Direct Injection of CRISPR/Cas9-Related mRNA into Cytoplasm of Parthenogenetically Activated Porcine Oocytes Causes Frequent Mosaicism for Indel Mutations

Some reports demonstrated successful genome editing in pigs by one-step zygote microinjection of mRNA of CRISPR/Cas9-related components. Given the relatively long gestation periods and the high cost of housing, the establishment of a single blastocyst-based assay for rapid optimization of the above system is required. As a proof-of-concept, we attempted to disrupt a gene (GGTA1) encoding the α-1,3-galactosyltransferase that synthesizes the α-Gal epitope using parthenogenetically activated porcine oocytes. The lack of α-Gal epitope expression can be monitored by staining with fluorescently labeled isolectin BS-I-B₄ (IB4), which binds specifically to the α-Gal epitope. When oocytes were injected with guide RNA specific to GGTA1 together with enhanced green fluorescent protein (EGFP) and human Cas9 mRNAs, 65% (24/37) of the developing blastocysts exhibited green fluorescence, although almost all (96%, 23/24) showed a mosaic fluorescent pattern. Staining with IB4 revealed that the green fluorescent area often had a reduced binding activity to IB4. Of the 16 samples tested, six (five fluorescent and one non-fluorescent blastocysts) had indel mutations, suggesting a correlation between EGFP expression and mutation induction. Furthermore, it is suggested that zygote microinjection of mRNAs might lead to the production of piglets with cells harboring various mutation types.     

Combination of heat treatment and chitosan coating to improve postharvest quality of wolfberry (Lycium barbarum)

The synergistic application of hot water dip at 42 °C for 30 min and 1% chitosan coating on differentiation in postharvest quality traits, microstructure as well as microbiological evolution of wolfberry fruits was investigated. Fresh wolfberry fruits were stored at 2 ± 0.5 °C and 90% relative humidity (RH) for 28 days. Results indicated the combination of prestorage heat treatment and chitosan coating maintained higher levels of ascorbic acid, total phenolic contents and antioxidant capacity as well as lower decay, compared with untreated wolfberry fruits. The possible mechanism was that the heat treatment almost sealed open stomata to limit the sites of pathogen penetration into fruits independently, followed by the biofilm formed by chitosan which controlled secondary infections as well as slowed changes in fruit respiration and metabolic activity in wolfberries. The synergistically treated fruit also exhibited a higher acceptability obtained by sensory analysis after cold storage. In this sense, the integrated application of heat treatment and chitosan coating could be regarded as an effective strategy to extend storage life and maintain the postharvest quality of wolfberry fruits.     

Flotation Behavior,Collector Adsorption Mechanism of Quartz and Feldspar-Quartz Systems Using PEA as a Novel Green Collector

Silicon - A novel green cationic surfactant Poly (propylene glycol) bis (2-aminopropyl ether) (PEA) with multiple amine groups was utilized as a collector for flotation separation of quartz from...     

Quantitative risk assessment using a Japanese hydrogen refueling station model

Although hydrogen refueling stations (HRSs) are becoming widespread across Japan and are essential for the operation of fuel cell vehicles, they present potential hazards. A large number of accidents such as explosions or fires have been reported, rendering it necessary to conduct a number of qualitative and quantitative risk assessments for HRSs. Current safety codes and technical standards related to Japanese HRSs have been established based on the results of a qualitative risk assessment and quantitative effectiveness validation of safety measures over ten years ago. In the last decade, there has been much development in the technologies of the components or facilities used in domestic HRSs and much operational experience as well as knowledge to use hydrogen in HRSs safely have been gained through years of commercial operation. The purpose of the present study is to conduct a quantitative risk assessment (QRA) of the latest HRS model representing Japanese HRSs with the most current information and to identify the most significant scenarios that pose the greatest risks to the physical surroundings in the HRS model. The results of the QRA show that the risk contours of 10^?3 and 10^?4 per year were confined within the HRS boundaries, whereas the risk contours of 10^?5 and 10^?6 per year are still present outside the HRS. Comparing the breakdown of the individual risks (IRs) at the risk ranking points, we conclude that the risk of jet fire demonstrates the highest contribution to the risks at all of the risk ranking points and outside the station. To reduce these risks and confine the risk contour of 10^?6 per year within the HRS boundaries, it is necessary to consider risk mitigation measures for jet fires.     

A dual-mode bandpass filter with enhanced capacitive perturbation

A dual-mode ring bandpass filter with two pairs of capacitors has been designed. The capacitors are used to control the location of the even- and odd-mode frequencies independently, allowing weak coupling for narrow-band filter design with realizable capacitance values. Theoretical expressions have been derived for these frequencies. A 4% bandwidth bandpass filter centered at 1.9 GHz was designed and tested with good agreement between theoretical and measured results.     

Laminated Reservoir Fracturing Technique

IntroductionOil production from laminated reservoirs is significant in Daqing Oilfield in the high water cut period. Hydraulic fracturing is a very efficient way to stimulate these laminated reservoirs. Daqing Oilfield is a multi-zone sandstone     

Transmittance and cathodoluminescence of AlN ceramics sintered with Ca3Al2O6 as sintering additive

Aluminum nitride ceramics were prepared by sintering with 0–4.8 mass% of Ca₃Al₂O₆ (C₃A) as a sintering additive. The transmittance in the range of 260–550 nm increased with increasing amount of C₃A. The cathodoluminescence intensity attributed to oxygen-induced defects decreased with increasing amount of C₃A. From the results, the increase of the transmittance in the range of 260–550 nm was considered to be related to the decrease of the oxygen-induced defect density.     

Formation of toxic aldehydes in cod liver oil after ultraviolet irradiation

Formaldehyde, acrolein, malonaldehyde (MA), acetaldehyde and propanal produced from cod liver oil upon ultraviolet irradiation (λ_max=300 nm) were derivatized into nitrogenor sulfur-containing compounds and then analyzed by capillary gas chromatography with a nitrogen-phosphorus detector or a flame photometric detector. Acrolein and MA were formed at levels of 10.9±3.06 and 190.2±38.4 nmol/mg of fish oil, respectively. Maximum levels of formaldehyde, acetaldehyde and propanal formed were 7.0±0.90, 49.1±4.5 and 35.8±4.0 nmol/mg of oil, respectively. Formation of propanal in large quantities corresponded to the high content of ɛ-3 fatty acids in cod liver oil.