The Emi2 Protein of Saccharomyces cerevisiae is a Hexokinase Expressed under Glucose Limitation

Hexokinases catalyze glucose phosphorylation at the first step in glycolysis in eukaryotes. In the budding yeast Saccharomyces cerevisiae , three enzymes for glucose phosphorylation have long been known: Hxk1, Hxk2, and Glk1. In this study, we focus on Emi2, a previously uncharacterized hexokinase-like protein of S. cerevisiae . Our data show that the recombinant Emi2 protein (rEmi2), expressed in Escherichia coli , possesses glucose-phosphorylating activity in the presence of ATP and Mg ²⁺ . It was also found that rEmi2 phosphorylates not only glucose but also fructose, mannose and glucosamine in vitro . In addition, we examined changes in the level of endogenous Emi2 protein in S. cerevisiae in the presence or absence of glucose and a non-fermentable carbon source. We found that the expression of Emi2 protein is tightly suppressed during proliferation in high glucose, while it is strongly upregulated in response to glucose limitation and the presence of a non-fermentable carbon source. Our data suggest that the expression of the endogenous Emi2 protein in S. cerevisiae is regulated under the control of Hxk2 in response to glucose availability in the environment.     

Ergonomics solution for crossing collisions based on field assessment of visual environment at urban intersections in Japan

This paper aims to assess quantitatively the actual visual environment of uncontrolled urban downtown intersections in Japan in relation to frequently occurring crossing collisions and to discuss the safety countermeasures for them. In Field Study 1 dealing with direct visibility, our ultra-wide-angle photograph analysis revealed that most of the right/left-ward visible range at 11 intersections were insufficient to check safety, and the quality of direct visibility was closely associated with causing crossing collisions. The countermeasures to reduce a blind area were determined to be a top priority. In Field Study 2 dealing with indirect visibility, more than half of the 25 traffic convex mirrors had marked shortcomings for preventive safety, and ergonomics guidelines ensuring indirect visibility were proposed for installing traffic convex mirrors. Low-cost/low-technology-oriented countermeasures are highly recommended to obtain clear/sufficient images of crucial information satisfying drivers' requirements on traffic convex mirrors in accordance with those ergonomics guidelines was highly recommended. Crossing collisions could be prevented by improvement of poor direct and indirect visibility.     

Physical indices for judging appearance harmony of materials

In visual design, harmony refers to the similarity among objects and the components of these objects that appear as if they belong together. Harmony is related to the body, mind, and emotions as we relate to our living space, which means that the harmony of real objects is an important characteristic. Although the appearance harmony in specific materials has been investigated, harmony among different materials has not received adequate attention. In a previous study, we investigated the appearance harmony among various materials by psychophysical experiments. In this study, we correlated the psychophysical evaluations obtained from the experiments and the physical properties of the materials. By analyzing the relationship between psychophysical evaluations and the physical measurements for each material pairs based on cluster analysis, we found that the property of texture represented by the anisotropy histogram and that of reflectance obtained from the data of the bidirectional reflectance distribution function are important indices that can be used to judge the harmony of materials. In particular, the texture properties were more effective in judging the appearance harmony of material pairs in the fixed condition for illumination and viewing angles, and the reflectance properties were superior for the material pairs in the variable condition for illumination and viewing angles.     

Investigation of metallic color perception using real‐world materials

Metallic colors have a unique appearance of glossiness with features such as highlights, contrast, and reflections on their surface, and therefore, metallic objects are very attractive to humans. Especially, gold, silver, and copper colors are familiar metals used as decorative materials, coins, and other furnishings. However, the mechanism and condition of metallic perception have not been fully investigated. There are a few studies for investigating metallic perception using rendered patches or images, but there is no study using real‐world objects. In our previous study, we developed a simple representation technique that made real objects appear to be made of gold by projecting a solid color onto a target nonmetallic object. By using the representation technique, in this study, we have further investigated the perception of metallic appearance such as gold, silver, and copper using real‐world materials, and analyzed the difference between these metallic perceptions. Our results indicate that the perception of the metallic object is different for gold, silver, and copper. Our new findings are as follows: the glitter required for the perception of gold and silver becomes an obstacle to the perception of copper; the metallic perception reveals that learning experience might be strongly affecting; and luminance adjustment is sensitive to the perception of metallic objects.     

Preparation of ultrafine polyurethane fiber web by laser‐electrospinning combined with air blowing

Thermoplastic polyurethane fiber webs were prepared using a laser‐heated electrospinning process combined with air blowing. The effect of spinning conditions such as air flow rate and air temperature on fiber diameter and molecular weight was investigated. Although the average fiber diameter decreased with increased air flow rate at each air temperature, the diameter increased when the air flow rate was >15 NL min^?1. In addition, the fiber was comparatively thicker with an increase in the air temperature. The variation in the fiber diameter tends to increase with the air flow rate, and a reduction in the molecular weight of the fiber by thermal degradation was suppressed. The thinnest and most uniform fiber with a diameter of 0.9 µm and a diameter coefficient variation of 15% was obtained at an air temperature of 25°C under an air flow rate of 15 NL min^?1. This fiber also had a minimum of decreased molecular weight. POLYM. ENG. SCI., 54:2605–2609, 2014. © 2013 Society of Plastics Engineers     

Characterization of alternative plasticizers in poly(vinyl chloride) sheets for blood containers

This study aimed to optimize the ratio of dioctyl 4‐cyclohexene‐1,2‐dicarboxylate (DOTH) and di‐isononyl‐cyclohexane‐1,2‐dicarboxylate (DINCH^®) for use as plasticizers in poly(vinyl chloride) (PVC) sheets. We also evaluated the biological safety of DOTH for its potential to be part of a safe PVC‐based blood container. The suppression of hemolysis in mannitol‐adenine‐phosphate / red cell concentrates (MAP/RCC) with DOTH/(DINCH^®‐PVC) sheets and the elution of plasticizers from the sheets increased with higher DOTH compositions. The properties of the PVC sheet containing DOTH and DINCH^® in the ratio of 25:33 parts against PVC 100 parts as a weight were almost identical to the PVC sheet made of di(2‐ethylhexyl) phthalate. From a subchronic toxicity test, DOTH did not show any adverse effects on all organs, including the testes, epididymis, liver, and kidneys. The no‐observed‐adverse‐effect level was 300 mg/kg body weight/day in a rat. These results suggest that DOTH/DINCH^® (25:33) is a promising candidate for the replacement of di(2‐ethylhexyl) phthalate in blood containers. J. VINYL ADDIT. TECHNOL., 22:520–528, 2016. © 2015 Society of Plastics Engineers     

Levels of Vibrio parahaemolyticus and thermostable direct hemolysin gene-positive organisms in retail seafood determined by the most probable number-polymerase chain reaction (MPN-PCR) method

The incidence and levels of Vibrio parahaemolyticus and thermostable direct hemolysin gene (tdh)-positive organisms in retail seafood were determined. The most probable number-polymerase chain reaction (MPN-PCR) method using a PCR procedure targeting the species-specific thermolabile hemolysin gene (tlh) and tdh was used to determine the levels of V. parahaemolyticus and tdh-positive organisms, respectively. In seafood for raw consumption, V. parahaemolyticus was found in four (13.3%) of 30 fish samples, 11 (55.0%) of 20 crustacean samples, and 29 (96.7%) of 30 mollusc samples. Levels of V. parahaemolyticus were below 10(4) MPN/100 g in all fish and crustacean samples tested. However, they were above 10(4) MPN/100 g in 11 (36.7%) of the 30 mollusc samples. In all seafood for raw consumption, the level of tdh-positive organisms was below the limit of detection (< 30 MPN/100 g). In seafood for cooking, V. parahaemolyticus was found in 15 (75.0%) of 20 fish samples, nine (45.0%) of 20 crustacean sample, and 20 (100%) of 20 mollusc samples. Levels of V. parahaemolyticus were above 10(4) MPN/100 g in only three (15.0%) and one (5.0%) of the 20 fish and 20 crustacean samples, respectively. However, they were above 10(4) MPN/100 g in 18 (90.0%) of the 20 mollusc samples. In seven (35.0%) of the 20 mollusc samples, tdh-positive organisms were found and their levels ranged from 3.6x10 to 1.1 x 103 MPN/100 g. From four of seven tdhpositive samples, tdh-positive V. parahaemolyticus was isolated.     

Computer-aided design of a factor Xa inhibitor by using MCSS functionality maps and a CAVEAT linker search

We have investigated a new approach to efficiently find a novel inhibitor against a serine protease (i.e. an activated coagulation factor X, FXa) by using de novo design programs and the X-ray crystal structure of the target enzyme. FXa is a coagulant enzyme that generates thrombin (a serine protease) and participates in both intrinsic and extrinsic coagulation pathways. We adopted multiple copy simultaneous search (MCSS) and CAVEAT linker search techniques, which disclosed a novel FXa inhibitor (T01312) consisting of two binding moieties (the benzamidinyl and adamantyl groups) and a linker unit (the carboxybenzylamine group). The inhibitory activity of T01312 against FXa was determined to be a small K(i)-value of 48nM, which is two orders of magnitude smaller than that against thrombin. An X-ray crystal analysis of T01312 complexed with trypsin (an analogue of FXa) and docking studies of T01312 with trypsin and FXa showed that: (i) the benzamidinyl group is a predominant binding moiety in the anionic pocket (S1 site) with an asparatic acid residue; (ii) a hydrophobic pocket (S4 site) is the binding site of the adamantyl group; (iii) the carboxylate group of the linker contributes to the selectivity for FXa against thrombin. Thus, the combination of the knowledge of the X-ray crystal structure of the target molecule with MCSS and CAVEAT linker search techniques proved to be an effective hit-finding method that does not require the screening of huge compound libraries.