). Most current guidelines recommend a period of triple therapy with OAT and dual antiplatelet (DAPT) agents over dual therapy with an OAT and DAPT but the clinical benefit is still unclear giving rise to limited evidence-based recommendations. AIM: To analyze the appropriateness of OAT and anti-platelet strategies post PCI in a real-world cohort of patients in 2 academic teaching hospitals. METHODS AND RESULTS: In total, 103 patients were retrospectively analyzed. As the initial 3 month stragey, 97 patients (94.2%) received triple therapy, 4 received Dual Therapy (3.9%) and 2 received DAPT (1.9%). Patients with predominant bleeding risk were significantly less likely to receive triple therapy (75% vs. 95.7% vs. 95.8%, P < 0.05). Apixaban was the most common OAT (65.7%) which in most cases was prescribed at an inappropriately reduced dose of 2.5 mg BD (80.4%). The MACE, bleeding and all-cause mortality rates did not differ significantly between groups. DISCUSSION: Our study demonstrates the widespread use of triple therapy for the first 3 months of treatment in 2 Irish academic hospitals. DOACs were prescribed in most cases at reduced doses not currently known to prevent stroke. This study highlights the need for more explicit management algorithms to improve evidence-based management in this area.

The Assessment of Cardiac Masses by Cardiac CT and CMR Including Pre-op 3D Reconstruction and Planning.

PURPOSE OF REVIEW: The purpose of this review is to (1) review the recent evidence examining the use of CT and CMR in the assessment of a suspected cardiac mass, (2) summarize the typical imaging features of the most common cardiac masses, and (3) examine the latest developments in the use of three-dimensional reconstructions and models in the preoperative assessment of a cardiac mass. RECENT FINDINGS: CMR can distinguish between tumors and non-tumor masses and between benign and malignant mass with a high degree of accuracy. CT and CMR are complementary tools in the evaluation of cardiac masses. CMR is the preferred initial imaging modality due to its versatile imaging planes and superior tissue characterization. CT better depicts calcification and has a higher spatial resolution compared with CMR, which is of particular importance in preoperative planning. CT also offers a valuable alternative in those with contraindications to CMR. Three-dimensional reconstructions, particularly of CT datasets, are a valuable adjunct in the preoperative assessment of a cardiac mass and may allow a better appreciation of the margins of the mass and its relationship with surrounding structures. Three-dimensional printing is an emerging technology which may be of additional value in selected patients with a cardiac mass.

Structural basis for reactivating the mutant TERT promoter by cooperative binding of p52 and ETS1.

Transcriptional factors ETS1/2 and p52 synergize downstream of non-canonical NF-κB signaling to drive reactivation of the -146C>T mutant TERT promoter in multiple cancer types, but the mechanism underlying this cooperativity remains unknown. Here we report the crystal structure of a ternary p52/ETS1/-146C>T TERT promoter complex. While p52 needs to associate with consensus κB sites on the DNA to function during non-canonical NF-κB signaling, we show that p52 can activate the -146C>T TERT promoter without binding DNA. Instead, p52 interacts with ETS1 to form a heterotetramer, counteracting autoinhibition of ETS1. Analogous to observations with the GABPA/GABPB heterotetramer, the native flanking ETS motifs are required for sustained activation of the -146C>T TERT promoter by the p52/ETS1 heterotetramer. These observations provide a unifying mechanism for transcriptional activation by GABP and ETS1, and suggest that genome-wide targets of non-canonical NF-κB signaling are not limited to those driven by consensus κB sequences.

Incidental cardiac findings on thoracic imaging.

The cardiac structures are well seen on nongated thoracic computed tomography studies in the investigation and follow-up of cardiopulmonary disease. A wide variety of findings can be incidentally picked up on careful evaluation of the pericardium, cardiac chambers, valves, and great vessels. Some of these findings may represent benign variants, whereas others may have more profound clinical importance. Furthermore, the expansion of interventional and surgical practice has led to the development and placement of new cardiac stents, implantable pacemaker devices, and prosthetic valves with which the practicing radiologist should be familiar. We present a collection of common incidental cardiac findings that can be readily identified on thoracic computed tomography studies and briefly discuss their clinical relevance.

Viability of Escherichia coli ATCC 8739 in Nutrient Broth, Luria-Bertani Broth and Brain Heart Infusion over 11 Weeks.

BACKGROUND: Escherichia coli is a widely studied prokaryotic system. A recent study had demonstrated that reduced growth of E. coli after extended culture in Luria-Bertani broth is a result of depletion of fermentable sugars but able to sustain extended cell culture due to the presence of amino acids, which can be utilized as a carbon source. However, this had not been demonstrated in other media. The study aimed to determine the growth and viability of E. coli ATCC 8739 in 3 different media, Nutrient Broth (NB), Brain Heart Infusion (BHI) and Luria-Bertani Broth (LB) over 11 weeks. METHODS: Growth of E. coli ATCC 8739 was determined by optical density. Viability was determined by serial dilution/spread-plate enumeration. After 11 weeks, the media were exhausted by repeated culture. Glucose was added to the exhausted media to determine whether glucose is the growth-limiting factor. RESULTS: Our results showed that cell density in all 3 media increased to about 1 × 10(9) cells/ml by the end of week 1, from the inoculation density of 2.67 × 10(5) cells/ml, peaked at about 1 × 10(13) cells/ml at week 4, before declining to about 5 × 10(7) cells/ml at week 7. Cell density is highly correlated to genomic DNA content (r(2) = 0.93) but poorly correlated to optical density (r(2)< 0.2). Our results also showed that the spent media were able to support further growth after glucose-supplementation. CONCLUSION: NB, LB and BHI are able to support extended periods of culture and glucose depletion is the likely reason for declining cell growth.

Atrial fibrillation in a primary care population: how close to NICE guidelines are we?

The National Institute for Health and Clinical Excellence (NICE) guidelines for the management of atrial fibrillation were published in June 2006. It was anticipated that they would potentially lead to increased demand for echocardiography (ECHO), increased access to secondary care services (for example for cardioversion), and require additional resources for monitoring anticoagulation. A primary care survey was therefore initiated in South Devon, in advance of publication of the guidelines as a snapshot of existing practice, to determine any additional resources and education required to meet the new standards. The main aim was to determine what proportion of patients were managed exclusively in primary care, how frequently patients were investigated by ECHO and whether anticoagulation was being appropriately targeted at patients at high risk of thromboembolic events.