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.     

Proteins in longissimus muscle of Korean native cattle and their relationship to meat quality

Proteomic profiling by two-dimensional gel electrophoresis and mass spectrometry of longissimus dorsi muscle tissue from Korean native cattle identified seven proteins that are differentially expressed in animals producing low and high quality grade beef. The expression level of alpha actin is increased in high quality grade beef and the expression levels of T-complex protein 1 (TCP-1), heat shock protein beta-1 (HSP27), and inositol 1,4,5-triphosphate receptor type1 (IP3R1), a new protein to be associated with meat quality, are increased in low quality grade beef. In particular, the quantitation of HSP27 and IP3R1 by both silver staining and immunoblotting correlated well with intramuscular fat content, meat tenderness, and free calcium levels. The data suggest that HSP27 and IP3R1 are potential meat quality biomarkers and their identification provides new insight into the molecular mechanisms and pathways associated with overall beef quality.     

Thermistor behavior of PEDOT:PSS thin film

The temperature-dependent resistance changing characteristics (thermistor behaviors) of a poly(3,4-ethylenedioxythophene):poly(4-styrenesulfonate) (PEDOT:PSS) thin film are investigated in the 30–100 °C range using Greek-cross and bar patterns. The PEDOT:PSS film was spin-coated onto a Si wafer passivated with a SiO₂ layer, and a conventional dry etching technique was used to pattern the PEDOT:PSS film in conjunction with a nitride etch mask layer. Cr/Au was used for the electrode material. It was found that the characteristic temperature (T₀) and resistivity of the PEDOT:PSS film have an inversely proportional relationship with the number of coatings and the number of interfaces between multiply coated PEDOT:PSS layers. It was also found that as the number of coatings and the number of the interfaces increase, lower temperature-dependent resistance changes are observed. The temperature coefficient of resistance (TCR) value of 60 nm thick PEDOT:PSS film was slightly larger than or comparable to that of a conventional metal (Au or Pt) thermistor. The possibility of utilizing the PEDOT:PSS thin film in thermistor applications is discussed.     

Field measurement and estimation of ventilation flow rates by using train-induced flow rate through subway vent shafts

This study measured and estimated the subway vent shaft air flow rate induced by moving trains in the tunnel. This work estimated the flow rate via the tunnel structure and train movement to determine the quantitative effect of vent shafts as air purification systems of natural ventilation to improve the air quality management of a subway. The amount of air suctioned into the tunnel is significantly larger than that vented from the tunnel. Thus, placing vent shafts near subway stations is desirable for natural ventilation systems. Experimental approaches to measure train-induced flow rates have not yet been published. Results of this study provide useful fundamental data to study the natural ventilation in a subway. Therefore, this study suggested the significant design factors required to control indoor air quality in a subway.     

Comparison of bioethanol production of simultaneous saccharification & fermentation and separation hydrolysis & fermentation from cellulose-rich barley straw

Cellulose rich barley straw, which has a glucan content of 62.5%, followed by dilute acid pretreatment, was converted to bioethanol by simultaneous saccharification and fermentation (SSF). The optimum fractionation conditions for barley straw were an acid concentration of 1% (w/v), a reaction temperature of 158 °C and a reaction time of 15 min. The maximum saccharification of glucan in the fractionated barley straw was 70.8% in 72 h at 60 FPU/gglucan, while the maximum digestibility of the untreated straw was only 18.9%. With 6% content WIS (water insoluble solid) for the fractionated barley straw, 70.5 and 83.2% of the saccharification yield were in SHF and SSF (representing with glucose equivalent), respectively, and a final ethanol concentration of 18.46 g/L was obtained under the optimized SSF conditions: 34 °C with 15 FPU/g-glucan enzyme loading and 1 g dry yeast cells/L. The results demonstrate that the SSF process is more effective than SHF for bioethanol production by around 18%.     

Direct stamping of silver nanoparticles toward residue-free thick electrode

Direct stamping of functional materials has been developed for cost-effective and process-effective manufacturing of nano/micro patterns. However, there remain several challenging issues like the perfect removal of the residual layer and realization of high aspect ratio. We have demonstrated facile fabrication of flexible strain sensors that have microscale thick interdigitated capacitors with no residual layer by a simple direct stamping with silver nanoparticles (AgNPs). Polyurethane (PU) prepolymer was utilized as an adhesive layer to transfer AgNPs more efficiently during the separation step of the flexible stamp from directly stamped AgNPs. Scanning electron microscopy images and energy dispersive x-ray spectroscopy analysis revealed residue-free transfer of microscale thick interdigitated electrodes onto two different flexible substrates (elastomeric and brittle) for the application to highly sensitive strain sensors.     

Application of in vitro gut fermentation models to food components: A review

This review introduces recent changes regarding measurement of preferences brought about by the discovery of false preferences that occur with paired preference testing. Testing protocols are examined critically. False preferences result from a response bias elicited by factors distinct from sensory properties of products being tested that can distort results. Statistical approaches to circumventing this problem are examined. Alternative methods are being developed to eliminate false preferences based on understanding information processing in the brain. This has led to development of disruptive experimental preference testing protocols based on hypothesized use of processing subroutines in the brain to protect the central processing system from overload. These disruptive protocols suppress response bias. Paired preference testing is designed to predict consumer behavior. A paucity of studies addressing whether they do is put into perspective.