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Purpose: The purpose of this study was to examine the location where the femoral artery contacts the vastus medialis at the adductor tendon hiatus, which is important when using the subvastus approach in medial closed wedge distal femoral osteotomy. We evaluated the correlation between differences in height, vastus medialis morphology, and lower limb alignment. Methods: Sixty knees (16 male, 44 female) that underwent plain computer tomography (CT) were included. Using the radiographic hip‐knee‐ankle (HKA) angle as a reference, the knees were divided into three groups of 20 knees: valgus, varus, and neutral. The mechanical lateral distal femoral angle (mLDFA) and distance from the medial femoral epicondyle to the centre of the femoral head (D1) were measured on full‐length weight‐bearing anteroposterior radiographs. The first cross‐section on CT where the vastus medialis muscle and femoral artery connect was defined as the cross‐sectional image for measurement. The direct distance from the medial epicondyle to the measured cross‐sectional image (D2) was measured in the coronal view. The ratio of the vastus medialis muscle width to the femoral posterior wall width was defined as the vastus medialis muscle coverage ratio (CR). Correlations between each measurement and group were evaluated. Results: There was a positive correlation between D1 and D2 in the overall, neutral, and varus groups; however, there was no correlation in the valgus group. A positive correlation was observed in terms of the relationship between CR and D2 in the overall and valgus groups. In addition, there was no statistically significant difference in the correlation between the mLDFA and D2, with patient height as a control variable overall and in all groups. Conclusion: In the valgus group, distance to the adductor hiatus was correlated with vastus medialis coverage. Overhang of the vastus medialis may be an important influencing factor of femoral and popliteus artery position. Level of Evidence: Level III, retrospective cohort study. [ABSTRACT FROM AUTHOR]

Proline is the most abundant amino acid in wine and beer, because the yeast Saccharomyces cerevisiae hardly assimilates proline during fermentation processes. Our previous studies showed that arginine induces endocytosis of the proline transporter Put4, resulting in inhibition of proline utilization. We here report a possible role of arginine sensing in the inhibition of proline utilization. We first found that two basic amino acids, ornithine, and lysine, inhibit proline utilization by inducing Put4 endocytosis in a manner similar to arginine, but citrulline does not. Our genetic screening revealed that the arginine transporter Can1 is involved in the inhibition of proline utilization by arginine. Intriguingly, the arginine uptake activity of Can1 was not required for the arginine‐dependent inhibition of proline utilization, suggesting that Can1 has a function beyond its commonly known function of transporting arginine. More importantly, our biochemical analyses revealed that Can1 activates signaling cascades of protein kinase A in response to extracellular arginine. Hence, we proposed that Can1 regulates proline utilization by functioning as a transceptor possessing the activity of both a transporter and receptor of arginine. Take‐away: Basic amino acids inhibit proline utilization by inducing Put4 endocytosis.Can1 is involved in the inhibition of proline utilization in yeast.Can1 is a transceptor acting as both a transporter and receptor of arginine. [ABSTRACT FROM AUTHOR]

Summary: In natural long days, the florigen gene FLOWERING LOCUS T (FT) shows a bimodal expression pattern with morning and dusk peaks in Arabidopsis. This pattern differs from the one observed in the laboratory, and little is known about underlying mechanisms. A red : far‐red (R : FR) ratio difference between sunlight and fluorescent light causes this FT pattern mismatch.We showed that bimodal FT expression patterns were induced in a day longer than 14 h with sunlight R : FR (= c. 1) conditions. By circadian gating experiments, we found that cumulative exposure of R : FR‐adjusted light (R : FR ratio was adjusted to 1 with FR supplement) spanning from the afternoon to the next morning required full induction of FT in the morning. Conversely, only 2 h of R : FR adjustment in the late afternoon was sufficient for FT induction at dusk.We identified that phytochrome A (phyA) is required for the morning FT expression in response to the R : FR adjustment on the previous day. As a part of this mechanism, we showed that PHYTOCHROME‐INTERACTING FACTOR 7 contributes to FT regulation.Our results suggest that phyA‐mediated high‐irradiance response and the external coincidence mechanism contribute to morning FT induction under natural long‐day conditions. [ABSTRACT FROM AUTHOR]

Nitric oxide (NO) is a ubiquitous signaling molecule in various organisms. In the yeast Saccharomyces cerevisiae, NO functions in both cell protection and cell death, depending on its concentration. Thus, it is important for yeast cells to strictly regulate NO concentration. The transcription factor Fzf1, containing five zinc fingers, is reportedly important for NO homeostasis by regulating the expression of the YHB1 gene, which encodes NO dioxygenase. However, the mechanism by which NO activates Fzf1 is still unclear. In this study, we showed that NO activated Fzf1 specifically at the protein level by RT‐qPCR and Western blotting. Our further transcriptional analyses indicated that cysteine residues in the fourth zinc finger (ZF4) are required for the NO‐responsive activation of Fzf1. Additionally, the present results suggest that ZF4 is important for the protein stability of Fzf1. From these results, we proposed possible mechanisms underlying Fzf1 activation. [ABSTRACT FROM AUTHOR]

The reduced form of nicotinamide adenine dinucleotide phosphate (NADPH), which is required for various redox systems involving antioxidative stress enzymes, is thus important for stress tolerance mechanisms. Here, we analyzed the stress response of the NADPH‐depleted cells of Saccharomyces cerevisiae. A cell viability assay showed that the NADPH depletion induced by disruption of the ZWF1 gene encoding glucose‐6‐phosphate dehydrogenase, which is the major determinant of the intracellular NADPH/NADP+ ratio, enhanced the tolerance of S. cerevisiae to both oxidative and nitrosative stresses. The subsequent analyses demonstrated that the antioxidative transcriptional factor Yap1 was activated and the cytosolic catalase Ctt1, whose expression is regulated by Yap1, was upregulated in zwf1Δ cells irrespective of the presence or absence of stress stimuli. Moreover, deletion of the YAP1 or CTT1 gene inhibited the increased stress tolerance of zwf1Δ cells, indicating that Ctt1 dominantly contributed to the higher stress tolerance of zwf1Δ cells. Our findings suggest that an NADPH‐independent mechanism enhances oxidative and nitrosative stress tolerance in ZWF1‐lacking yeast cells. [ABSTRACT FROM AUTHOR]

Proline is a predominant amino acid in grape must, but it is poorly utilized by the yeast Saccharomyces cerevisiae in wine‐making processes. This sometimes leads to a nitrogen deficiency during fermentation and proline accumulation in wine. Although the presence of other nitrogen sources under fermentation conditions is likely to interfere with proline utilization, the inhibitory mechanisms of proline utilization remain unclear. In this study, we examined the effect of arginine on proline utilization in S. cerevisiae. We first constructed a proline auxotrophic yeast strain and identified an inhibitory factor by observing the growth of cells when proline was present as a sole nitrogen source. Intriguingly, we found that arginine, and not ammonium ion, clearly inhibited the growth of proline auxotrophic cells. In addition, arginine prevented the proline consumption of wild‐type and proline auxotrophic cells, indicating that arginine is an inhibitory factor of proline utilization in yeast. Next, quantitative polymerase chain reaction (PCR) analysis showed that arginine partially repressed the expression of genes involved in proline degradation and uptake. We then observed that arginine induced the endocytosis of the proline transporters Put4 and Gap1, whereas ammonium induced the endocytosis of only Gap1. Hence, our results may involve an important mechanism for arginine‐mediated inhibition of proline utilization in yeast. The breeding of yeast that utilizes proline efficiently could be promising for the improvement of wine quality. [ABSTRACT FROM AUTHOR]

Ricinoleic acid (RA) is the main fatty acid component of castor oil and was found to inhibit Ca2+‐signal transduction pathway‐mediated cell cycle regulation in a yeast‐based drug screening assay. RA is expected to have antidiabetic, antiallergy, and/or anticancer properties but its target molecule is unknown. To identify a novel pharmacological effect of RA, we investigated its target molecule in the Ca2+‐signal transduction pathway. RA inhibition of calcineurin (CN) was examined in a yeast‐based CN inhibitor screening assay using the rsp5A401E mutant and in a phosphatase assay using recombinant human CN. RA showed growth‐restoration activity at 5 μg/spot in the CN inhibitor screening assay with the rsp5A401E yeast strain. Furthermore, it directly inhibited CN without immunophilins at Ki = 33.7 μM in a substrate‐competitive manner. The effects of RA on CN in mammalian cells were further evaluated by measuring β‐hexosaminidase (β‐HEX) release in RBL‐2H3 cells. RA at 50 μM suppressed the release of β‐HEX from RBL‐2H3 cells. Moreover, this compound was found to inhibit glycogen synthase kinase‐3β (GSK‐3β), as determined by a kinase assay using recombinant human GSK‐3β. RA inhibited GSK‐3β at Ki = 1.43 μM in a peptide substrate‐competitive manner. The inhibition of GSK‐3β by this molecule was further assessed in mammalian cells by measuring the inhibition of glucose production in H4IIE rat hepatoma cells. RA at 25 μM suppressed glucose production in these cells. These findings indicate that RA and/or castor oil could be a useful functional fatty acid to treat allergy or type 2 diabetes. [ABSTRACT FROM AUTHOR]

The 14th International Congress on Yeasts ( ICY14) was held at Awaji Yumebutai International Conference Center (Awaji, Hyogo) in Japan from 11 to 15 September 2016. The main slogan of ICY14 was 'Yeasts for Global Happiness', which enabled us to acknowledge the high-potential usefulness of yeasts contributing to the global happiness in terms of food/beverage, health/medicine and energy/environment industries, as well as to basic biosciences. In addition, two more concepts were introduced: 'from Japan to the world' and 'from senior to junior'. As it was the first ICY meeting held in Japan or other Asian countries, ICY14 provided a good opportunity to widely spread the great achievements by Japanese and Asian yeast researchers, such as those by the 2016 Nobel Laureate Dr. Yoshinori Ohsumi, and also, to convey the fun and importance of yeasts to the next generation of researchers from Asia and all over the world. As a result, a total of 426 yeast lovers from 42 countries (225 overseas and 201 domestic participants) with different generations attended ICY14 to share the latest knowledge of a wide range of yeast research fields and to join active and constructive scientific discussions. [ABSTRACT FROM AUTHOR]

γ-Glutamyl kinase ( GK; the PRO1 gene product) is a key enzyme in the Saccharomyces cerevisiae proline biosynthesis pathway. Δpro1 cells are more sensitive to various stresses than wild-type cells, suggesting that GK has an alternative function independent of proline biosynthesis. We show that PRO1 genetically interacts with UBP3, which encodes ubiquitin-specific protease, and is required for selective autophagy of ribosomes (ribophagy). Interestingly, yeast cells with PRO1 deletion or expressing inactive GK display a defect for ribophagy but not for nonselective autophagy, indicating that GK activity is indispensable for ribophagy. Gene disruption analysis suggests that ribophagy is important for cell survival during nitrogen starvation. [ABSTRACT FROM AUTHOR]

During fermentation processes, Saccharomyces cerevisiae cells are exposed to multiple stresses, including a high concentration of ethanol that represents toxicity through intracellular reactive oxygen species (ROS) generation. We previously reported that proline protected yeast cells from damage caused by various stresses, such as freezing and ethanol. As an anti-oxidant, proline is suggested to scavenge intracellular ROS. In this study, we examined the role of intracellular proline during ethanol treatment in S. cerevisiae strains that accumulate different concentrations of proline. When cultured in YPD medium, there was a significant accumulation of proline in the put1 mutant strain, which is deficient in proline oxidase, in the stationary phase. Expression of the mutant PRO1 gene, which encodes the γ-glutamyl kinase variant (Asp154Asn or Ile150Thr) with desensitization to feedback inhibition by proline in the put1 mutant strain, showed a prominent increase in proline content as compared with that of the wild-type strain. The oxidation level was clearly increased in wild-type cells after exposure to ethanol, indicating that the generation of ROS occurred. Interestingly, proline accumulation significantly reduces the ROS level and increases the survival rate of yeast cells in the stationary phase under ethanol stress conditions. However, there was not a clear correlation between proline content and survival rate in yeast cells. An appropriate level of intracellular proline in yeast might be important for its stress-protective effect. Hence, the engineering of proline metabolism could be promising for breeding stress-tolerant industrial yeast strains. Copyright © 2016 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

Purine catabolism is regarded as a housekeeping function that remobilizes nitrogen for plant growth and development. However, emerging evidence suggests that certain purine metabolites might contribute to stress protection of plants. Here, we show that in Arabidopsis, the intermediary metabolite allantoin plays a role in abiotic stress tolerance via activation of abscisic acid ( ABA) metabolism. The aln loss-of-function of ALN, encoding allantoinase, results in increased allantoin accumulation, genome-wide up-regulation of stress-related genes and enhanced tolerance to drought-shock and osmotic stress in aln mutant seedlings. This phenotype is not caused by a general response to purine catabolism inhibition, but rather results from a specific effect of allantoin. Allantoin activates ABA production both through increased transcription of NCED3, encoding a key enzyme in ABA biosynthesis, and through post-translational activation via high-molecular-weight complex formation of BG1, a β-glucosidase hydrolysing glucose-conjugated ABA. Exogenous application of allantoin to wild-type plants also activates the two ABA-producing pathways that lead to ABA accumulation and stress-responsive gene expression, but this effect is abrogated in ABA-deficient and BG1-knockout mutants. We propose that purine catabolism functions not only in nitrogen metabolism, but also in stress tolerance by influencing ABA production, which is mediated by the possible regulatory action of allantoin. [ABSTRACT FROM AUTHOR]

doi: 10.1111/j.1741-2358.2012.00666.x Chewing number is related to incremental increases in body weight from 20 years of age in Japanese middle-aged adults Background: Eating habits are associated with both current obesity and incremental increases in body weight from young adulthood, but no study has focused on chewing number during meals among community residents. Objective: This study focused on the relationship between chewing number and incremental increases in body weight from 20 years of age. Methods: A total of 93 persons aged 35-61 years participated. The subjects were asked to set the device and record their chewing number during each meal on a particular day. They were also asked whether their body weight had increased by 10 kg or more since they were 20 years old. Results: The body weight of 28 subjects (30%) had increased more than 10 kg since the age of 20 years. Total chewing number showed a relationship with such body weight increases. The odds ratio of weight increments of more than 10 kg for the lowest tertile group was 4.6 [95% confidence interval (CI), 1.3-16.2] relative to the highest tertile group (Model 1). The odds ratio of weight increments for the lowest tertile group increased to 6.3 (95% CI, 1.6-25.4) in Model 2 and to 9.1 (95% CI, 1.7-49.8) in Model 3. Conclusion: Although this study was limited because it did not consider all risk factors, categorical chewing number was related independently to body weight increments of more than 10 kg from 20 years of age. [ABSTRACT FROM AUTHOR]

Rsp5, an essential HECT-type ubiquitin ligase, is the only yeast Saccharomyces cerevisiae member of the Nedd4 family. Rsp5 triggers the ubiquitination-dependent endocytosis of the general amino acid permease Gap1 in response to a good nitrogen source. Previously, we showed that the Thr357Ala/Lys764Glu variant Rsp5 induces the constitutive inactivation of Gap1, which is mainly involved in uptake of the toxic proline analogue, l-azetidine-2-carboxylate ( AZC). Here, our experimental results indicated that the Thr357Ala substitution in the substrate-recognizing WW2 domain of Rsp5 constitutively causes the down-regulation of four proline permeases (Gap1, Put4, Agp1 and Gnp1), leading to AZC tolerance to yeast cells. In RSP5T357A cells, Gap1 was highly ubiquitinated and constantly delivered to the vacuole from the Golgi without sorting to the plasma membrane. Analyses of RSP5 mutants using antiphosphopeptide antibody suggest that Thr phosphorylation occurred in all three WW domains and, interestingly, that Thr357 in the WW2 domain was phosphorylated, in agreement with the in vitro result for the mouse Rsp5 orthologue. Furthermore, the phosphorylation-mimic mutant (Thr357Asp) showed strong sensitivity to AZC. From these results, we propose a possible mechanism involved in the regulation of Rsp5 activity for Gap1 down-regulation via the phosphorylation of a conserved Thr357 in the Nedd4 family. [ABSTRACT FROM AUTHOR]

We investigated the optical properties of La2 Zr2 O7 transparent ceramics and found that it showed abnormal partial dispersion. The origin of the abnormal partial dispersion was also investigated by measuring the vacuum ultraviolet reflectance and infrared absorption and by performing first-principle calculations. All the measurements and calculations were also performed on LaAlO3- Sr( Al, Ta) O3 and Ba( Zr, Mg, Ta) O3 transparent ceramics, which showed normal dispersion, for comparison purposes. A comparison of the measurements and calculations with the numerical investigations carried out using a dispersion equation indicated that a weak intensity of absorption of La2 Zr2 O7 in the high energy region (20-30 eV) was thought to be the predominant reason for the abnormality. In addition, a narrower bandwidth of the O-2 p band of La2 Zr2 O7 resulted in a high intensity of absorption around 10 eV, which in turn resulted in a relatively low ratio of the intensity of absorption between 20 and 30 eV to the intensity of absorption around 10 eV. This narrower bandwidth of O-2 p was thought to augment the abnormality of La2 Zr2 O7. [ABSTRACT FROM AUTHOR]

Abstract: The proline metabolism intermediate Δ1-pyrroline-5-carboxylate (P5C) induces cell death in animals, plants and yeasts. To elucidate how P5C triggers cell death, we analyzed P5C metabolism, mitochondrial respiration and superoxide anion generation in the yeast Saccharomyces cerevisiae. Gene disruption analysis revealed that P5C-mediated cell death was not due to P5C metabolism. Interestingly, deficiency in mitochondrial respiration suppressed the sensitivity of yeast cells to P5C. In addition, we found that P5C inhibits the mitochondrial respiration and induces a burst of superoxide anions from the mitochondria. We propose that P5C regulates cell death via the inhibition of mitochondrial respiration. [Copyright &y& Elsevier]

Epitaxially grown (100) three-axis-oriented (Ba0.7Sr0.3)TiO3 thin film on (100) platinum (Pt) coated (100) magnesium oxide (MgO) substrate and (100) three-axis-oriented BaTiO3 thin film with BaZrO3 buffer layer were prepared on an MgO(100) substrate using the chemical solution deposition method. The growth of the film was found to depend on the annealing condition. Transmission electron microscope revealed cube-on-cube epitaxial growth. The thin films exhibited a (100) three-axis-orientation that followed the (100) orientation of the MgO substrate and the (100) orientation of the Pt or BaZrO3 bottom layer, as observed from an X-ray pole figure measurement and the selected area electron diffraction patterns. [ABSTRACT FROM AUTHOR]

We fabricated a new series of transparent ceramics based on (1- x) LaAlO3- x Sr(Al0.5Ta0.5)O3 solid solution. These transparent ceramics were obtained by a conventional solid-state reaction, and the crystal structure and its optical properties were investigated. The X-ray diffraction ( XRD) analyses of the ceramics revealed a cubic perovskite structure with a higher Sr(Al0.5Ta0.5)O3 content ( Pm-3 m for x = 0.4-0.6 and Fm-3 m for x = 0.7-1.0), whereas the ceramics contained rhombohedral perovskite phase at x = 0.0-0.3. However, the Raman scattering study revealed that an Fm-3 m ordered structure was contained above x = 0.2. High transmittance was obtained at the cubic composition ( x = 0.4-1.0), and was particularly high at x = 0.5-0.7. It was demonstrated that these transparent ceramics had both a high refractive index and a high Abbe number (low dispersion). The Abbe number of the x = 0.5 composition, 37.8, was higher than that of Ba(Mg,Ta)O3 system transparent ceramics (~30). [ABSTRACT FROM AUTHOR]

Well-crystallized cuprous oxide (Cu2O) nanoparticles were successfully fabricated using a microemulsion (ME) method. A solution containing copper (I) element for hydrolysis reaction in small water droplets was obtained using copper (I) acetate as the raw material with the appropriate addition of chelating agents ( n-propylamine or benzylamine). A transparent yellowish suspension containing Cu2O nanoparticles (ME-Cu2O suspension) was prepared at room temperature, using cyclohexane or benzene as a hydrophobic solvent. The average sizes of the Cu2O nanoparticles obtained in this case were estimated to be 3.9 and 6.2 nm, respectively. Cu2O thin film (average size=24.6 nm) and a hybrid composite of Cu2O nanoparticles with a polymer matrix (average size=5.5 nm) were also prepared from a ME-Cu2O suspension. Optical absorption study revealed that direct band gaps of the Cu2O thin film (2.50 eV) and the hybrid composite (2.65 eV) were much greater than that of bulk Cu2O (2.17 eV), as a result of quantum size effect in Cu2O. The photoluminescence of the obtained Cu2O nanoparticles was measured at 12 K. The present study provides a new promising method to fabricate well-crystallized Cu2O nanoparticles of single nanometer sizes. [ABSTRACT FROM AUTHOR]

We investigated the crystal structures and optical properties of Ba(M,Mg,Ta)O3, Ba(M,Zn,Ta)O3, and Ba(M,Mg,Nb)O3 (M=Ti, Sn, Zr, and Hf) transparent ceramics. Tetravalent cation substitution resulted in a reduction in the 1:2 order structure of the B-site of the parent perovskites; this in turn led to an increase in the transmittance of the ceramics. There was an increase in the refractive index of Ba(Mg1/3Ta2/3)O3 due to Ti, Zr, and Hf substitution at the B-site; the refractive index was particularly high in the case of Ti substitution. Furthermore, Ti substitution resulted in a significant decrease in the Abbe number of Ba(Mg1/3Ta2/3)O3. The refractive index of the perovskites was calculated with high accuracy using the Gladstone–Dale relation. We hypothesize that the Abbe number of perovskites is dependent on its band-gap energy. [ABSTRACT FROM AUTHOR]

(100) three-axis-oriented BaTiO3 thin film with BaZrO3 buffer layer was prepared on an MgO(100) substrate by the chemical solution deposition method. The epitaxial growth of the (100) BaTiO3 layer was enabled by inducing a BaZrO3 bottom layer. Transmission electron microscopy revealed cube-on-cube epitaxial growth. The BaTiO3 thin film exhibited a (100) three-axis-orientation that followed the (100) orientation of the MgO substrate and the (100) orientation of the BaZrO3 bottom layer, as observed from an X-ray pole figure measurement and the selected area electron diffraction patterns. [ABSTRACT FROM AUTHOR]

L-Azetidine-2-carboxylic acid (AZC) is a toxic four-membered ring analogue of L-proline that is transported into cells by proline transporters. AZC and L-proline in the cells are competitively incorporated into nascent proteins. When AZC is present in a minimum medium, misfolded proteins are synthesized in the cells, thereby inhibiting cell growth. The MPR1 gene has been isolated from the budding yeast Saccharomyces cerevisiae Σ1278b as a multicopy suppressor of AZC-induced growth inhibition. MPR1 encodes a novel acetyltransferase that detoxifies AZC via N-acetylation. Since MPR1 is absent in the laboratory strain of S. cerevisiae S288C, it could be a positive selection marker that confers AZC resistance in the S288C background strains. To examine the usefulness of MPR1, we constructed some plasmid vectors that harboured MPR1 under the control of various promoters and introduced them into the S288C-derived strains. The expression of MPR1 conferred AZC resistance that was largely dependent on the expression level of MPR1. In an additional experiment, the galactose-inducible MPR1 and ppr1+, the fission yeast Schizosaccharomyces pombe homologue of MPR1, were used for gene disruption by homologous recombination, and here AZC-resistant colonies were also successfully selected. We concluded that our MPR1-AZC system provides a powerful tool for yeast transformation. Copyright © 2009 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

The ubiquitination and subsequent degradation of stress-induced abnormal proteins are indispensable to cell survival. We previously showed that a yeast ( Saccharomyces cerevisiae) mutant carrying a single amino acid change, Ala401Glu, in RSP5, which encodes an essential E3 ubiquitin ligase, is hypersensitive to various stresses. To identify the protein substrates of Rsp5, we performed a comparative proteome analysis of the wild-type and rsp5 mutant strains under stress conditions. The results we obtained indicate that several proteins, including the α-subunit of nascent polypeptide-associated complex (αNAC, Egd2) accumulated in the rsp5 mutant. To investigate whether or not Rsp5 ubiquitinates these proteins in a stress-dependent manner, cell extracts were analyzed by immunoprecipitation followed by western blotting after exposure to temperature upshift. Interestingly, Egd2 was ubiquitinated in the wild-type cells but not in the rsp5 mutant cells under these stress conditions. We also detected in vitro ubiquitination of Egd2 by Rsp5 at elevated temperature. Moreover, Egd2 was ubiquitinated in the egd1 and not4 deletion mutants lacking βNAC and the RING-type ubiquitin ligase Not4, respectively, indicating that ubiquitination of Egd2 is independent of Egd1 and Not4. We also showed that, under stress conditions, Egd2 was mainly degraded via the proteasome pathway. These results strongly suggest that Rsp5 is involved in selective ubiquitination and degradation of stress-induced unstable proteins, such as Egd2. Structured digital abstract • : EGD2 (uniprotkb: ) physically interacts ( ) with ubiquitin (uniprotkb: ) by anti-tag co-immunoprecipitation ( ) [ABSTRACT FROM AUTHOR]

Mpr1 is an enzyme that catalyzes the N-acetylation of the toxic l-azetidine-2-carboxylic acid (AZC). Recently, Mpr1 has been shown to reduce levels of intracellular reactive oxygen species (ROS) under oxidative stress. The natural substrate involved in the ROS elimination in vivo is still unknown. Mpr1 has been purified and crystallized in space groups P1 and P3112. X-ray data were collected to 1.9 Å resolution from a trigonal crystal soaked with AZC. [ABSTRACT FROM AUTHOR]

Monodispersed and spherically shaped zinc oxide (ZnO) nanoparticles with particle sizes of <5 nm were successfully fabricated by the microemulsion (ME) method. In this process, ZnO nanoparticles were formed at room temperature by the hydrolysis reaction of zinc diethoxide in the small water droplets in reverse micelles. Electron diffraction patterns and lattice images demonstrated that the obtained ZnO nanoparticles were composed of well-crystallized single crystallites having a wurtzite structure. Further, we succeeded in controlling the average particle sizes in the range of 3.9–2.4 nm by changing the surfactants. The bandgap energies of the ZnO nanoparticles estimated from the derivative curve of the optical absorption spectra were 3.65 eV (3.9 nm), 3.66 eV (2.9 nm), 3.69 eV (2.7 nm), and 3.72 eV (2.4 nm). The bandgap energies were found to be considerably higher than the bandgap energy of the ZnO bulk (=3.37 eV). Thus, the quantum size effect in the ME-derived ZnO nanoparticles was successfully observed in this study. The photoluminescence spectra of the ZnO nanoparticles exhibited intense green luminescence at an energy of around 2.5 eV. [ABSTRACT FROM AUTHOR]

One-dimensional (1D) photonic crystals composed of a center polycrystalline yttrium iron garnet (YIG) plate sandwiched with alternate alumina and resin plates were fabricated using stereolithography. The formed crystals showed a transmission mode of microwave in band gap as a result of resonance at the YIG plate. The transmission frequency and intensity were controlled by applying an external magnetic field. These phenomena can be explained by Faraday and Voigt effects of YIG in magnetic field. The propagation characteristics of microwaves agreed well with the calculated results using the finite element method. [ABSTRACT FROM AUTHOR]

Pulsed laser ablation (PLA) was used to prepare nanoparticles of barium titanate (BaTiO3). PLA using a titanium-rich Ba–Ti–O ceramic target in oxygen ambience at 400 Pa and following online annealing at 1073 K enabled the production of BaTiO3 nanoparticles (BTNPs) with a perovskite structure having nearly stoichiometric composition. These nanoparticles were directly introduced into a differential mobility analyzer for size classification, leading to the production of monodispersed and nonagglomerated BTNPs with an average particle size of 13.5 nm. The size distribution of the size-classified BTNPs was found to be reasonably narrow with a geometric standard deviation of 1.07. [ABSTRACT FROM AUTHOR]

Yeasts used in bread making are exposed to air-drying stress during dried yeast production processes. To clarify the genes required for air-drying tolerance, we performed genome-wide screening using the complete deletion strain collection of diploid Saccharomyces cerevisiae. The screening identified 278 gene deletions responsible for air-drying sensitivity. These genes were classified based on their cellular function and on the localization of their gene products. The results showed that the genes required for air-drying tolerance were frequently involved in mitochondrial functions and in connection with vacuolar H+-ATPase, which plays a role in vacuolar acidification. To determine the role of vacuolar acidification in air-drying stress tolerance, we monitored intracellular pH. The results showed that intracellular acidification was induced during air-drying and that this acidification was amplified in a deletion mutant of the VMA2 gene encoding a component of vacuolar H+-ATPase, suggesting that vacuolar H+-ATPase helps maintain intracellular pH homeostasis, which is affected by air-drying stress. To determine the effects of air-drying stress on mitochondria, we analysed the mitochondrial membrane potential under air-drying stress conditions using MitoTracker. The results showed that mitochondria were extremely sensitive to air-drying stress, suggesting that a mitochondrial function is required for tolerance to air-drying stress. We also analysed the correlation between oxidative-stress sensitivity and air-drying-stress sensitivity. The results suggested that oxidative stress is a critical determinant of sensitivity to air-drying stress, although ROS-scavenging systems are not necessary for air-drying stress tolerance. Copyright © 2008 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

Rsp5 is an essential and multi-functional E3 ubiquitin ligase in Saccharomyces cerevisiae. We previously isolated the Ala401Glu rsp5 mutant that is hypersensitive to various stresses. In rsp5A401E cells, the transcription of the stress protein genes was defective. To understand the mechanism by which Rsp5 regulates the expression of stress proteins, we analyzed the expression and localization of two major transcription factors, Hsf1 and Msn2/4, required for stress protein gene expression in S. cerevisiae. The mRNA levels of HSF1 and MSN2/4 in rsp5A401E cells were slightly lower than those of wild-type cells. An interesting finding is that the protein levels of HSF1 and Msn2/4 were remarkably defective in rsp5A401E cells after exposure to temperature up-shift and ethanol, although these proteins are mainly localized in the nucleus under these stress conditions. We also showed that the mRNAs of HSF1 and MSN2/4 were accumulated in the nucleus of rsp5A401E cells after exposure to temperature up-shift and ethanol, and even under non-stress conditions, suggesting that Rsp5 is required for the nuclear export of these mRNAs. These results indicate that, in response to environmental stresses, Rsp5 primarily regulates the expression of Hsf1 and Msn2/4 at the post-transcriptional level and is involved in the repair system of stress-induced abnormal proteins. [ABSTRACT FROM AUTHOR]

In the modern baking industry, high-sucrose-tolerant (HS) and maltose-utilizing (LS) yeast were developed using breeding techniques and are now used commercially. Sugar utilization and high-sucrose tolerance differ significantly between HS and LS yeasts. We analysed the gene expression profiles of HS and LS yeasts under different sucrose conditions in order to determine their basic physiology. Two-way hierarchical clustering was performed to obtain the overall patterns of gene expression. The clustering clearly showed that the gene expression patterns of LS yeast differed from those of HS yeast. Quality threshold clustering was used to identify the gene clusters containing upregulated genes (cluster 1) and downregulated genes (cluster 2) under high-sucrose conditions. Clusters 1 and 2 contained numerous genes involved in carbon and nitrogen metabolism, respectively. The expression level of the genes involved in the metabolism of glycerol and trehalose, which are known to be osmoprotectants, in LS yeast was higher than that in HS yeast under sucrose concentrations of 5-40%. No clear correlation was found between the expression level of the genes involved in the biosynthesis of the osmoprotectants and the intracellular contents of the osmoprotectants. The present gene expression data were compared with data previously reported in a comprehensive analysis of a gene deletion strain collection. Welch's t-test for this comparison showed that the relative growth rates of the deletion strains whose deletion occurred in genes belonging to cluster 1 were significantly higher than the average growth rates of all deletion strains. Copyright © 2007 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

The extraction of the fundamental frequency of speech is an important issue in many speech processing fields. In this paper, we propose a fundamental frequency extraction method based on the power of the amplitude spectrum in the bandwidth where the fundamental frequency is present. This method varies the power of the amplitude spectrum depending on the signal-to-noise ratio (SNR) of the input speech signal in each analysis frame. Therefore, in a white noise environment, this method is applicable to a wide range of speech signals from a high signal-to-noise ratio to a low signal-to-noise ratio. By conducting preliminary tests using true vowels, we derive the equation representing the relationship between the power of the amplitude spectrum and the signal-to-noise ratio. This is also applied to real continuous speech, and the superiority of the proposed method is clearly demonstrated in comparative tests with conventional methods. © 2007 Wiley Periodicals, Inc. Electron Comm Jpn Pt 3, 90(5): 19–30, 2007; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/ecjc.20069 [ABSTRACT FROM AUTHOR]

Abstract: Rsp5 is an essential E3 ubiquitin ligase in Saccharomyces cerevisiae and is known to ubiquitinate plasma membrane permeases followed by endocytosis and vacuolar degradation. We previously isolated the rsp5 mutant that is hypersensitive to various stresses, suggesting that Rsp5 is involved in degradation of stress-induced abnormal proteins. Here, we analyzed the ability to refold the proteins by stress proteins in the rsp5 mutant. The transcription of stress protein genes in the rsp5 mutant was significantly lower than that in the wild-type strain when exposed to temperature up-shift, ethanol or sorbitol. Interestingly, the amounts of transcription factors Hsf1 and Msn4 were remarkably defective in the rsp5 mutant. These results suggest that expression of stress proteins are mediated by Rsp5 and that Rsp5 primarily regulates post-translational modification of Hsf1 and Msn4. [Copyright &y& Elsevier]

To understand endocytic trafficking in Schizosaccharomyces pombe, we constructed an end4 disruption mutant. The end4+ gene encodes a protein homologous to Sla2p/End4p, which is essential for the assembly and function of the cytoskeleton and endocytosis in Saccharomyces cerevisiae. We characterized the fission yeast mutant end4Δ as well as ypt7Δ, which is deficient in vacuolar fusion and, hence, endocytosis. The delivery of FM4-64 to the vacuolar membrane, accumulation of Lucifer yellow CH and internalization of plasma membrane protein Map3-GFP were inhibited in the end4 mutant. Deletion of end4 resulted in pleiotropic phenotypes consistent with F-actin depolarization, including high temperature sensitivity, abnormal morphology and mating defects. Extensive missorting of carboxypeptidase Y was detected in the ypt7 mutant; however, little missorting was detected in the end4 mutant. These results indicate that End4p is essential for the internalization process and Ypt7p affects endocytosis at a post-internalization step after the intersection of the endocytic and the vacuolar protein-sorting pathways in fission yeast. Copyright © 2004 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

In the surgical treatment of secondaryhyperparathyroidism (2HPT) due to uremia, it is considered necessary toremove all parathyroid glands from the neck to prevent persistent andrecurrent parathyroid hyperfunction. However, in some cases fewer thanfour parathyroid glands can be recognized at initial operation; in thepresent study, we evaluated the long-term prognosis and estimatedsurgical strategy in such cases. Between March 1981 and January 1999,822 patients underwent total parathyroidectomy (PTx) with forearmautograft for advanced 2HPT at the Department of Transplant Surgery of Nagoya Second Red Cross Hospital. In 21 cases (2.6%) fewer than fourparathyroid glands were macroscopically found at the initial operation.These cases were followed up and their parathyroid function wasevaluated by measurement of intact parathyroid hormone (PTH). In 20 ofthe 21 cases three glands were found, in 1 patient only two glands. In5 of these cases the fourth gland was identified first afterpostoperative histopathologic evaluation. In all these cases the intactPTH level was normalized. In 8 of the remaining 16 cases high PTHlevels persisted after the initial operation, including 3 patients who underwent neck reexploration. However, in the other 7 patients PTHlevels dropped within normal range immediately after PTx and a fourthgland has never been recognized. One patient was lost to follow-up. Thus, using our operative strategy, 12 of 822 cases (0.85%) did notdevelop persistent or recurrent HPT even though only three glands wereidentified at the operation. To avoid postoperative hypoparathyroidism,autotransplantation should be performed when fewer than four parathyroid glands are found at the initial operation. [ABSTRACT FROM AUTHOR]

The significance of non-αgalactosyl antigens remains unclear in pig-to-primate xenotransplantation. Hanganutziu-Deicher (H-D) antigens with terminal N-glycolylneuraminic acid (NeuGc) are widely expressed on endothelial cells of mammalian species, with the exception of humans. As baboons and monkeys also express H-D antigens, a pig-to-non-human primate experimental model cannot resolve the question of whether H-D antigens can elicit a potent humoral response in human recipients. The purpose of this study was to elucidate the clinical significance of H-D antigens by examining the sera from patients who have been previously exposed to porcine tissue. After the digestion of porcine aortic endothelial cells (PAEC) by neuraminidase, NeuGc and N-acetylneuraminic acid (NeuAc) were quantitated by HPLC. IgG and IgM antibody levels against H-D antigens were measured by NeuGc-GM3-coated ELISA plates in the sera of patients who had undergone ex vivo kidney perfusion 1 to 3 weeks and 2 years previously (n = 2) or had been injected with fetal porcine islets 2 months previously (n = 10). HPLC determined that 9.7×107 NeuAc and 6.3×107 NeuGc residues per cell were released from PAEC by neuraminidase, while 25.7×107 NeuAc and an undetectable level of NeuGc were released from human aortic endothelial cells (HAEC). No significant elevation of IgG or IgM antibody levels against NeuGc-GM3 was observed in sera from patients with a history of porcine exposure. Considering the active production of antibody against the foreign galactosyl antigens after pig-to-human xenotransplantation, some production of antibodies against the equally foreign H-D antigens would be expected, because large amounts of NeuGc terminated saccharides are present in the pig endothelial cell surface. However, no production of antibodies directed to H-D antigens could be found in patients exposed to porcine tissue. Further studies are warranted to explain why H-D antigens do not elicit a significant antibody production. [ABSTRACT FROM AUTHOR]

Successful clinical ABO-incompatible renal transplantation has been achieved by the removal of anti-A or anti-B antibodies using double filtration plasmapheresis (DFPP). We have compared changes in the levels of anti-donor antibodies and the histopathology of the renal grafts following human ABO-incompatible allotransplantation and pig-to-baboon xenotransplantation using pretransplant DFPP. DFPP was performed on days -6, -4, -2 and -1 before the ABO-incompatible transplants (n=25) and on days -2 and 0 (immediately before reperfusion) in the xenotransplants (n=4). In two baboons (XenoTx Group I) the extent of antibody removal was comparable to that in the ABO-incompatible patients, and an even greater level of removal was achieved in another two baboons (XenoTx Group II). Anti-A and anti-B and anti-pig IgM and IgG antibodies were measured by flow cytometry. All clinical ABO-incompatible renal grafts are functioning, except two which were lost from recurrence of the original disease or from chronic rejection. Three other grafts underwent humoral rejection episodes, which were successfully treated. DFPP reduced the mean anti-A/B IgM and IgG antibody levels to 8% and 13% of pre-treatment levels, respectively. After kidney transplantation, they were maintained at 37% and 46% of pre-DFPP level. No antibody binding to the transplanted kidney was detected at any time (1 h to 2 yr) after ABO-incompatible allotransplantation. In contrast, in XenoTx Group I, the same extent of antibody removal (90%) prevented hyperacute rejection, but the two grafts were rejected on days 6 and 7, respectively, from acute vascular rejection. In XenoTx Group II, the additional DFPP that was required to deplete the remaining 10% of anti-pig antibody was poorly tolerated and the two baboons died 4 h and 2 days, respectively, after renal transplantation. Although anti-pig IgM antibodies were reduced to 2% of pre-treatment level, IgM and 1 binding were detected in the graft as early as 1 h... [ABSTRACT FROM AUTHOR]