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.     

Chemokines Up-Regulated in Epithelial Cells Control Senescence-Associated T Cell Accumulation in Salivary Glands of Aged and Sjögren’s Syndrome Model Mice

Immunosenescence is characterized by age-associated changes in immunological functions. Although age- and autoimmune-related sialadenitis cause dry mouth (xerostomia), the roles of immunosenescence and cellular senescence in the pathogenesis of sialadenitis remain unknown. We demonstrated that acquired immune cells rather than innate immune cells infiltrated the salivary glands (SG) of aged mice. An analysis of isolated epithelial cells from SG revealed that the expression levels of the chemokine CXCL13 were elevated in aged mice. Senescence-associated T cells (SA-Ts), which secrete large amounts of atypical pro-inflammatory cytokines, are involved in the pathogenesis of metabolic disorders and autoimmune diseases. The present results showed that SA-Ts and B cells, which express the CXCL13 receptor CXCR5, accumulated in the SG of aged mice, particularly females. CD4⁺ T cells derived from aged mice exhibited stronger in vitro migratory activity toward CXCL13 than those from young mice. In a mouse model of Sjögren’s syndrome (SS), SA-Ts also accumulated in SG, presumably via CXCL12-CXCR4 signaling. Collectively, the present results indicate that SA-Ts accumulate in SG, contribute to the pathogenesis of age- and SS-related sialadenitis by up-regulating chemokines in epithelial cells, and have potential as therapeutic targets for the treatment of xerostomia caused by these types of sialadenitis.     

Single-polarization single-mode photonic crystal fibers

A new structure of single-polarization single-mode (SPSM) photonic crystal fiber (PCF) is proposed and analyzed by using a full-vector finite element method with anisotropic perfectly matched layers. From the numerical results it is confirmed that the proposed fiber is low-loss SPSM-PCF within the wavelengths ranging from 1.48 to 1.6 /spl mu/m, where only the slow-axis mode exists and the confinement loss is less than 0.1 dB/km.     

Effect of Electron Correlation on Phonon Spectra in Cuprates

We examine theoretically the phonon dynamics in the electron-phonon-coupled systems. The model Hamiltonian is a one-dimensional Hubbard model where the hopping of electrons induces the electron-phonon coupling. The numerically exact diagonalization of the Hamiltonian with truncation of the total number of phonons is performed. We calculate the phonon excitation spectra and find that the softening of phonons occurs in insulating case, and a diffusive character appears in the metallic case. We also calculate the charge and spin excitation spectra and find that the dynamics of the phonon coupled with the hopping of electrons is affected by the low energy charge and spin excitations.     

The apolipoprotein E allele epsilon 4 is overrepresented in patients with the Lewy body variant of Alzheimer's disease

We determined apolipoprotein E (ApoE) genotypes in 122 autopsied demented patients. The frequency of the ApoE epsilon 4 allele was 39.6% in Alzheimer's disease (AD), 29.0% in the Lewy body variant of AD (LBV), and 6.25% in diffuse Lewy body disease. For AD and LBV patients, the epsilon 4 frequency was significantly higher than that reported in nondemented controls (10 to 15%). Therefore, LBV and AD share ApoE epsilon 4 as a genetic risk factor, providing further evidence that these conditions overlap.     

Long-term follow-up (8 to 17 years) after thromboexclusion operation for thoracic aortic aneurysms

Forty-eight Russell's viper bite patients (40 males, 8 females), age ranging from 16-76 years were studied. Out of 48 patients, 14 were found to have a prolonged whole blood clotting time test (WBCT) (i.e. incoagulable blood) (Group 1); 23 had a normal WBCT (i.e. clotted blood) (Group II); and 11 patients had a normal WBCT on admission which changed to non-clotting during the clinical course (Group III). Four patients from group I developed hypotension and 2 expired. The serum cortisol concentration (mean +/- SEM) on admission among groups I and II were 639 +/- 45.6 and 424 +/- 33.2 nmol/l respectively. The blood cortisol level in 35 subjects (controls) were 370.7 +/- 17.7 nmol/l (mean +/- SEM). There was a significant rise of blood cortisol in patients with incoagulable blood when compared to controls at the time of admission to the hospital (p < 0.05); but there was no significant difference among those patients with clotted blood. A much higher mean serum cortisol level was observed in 4 patients with hypotension as compared to 10 patients without shock. These patients with hypotension according to our study shown to have a favorable response to steroid therapy and eventually recovered. Whether higher doses of steroid in addition to antiserum confer extra benefit in suppressing nonspecific venom effects on the pituitary and/or adrenal is not known.     

a-Si alloy three-stacked solar cells with high stabilized-efficiency

a-Si alloy three-stacked solar cells have been studied to improve the stabilized efficiency of a-Si: H based solar cells. Based on the analysis by the individual characterization method of the component cells in stacked type cells, the a-Si :H middle cell was replaced with an a-SiGe :H cell. Furthermore, the optical confinement technology was improved to obtain a high-output current with thin i-layer thickness in the a-SiGe :H bottom cell. By this device design, the initial conversion efficiency was improved up to 12.4% and more than a 10% stabilized efficiency was obtained in a-SiC :H/a-SiGe :H/a-SiGe :H three-stacked cells. These cell characteristics were confirmed by measurements at the JQA Organization (the former JMI Institute).     

Large Coulomb blockade oscillations at room temperature in ultranarrow wire channel MOSFETs formed by slight oxidation process

We propose a new fabrication technique of room-temperature operating silicon single-electron transistors (SETs). The devices are in the form of ultranarrow wire channel MOSFETs, where a sub-10-nm channel is formed by wet etching and slight thermal oxidation. Large Coulomb blockade (CB) oscillations whose peak-to-valley current ratio at room temperature is as high as 6.8 are observed in the fabricated ultranarrow wire channel MOSFETs. It is found that larger CB oscillations are obtained in the ultranarrow wire channel SETs than in the point-contact channel SETs. It is considered that the potential fluctuations induced during the channel formation processes give rise to multiple-dot SET structures in the ultranarrow wire channel MOSFETs.     

Markedly compressible behaviors of gellan hydrogels in a constrained geometry at ultraslow strain rates

The fracture behaviors of gellan hydrogels under compression remarkably depend on the strain rate as well as the boundary conditions for lateral expansion. In the geometry with no constraint for lateral expansion (conventional uniaxial compression), the gels rupture at relatively small strains independently of the compression rates. In contrast, when the gels are compressed extremely slowly (at a strain rate of ca. 10⁻⁵ s⁻¹) in the geometry prohibiting the lateral expansion at their top and bottom surfaces, they are remarkably compressible down to 2% of the initial height without macroscopic fracture and they are accompanied by a large amount of water release. In such markedly compressed gels, many microscopic cracks are formed around the central layer, where strain concentration occurs due to the nonuniform deformation arising from the constrained geometry. In the highly compressible case, the formation of macroscopic cracks is prevented by the localization of microscopic cracks as well as the enhancement in mechanical toughness by a significant increase in polymer concentration due to water release.     

Modulation characteristic of waveguide-type optical frequency combgenerator

We report the modulation characteristics of a waveguide-type optical frequency comb generator (WG-OFCG) with the advantages of compactness, high modulation index and low driving power. The characteristics of the optical frequency comb (OFC) generated when the modulation index exceeds 2π are discussed. The power of the modulation sidebands was measured by the optical heterodyne method with an external-cavity laser as a local oscillator. The dependence of the modulation index of the WG-OFCG on the modulation frequency up to 40 GHz was measured. The generation span of the OFC at the modulation frequency detuned from an integer multiple of the free spectral range (FSR) is discussed