Identification and Characterization of PSEUDO-RESPONSE REGULATOR (PRR) 1a and 1b Genes by CRISPR/Cas9-Targeted Mutagenesis in Chinese Cabbage (Brassica rapa L.)

The CRISPR/Cas9 site-directed gene-editing system offers great advantages for identifying gene function and crop improvement. The circadian clock measures and conveys day length information to control rhythmic hypocotyl growth in photoperiodic conditions, to achieve optimal fitness, but operates through largely unknown mechanisms. Here, we generated core circadian clock evening components, Brassica rapa PSEUDO-RESPONSE REGULATOR (BrPRR) 1a, 1b, and 1ab (both 1a and 1b double knockout) mutants, using CRISPR/Cas9 genome editing in Chinese cabbage, where 9–16 genetic edited lines of each mutant were obtained. The targeted deep sequencing showed that each mutant had 2–4 different mutation types at the target sites in the BrPRR1a and BrPRR1b genes. To identify the functions of BrPRR1a and 1b genes, hypocotyl length, and mRNA and protein levels of core circadian clock morning components, BrCCA1 (CIRCADIAN CLOCK-ASSOCIATED 1) and BrLHY (LATE ELONGATED HYPOCOTYL) a and b were examined under light/dark cycles and continuous light conditions. The BrPRR1a and 1ab double mutants showed longer hypocotyls, lower core circadian clock morning component mRNA and protein levels, and a shorter circadian rhythm than wildtype (WT). On the other hand, the BrPRR1b mutant was not significantly different from WT. These results suggested that two paralogous genes may not be associated with the same regulatory function in Chinese cabbage. Taken together, our results demonstrated that CRISPR/Cas9 is an efficient tool for achieving targeted genome modifications and elucidating the biological functions of circadian clock genes in B. rapa, for both breeding and improvement.     

A Novel Neurofilament Light Chain ELISA Validated in Patients with Alzheimer’s Disease,Frontotemporal Dementia,and Subjective Cognitive Decline,and the Evaluation of Candidate Proteins for Immunoassay Calibration

Neurofilament light chain (Nf-L) is a well-known biomarker for axonal damage; however, the corresponding circulating Nf-L analyte in cerebrospinal fluid (CSF) is poorly characterized. We therefore isolated new monoclonal antibodies against synthetic peptides, and these monoclonals were characterized for their specificity on brain-specific intermediate filament proteins. Two highly specific antibodies, ADx206 and ADx209, were analytically validated for CSF applications according to well-established criteria. Interestingly, using three different sources of purified Nf-L proteins, a significant impact on interpolated concentrations was observed. With a lower limit of analytical sensitivity of 100 pg/mL using bovine Nf-L as the calibrator, we were able to quantify the Nf-L analyte in each sample, and these Nf-L concentrations were highly correlated to the Uman diagnostics assay (Spearman rho = 0.97, p < 0.001). In the clinical diagnostic groups, the new Nf-L ELISA could discriminate patients with Alzheimer’s disease (AD, n = 20) from those with frontotemporal lobe dementia (FTD, n = 20) and control samples with subjective cognitive decline (SCD, n = 20). Henceforth, this novel Nf-L ELISA with well-defined specificity and epitopes can be used to enhance our understanding of harmonizing the use of Nf-L as a clinically relevant marker for neurodegeneration in CSF.     

Cavity length and absorption dependent longitudinal beat in iodine photodissociation laser

As the cavity length of the iodine photodissociation laser (iodine PDL) is varied, the number of neighboring coherent modes increases and the time-dependent mode-competition effect gives rise to typical changes in the pulse shape. The ratio of the number of all coherent modes to that of all oscillating modes is found to be as high as 80 percent when the cavity length is 450 cm. The frequency width in which the neighboring modes are coherent is approximately 0.1 GHz for all cavity lengths used. The effect of a BDN absorber placed in the cavity is distinct in the shorter cavities.     

Determinants of Corporate R&D Investment: An Empirical Study Comparing Korea's IT Industry with Its Non‐IT Industry

In our study, we extracted the market, finance, and government factors determining R&D investment of individual firms in the IT industry in Korea. We collected the financial data of 515 individual firms belonging to IT and non‐IT industries between 1980 and 1999 from the Korea Investors Service's database and investigated the empirical relationship between the factors using an ordinary regression model, a fixed effects model, and a random effects model. The main findings of our study are as follows: i) The Herfindahl Index variable representing the degree of market concentration is statistically insignificant in explaining R&D expenditures in the IT manufacturing industry. ii) Assets, which is used as a proxy variable for firm size, have a positive and statistically significant coefficient. These two results suggest that the Schumpeterian Hypothesis may be only partially applied to the IT manufacturing industry in Korea. iii) The dividend variable has a negative value and is statistically significant, indicating that a tendency of high dividends can restrict the internal cash flow for R&D investment. iv) The sales variable representing growth potential shows a positive coefficient. v) The subsidy as a proxy variable for governmental R&D promotion policies is positively correlated with R&D expenditure. This suggests that government policy has played a significant role in promoting R&D activities of IT firms in Korea since 1980. vi) Using a dummy variable, we verified that firms reduced their R&D investments to secure sufficient liquidity under the restructuring pressure during Korea's 1998 and 1999 economic crisis.     

Miniaturized Flexible Electronic Systems with Wireless Power and Near‐Field Communication Capabilities

A class of thin, lightweight, flexible, near‐field communication (NFC) devices with ultraminiaturized format is introduced, and systematic investigations of the mechanics, radio frequency characteristics, and materials aspects associated with their optimized construction are presented. These systems allow advantages in mechanical strength, placement versatility, and minimized interfacial stresses compared to other NFC technologies and wearable electronics. Detailed experimental studies and theoretical modeling of the mechanical and electromagnetic properties of these systems establish understanding of the key design considerations. These concepts can apply to many other types of wireless communication systems including biosensors and electronic implants.     

Steam reforming of liquefied natural gas (LNG) for hydrogen production over nickel–boron–alumina xerogel catalyst

A series of mesoporous nickel–boron–alumina xerogel (x-NBA) catalysts with different boron/nickel molar ratio (x = 0–1) were prepared by an epoxide-driven sol–gel method. The effect of boron/nickel molar ratio on the catalytic activities and physicochemical properties of nickel–boron–alumina xerogel catalysts was investigated in the steam reforming of liquefied natural gas (LNG). All the mesoporous x-NBA catalysts showed similar surface area. Introduction of boron increased interaction between nickel and support. In addition, introduction of boron into x-NBA catalysts reduced methane activation energy and increased nickel surface area. Promotion of boron had a positive effect on the catalytic activity due to the increase of adsorbed methane and nickel surface area. The amount of adsorbed methane and nickel surface area exhibited volcano-shaped trends with respect to boron/nickel molar ratio. LNG conversion and hydrogen yield increased with increasing the amount of adsorbed methane and with increasing nickel surface area. Among the catalysts, 0.3-NBA, which retained the largest amount of adsorbed methane and the highest nickel surface area, showed the best catalytic performance. It was also revealed that x-NBA catalysts showed strong coke resistance during the steam reforming reaction.     

Hyperbaric storage at and above room temperature of a highly perishable food

Hyperbaric storage at naturally variable room temperature (RT) conditions (18–21 °C) and above (30 °C) was evaluated as a possible new food preservation method, regardless of temperature. Preservation of watermelon juice (used as a case study of a highly perishable food) at RT and 5 °C at atmospheric pressure was compared to preservation under 100 MPa at RT. After 8 h of hyperbaric storage at 100 MPa, the initial microbial loads of the watermelon juice were reduced by 1 log unit for total aerobic mesophiles, and 1–2 log units for Enterobacteriaceae and yeasts and moulds, to levels of about 3 log units for the former and below the detection limit for the latter, and remained thereafter unchanged up to 60 h. Similar results were obtained at 30 °C at 100 MPa after 8 h. At atmospheric pressure at RT (24 h) and 30 °C (8 h), microbial levels were already above quantification limits and unacceptable for consumption. Furthermore, pressure attenuated the increase in titratable acidity verified at atmospheric pressure, but caused higher colour changes, especially a higher lightness and a lower browning degree. Post-hyperbaric storage at 5 °C revealed an extended shelf life, as an additional benefit of hyperbaric storage. These results show that hyperbaric storage is a very promising food preservation methodology.     

Effect of a cold‐active pectinolytic system on colour development of Malbec red wines elaborated at low temperature

The effect of a new cold‐active pectinolytic system on colour of Malbec wines was studied under the following winemaking conditions: (i) fermentation at low temperature (20 °C) and (ii) prefermentative cold maceration (PCM) (5 °C–7 days) followed by traditional fermentation (28 °C). The pectinolytic system was mainly composed of polymethylgalacturonase and pectin lyase activities, detected under similar conditions to those in winemaking (pH 3.6–20 °C). The results show that the enzyme system significantly accelerated colour extraction by reducing the maceration time necessary for vinification at low temperature and shortening the PCM stage. Enzyme‐treated wines exhibited better chromatic parameters than their controls at devatting and after 6 months of storage. The cold‐active enzyme compensated the decrease in colour extraction due to the low maceration temperature, achieving high‐quality wines with chromatic characteristics similar to those of traditional wines.     

On the approximate frequentist validity of the posterior quantiles of a parametric function: results based on empirical and related likelihoods

With reference to a wide class of empirical and related likelihoods, we study priors which ensure approximate frequentist validity of the posterior quantiles of a general parametric function. It is seen that no data-free prior entails such frequentist validity but, at least for the usual empirical likelihood, a data-dependent prior serves the purpose. Accounting for the nonlinearity of the parametric function of interest requires special attention in the derivation. A simulation study is seen to provide support, in finite samples, to our asymptotic results.