3′-UTR engineering to improve soluble expression and fine-tuning of activity of cascade enzymes in Escherichia coli

3'-Untranslated region (3'UTR) engineering was investigated to improve solubility of heterologous proteins (e.g., Baeyer-Villiger monooxygenases (BVMOs)) in Escherichia coli. Insertion of gene fragments containing putative RNase E recognition sites into the 3'UTR of the BVMO genes led to the reduction of mRNA levels in E. coli. Importantly, the amounts of soluble BVMOs were remarkably enhanced resulting in a proportional increase of in vivo catalytic activities. Notably, this increase in biocatalytic activity correlated to the number of putative RNase E endonucleolytic cleavage sites in the 3'UTR. For instance, the biotransformation activity of the BVMO BmoF1 (from Pseudomonas fluorescens DSM50106) in E. coli was linear to the number of RNase E cleavage sites in the 3'UTR. In summary, 3'UTR engineering can be used to improve the soluble expression of heterologous enzymes, thereby fine-tuning the enzyme activity in microbial cells.1184Ysciescopu

The importance of sonographic landmarks by transcutaneous laryngeal ultrasonography in post-thyroidectomy vocal cord assessment

During examination of the vocal cords (VC) using transcutaneous laryngeal ultrasonography (TLUSG), 3 sonographic landmarks (namely, false VC [FC], true VC [TC], and arytenoids [AR]) are often seen. However, it remains unclear which landmark provides a more reliable assessment and whether seeing more landmarks improves the diagnostic accuracy and reliability. METHODS: We evaluated prospectively 245 patients from 2 centers. One assessor from each center performed all TLUSG examinations and their findings were validated by direct laryngoscopy. All 3 sonographic landmarks were routinely visualized whenever possible. The rate of visualization and diagnostic accuracy between the 3 landmarks were compared. RESULTS: Eighteen patients suffered postoperative VC palsy (VCP). Both centers had comparable visualization or assessability rate of ≥ 1 sonographic landmark (94.9 and 95.3%; P = 1.000) and 100% sensitivity on postoperative TLUSG. The rates of FC, TC, and AR visualization were 92.7%, 36.7%, and 89.8%, respectively. The sensitivity, specificity, and diagnostic accuracy and the proportion of true positives, false positives, and true negatives between using 1, 2, landmarks and 3 landmarks were comparable (P > .05). CONCLUSION: Each sonographic landmark had similar reliability and diagnostic accuracy. Identifying all 3 sonographic landmarks was not mandatory and visualizing normal movement in one of the sonographic landmarks would be sufficient to exclude VCP. Copyright © 2014 Elsevier Inc. All rights reserved.postprin

An adult zebrafish model for Laribacter hongkongensis infection: Koch’s postulates fulfilled

published_or_final_versio

Studying the effect of sonographic landmarks imaged on transcutaneous laryngeal ultrasonography on perioperative vocal cord assessment

Scientific Session 8INTRODUCTION: Transcutaneous laryngeal ultrasound (TLUSG) is a non-invasive way of assessing vocal cord (VC) function. During examination, the assessor often looks at 3 sonographic landmarks (namely, false VC (FC), true VC (TC) and arytenoids (AR)) to ascertain VC movement. However, it is unclear among these landmarks, which one provides the most reliable VC assessment as not all patients would have all three landmarks identified on the same examination. We postulated that perhaps finding all three sonographic landmarks may further improve diagnostic accuracy. To address these questions, we prospectively evaluated consecutive patients over two institutions …published_or_final_versio

Dynamic Failure Properties of the Porcine Medial Collateral Ligament-Bone Complex for Predicting Injury in Automotive Collisions

The goal of this study was to model the dynamic failure properties of ligaments and their attachment sites to facilitate the development of more realistic dynamic finite element models of the human lower extremities for use in automotive collision simulations. Porcine medial collateral ligaments were chosen as a test model due to their similarities in size and geometry with human ligaments. Each porcine medial collateral ligament-bone complex (n = 12) was held in a custom test fixture placed in a drop tower to apply an axial impulsive impact load, applying strain rates ranging from 0.005 s-1 to 145 s-1. The data from the impact tests were analyzed using nonlinear regression to construct model equations for predicting the failure load of ligament-bone complexes subjected to specific strain rates as calculated from finite element knee, thigh, and hip impact simulations. The majority of the ligaments tested failed by tibial avulsion (75%) while the remaining ligaments failed via mid-substance tearing. The failure load ranged from 384 N to 1184 N and was found to increase with the applied strain rate and the product of ligament length and cross-sectional area. The findings of this study indicate the force required to rupture the porcine MCL increases with the applied bone-to-bone strain rate in the range expected from high speed frontal automotive collisions

Disrupted-in-schizophrenia 1 (DISC1) and Syntaphilin collaborate to modulate axonal mitochondrial anchoring

In neuronal axons, the ratio of motile-to-stationary mitochondria is tightly regulated by neuronal activation, thereby meeting the need for local calcium buffering and maintaining the ATP supply. However, the molecular players and detailed regulatory mechanisms behind neuronal mitochondrial movement are not completely understood. Here, we found that neuronal activation-induced mitochondrial anchoring is regulated by Disrupted-in-schizophrenia 1 (DISC1), which is accomplished by functional association with Syntaphilin (SNPH). DISC1 deficiency resulted in reduced axonal mitochondrial movement, which was partially reversed by concomitant SNPH depletion. In addition, a SNPH deletion mutant lacking the sequence for interaction with DISC1 exhibited an enhanced mitochondrial anchoring effect than wild-type SNPH. Moreover, upon neuronal activation, mitochondrial movement was preserved by DISC1 overexpression, not showing immobilized response of mitochondria. Taken together, we propose that DISC1 in association with SNPH is a component of a modulatory complex that determines mitochondrial anchoring in response to neuronal activation.117Ysciescopu

Novel IL10 gene family associations with systemic juvenile idiopathic arthritis

Juvenile idiopathic arthritis (JIA) is the most common cause of chronic childhood disability and encompasses a number of disease subgroups. In this study we have focused on systemic JIA (sJIA), which accounts for approximately 11% of UK JIA cases. This study reports the investigation of three members of the IL10 gene family as candidate susceptibility loci in children with sJIA. DNA from 473 unaffected controls and 172 patients with sJIA was genotyped for a single nucleotide polymorphism (SNP) in IL19 and IL20 and two SNPs in IL10. We examined evidence for association of the four SNPs by single marker and haplotype analysis. Significant differences in allele frequency were observed between cases and controls, for both IL10-1082 (p = 0.031) and IL20-468 (p = 0.028). Furthermore, examination of the haplotypes of IL10-1082 and IL20-468 revealed greater evidence for association (global p = 0.0006). This study demonstrates a significant increased prevalence of the low expressing IL10-1082 genotype in patients with sJIA. In addition, we show a separate association with an IL20 polymorphism, and the IL10-1082A/IL20-468T haplotype. The two marker 'A-T' haplotype confers an odds ratio of 2.24 for sJIA. This positive association suggests an important role for these cytokines in sJIA pathogenesis

Thumb reconstruction by grafting skeletonized amputated phalanges and soft tissue cover – A new technique: A case series

This study reports five cases of crush-avulsion injury to the thumb at different levels presented at our plastic and hand surgery unit between 2005 and 2007. All of the patients were male labors with machine injuries to the thumb with non-replantable amputations. Distal phalanx or proximal phalanx, or both, were used as a free cortical bone graft. The amputated part was skeletonized keeping the periosteum attached to the cortical bone of the phalanx fixing it to the stump and covering it by either local flap like dorsal metacarpal flap or regional flaps like the distally based pedicled radial forearm flap and neurovascular island sensate flap or groin flap. The results were functionally and cosmetically good and follow up X rays showed no osteoporotic resorption after one year

Prevalence and effects of emphysema in never-smokers with rheumatoid arthritis interstitial lung disease

AIMS: Autoimmune conditions such as rheumatoid arthritis-related interstitial lung disease (RA-ILD) have been linked to the existence of emphysema in never-smokers. We aimed to quantify emphysema prevalence in RA-ILD never-smokers and investigate whether combined pulmonary fibrosis and emphysema (CPFE) results in a worsened prognosis independent of baseline disease extent. METHODS: RA-ILD patients presenting to the Royal Brompton Hospital (n=90) and Asan Medical Center (n=155) had CT's evaluated for a definite usual interstitial pneumonia (UIP) pattern, and visual extents of emphysema and ILD. RESULTS: Emphysema, identified in 31/116 (27%) RA-ILD never-smokers, was associated with obstructive functional indices and conformed to a CPFE phenotype: disproportionate reduction in gas transfer (DLco), relative preservation of lung volumes. Using multivariate logistic regression, adjusted for patient age, gender and ILD extent, emphysema presence independently associated with a CT-UIP pattern in never-smokers (0.009) and smokers (0.02). On multivariate Cox analysis, following adjustment for patient age, gender, DLco, and a CT-UIP pattern, emphysema presence (representing the CPFE phenotype) independently associated with mortality in never-smokers (p=0.04) and smokers (p<0.05). CONCLUSION: 27% of RA-ILD never-smokers demonstrate emphysema on CT. Emphysema presence in never-smokers independently associates with a definite CT-UIP pattern and a worsened outcome following adjustment for baseline disease severity