High-Throughput Screening of Acyl-CoA Thioesterase I Mutants Using a Fluid Array Platform

Screening target microorganisms from a mutated recombinant library plays a crucial role in advancing synthetic biology and metabolic engineering. However, conventional screening tools have several limitations regarding throughput, cost, and labor. Here, we used the fluid array platform to conduct high-throughput screening (HTS) that identified Escherichia coli ???TesA thioesterase mutants producing elevated yields of free fatty acids (FFAs) from a large (106) mutant library. A growth-based screening method using a TetA-RFP fusion sensing mechanism and a reporter-based screening method using high-level FFA producing mutants were employed to identify these mutants via HTS. The platform was able to cover >95% of the mutation library, and it screened target cells from many arrays of the fluid array platform so that a post-analysis could be conducted by gas chromatography. The ???TesA mutation of each isolated mutant showing improved FFA production in E. coli was characterized, and its enhanced FFA production capability was confirmed

Silicon improves growth and antioxidative defense system in salt-stressed Kentucky bluegrass (Poa pratensis L.), ‘Perfection’ and ‘Midnight’

After 400 mM of NaCl treatment, 0.1 and 1.0 mM of silicon (Si) were added to Kentucky bluegrass, ‘Perfection’ and ‘Midnight,’ to identify the effect of Si on the antioxidant defense mechanisms and inorganic ions as a parameter in the salt stress on grass. Compared to the control, the NaCl treatment caused a significant decrease in the shoot length and the fresh and dry weight of shoot and root of the both types of Kentucky bluegrass. Adding Si after the NaCl treatment increased the growth and dry and fresh weight of shoot and root in the grass. In both types, the Na+ concentration significantly increased after the NaCl-only treatment compared to control and decreased dramatically after 0.1 and 1.0 mM Si were added following the NaCl treatment compared to the NaCl-only treatment. K+ and Si concentrations remarkably increased in the shoot and root when Si was added after NaCl treatment. There was a significant reduction in the oxygen radical absorption capacity and the total phenolic compounds in the both types. Compared to the NaCl-only treatment, higher glutathione and lower proline concentrations were observed in the plant treated with Si after NaCl treatment. These results suggest that, even though Si is not generally classified as ‘essential element’, Si may have a significant involvement in the antioxidant defense mechanisms and inorganic ions in the salt stress on grass.Key words: diphenyl-1-picrylhydrazyl (DPPH), total phenolic concentration, proline, salt stress

THE EFFECTS OF WHEELCHAIR CAMBER AND HANDRIM SIZE IN WHEELCHAIR BASKETBALL MOVEMENT

Improving the wheelchair design can be an important determinant of high performance in the wheelchair basketball. Researchers have focused on increasing the efficiency of the wheelchair, looking at the factors such as the seat position, handrim size, and wheel camber. Most handrim and wheel chamber studies, however, have focused only on the propulsive phase. Quick turn as well as fast propulsion is essential to the success in a wheelchair basketball game and the energy efficiency becomes particularly important in a prolonged wheelchair use. The purpose of this study was to investigate the effects of wheelchair camber and handrim size on the linear propulsion, turn velocity, and efficiency

Valproic Acid Downregulates the Expression of MGMT and Sensitizes Temozolomide-Resistant Glioma Cells

Temozolomide (TMZ) has become a key therapeutic agent in patients with malignant gliomas; however, its survival benefit remains unsatisfactory. Valproic acid (VPA) has emerged as an anticancer drug via inhibition of histone deacetylases (HDACs), but the therapeutic advantages of a combination with VPA and TMZ remain poorly understood. The main aim of the present study was to determine whether an antitumor effect could be potentiated by a combination of VPA and TMZ, especially in TMZ-resistant cell lines. A combination of VPA and TMZ had a significantly enhanced antitumor effect in TMZ-resistant malignant glioma cells (T98 and U138). This enhanced antitumor effect correlated with VPA-mediated reduced O6-methylguanine-DNA methyltransferase (MGMT) expression, which plays an important role in cellular resistance to alkylating agents. In vitro, the combination of these drugs enhanced the apoptotic and autophagic cell death, as well as suppressed the migratory activities in TMZ-resistant cell lines. Furthermore, in vivo efficacy experiment showed that treatment of combination of VPA and TMZ significantly inhibited tumor growth compared with the monotherapy groups of mice. These results suggest that the clinical efficacy of TMZ chemotherapy in TMZ-resistant malignant glioma may be improved by combination with VPA

Predictors of Successful Trial without Catheter for Postoperative Urinary Retention Following Non-Urological Surgery

Purpose To investigate the success rate of trial without catheter (TWOC) for postoperative urinary retention (POUR) after non-urological surgery and to determine predictors of successful TWOC. Methods A total of 104 patients who underwent non-urological surgery and were referred to the department of urology for POUR were included in this retrospective study. All eligible patients underwent indwelling catheterization as an initial treatment and then TWOC was performed 3 to 7 days later. POUR was defined as micturition difficulty with greater than 400 mL of postvoid residual (PVR) urine volume measured by catheterization after non-urological surgery. Successful TWOC was defined as voiding with less than 100 mL of PVR urine volume. Predictive factors were identified by multivariate regression analysis. All definitions corresponded to recommendations of the International Continence Society. Results The mean age of the patients was 65.2 (range, 23 to 92) years. There were 45 male and 59 female patients. Intraoperative indwelling catheterization was performed in 69 (66.3%) patients. Mean duration of indwelling catheterization for POUR was 5.0 (range, 3.0 to 7.0) days and 83 (79.8%) patients received medication with an alpha-blocker. A successful TWOC was observed in 70 (67.4%) patients. The mean age of the patients with failure of TWOC was significantly higher than that of the patients with successful TWOC. The percentages of female patients, spinal surgery, and prone position during surgery in patients with unsuccessful TWOC were higher than in those with successful TWOC. In the multivariate logistic regression analysis, age and location of surgery (spine vs. non-spine) were the independent predictors of successful TWOC for POUR. Conclusions Our data suggest that older age and spinal surgery may be important risk factors for failure of TWOC for POUR after non-urological surgery. Thus, adequate prevention measures may be necessary for POUR after non-urological surgery, especially in patients with these risk factors

Effect of Growth Parameters and Annealing Atmosphere on the Properties of Cu 2

Cu2ZnSnS4 (CZTS) thin films were deposited using the cosputtering technique. The growth parameters, such as working pressure, target powers, and postannealing atmosphere, were optimized for CZTS films deposition. A comparative study between post annealing using sulfur vapor in a quartz tube furnace and sulfurization chamber using H2S gas was carried out to optimize the kesterite Cu2ZnSnS4 phase. 10 min annealing at 530°C in the furnace in sulfur vapor eliminated all the secondary phases and formed kesterite Cu2ZnSnS4. The diffusion of sulfur in the film during the annealing process enhanced the crystallinity of the film. The kesterite Cu2ZnSnS4 phase was confirmed by X-ray diffraction, Raman scattering, and optical measurements. The film showed phonon peaks corresponding to the kesterite CZTS, high-absorption coefficient (1.1 × 105 cm−1), and desired optical direct band gap (1.5 eV)