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.     

Bread quality of frozen dough substituted with modified tapioca starches

Rheological properties of dough and bread quality of frozen dough-bread containing 18.4% of hydroxypropylated (HTS), acetylated (ATS), and phosphorylated cross-linked (PTS) tapioca starch with different degrees of modification and 1.6% of dried powdered gluten were compared to the same amount of native tapioca starch (NTS) or wheat flour-bread. Doughs substituted with native or modified tapioca starches had the same mixing tolerance as 100% wheat flour. The dough was frozen and stored for 1 week at −18°C, and thawed (one freeze-cycle). The amount of freezable water in the dough substituted with native or modified tapioca starches was not significantly different from that of wheat flour. Frozen dough-bread substituted with highly modified HTS (degree of substitution; DS 0.09–0.11) retarded bread staling, while lowly modified HTS (DS 0.06–0.07) or ATS (DS 0.02–0.04), and PTS (0.004–0.020% phosphoryl content) substitution fastened bread staling as compared with frozen dough-bread baked from wheat flour. The breadcrumbs containing HTS and ATS felt tacky, whereas the bread containing PTS was dry feel. HTS and ATS swelled and collapsed easily during heating, while PTS was difficult to swell and disperse as compared with NTS, therefore the gelatinization properties seemed to affect the texture of bread. Breadcrumb containing HTS showed small firmness during storage, and highly modified HTS-h (DS 0.1) was the smallest. This means highly hydroxypropylated tapioca starch significantly retards bread staling. Staling properties and texture of frozen dough-bread with various tapioca starches were the same as conventional bread baked with the same amount of tapioca starches. These results suggest that a one freeze–thaw cycle and a 1-week frozen period do not change characteristics of starch, gelatinization and retrogradation properties as compared with the conventional method, and the highly modified HTS-h is prominent anti-staling food-stuff in frozen dough.     

Evolution of microhardness and microstructure in a cast Al–7 % Si alloy during high-pressure torsion

Disks of a cast Al–7 % Si alloy were processed through high-pressure torsion (HPT) for 1/4, 1/2, 1, 5, and 10 revolutions under a pressure of 6.0 GPa and at temperatures of 298 and 445 K. The hardness of the samples after processing was significantly higher than in the cast sample, and the hardness profiles across the samples became more uniform with increasing numbers of turns. Processing at higher temperature gave lower hardness values. Experiments were conducted to examine the effects of HPT processing on various microstructural aspects of the cast Al–7 % Si alloy such as the grain size, the Taylor factor, and the fraction of high-angle grain boundaries. The results demonstrate that there is a correlation between trends in the microhardness values and the observed microstructures.     

The metabolism and distribution of sesame lignans (sesamin and episesamin) in rats

In this study’ we examined the distribution and metabolism of refined sesame oil lignans (sesamin and episesamin) in rat. For 8 wk rats were fed the diet including 0.5% (w/w) sesame lignans (sesamin and episesamin) with 5% (w/w) corn oil or eicosapentaenoic acid (EPA)-rich oil. The concentrations of sesamin and episesamin in rat liver after their administration for 8 wk were very low; both of them were less than 0.5 μg/g liver. These were observed in both oil groups although the fatty acid compositions of dietary oils were completely different. No significant difference existed in lymphatic absorption between sesamin and episesamin. To investigate the distribution of sesamin and episesamin in rats’ the concentrations of sesamin and episesamin were determined in tissues and serum within 24h after administration to rats. Sesamin and episesamin may be’ at first’ incorporated into the liver and then transported to the other tissues (lung’ heart’ kidney’ and brain). They are lost from the body within 24h after administration. There was no significant difference in lymphatic absorption between sesamin and episesamin’ but the amount of sesamin was significantly lower than that of episesamin in all tissues and serum. These results suggest that sesamin is absorbed in lymph the same as episesamin’ but that sesamin is subsequently metabolized faster by the liver.     

G-Protein-Coupled Receptors Mediate Modulations of Cell Viability and Drug Sensitivity by Aberrantly Expressed Recoverin 3 within A549 Cells

To elucidate the currently unknown molecular mechanisms responsible for the aberrant expression of recoverin (Rec) within cancerous cells, we examined two-dimensional (2D) and three-dimensional (3D) cultures of Rec-negative lung adenocarcinoma A549 cells which had been transfected with a plasmid containing human recoverin cDNA (A549 Rec) or an empty plasmid as a mock control (A549 MOCK). Using these cells, we measured cytotoxicity by several anti-tumor agents (2D), cellular metabolism including mitochondrial and glycolytic functions by a Seahorse bio-analyzer (2D), the physical properties, size and stiffness of the 3D spheroids, trypsin sensitivities (2D and 3D), and RNA sequencing analysis (2D). Compared with the A549 MOCK, the A549 Rec cells showed (1) more sensitivity toward anti-tumor agents (2D) and a 0.25% solution of trypsin (3D); (2) a metabolic shift from glycolysis to oxidative phosphorylation; and (3) the formation of larger and stiffer 3D spheroids. RNA sequencing analysis and bioinformatic analyses of the differentially expressed genes (DEGs) using Gene Ontology (GO) enrichment analysis suggested that aberrantly expressed Rec is most likely associated with several canonical pathways including G-protein-coupled receptor (GPCR)-mediated signaling and signaling by the cAMP response element binding protein (CREB). The findings reported here indicate that the aberrantly expressed Rec-induced modulation of the cell viability and drug sensitivity may be GPCR mediated.     

Using deformation mechanism maps to depict flow processes in superplastic ultrafine-grained materials

Deformation mechanism maps are a very useful tool for displaying deformation mechanisms as a function of the three fundamental parameters of high temperature flow: the applied stress, the testing temperature and the grain size of the material. These maps are used extensively in the field of high temperature creep but there has been very little use with ultrafine-grained (UFG) metals. This article reviews the principles of deformation mechanism maps, presents examples of maps for some representative metals processed by equal-channel angular pressing or high-pressure torsion and then describes a simple procedure for constructing maps for UFG materials.     

Note: In vivo pH imaging system using luminescent indicator and color camera

Microscopic in vivo pH imaging system is developed that can capture the luminescent- and color-imaging. The former gives a quantitative measurement of a pH distribution in vivo. The latter captures the structural information that can be overlaid to the pH distribution for correlating the structure of a specimen and its pH distribution. By using a digital color camera, a luminescent image as well as a color image is obtained. The system uses HPTS (8-hydroxypyrene-1,3,6-trisulfonate) as a luminescent pH indicator for the luminescent imaging. Filter units are mounted in the microscope, which extract two luminescent images for using the excitation-ratio method. A ratio of the two images is converted to a pH distribution through a priori pH calibration. An application of the system to epidermal cells of Lactuca Sativa L is shown.     

Observation of fracture sealing in high-strength and ultra-low-permeability concrete by micro-focus X-ray CT and SEM/EDX

Fracture sealing by precipitation is known to occur in high-strength and ultra-low-permeability concrete (HSULPC) immersed in water. Because a high ability to retard radionuclide migration is required for HSULPC, understanding both the sealing process and the composition of sealing deposits is important to identify optimum conditions for significant sealing. In this study, sealing of a macro-fractured HSULPC specimen with initial aperture of approximately 0.1 mm was investigated in simulated seawater over 49 days. The composition of sealing deposits at 49 days after immersion was clarified by scanning electron microscopy and energy dispersive X-ray spectroscopy (SEM/EDX), and the progress of sealing during the 49 days was clarified by image analysis with micro-focus X-ray computed tomography (X-rayCT). Both the SEM/EDX and X-rayCT results showed that significant sealing was attained only near the outermost part of the specimen. The SEM/EDX results showed that a thin brucite layer formed on the entire specimen surface over which significant precipitation of calcium carbonate occurred and sealed the macro-fracture only near the outermost part of the specimen. The X-rayCT results indicated that the amount of sealing deposits in the macro-fracture ( \(P_\mathrm{seal})\) reached 70 % in the mostly sealed region at 49 days and the rate of change in \(P_\mathrm{seal}\) became maximum \((3.7\,\% \hbox { day}^{-1})\) during 7–21 days after immersion, then decreased. In conclusion, the findings in this study represent an important clue in the search for optimum conditions to achieve fracture sealing in HSUPLC.     

Synthesis of the Right-Side Structure of Type B Physalins

We present a full account of our synthetic studies on the racemic DEFGH-ring moiety of physalins, featuring domino ring transformation of a tricyclic key intermediate. We also report the results of a detailed mechanistic examination of the domino ring transformation, as well as a reoptimization of the 2,3-Wittig rearrangement and methylation steps. Furthermore, we have newly established a method for the preparation of an optically active synthetic intermediate by enzymatic kinetic resolution. Our work provides access to both natural and nonnatural right-side physalin structures.