Functional roles of arginine residues in mung bean vacuolar H+-pyrophosphatase

AbstractPlant vacuolar H+-translocating inorganic pyrophosphatase (V-PPase EC 3.6.1.1) utilizes inorganic pyrophosphate (PPi) as an energy source to generate a H+ gradient potential for the secondary transport of ions and metabolites across the vacuole membrane. In this study, functional roles of arginine residues in mung bean V-PPase were determined by site-directed mutagenesis. Alignment of amino-acid sequence of K+-dependent V-PPases from several organisms showed that 11 of all 15 arginine residues were highly conserved. Arginine residues were individually substituted by alanine residues to produce R→A-substituted V-PPases, which were then heterologously expressed in yeast. The characteristics of mutant variants were subsequently scrutinized. As a result, most R→A-substituted V-PPases exhibited similar enzymatic activities to the wild-type with exception that R242A, R523A, and R609A mutants markedly lost their abilities of PPi hydrolysis and associated H+-translocation. Moreover, mutation on these three arginines altered the optimal pH and significantly reduced K+-stimulation for enzymatic activities, implying a conformational change or a modification in enzymatic reaction upon substitution. In particular, R242A performed striking resistance to specific arginine-modifiers, 2,3-butanedione and phenylglyoxal, revealing that Arg242 is most likely the primary target residue for these two reagents. The mutation at Arg242 also removed F− inhibition that is presumably derived from the interfering in the formation of substrate complex Mg2+–PPi. Our results suggest accordingly that active pocket of V-PPase probably contains the essential Arg242 which is embedded in a more hydrophobic environment

Evolution of carbapenem resistance in Acinetobacter baumannii: An 18-year longitudinal study from a medical center in northern Taiwan

BackgroundCarbapenem-resistant Acinetobacter baumannii has emerged as an important cause of nosocomial infections with high morbidity and mortality. The carbapenemases, especially class D carbapenem-hydrolyzing oxacillinases (CHDLs), play an important role, but the relationship between their prevalence trend and carbapenem resistance remains unclear.Materials and methodsBetween 1995 and 2012, we collected 667 isolates of A. baumannii from a single medical center in northern Taiwan. Pulsed-field gel electrophoresis (PFGE) was used to determine clonality. Antimicrobial susceptibility was determined. Carbapenemase genes and associated genetic structures were detected by polymerase chain reaction.ResultsIsolates were heterogeneous on PFGE. Susceptibility to carbapenem decreased steadily over the study period from 88.1% (2001–2003) to <25% (2010–2012), whereas the isolates remained susceptible to colistin (nearly 100%) and partially susceptible to tigecycline (80%). Starting in 2001, isolates carrying the ISAba1-blaOXA-51-like allele were consistently identified. Isolates containing the transposons Tn2006 or Tn2008 first appeared in 2007 with increasing carriage rates from 17.5% (2007–2009) to 50.0% (2010–2012). The IS1008-ΔISAba3-blaOXA-58-like, blaOXA-72 and metallo-β-lactamase genes were detected only sporadically. Isolates carrying CHDL genes were resistant to multiple drugs, including carbapenem, but remained susceptible to colistin (100.0%).ConclusionIncreased carbapenem resistance in A. baumannii may be caused by the increased prevalence of isolates containing the ISAba1-blaOXA-51-like allele and the transposons Tn2006 and Tn2008

miRTarBase update 2014: an information resource for experimentally validated miRNA-target interactions

MicroRNAs (miRNAs) are small non-coding RNA molecules capable of negatively regulating gene expression to control many cellular mechanisms. The miRTarBase database (http://mirtarbase.mbc.nctu.edu.tw/) provides the most current and comprehensive information of experimentally validated miRNA-target interactions. The database was launched in 2010 with data sources for >100 published studies in the identification of miRNA targets, molecular networks of miRNA targets and systems biology, and the current release (2013, version 4) includes significant expansions and enhancements over the initial release (2010, version 1). This article reports the current status of and recent improvements to the database, including (i) a 14-fold increase to miRNA-target interaction entries, (ii) a miRNA-target network, (iii) expression profile of miRNA and its target gene, (iv) miRNA target-associated diseases and (v) additional utilities including an upgrade reminder and an error reporting/user feedback system

Erythropoietin improves long-term neurological outcome in acute ischemic stroke patients: a randomized, prospective, placebo-controlled clinical trial.

Mortality and disability following ischemic stroke (IS) remains unacceptably high with respect to the conventional therapies. This study tested the effect of erythropoietin (EPO) on long-term neurological outcome in patients after acute IS. This study aimed to evaluate the safety and efficacy of two consecutive doses of EPO (5,000 IU/dose, subcutaneously administered at 48 hours and 72 hours after acute IS) on improving the 90-day combined endpoint of recurrent stroke or death that has been previously reported. A secondary objective was to evaluate the long-term (that is, five years) outcome of patients who received EPO.This article is freely available via Open Access. Click on the 'Additional Link' above to access the full-text via the publisher's site.Published (Open Access

High-Mass Loading Hierarchically Porous Activated Carbon Electrode for Pouch-Type Supercapacitors with Propylene Carbonate-Based Electrolyte

Rational design and development of the electrodes with high-mass loading yet maintaining the excellent electrochemical properties are significant for a variety of electrochemical energy storage applications. In comparison with the slurry-casted electrode, herein, a hierarchically porous activated carbon (HPAC) electrode with higher mass loading (8.3 ± 0.2 mg/cm2) is successfully prepared. The pouch-type symmetric device (1 cell) with the propylene carbonate-based electrolyte shows the rate capability (7.1 F at 1 mA/cm2 and 4.8 F at 10 mA/cm2) and the cycling stability (83% at 12,000 cycles). On the other hand, an initial discharge capacitance of 32.4 F and the capacitance retention of 96% after 30,000 cycles are delivered from a pouch-type symmetric supercapacitor (five cells). The corresponding electrochemical performances are attributed to the fascinating properties of the HPAC and the synergistic features of the resulting electrode

ATF3 Protects against LPS-Induced Inflammation in Mice via Inhibiting HMGB1 Expression

Lipopolysaccharide (LPS) triggers innate immunity mainly via TLR4 signaling. ATF3 is a negative regulator of TLR4 signaling. HMGB1 plays a critical role in the final step of sepsis. However, the mechanisms of ATF3 and the role of HMGB1 in regulating innate immunity-induced sepsis are incompletely understood. In this study, we found that serum HMGB1 levels were 10-fold higher in patients with sepsis than normal controls. We further demonstrated that ATF3 gene knockout in mice subjected to LPS-induced endotoxemia correlates with an increase in the mortality rate and the elevated expression of IL-6, TNF-α, NO, MCP-1, and HMGB1 in the lung tissues or serum. The biochemical effects of ATF3 were observed in in vitro macrophages and blocked by ATF3 siRNA treatment. We have also shown that adeno-associated virus-mediated ATF3 gene transfer protected ATF3 knockout mice from LPS-induced mortality. In addition, ATF3 knockdown increased LPS-induced release of HMGB1. In conclusion, upregulation of ATF3 contributes to the reduced release of inflammatory molecules, especially HMGB1, which induced lung injury and increased the survival rate of mice after LPS challenge. Therefore, suppressing LPS-induced inflammation with ATF3 induction or ATF3 mimetics may be an important strategy for sepsis therapy

Body Weight Effects on Extra-Osseous Subtalar Arthroereisis

Implant extrusion in subtalar arthroereisis is a common complication for pediatric flexible flatfoot. However, there were a limited number of articles addressing the body weight effects on implant extrusion after the procedure. We conducted a 24-month follow-up assessment after subtalar arthroereisis. Surgical patients who underwent the Vulpius procedure were retrospectively collected from May 2010 to January 2017, including 59 cases of both feet having implants in situ and 43 cases of both feet having implant extrusion. The average age of 102 patients was 9 years old. The mean body mass index (BMI) of the implant in situ group was 19.5, whilst the extrusion group was 21.2 (p = 0.035). The inter-observer correlation was excellent. There were 11 cases (39.3%) of bilateral extrusion in the overweight group (BMI &ge; 24) and 13 cases (23.2%) in the low body weight group (BMI &le; 18.5) (p &lt; 0.0004). Postoperative radiographic angles were corrected in both the implant in situ group and the extrusion group. Nonetheless, the implant in situ group revealed better postoperative outcomes of Meary&rsquo;s angle and the talonavicular angle from an anterior-posterior view, and the talar inclination angle from a lateral view. We conclude that a higher BMI is related to implant extrusion and worse results after subtalar arthroereisis. Further prospective study to investigate whether preoperative weight loss results in improved surgical outcomes is warranted in the future

DoA Estimation for FMCW Radar by 3D-CNN

A method of direction-of-arrival (DoA) estimation for FMCW (Frequency Modulated Continuous Wave) radar is presented. In addition to MUSIC, which is the popular high-resolution DoA estimation algorithm, deep learning has recently emerged as a very promising alternative. It is proposed in this paper to use a 3D convolutional neural network (CNN) for DoA estimation. The 3D-CNN extracts from the radar data cube spectrum features of the region of interest (RoI) centered on the potential positions of the targets, thereby capturing the spectrum phase shift information, which corresponds to DoA, along the antenna axis. Finally, the results of simulations and experiments are provided to demonstrate the superior performance, as well as the limitations, of the proposed 3D-CNN