Conformational Analysis of a High‐Mannose‐Type Oligosaccharide Displaying Glucosyl Determinant Recognised by Molecular Chaperones Using NMR‐Validated Molecular Dynamics Simulation

Exploration of the conformational spaces of flexible oligosaccharides is essential to gain deeper insights into their functional mechanisms. Here we characterised dynamic conformation of a high‐mannose‐type dodecasaccharide with a terminal glucose residue, a critical determinant recognised by molecular chaperones. The dodecasaccharide was prepared by our developed chemoenzymatic technique, which uses ¹³C labelling and lanthanide tagging to detect conformation‐dependent paramagnetic effects by NMR spectroscopy. The NMR‐validated molecular dynamics simulation produced the dynamic conformational ensemble of the dodecasaccharide. This determined its spatial distribution as well as the glycosidic linkage conformation of the terminal glucose determinant. Moreover, comparison of our results with previously reported crystallographic data indicates that the chaperone binding to its target oligosaccharides involves an induced‐fit mechanism.     

A method of using the incident beam tilt as a eucentric goniometer in transmission electron microscopy

The principal difficulties in constructing and operating a eucentric specimen tilting goniometer in a transmission electron microscope are discussed, together with the goniometric function of the incident beam tilt. The latter function is found easy to operate in a eucentric manner. The imaging beam then will have a non-axial path, which will increase particularly the field chromatic aberration. Earlier, however, a technique for the compensation of the chromatic aberration during displaced aperture dark field image formation has been developed. In combination with this technique, it proved possible to use the ordinary incident beam tilt as a eucentric goniometer. Image sequences were obtained, with accurately varied diffraction conditions. The tilt angles and the direction of the tilt axis can be very accurately determined from the displacements of the diffraction pattern.     

On dark field techniques in transmission electron microscopy.

Different dark field techniques in transmission electron microscopy are investigated. In particular, a displaced aperture technique is developed, which produces high quality off-axis dark field images. The essential feature of the method is that the chromatic aberration of the objective and intermediate lenses is compensated for with the aid of a one-stage beam deflector in the object plane of the intermediate lens. In addition, the image distortion and the effect of the curvature of the image field are minimized by reducing the coefficient of sperical aberration of the intermediate lens. Diffracted beams, forming angles alpha up to 2.5 degrees with the optical axis, can be used for dark field imaging. Resolutions better than 30 A for alpha = 2 degrees, and 17 A for alpha = 1 degree, are obtained.     

Concurrent subarachnoid hemorrhage due to ruptured aneurysm and hypertensive intracerebral hemorrhage--case report

A 64-year-old female presented with hypertensive thalamic hemorrhage concurrent with subarachnoid hemorrhage (SAH) due to a ruptured aneurysm manifesting as sudden onset of right hemiparesis followed by severe headache. The aneurysm was located in the basilar artery at the origin of the superior cerebellar artery, remote from the thalamic hematoma. The aneurysm was clipped 3 weeks after SAH. She was discharged with slight right hemiparesis. The method and timing of surgery for such patients depend on hematoma size, location of the aneurysm and hematoma, and neurological status. The intracerebral hemorrhage remote from the ruptured aneurysm should be treated initially if necessary, and the aneurysm clipped after the brain swelling has reduced.     

Measurement of acoustic reflection coefficients by an ultrasonicmicrospectrometer

An ultrasonic microspectrometer (UMSM) was developed in order to evaluate the elastic properties of a solid specimen at a small spot on its surface. In this system, spherical-planar-pair (SPP) lenses were used, by which the acoustic reflection coefficient of a liquid/solid interface was measured as a function of the incident angle in the frequency range from 20 to 140 MHz. Using a specimen of fused quartz whose material constants were well known, the measurement accuracy was examined. The phase velocity of a leaky Rayleigh wave was obtained from the phase change of the reflection coefficient with 0.4% accuracy in this frequency range. For a specimen of steel with a large acoustic attenuation, bulk attenuation factors and their frequency dependence were successfully estimated by computer-fitting of the reflection coefficient. As an example of anisotropic materials, the reflection coefficient of X-cut quartz was also measured. Measured phase of the reflection coefficient was in good agreement with numerical calculation     

Ferroelectric switching in spin-coated nylons 11 and 12

Here, we investigate the correlation between the crystal structures and the ferroelectric switching by a sinusoidal alternative electric field for spin-coated nylon 11 films as an odd nylon and for nylon 12 as an even nylon. These spin-coated nylons afforded thin films with thicknesses ranging from 101 to 125 nm. The obtained thin films were subjected to melt-quenching, melt-cooling down, annealing-quenching, or annealing-cooling down. These processes were notably related to the resulting crystal structures. In particular, the crystal structures involving twisted bonds in the molecular chains were significantly related to ferroelectricity in both nylon 11 and nylon 12. Namely, the vector component of the amide dipole moments is transverse to the direction of the molecular chains, which is induced by the presence of more twisted bonds and is significantly related to the remanent polarization P_r for both nylons. In nylon 11, the hydrogen bonding interaction between the intermolecular amide dipole moments in the α and δ crystal forms was weakened by the existence of more twisted bonds. In nylon 12, the nonpolar γ crystal form was transformed to a polarizable γ’ crystal form because of the existence of more twisted bonds. © 2019 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2020 , 137, 48438.     

Determination of the orthotropic elastic constants of thin PET film by an ultrasonic micro-spectrometer

The elastic constants of a thin polyethylene terephthalate (PET) film, whose thickness was 12 mum, were determined from the acoustic reflection coefficients. The reflection coefficients were measured by an ultrasonic micro-spectrometer (UMSM), and all of the elastic constants were obtained through the fitting of a theory to the dip points of the magnitude of the reflection constants where leaky Lamb waves were excited. As a result, the film was found to be an orthotropic material, and the observed elastic constants were consistent with those measured by tensile tests. Besides, the in-plane shear modulus was found to be much larger than the out-of-plane shear moduli. Then, the reflection coefficients were calculated using the obtained stiffness matrix. The calculated values reproduced not only the dip points but also the change in magnitude of measured reflection coefficients depending on the frequency and the incident angle. These agreements suggest that the stiffness matrix components were determined accurately.     

Establishment of a Rapid Detection System for ISG20-Dependent SARS-CoV-2 Subreplicon RNA Degradation Induced by Interferon-α

Inhaled nebulized interferon (IFN)-α and IFN-β have been shown to be effective in the management of coronavirus disease 2019 (COVID-19). We aimed to construct a virus-free rapid detection system for high-throughput screening of IFN-like compounds that induce viral RNA degradation and suppress the replication of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). We prepared a SARS-CoV-2 subreplicon RNA expression vector which contained the SARS-CoV-2 5′-UTR, the partial sequence of ORF1a, luciferase, nucleocapsid, ORF10, and 3′-UTR under the control of the cytomegalovirus promoter. The expression vector was transfected into Calu-3 cells and treated with IFN-α and the IFNAR2 agonist CDM-3008 (RO8191) for 3 days. SARS-CoV-2 subreplicon RNA degradation was subsequently evaluated based on luciferase levels. IFN-α and CDM-3008 suppressed SARS-CoV-2 subreplicon RNA in a dose-dependent manner, with IC50 values of 193 IU/mL and 2.54 μM, respectively. HeLa cells stably expressing SARS-CoV-2 subreplicon RNA were prepared and treated with the IFN-α and pan-JAK inhibitor Pyridone 6 or siRNA-targeting ISG20. IFN-α activity was canceled with Pyridone 6. The knockdown of ISG20 partially canceled IFN-α activity. Collectively, we constructed a virus-free rapid detection system to measure SARS-CoV-2 RNA suppression. Our data suggest that the SARS-CoV-2 subreplicon RNA was degraded by IFN-α-induced ISG20 exonuclease activity.     

Display Method for Online Bookstore Emulating Real Bookstore by WebGL

Thanks to the spread of the Internet, opportunities to use online bookstores have been increasing in recent years. However, it is difficult for the typical customer to intuitively grasp which book sells well or which book is recommended by the shop. We have therefore developed a display system for virtual bookstores in which an image of books arranged in a bookshelf or stacked on a table is reproduced by using WebGL. Since the amount of book stock that is managed by the server side needs to be changed as orders come in from clients, we introduce a server side technology using a hypertext preprocessor (PHP).