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.     

Identification of stress-induced leakage current components and thecorresponding trap models in SiO2 films [MOStransistors]

Time-decay stress-induced leakage current (SILC) has been systematically investigated for the cases of both Fowler-Nordheim (FN) stress and substrate hot-hole stress. From the three viewpoints of the reproducibility of the current component for the gate voltage scan, the change of oxide charge during the gate voltage scan, and the resistance of the current component to thermal annealing, it has been found that time-decay stress-induced leakage current is composed of five current components, regardless of stress type. Trap models corresponding to each current component have been proposed. In addition, it has also been proven that holes generate the electron traps related to one of those current components     

High-dose intravenous steroid pulse therapy in thyroid-associated ophthalmopathy

MIcrospheres containing diclofenac sodium (DS) were prepared using carboxymethylcellulose (CMC) as the main support material (1.0, 2.0, 3.0% (w/v)) and aluminum chloride as the crosslinker. Drug to polymer ratios of 1:1, 1:2 and 1:4 were used to obtain a range of microspheres. The microspheres were then coated with an enteric coating material, Eudragit S-100, efficiency, % yield value, particle sizes an in-vitro dissolution behaviour were investigated. The surface of the enteric coated microspheres seemed to be all covered with Eudragit S-100 from scanning electron microscopy observation. It was also observed that increasing the CMC concentration led to an increase in the encapsulation efficiency, % yield value and particle size and decreased the release rate. Eudragit S-100 coating did not significantly alter the size but the release rate was significantly lower even when the lower concentration solution was used.     

Kinetic analysis for effect of initial substrate concentration on growth and secondary metabolite production in cultures of Nicotiana tabacum

Suspension cultures of tobacco cells were studied using airlift and rotary-drum bioreactors. The effect of initial concentrations of a major substrate, sucrose, on the growth and production of a secondary metabolite, phenolic compounds, was investigated. The dry weights and total concentrations of the phenolic compounds increased with the initial sucrose concentration in both bioreactors. Both bioreactors were found to have the same tendency for the effect of initial sucrose concentration. The structured model, presented previously was modified by considering that sucrose was hydrolyzed to glucose and fructose by an enzymatic reaction. The previous and the new models were applied to the above two sets of experimental data obtained with two bioreactors, independently. The hydrolysis of sucrose was elucidated to contribute slightly to the overall kinetics of growth and secondary metabolite production in these cultures. Furthermore, the levels of shear damage in each bioreactor were quantitatively compared based on the death rate constant, k_i, which is one of the model constants.     

Reduction of higher harmonic sequences in terminal voltage of an AC exciting synchronous generator using magnetically anisotropic slot wedges

This paper describes relationships between characteristics of a magnetically anisotropic slot wedge and voltage higher harmonics in an ac exciting synchronous generator terminal due to permeance ripple caused by an open-slot-type rotor. From the relationships obtained, practical guidelines were formulated for machine designs applying the magnetically anisotropic slot wedge. The results are as follows.

• 1 (1) Equivalent permeability of the magnetically anisotropic slot wedge became isotropic wedge permeability for air-gap flux ripple.
• 2 (2) Two ways were identified to reduce voltage-higher harmonics by using the slot wedges.
• 3 (3) The level of leakage flux passing through the anisotropic slot wedge was determined only by its permeability in the width direction.
• 4 (4) Effectiveness of the magnetic anisotropic slot wedges inserted into the stator slots was verified from experimental results using a model machine of the open- slot-type rotor.

     

Isolation and characterization of a novel human pancreas-specific gene, pancpin, that is down-regulated in pancreatic cancer cells

We reviewed 19 children and adolescents with cervical spine congenital synostosis as in Klippel-Feil syndrome (KFS), with an average follow-up of 12.5 years. We paid particular attention to neurologic complications associated with cervical spine abnormalities. Five patients were affected by neurologic complications; four underwent a surgical procedure; and 14 had no neurologic finding. Two had hypermobility at one level, and one had hypermobility at two levels. We found that the more numerous the occipito-C1 abnormalities, the more significant the neurologic risk. In contrast, this risk was not related to the number of "mobile blocks" or to age. Various mechanisms of neural complications have been studied in the literature: medullary abnormality, spinal instability, narrowing of the cervical canal, and vascular dysfunction. Surgery is usually thought to be required in cases with neurologic complications. The indication for surgery is, however, less clear in cases of pure instability without neurologic involvement because surgery is likely to increase the future risks at mobile disks either above or below the fuse level. Careful clinical and radiologic observation is necessary in such patients. Magnetic resonance imaging (MRI) with lateral views in flexion and extension seem to be the best method for detecting impingement of the spine on the cord.     

Effect of Elastic Energy due to Atomic Size Factor on Ordering and Decomposition Behaviour of Binary Solid Solutions

A theory recently developed by the present authors is applied to the study of the effect of elastic energy due to atomic size factor on the transformation behaviour of binary solid solutions. lt is found that elastic interaction energy (EIE), which is a part of the total elastic energy plays a key role in both ordering elastic interaction ordering (EIO) and spinodal decomposition. The present study gives a reasonable explanation to the historical dilemmas, "elastic energy paradox" and "atomic size factor paradox . By solving these confusing problems, the coexistence of ordering (EIO) and decomposition, which has been regarded as impossible by conventional theories. can be well understood. The mechanism is as follows: lowering of elastic energy demands EIO, and such an ordering provides a driving force for spinodal decomposition. Therefore, in alloys with large atomic size factor, spinodal decomposition is preceded and induced by ordering. Ordering and spinodal decomposition are thus closely related processes to each other     

Theory of one-tape linear-time Turing machines

A theory of one-tape two-way one-head off-line linear-time Turing machines is essentially different from its polynomial-time counterpart since these machines are closely related to finite state automata. This paper discusses structural-complexity issues of one-tape Turing machines of various types (deterministic, nondeterministic, reversible, alternating, probabilistic, counting, and quantum Turing machines) that halt in linear time, where the running time of a machine is defined as the length of any longest computation path. We explore structural properties of one-tape linear-time Turing machines and clarify how the machines’ resources affect their computational patterns and power.     

Scale free interval graphs

Scale free graphs have attracted attention by their non-uniform structure that can be used as a model for various social and physical networks. In this paper, we propose a natural and simple random model for generating scale free interval graphs. The model generates a set of intervals randomly under a certain distribution, which defines a random interval graph. The main advantage of the model is its simpleness. The structure/properties of generated graphs are analyzable by relatively simple probabilistic and/or combinatorial arguments, which is different from many other models. Based on such arguments, we show for our random interval graph that its degree distribution follows a power law, and that it has a large average clustering coefficient.     

Quantum Monte Carlo Device Simulation of Nano-Scaled SOI-MOSFETs

Quantum transport properties of nano-scaled SOI-MOSFETs are investigated based on a quantum Monte Carlo (MC) device simulation. The quantum mechanical effects are incorporated in terms of a quantum correction of potential in the well-developed particle MC computational techniques. The ellipsoidal multi-valleys of silicon conduction band are also considered in the simulation. First, the validity of the quantum MC technique is verified by comparing the simulated results with a self-consistent Schrödinger-Poisson solution at thermal equilibrium. Then, we apply the technique to non-equilibrium and quasi-ballistic quantum transport in nano-scaled SOI-MOSFETs.