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.     

Glibenclamide for the Treatment of Ischemic and Hemorrhagic Stroke

Ischemic and hemorrhagic strokes are associated with severe functional disability and high mortality. Except for recombinant tissue plasminogen activator, therapies targeting the underlying pathophysiology of central nervous system (CNS) ischemia and hemorrhage are strikingly lacking. Sur1-regulated channels play essential roles in necrotic cell death and cerebral edema following ischemic insults, and in neuroinflammation after hemorrhagic injuries. Inhibiting endothelial, neuronal, astrocytic and oligodendroglial sulfonylurea receptor 1–transient receptor potential melastatin 4 (Sur1–Trpm4) channels and, in some cases, microglial K_ATP (Sur1–Kir6.2) channels, with glibenclamide is protective in a variety of contexts. Robust preclinical studies have shown that glibenclamide and other sulfonylurea agents reduce infarct volumes, edema and hemorrhagic conversion, and improve outcomes in rodent models of ischemic stroke. Retrospective studies suggest that diabetic patients on sulfonylurea drugs at stroke presentation fare better if they continue on drug. Additional laboratory investigations have implicated Sur1 in the pathophysiology of hemorrhagic CNS insults. In clinically relevant models of subarachnoid hemorrhage, glibenclamide reduces adverse neuroinflammatory and behavioral outcomes. Here, we provide an overview of the preclinical studies of glibenclamide therapy for CNS ischemia and hemorrhage, discuss the available data from clinical investigations, and conclude with promising preclinical results that suggest glibenclamide may be an effective therapeutic option for ischemic and hemorrhagic stroke.     

Behavior of 316L stainless steel containing corrosion pits under cyclic loading

The environmental performance of 316L grade stainless steel, in the form of tensile specimens containing a single corrosion pit with various aspect ratios, under cyclic loading in aerated chloride solutions is investigated in this study. Results from environmental tests were compared and contrasted with those obtained using finite element analysis (FEA). Fractography of the failed specimens obtained from experiments revealed that fatigue crack initiation took place at the base of the shallow pit. The crack initiation shifted towards the shoulder and the mouth of the pit for pits of increasing depth. This process is well predicted by FEA, as the strain contour maps show that strain is the highest around the centric strip of the pit. However, for shallow pits, local strain is uniformly distributed around that strip but begins to concentrate more towards the shoulder and the mouth region for increasingly deep pits.     

Hot corrosion Type II of FeCr‐based model alloys for boiler and heat exchanger applications

The hot corrosion Type II of the alloys FeCr20, FeCr20Ni10, FeCr20Ni20, and FeCr20Co10 is investigated at 700°C in air + 0.5% SO₂ with deposits consisting of Na₂SO₄ and a eutectic mixture of Na₂SO₄ and MgSO₄ for 24, 100, and 300 h. The alloying elements nickel and cobalt have a positive influence when tests are conducted using a MgSO₄‐Na₂SO₄ deposit. In this case, they reduce the metal loss and increase the time to the propagation stage. In contrast, when the alloys are exposed with a Na₂SO₄ deposit, these alloying elements increase the metal loss and allow for the transition to the propagation stage because they can form molten phases with the Na₂SO₄. During the incubation stage an oxide scale forms on the FeCr20 alloy, which is thicker than the one formed during exposure without a deposit, and iron oxides are observed, which precipitate in the deposit. The propagation stage occurs by a dissolution and precipitation mechanism forming localized pitting attack. Iron is the main species that dissolves and precipitates, while chromium remains mainly as an oxide beneath the initial surface. The additional elements are found in the pit and in the salt deposit.     

Investigation on phase shifting for a 4 K Stirling pulse tube cryocooler with He-3 as working fluid

He-3 is generally recognized for its ability to provide more excellent thermophysical performance than He-4, especially in the 4 K temperature range. However, this was not always the case in our preliminary experiments on a three-stage Stirling-type pulse tube cryocooler (SPTC). Our ongoing studies, as reported in this paper, demonstrate that the different working fluids also affect the performance through their phase shifting capability. This feature has been passed over in large part by researchers considering refrigerant substitution. Unlike previous theoretical analyses that focus primarily on regenerator losses, this report investigates the effects of the working fluid on the phase angle at the cold end in order to quantitatively reveal the relationship between the lowest attainable temperature and the cooling capacity. The analysis agrees well with our experimental results on a three-stage SPTC. While running with the operating parameters optimized for He-3, the lowest temperature of the SPTC decreased from 5.4 K down to 4.03 K. This is the lowest refrigeration temperature ever achieved with a three-stage SPTC.     

300M超高强钢在中性盐雾环境中的腐蚀行为及机制

摘要：为了研究300M超高强钢在中性盐雾环境中的腐蚀行为及腐蚀机制，采用失重法，宏观、微观腐蚀形貌分析，三维表面轮廓分析及电化学分析的研究方法，来表征腐蚀实验现象并进行分析。结果表明：300M超高强钢在中性盐雾环境中的腐蚀产物为FeOOH、Fe2O3、Fe(OH)3和Fe3O4；腐蚀速率随着腐蚀时间逐渐降低，腐蚀后期（72h）腐蚀速率降低50%；腐蚀初期以点蚀为主，点蚀坑通过横向扩展，逐渐发展为后期的均匀腐蚀，腐蚀表面形貌呈沟壑状；外腐蚀层对基体的保护能力很弱，Cr元素在锈层靠近基体的一侧偏聚使内腐蚀层具有一定的抗腐蚀性。     

Interlayer Engineering of Molybdenum Trioxide toward High‐Capacity and Stable Sodium Ion Half/Full Batteries

Orthorhombic molybdenum trioxide (MoO₃) is one of the most promising anode materials for sodium‐ion batteries because of its rich chemistry associated with multiple valence states and intriguing layered structure. However, MoO₃ still suffers from the low rate capability and poor cycle induced by pulverization during de/sodiation. An ingenious two‐step synthesis strategy to fine tune the layer structure of MoO₃ targeting stable and fast sodium ionic diffusion channels is reported here. By integrating partially reduction and organic molecule intercalation methodologies, the interlayer spacing of MoO₃ is remarkably enlarged to 10.40 Å and the layer structural integration are reinforced by dimercapto groups of bismuththiol molecules. Comprehensive characterizations and density functional theory calculations prove that the intercalated bismuththiol (DMcT) molecules substantially enhanced electronic conductivity and effectively shield the electrostatic interaction between Na⁺ and the MoO₃ host by conjugated double bond, resulting in improved Na⁺ insertion/extraction kinetics. Benefiting from these features, the newly devised layered MoO₃ electrode achieves excellent long‐term cycling stability and outstanding rate performance. These achievements are of vital significance for the preparation of sodium‐ion battery anode materials with high‐rate capability and long cycling life using intercalation chemistry.     

Corrosion of hot-dip-galvanised steel and zinc alloy-coated steel in ammonia and ammonium chloride

There are many potential causes of corrosion in animal buildings. Animals exhale large quantities of moisture into the air creating high relative humidity in the building if the moisture is not properly vented. High humidity increases the potential for condensation. In addition, ammonia may be found in large quantities in animal buildings. Ammonia is released from manure and urine. In addition, ammonium chloride is used as a nitrogen source in fertilisers. In this study, the atmospheric corrosion of hot-dip-galvanised steel and zinc alloy-coated steel such as zinc–aluminium and zinc–aluminium–magnesium has been studied in atmospheres containing different levels of ammonia. Investigations have also been conducted at different levels of ammonium chloride. The results are discussed in view of the mechanisms of corrosion of zinc and zinc alloy-coated steel in ammonia and ammonium chloride-containing environments.