The initial step of the thermal unfolding of 3-isopropylmalate dehydrogenase detected by the temperature-jump Laue method

A temperature-jump (T-jump) time-resolved X-ray crystallographictechnique using the Laue method was developed to detect small,localized structural changes of proteins in crystals exposedto a temperature increase induced by laser irradiation. In achimeric protein between thermophilic and mesophilic 3-isopropylmalatedehydrogenases (2T2M6T), the initial structural change uponT-jump to a denaturing temperature (~90°C) was found tobe localized at a region which includes a ß-turn and aloop located between the two domains of the enzyme. A mutant,2T2M6T-E110P/S111G/S113E, having amino acid replacements inthis ß-turn region with the corresponding residues ofthe thermophilic enzyme, showed greater stability than the originalchimera (increase of T_m by ~10°C) and no T-jump-inducedstructural change in this region was detected by our method.These results indicate that thermal unfolding of the originalchimeric enzyme, 2T2M6T, is triggered in this ß-turn region.     

Identification of Tumor Suppressive Genes Regulated by miR-31-5p and miR-31-3p in Head and Neck Squamous Cell Carcinoma

We identified the microRNA (miRNA) expression signature of head and neck squamous cell carcinoma (HNSCC) tissues by RNA sequencing, in which 168 miRNAs were significantly upregulated, including both strands of the miR-31 duplex (miR-31-5p and miR-31-3p). The aims of this study were to identify networks of tumor suppressor genes regulated by miR-31-5p and miR-31-3p in HNSCC cells. Our functional assays showed that inhibition of miR-31-5p and miR-31-3p attenuated cancer cell malignant phenotypes (cell proliferation, migration, and invasion), suggesting that they had oncogenic potential in HNSCC cells. Our in silico analysis revealed 146 genes regulated by miR-31 in HNSCC cells. Among these targets, the low expression of seven genes (miR-31-5p targets: CACNB2 and IL34; miR-31-3p targets: CGNL1, CNTN3, GAS7, HOPX, and PBX1) was closely associated with poor prognosis in HNSCC. According to multivariate Cox regression analyses, the expression levels of five of those genes (CACNB2: p = 0.0189; IL34: p = 0.0425; CGNL1: p = 0.0014; CNTN3: p = 0.0304; and GAS7: p = 0.0412) were independent prognostic factors in patients with HNSCC. Our miRNA signature and miRNA-based approach will provide new insights into the molecular pathogenesis of HNSCC.     

Subcooled Flow Boiling in a Minichannel

It has been considered that dry-out occurs easily in boiling heat transfer for a small channel, a mini- or microchannel, because the channel was easily filled with coalescing vapor bubbles. In the present study, the experiments of subcooled flow boiling of water were performed under atmospheric conditions for a horizontal rectangular channel for which the size is 1 mm height and 1 mm width, with a flat heating surface of 10 mm length and 1 mm width placed on the bottom of the channel. The heating surface has a top of copper heating block and is heated by ceramic heaters. In the high heat flux region of nucleate boiling, about 70–80% of the heating surface was covered with a large coalescing bubble and the boiling reached critical heat flux as observed by high-speed video. In the beginning of transition boiling, coalescing bubbles were collapsed to many fine bubbles and microbubble emission boiling was observed at liquid subcooling higher than 30 K. The maximum heat flux obtained was 8 MW/m² (800 W/cm²) at liquid subcooling of higher than 40 K and a liquid velocity of 0.5 m/s. However, the surface temperature was very much higher than that of a centimeter-scale channel. The high-speed video photographs indicated that microbubble emission boiling occurs in the deep transition boiling region.     

Electron microscopy at a sub-50 pm resolution

An aberration-corrected electron microscope developed in CREST project has been applied for imaging atoms and clusters buried inside crystals. The resolution of the microscope in scanning transmission electron microscopy (STEM) has experimentally proved to be better than 47 pm by use of a cold-field emission gun at 300 kV. The high resolution has given an advantage for imaging light elements such as lithium atoms discriminating one by one. Moreover, a three-dimensional structure imaging has been demonstrated for dopant clusters by a sub-50 pm STEM, using its high depth resolution.     

Verification of a model for estimating the void fraction in a three-component randomly packed bed

The void fraction in a three-component randomly packed bed was calculated from the authors' model, and the calculated values were compared with published experimental data for spherical and irregularly shaped particles and with results from computer simulations. Results from the model were in good agreement with simulated and published experimental data.     

Quantitative bioimaging analysis of gonads in olvas-GFP/ST-II YI Medaka (transgenic Oryzias latipes) exposed to ethinylestradiol

This study investigates the adverse and persistent effects of ethinylestradiol (EE2) on mature gonads of transgenic olvas-GFPIST II-YI medaka (Oryzias latipes). The measurement of gonadal size calculating the GFP-fluorescent area was used as a technique that enabled monitoring gonads in living specimens by GFP fluorescence. First, mature medaka were exposed to EE2 (47.8-522 ng/L) for 4 weeks. The gonads showed a significant reduction of the GFP-fluorescent area and Gonadosomatic Index in males exposed to EE2 at >216 ng/L and females exposed at 522 ng/L. Histologically, males at all treatments exhibited testis-ova and additionally, high connective tissue prevalence at > or =216 ng/L. Next, mature male medaka were exposed to EE2 (43.7-473 ng/L) for 3 weeks and allowed to depurate for 6 weeks, to investigate persistent effects of EE2. Continuous gonad observation showed that GFP began to decline 3 weeks after initial exposure to > or =215 ng/L. After depuration, the gonad's fluorescent areas gradually recovered, with no statistical difference at the end of the depuration period; normal spermatogenesis was present in these individuals. Alterations in GFP fluorescence clearly indicate the condition of the gonad in transgenic medaka and this strain showed a facilitated screening fish model to detect the adverse effects on the gonad by estrogenic chemicals.     

Beneficial Effects of Supplementation of the Rare Sugar “D‐allulose” Against Hepatic Steatosis and Severe Obesity in Lepob/Lepob Mice

A rare sugar, D‐allulose (also called D‐psicose), has recently been applied as a food supplement in view of controlling diabetes and obesity in Japan. D‐allulose has been proven to have unique effects against hyperglycemia and hyperlipidemia in a number of studies using several species of rats and mice. However, the antiobesity effects of D‐allulose have not yet been assessed in Lep^ob/Lep^ob (ob/ob) mice. Therefore, this study explored the dietary supplemental effects of this sugar in leptin‐deficient ob/ob mice. Consequently, the subchronic ingestion of D‐allulose in ob/ob mice for 15 wk significantly decreased the body and liver weights, and the loss of body weight was involved in the reduction of the total fat mass, including abdominal visceral fat, and not fat‐free body mass, including muscle. Furthermore, D‐allulose improved hepatic steatosis, as evaluated using hepatic histological studies and MRI. In the normal mice, none of these parameters were influenced by the single or long‐term ingestion of D‐allulose. These results indicate that dietary supplementation of D‐allulose especially influences postprandial hyperglycemia and obesity‐related hepatic steatosis, without exercise therapy or dietary restriction. Therefore, D‐allulose may be useful as a supplement for preventing and improving obesity and obesity‐related disorders.     

High Magnetic Field Micro-Calorimeter and Application for BE-Condensation of Cr5+ Dimer System

A micro-calorimeter working at ultra-low temperature down to 75 mK and high magnetic field up to 17.5 T has been developed. A typical sample weight is as small as 100 μg. A magnetic phase diagram of magnon Bose-Einstein condensation (BEC) in Ba₃Cr₂O₈ is determined. The scaling index of the phase boundary is very close to that of the isotropic three dimensional BEC at low temperature, in spite of the existence of Dzyaloshinskii-Moriya (DM) interaction which is confirmed in the previous ESR measurement.     

Impact of Oncogenic Targets by Tumor-Suppressive miR-139-5p and miR-139-3p Regulation in Head and Neck Squamous Cell Carcinoma

We newly generated an RNA-sequencing-based microRNA (miRNA) expression signature of head and neck squamous cell carcinoma (HNSCC). Analysis of the signature revealed that both strands of some miRNAs, including miR-139-5p (the guide strand) and miR-139-3p (the passenger strand) of miR-139, were downregulated in HNSCC tissues. Analysis of The Cancer Genome Atlas confirmed the low expression levels of miR-139 in HNSCC. Ectopic expression of these miRNAs attenuated the characteristics of cancer cell aggressiveness (e.g., cell proliferation, migration, and invasion). Our in silico analyses revealed a total of 28 putative targets regulated by pre-miR-139 (miR-139-5p and miR-139-3p) in HNSCC cells. Of these, the GNA12 (guanine nucleotide-binding protein subunit alpha-12) and OLR1 (oxidized low-density lipoprotein receptor 1) expression levels were identified as independent factors that predicted patient survival according to multivariate Cox regression analyses (p = 0.0018 and p = 0.0104, respectively). Direct regulation of GNA12 and OLR1 by miR-139-3p in HNSCC cells was confirmed through luciferase reporter assays. Moreover, overexpression of GNA12 and OLR1 was detected in clinical specimens of HNSCC through immunostaining. The involvement of miR-139-3p (the passenger strand) in the oncogenesis of HNSCC is a new concept in cancer biology. Our miRNA-based strategy will increase knowledge on the molecular pathogenesis of HNSCC.