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• One-hundred and five spine tumor patients were examined • The complication rate was 36.2% and the major complication rate was 19.1% • 3 scoring systems were compared to assess their ability to predict complications • The NESMS had the highest accuracy but did not meet the threshold for utility Estimating complications in oncological spine surgery is challenging. The objective of this study was to compare the accuracy of three scoring systems for predicting perioperative morbidity after surgery for spinal metastases. One-hundred and five patients who underwent surgery between 2013 and 2019 were included in this study. All patients had scores retrospectively calculated using the New England Spinal Metastasis Score (NESMS), Metastatic Spinal Tumor Frailty Index (MSTFI), and Anzuategui scoring systems. The main outcome measure was development of a medical complication (minor or major) within 30 days of surgery. The predictive ability for each system was assessed using receiver operating characteristic analysis and calculations of the area under the curve (AUC). The average age for all patients was 61 years and 61/105 patients (58.1%) were male. The most common primary tumor origins were hematologic (23.8%), prostate (16.2%), breast (14.3%), and lung (13.3%). The overall 30-day complication rate was 36.2% and the rate of major complications was 21.9%. Among all patients who underwent oncological spine surgery, the NESMS score had the highest AUC for 30-day overall (AUC 0.64; 95% CI, 0.53 – 0.75) and major morbidity (AUC 0.68; 95% CI, 0.54– 0.81) in our population. However, the accuracy did not meet the threshold for clinical utility. Future prospective validation of these systems in other populations is encouraged. [ABSTRACT FROM AUTHOR]

Abstract: Dissolved oxygen (DO)-controlled nar promoter expression vectors were constructed, and their expression efficiency was compared with that of the T7 promoter pET22 expression vector by expressing human growth hormone (hGH), enhanced green fluorescence protein (EGFP), and β-tyrosinase in Escherichia coli cells. The nar promoter expression vector pRBS, which was engineered with a 5′-untranslated region and ribosomal binding site for the T7 promoter, expressed hGH at a rate of up to 32% of the total cellular proteins (TCP) in E. coli W3110narL−. The expression level of hGH was further enhanced, up to ∼42% of the TCP, by adding the N-terminal peptide tag of β-galactosidase to hGH, which was comparable to the expression of ∼43% of the TCP in pET-lac:hGH/BL21(DE3). A further engineered expression vector, pRBS(fnr), which coexpressed fumarate/nitrate reductase (fnr), expressed more EGFP than pET22 in BL21(DE3). In addition, recombinant β-tyrosinase was successfully expressed at a rate of up to ∼45% of the TCP in pRBS(fnr) in W3110narL−. From these results, the DO-controlled nar promoter system developed in this study can be considered a reliable and cost-effective expression system for protein production, especially in large-scale fermentation, as an alternative to the pET/BL(DE3) system. [ABSTRACT FROM AUTHOR]

Abstract: Escherichia coli has been the most widely used host for the production of recombinant proteins because it is the best characterized system in every aspect. Furthermore, the high cell density culture of recombinant E. coli has allowed production of various proteins with high yield and high productivities. Various cultivation strategies employing different host strains and expression systems have been successfully employed for the production of recombinant proteins. New strategies for strain improvement towards the goal of enhanced protein production are actively being developed based on high-throughput omics approaches such as transcriptomics and proteomics. This paper reviews recent advances in the production of recombinant proteins by high cell density culture of E. coli. [Copyright &y& Elsevier]

Recombinant Escherichia coli (E. coli) displaying lipase on the cell surface was examined as a whole cell biocatalyst for enantioselective resolution of racemic compounds. The Pseudomonas fluorescens lipase was displayed on the cell surface of E. coli by fusing the lipase gene to the Salmonella typhimurium outer membrane protein C (OmpC) gene by C-terminal deletion–fusion strategy. The localization of truncated OmpC-lipase fusion protein was confirmed by confocal microscopy and whole cell lipase activity. As an application for enantioselective biocatalyst, whole cell bioconversion was examined using cell surface displayed lipase. When racemic ethyl 3-hydroxybutyrate, methyl mandelate and cis-3-acetoxy-4-phenylazetidin-2-one were used as substrates, (R)-3-hydroxybutyric acid, (S)-mandelic acid and (3S, 4R)-cis-3-hydroxy-4-phenylazetidin-2-one were successfully obtained with the enantiomeric excess of greater than 99%. The cell surface displayed lipase was found to be maintained without much loss of activity and selectivity during 10 repeated reactions over 120h. These results suggest that cell surface displayed lipase has a high enantioselectivity, broad substrate specificity and reusability, which makes it an excellent system for the production of various optically active compounds. [Copyright &y& Elsevier]

Cell-surface display allows peptides and proteins to be displayed on the surface of microbial cells by fusing them with the anchoring motifs. The protein to be displayed – the passenger protein – can be fused to an anchoring motif – the carrier protein – by N-terminal fusion, C-terminal fusion or sandwich fusion. The characteristics of carrier protein, passenger protein and host cell, and fusion method all affect the efficiency of surface display of proteins. Microbial cell-surface display has many potential applications, including live vaccine development, peptide library screening, bioconversion using whole cell biocatalyst and bioadsorption. [Copyright &y& Elsevier]

Amylose microparticles can be produced by self-assembly of amylose molecules through an amylosucrase-mediated synthesis. Here we investigated the role of short-chain fatty acids in the formation of amylose microparticles and the fate of these fatty acids at the end of the reaction. The rate of self-assembly and production yields of amylose microparticles were significantly enhanced in the presence of fatty acids. The effect was dependent on the length of the fatty acid carbon tail; butanoic acid (C4) was the most effective, followed by hexanoic acid (C6) and octanoic acid (C8). The amylose microparticles were investigated by carrying out SEM, XRD, Raman, NMR, FT-IR and DSC analysis. The size, morphology and crystal structure of the resulting amylose microparticles were comparable with those of amylose microparticles produced without fatty acids. The results indicated the carboxyl group of the fatty acid to be responsible for promoting the self-assembly of amylose chains to form microparticles. The fatty acids were eventually removed from the microstructure through the tight association of amylose double helices to form the amylose microparticles. [ABSTRACT FROM AUTHOR]

The transradial approach is increasingly used for percutaneous coronary intervention (PCI), and we therefore aimed to compare the clinical outcomes after transradial intervention (TRI) and transfemoral intervention (TFI) in all patients undergoing PCI. Among 6,973 patients enrolled in a nationwide, prospective, multicenter registry (February 2013 to September 2013), 1,860 underwent TRI (n = 1,445, 77.7%) and TFI (n = 415, 22.3%). Bleeding and major adverse cardiac events (MACE; death, myocardial infarction, revascularization, or stent thrombosis) were compared. Bleeding occurred in 42 patients (2.3%) and was significantly less likely in the TRI versus TFI group (overall cohort: 1.5% vs 4.8%, p = 0.001; propensity score-matched: n = 728, 2.7% vs 5.2%, p = 0.048). Multivariate regression revealed that TRI was negatively associated with bleeding (odds ratio 0.42, 95% CI 0.21 to 0.83, p = 0.013). MACE occurred in 152 patients (8.2%). Kaplan-Meier estimates showed higher MACE-free survival rates in the TRI versus TFI group (overall cohort: 93.3% vs 86.7%, log-rank p = 0.026; propensity score-matched: 91.8% vs 86.5%, log-rank p = 0.04). Cox proportional analysis demonstrated that TRI independently predicted improved MACE (hazard ratio 0.64, 95% CI 0.43 to 0.91, p = 0.024). In conclusion, TRI is associated with reduced bleeding rates and better clinical outcomes than TFI in all patients undergoing PCI. [ABSTRACT FROM AUTHOR]

Rheumatoid arthritis (RA) is a chronic immune-mediated inflammatory disease characterized by persistent inflammation and joint destruction. A lipid mediator (LM, namely, 17S-monohydroxy docosahexaenoic acid, resolvin D5, and protectin DX in a ratio of 3:47:50) produced by soybean lipoxygenase from DHA, exhibits anti-inflammatory activity. In this study, we determined the effect of LM on collagen antibody-induced arthritis (CAIA) in mice and receptor activator of nuclear factor-κB ligand (RANKL)-induced osteoclast formation in RAW264.7 cells. LM effectively downregulated the expression of tartrate-resistant acid phosphatase (TRAP) and cathepsin K, inhibited osteoclast formation, and suppressed the NF-κB signaling pathway in vitro. In vivo , LM at 10 μg/kg/day significantly decreased paw swelling and inhibited progression of arthritis in CAIA mice. Moreover, proinflammatory cytokine (tumor necrosis factor-α, interleukin (IL)-6, IL-1β, IL-17, and interferon-γ) levels in serum were decreased, whereas IL-10 levels were increased following LM treatment. Furthermore, LM alleviated joint inflammation, cartilage erosion, and bone destruction in the ankles, which may be related to matrix metalloproteinase and Janus kinase (JAK)–signal transducer and activators of transcription (STAT) signaling pathway. Our findings suggest that LM attenuates arthritis severity, restores serum imbalances, and modifies joint damage. Thus, LM represents a promising therapy for relieving RA symptoms. [Display omitted] • Lipid mediators (LM) produced by soybean lipoxygenase from DHA attenuated RANKL- induced osteoclast formation. • LM improved RA symptoms and inflammatory situation in CAIA mice. • LM alleviated cartilage degradation and bone erosion in CAIA mice. • LM regulated the synovial inflammation by inhibiting the JAK/STAT signaling pathway. [ABSTRACT FROM AUTHOR]

• Self-assembly kinetics of SCGA were temperature-dependent. • Yield and particle size were decreased with increasing temperature. • Fastest formation and crystallization rates were observed at 4 and 20℃, respectively. • Nucleation enabled the formation of SCGA at 60℃. • Temperature and nucleation changed the physicochemical properties of SCGA. Self-assembly (formation and crystallization) kinetics of short-chain glucan aggregates (SCGAs) prepared at isothermal conditions (4, 20, 40, and 60℃) with or without nucleation (4℃, 1 h) were investigated. The fastest formation and crystallization rates of SCGA were observed when short-chain glucan was stored at 4℃ and 20℃, respectively. SCGA was not formed at 60℃. However, nucleation resulted in SCGA forming-ability at 60℃. Moreover, nucleation increased the yield in all temperature conditions. SCGA with nucleation decreased the crystal melting transition temperature range. All SCGAs had nanosized particles (<500 nm) with B-type crystal patterns regardless of temperature and nucleation. Consequently, self-assembly temperature and presence of nucleation step could change the physicochemical characteristics of SCGA, and manipulation of the nucleation step is expected to be an effective method to increase the yield of SCGA and produce SCGA at high temperature. [ABSTRACT FROM AUTHOR]

Background The transradial (TR) approach for percutaneous coronary intervention (PCI) is challenging and associated with failure in elderly patients. We compared the TR and transfemoral (TF) approaches in patients > 80 years with acute myocardial infarction (MI) undergoing PCI. Methods A total of 1945 (7.2%) octogenarians were enrolled from among 27,129 patients in the Korea Acute Myocardial Infarction Registry. The TR group (n = 336, 17.3%) was compared with the TF group (n = 1609, 82.7%) in the overall and propensity-matched cohorts with respect to procedural success, complications, in-hospital mortality, and one-year mortality and total major adverse cardiac event (MACE; death, MI, and revascularization) rate. Results In the overall cohort, the TR group had lower incidence of Killip class III or IV compared to the TF group. The disease extent and lesion severity were similar between groups, as was the procedural success rate (97.7% vs. 98.3%); however, in-hospital complications were significantly lower in the TR group (8.1% vs. 20.3%). In-hospital mortality was significantly lower in the TR group than the TF group (3.4% vs. 11.4%), as were the one-year mortality and total MACE (9.8% vs. 18.4% and 13% vs. 21.9%, respectively). These outcomes were consistent in the propensity-matched cohort. The TR approach was found to be a significant predictor of low in-hospital mortality (OR 0.355, 95% CI 0.139–0.907), but not of one-year mortality (OR 0.644, 95% CI 0.334–1.240). Conclusions In octogenarians with acute MI undergoing PCI, the TR approach was more effective than the TF approach as it had lower complication rate and better clinical outcomes with comparable procedural success. [ABSTRACT FROM AUTHOR]

DNA based synthetic molecular walkers are reminiscent of biological protein motors. They are powered by hybridization with fuel strands, environment induced conformational transitions, and covalent chemistry of oligonucleotides. Recent developments in experimental techniques enable direct observation of individual walkers with high temporal and spatial resolution. The functionalities of state-of-the-art DNA walker systems can thus be analyzed for various applications. Herein we review recent progress on DNA walker principles and characterization methods, and evaluate various aspects of their functions for future applications. [ABSTRACT FROM AUTHOR]

Abstract: Exocellulases play a key role in cleaving the accessible ends of cellulose molecules to release soluble glucose and cellobiose. To date, there have been no screens for exocellulase owing to assay protocol limitations, the high cost of substrates, and low activity of exocellulases compared with endocellulases. This study is the first to demonstrate direct screening for exocellulase activity using a robotic, high-throughput screening (HTS) system. Cell growth in 96-well plates was measured by monitoring optical density over 11–14h at 37°C with agitation. Fluorescence of methylumbelliferyl groups released from 4-methylumbelliferyl-β-D-cellobioside was determined using a VICTOR3 microplate reader. This new HTS system enabled activity verification of more than 104 clones per day. As a result, we obtained four exocellulases clones (CelEx-SF301, CelEx-SF309, CelEx-BR12 and CelEx-BR15) from 29,006 metagenomic fosmid clones that had previously been prepared from sweet potato field soil microbes and rumen fluid. This powerful approach could be effectively applied to screen various metagenomic resources for new enzymes. [Copyright &y& Elsevier]

Based on the previous study for the water-air mixture, the characteristics of AC electromagnetic flowmeter were investigated in the liquid metal two-phase flow. Two-phase flow experiments, encompassing bubbly to slug flow regimes, were conducted with the liquid sodium–nitrogen mixture. The simple relation ΔUTP=ΔUSP(1−α), relating flowmeter signal between single-phase flow and two-phase flow, was verified with measurements of the potential differences and the void fraction α for a bubbly flow regime in water-air loop. Whereas there was no difference in the shape of the raw signal between single-phase flow and bubbly flow, the signal amplitude of bubbly flow was greater than that of single-phase flow under the same liquid flow (sodium or water) rates, since the passage area of the liquid flow was reduced. In the case of slug flow, the phase and the amplitude of the flowmeter output showed a change in the flow characteristics around each slug bubble and the position of a slug bubble itself. Therefore, the electromagnetic flowmeter shows a good potential as a useful device for identifying the flow regimes. The void fraction can be simply measured with two electromagnetic flowmeters for a low-void fraction flow such as bubbly flow. Thus, the flowmeters can be used to measure the void fraction for a liquid sodium. [Copyright &y& Elsevier]