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Type 2 diabetes (T2D) is a heterogeneous disease that develops through diverse pathophysiological processes1,2and molecular mechanisms that are often specific to cell type3,4. Here, to characterize the genetic contribution to these processes across ancestry groups, we aggregate genome-wide association study data from 2,535,601 individuals (39.7% not of European ancestry), including 428,452 cases of T2D. We identify 1,289 independent association signals at genome-wide significance (P< 5 × 10−8) that map to 611 loci, of which 145 loci are, to our knowledge, previously unreported. We define eight non-overlapping clusters of T2D signals that are characterized by distinct profiles of cardiometabolic trait associations. These clusters are differentially enriched for cell-type-specific regions of open chromatin, including pancreatic islets, adipocytes, endothelial cells and enteroendocrine cells. We build cluster-specific partitioned polygenic scores5in a further 279,552 individuals of diverse ancestry, including 30,288 cases of T2D, and test their association with T2D-related vascular outcomes. Cluster-specific partitioned polygenic scores are associated with coronary artery disease, peripheral artery disease and end-stage diabetic nephropathy across ancestry groups, highlighting the importance of obesity-related processes in the development of vascular outcomes. Our findings show the value of integrating multi-ancestry genome-wide association study data with single-cell epigenomics to disentangle the aetiological heterogeneity that drives the development and progression of T2D. This might offer a route to optimize global access to genetically informed diabetes care.

Type 2 diabetes (T2D) shows heterogeneous body mass index (BMI) sensitivity. Here, we performed stratification based on BMI to optimize predictions for BMI-related diseases. We obtained BMI-stratified datasets using data from more than 195,000 individuals (nT2D= 55,284) from BioBank Japan (BBJ) and UK Biobank. T2D heritability in the low-BMI group was greater than that in the high-BMI group. Polygenic predictions of T2D toward low-BMI targets had pseudo-R2values that were more than 22% higher than BMI-unstratified targets. Polygenic risk scores (PRSs) from low-BMI discovery outperformed PRSs from high BMI, while PRSs from BMI-unstratified discovery performed best. Pathway-specific PRSs demonstrated the biological contributions of pathogenic pathways. Low-BMI T2D cases showed higher rates of neuropathy and retinopathy. Combining BMI stratification and a method integrating cross-population effects, T2D predictions showed greater than 37% improvements over unstratified-matched-population prediction. We replicated findings in the Tohoku Medical Megabank (n= 26,000) and the second BBJ cohort (n= 33,096). Our findings suggest that target stratification based on existing traits can improve the polygenic prediction of heterogeneous diseases.

The object of this paper is to develop numerical procedures for creep-fatigue-oxidation life prediction and damage assessments based on engineering damage approach, in response to high-reliability and long-life design requirement that supports low-carbon and new-energy technologies. In order to achieve the prediction results in terms of cycle-dependent stress–strain responses, crack initiation life prediction and multi-damage evolutions, the generalized creep-fatigue loading conditions including tension-hold-only, compression-hold-only and tension-compression-hold-both, which are abbreviated as CP, PC, and CC types, are conducted for IN 718 at 650 °C. Results show that creep-fatigue deformation behaviors are well depicted through the evolutions of hysteresis loops, cyclic softening curves and stress responses. With incorporating into oxidation damage especially under long-life conditions, both error band and probability density function for life prediction are quantitatively improved. In addition, the cycle-dependent roles in multi-damage evolutions are clearly observed in a set of radar graphs, where fatigue, creep and oxidation damage are manifested as different evolutionary features. Finally, the technical route in the transition from deterministic to probabilistic multi-damage assessments is discussed based on the established creep-fatigue-oxidation diagram.

According to quantum chromodynamics, vacuum is not an empty space, because it is filled with quark–antiquark pairs. The pair has the same quantum numbers as the vacuum and forms a condensate because the strong interaction of the quantum chromodynamics is too strong to leave the vacuum empty. This quark–antiquark condensation, the chiral condensate, breaks the chiral symmetry of the vacuum. The expectation value of the chiral condensate is an order parameter of the chiral symmetry, which is expected to decrease at high temperatures or high matter densities where the chiral symmetry is partially restored. Head-on collisions of nuclei at ultra-relativistic energies have explored the high-temperature regime, but experiments at high densities are rare. Here we measure the spectrum of pionic 121Sn atoms and study the interaction between the pion and the nucleus. We find that the expectation value of the chiral condensate is reduced at finite density compared to the value in vacuum. The reduction is linearly extrapolated to the nuclear saturation density and indicates that the chiral symmetry is partially restored due to the extremely high density of the nucleus.

Identifying the host genetic factors underlying severe COVID-19 is an emerging challenge1–5. Here we conducted a genome-wide association study (GWAS) involving 2,393 cases of COVID-19 in a cohort of Japanese individuals collected during the initial waves of the pandemic, with 3,289 unaffected controls. We identified a variant on chromosome 5 at 5q35 (rs60200309-A), close to the dedicator of cytokinesis 2 gene (DOCK2), which was associated with severe COVID-19 in patients less than 65 years of age. This risk allele was prevalent in East Asian individuals but rare in Europeans, highlighting the value of genome-wide association studies in non-European populations. RNA-sequencing analysis of 473 bulk peripheral blood samples identified decreased expression of DOCK2associated with the risk allele in these younger patients. DOCK2expression was suppressed in patients with severe cases of COVID-19. Single-cell RNA-sequencing analysis (n= 61 individuals) identified cell-type-specific downregulation of DOCK2and a COVID-19-specific decreasing effect of the risk allele on DOCK2expression in non-classical monocytes. Immunohistochemistry of lung specimens from patients with severe COVID-19 pneumonia showed suppressed DOCK2 expression. Moreover, inhibition of DOCK2 function with CPYPP increased the severity of pneumonia in a Syrian hamster model of SARS-CoV-2 infection, characterized by weight loss, lung oedema, enhanced viral loads, impaired macrophage recruitment and dysregulated type I interferon responses. We conclude that DOCK2 has an important role in the host immune response to SARS-CoV-2 infection and the development of severe COVID-19, and could be further explored as a potential biomarker and/or therapeutic target.

We assembled an ancestrally diverse collection of genome-wide association studies (GWAS) of type 2 diabetes (T2D) in 180,834 affected individuals and 1,159,055 controls (48.9% non-European descent) through the Diabetes Meta-Analysis of Trans-Ethnic association studies (DIAMANTE) Consortium. Multi-ancestry GWAS meta-analysis identified 237 loci attaining stringent genome-wide significance (P< 5 × 10−9), which were delineated to 338 distinct association signals. Fine-mapping of these signals was enhanced by the increased sample size and expanded population diversity of the multi-ancestry meta-analysis, which localized 54.4% of T2D associations to a single variant with >50% posterior probability. This improved fine-mapping enabled systematic assessment of candidate causal genes and molecular mechanisms through which T2D associations are mediated, laying the foundations for functional investigations. Multi-ancestry genetic risk scores enhanced transferability of T2D prediction across diverse populations. Our study provides a step toward more effective clinical translation of T2D GWAS to improve global health for all, irrespective of genetic background.

Current genome-wide association studies do not yet capture sufficient diversity in populations and scope of phenotypes. To expand an atlas of genetic associations in non-European populations, we conducted 220 deep-phenotype genome-wide association studies (diseases, biomarkers and medication usage) in BioBank Japan (n= 179,000), by incorporating past medical history and text-mining of electronic medical records. Meta-analyses with the UK Biobank and FinnGen (ntotal= 628,000) identified ~5,000 new loci, which improved the resolution of the genomic map of human traits. This atlas elucidated the landscape of pleiotropy as represented by the major histocompatibility complex locus, where we conducted HLA fine-mapping. Finally, we performed statistical decomposition of matrices of phenome-wide summary statistics, and identified latent genetic components, which pinpointed responsible variants and biological mechanisms underlying current disease classifications across populations. The decomposed components enabled genetically informed subtyping of similar diseases (for example, allergic diseases). Our study suggests a potential avenue for hypothesis-free re-investigation of human diseases through genetics.

Byline: Yoshiki Sawa, Keishi Kadoba, Ken Suzuki, Hong-Zhi Bai, Yasufumi Kaneda, Ryota Shirakura, Hikaru Matsuda Abstract: Objective: To confirm gene transfer techniques especially into the whole heart, we tried out a gene transfer method involving liposome with the viral envelope hemagglutinating virus of Japan liposome as an alternative to existing techniques such as cationic lipofection or other viral vectors. Method: For this study, hemagglutinating virus of Japan liposome (H group) or cationic liposome(L group) was used to compare the efficacy of gene transfection of oligonucleotide labeled with fluorescein isothiocyanate and cDNA of [beta]-galactosidase and human manganese-superoxide dismutase. Fluorescein-labeled oligonucleotide, cDNA of [beta]-galactosidase, or manganese-superoxide dismutase was complexed with liposomes, DNA-binding nuclear protein, and the viral protein coat of hemagglutinating virus of Japan. After donor rat hearts arrested by cardioplegia had been harvested, the coronary artery during cardioplegic arrest was infused via an aortic cannula with the liposome-gene complex. Next, the hearts were transplanted into the abdomen of recipient rats of the same strain, and all recipients were put to death after 3 days of transfection. Results: Fluorescein isothiocyanate was detected in the nuclei of more than 70% of the myocytes (75% [+ or -] 14%, n = 5) in the H group compared with fewer than 10% in the L group (7% [+ or -] 5%, n = 5). The intensity of fluorescein isothiocyanate was significantly higher in the H group (979 [+ or -] 112 FI) than in the L group (116 [+ or -] 68 FI). [beta]-Galactosidase was expressed in the cytosol of more than 50% of the myocytes in the H group (61% [+ or -] 7%, n = 5) compared with none in the L group (0%, n = 5). After 3 days of gene transfection, and when exposed to ischemia (30 minutes, 37[degrees] C) and reperfusion (30 minutes, 37[degrees] C) with Langendorff apparatus, the hearts transfected with manganese-superoxide dismutase (S group, n = 5) showed a significantly higher percentage of recovery of left ventricular end-diastolic pressure (S vs C, 86% [+ or -] 3% vs 54% [+ or -] 12%) and coronary flow (98% [+ or -] 2% vs 66% [+ or -] 12%) than did the control hearts (C group, n = 5). Western blotting analysis showed an apparent increased expression of manganese-superoxide dismutase in the hearts transfected with manganese-superoxide dismutase compared with the control hearts. These results clearly demonstrated that the donor hearts were transfected with fluorescein-labeled oligonucleotide and the [beta]-galactosidase gene as a result of coronary infusion of the hemagglutinating virus of Japan liposome during cardioplegic arrest at the time of harvest. Furthermore, the hearts transfected with manganese-superoxide dismutase showed significant improvement in tolerance against ischemia-reperfusion injury. Conclusion: We believe that this method represents a novel in vivo gene transfer technique for the heart and thus may provide a new tool for research and therapy of heart transplantation. (J Thorac Cardiovasc Surg 1997;113:512-9) Article History: Received 6 May 1996; Revised 12 June 1996; Revised 11 October 1996; Accepted 21 October 1996 Article Note: (footnote) [star] From the First Department of Surgery,a Department of Institute for Molecular and Cellular Biology,b and Department of Organ Transplantation,c Osaka University Medical School, Suita, Osaka 565, Japan., [star][star] Read at the Seventy-sixth Annual Meeting of The American Association for Thoracic Surgery, San Diego, Calif., April 28-May 1, 1996., a Address for reprints: Hikaru Matsuda MD, First Department of Surgery, Osaka University Medical School, 2-2 Yamada-oka, Suita, Osaka 565, Japan., aa 0022-5223/97 $5.00 + 0 12/6/78770

Molecular aggregation structure and mechanical as well as thermal properties of novel well-defined multiblock copolymers consisting of crystalline syndiotactic polystyrene (sPS) and rubbery hydrogenated polyisoprene (hPIp) were investigated. The morphology and crystalline ordered structure of the multiblock copolymer films prepared by solvent casting from 1,2-dichlorobenzene solution depended on the volume fraction of sPS (VFsPS) and number of blocks. The multiblock copolymer films exhibited ordered morphology with low crystallinity. The crystallinity of the sPS reduced with decreasing the VFsPS. The pentablock copolymer produced more ordered morphology and less crystallinity than the triblock copolymers. The anisotropic orientation and mechanical stability of the δ form sPS crystals in the spherical sPS domains during uniaxial stretching were demonstrated. Tensile testing and dynamic mechanical analysis indicated that these multiblock copolymer films with appropriate sPS fraction are strong, tough, and elastic and thus could be potential candidates for a new type of thermoplastic elastomer with discrete crystalline hard domains.

The overwhelming majority of participants in current genetic studies are of European ancestry. To elucidate disease biology in the East Asian population, we conducted a genome-wide association study (GWAS) with 212,453 Japanese individuals across 42 diseases. We detected 320 independent signals in 276 loci for 27 diseases, with 25 novel loci (P< 9.58 × 10−9). East Asian–specific missense variants were identified as candidate causal variants for three novel loci, and we successfully replicated two of them by analyzing independent Japanese cohorts; p.R220W of ATG16L2(associated with coronary artery disease) and p.V326A of POT1(associated with lung cancer). We further investigated enrichment of heritability within 2,868 annotations of genome-wide transcription factor occupancy, and identified 378 significant enrichments across nine diseases (false discovery rate < 0.05) (for example, NKX3-1for prostate cancer). This large-scale GWAS in a Japanese population provides insights into the etiology of complex diseases and highlights the importance of performing GWAS in non-European populations.

Meta-analyses of genome-wide association studies (GWAS) have identified more than 240 loci that are associated with type 2 diabetes (T2D)1,2; however, most of these loci have been identified in analyses of individuals with European ancestry. Here, to examine T2D risk in East Asian individuals, we carried out a meta-analysis of GWAS data from 77,418 individuals with T2D and 356,122 healthy control individuals. In the main analysis, we identified 301 distinct association signals at 183 loci, and across T2D association models with and without consideration of body mass index and sex, we identified 61 loci that are newly implicated in predisposition to T2D. Common variants associated with T2D in both East Asian and European populations exhibited strongly correlated effect sizes. Previously undescribed associations include signals in or near GDAP1, PTF1A, SIX3, ALDH2, a microRNA cluster, and genes that affect the differentiation of muscle and adipose cells3. At another locus, expression quantitative trait loci at two overlapping T2D signals affect two genes—NKX6-3and ANK1—in different tissues4–6. Association studies in diverse populations identify additional loci and elucidate disease-associated genes, biology, and pathways.

ABSTRACTThe basidiomycetous yeast Pseudozyma antarcticaGB-4(0) esterase (PaE) is a promising candidate for accelerating degradation of used biodegradable plastics (BPs). To increase safety and reduce costs associated with the use of PaE, we constructed a self-cloning strain with high-PaE productivity. A Lys12 gene (PaLYS12)-deleted lysine auxotroph strain GB4-(0)-L1 was obtained from GB-4(0) by ultraviolet mutagenesis and nystatin enrichment. Subsequently, the PaE gene (PaCLE1) expression cassette consisting of GB-4(0)-derived PaCLE1, under the control of a xylose-inducible xylanase promoter with PaLYS12, was randomly introduced into the GB4-(0)-L1 genome. A PaE high-producing strain, PGB474, was selected from among the transformants by high throughput double-screening based on its ability to degrade emulsified polybutylene succinate-co-adipate. Quantitative PCR revealed that four copies of the PaE gene expression cassette were introduced into the PGB474 genome. PGB474 produced 2.0 g/L of PaE by xylose-fed-batch cultivation using a 3-L jar fermentor for 72 h.

To perform detailed fine-mapping of the major-histocompatibility-complex region, we conducted next-generation sequencing (NGS)-based typing of the 33 human leukocyte antigen (HLA) genes in 1,120 individuals of Japanese ancestry, providing a high-resolution allele catalog and linkage-disequilibrium structure of both classical and nonclassical HLA genes. Together with population-specific deep-whole-genome-sequencing data (n= 1,276), we conducted NGS-based HLA, single-nucleotide-variant and indel imputation of large-scale genome-wide-association-study data from 166,190 Japanese individuals. A phenome-wide association study assessing 106 clinical phenotypes identified abundant, significant genotype–phenotype associations across 52 phenotypes. Fine-mapping highlighted multiple association patterns conferring independent risks from classical HLA genes. Region-wide heritability estimates and genetic-correlation network analysis elucidated the polygenic architecture shared across the phenotypes.

To understand the genetics of type 2 diabetes in people of Japanese ancestry, we conducted A meta-analysis of four genome-wide association studies (GWAS; 36,614 cases and 155,150 controls of Japanese ancestry). We identified 88 type 2 diabetes–associated loci (P< 5.0 × 10−8) with 115 independent signals (P< 5.0 × 10−6), of which 28 loci with 30 signals were novel. Twenty-eight missense variants were in linkage disequilibrium (r2> 0.6) with the lead variants. Among the 28 missense variants, three previously unreported variants had distinct minor allele frequency (MAF) spectra between people of Japanese and European ancestry (MAFJPN> 0.05 versus MAFEUR< 0.01), including missense variants in genes related to pancreatic acinar cells (GP2) and insulin secretion (GLP1R). Transethnic comparisons of the molecular pathways identified from the GWAS results highlight both ethnically shared and heterogeneous effects of a series of pathways on type 2 diabetes (for example, monogenic diabetes and beta cells).

Modified 9Cr-1Mo steel is a heat-resistant steel developed for a steam generator in a FBR (Fast Breeder Reactor) and it has been applied to various thermal power plants. Recently, it was found that the fatigue limits did not appear up to 108 cycles at temperatures higher than 500oC. The reason for the decrease of the fatigue life was attributed to the change of the initially designed microstructure of the alloy. The initially dispersed fine lath martensitic texture disappeared at temperatures higher than 500°C, when the magnitude of the applied stress exceeded a certain critical value. In order to explicate the dominant factors of the change quantitatively, the change of the microstructure and the strength of the alloy were continuously observed by applying an intermittent fatigue and creep tests at elevated temperatures and EBSD analysis. It was found that there was a critical stress which caused the microstructure change at each test temperature higher than 500°C, and the activation energy of the change was determined as a function of temperature and the applied tensile stress. The dominant factor of the micro structure change was the stress-induced acceleration of the atomic diffusion of the component element of the alloy.

The degradation process of the micro texture of Ni-base superalloys was observed under fatigue and creep-fatigue loading conditions at elevated temperatures higher than 700oC by applying electron back-scatter diffraction (EBSD) analyses. The local distribution of the crystallinity of a grain and a grain boundary was defined quantitatively by analyzing the Kikuchi pattern obtained from each electron-beam-irradiated area with a diameter of 50 nm. The calculated image quality value was used for the analysis. It was found that the crystallinity of grain boundaries degraded seriously under creep-fatigue loading conditions due to the acceleration of anisotropic strain-induced diffusion of component elements. Since the initial finely-controlled strengthened micro texture disappeared due to the anisotropic diffusion of component elements, this degradation was found to cause the drastic decrease of the strength of grains and grain boundaries and thus, lifetime of the material. The decrease of the strength of both grains and grain boundaries was measured by micro tensile test system in a scanning electron microscope by making a small sample from bulk specimen using focused ion beam. The strength of a grain and a grain boundary varied drastically depending on their crystallinity, in other words, the image quality value.

Brown fat dissipates energy as heat and protects against obesity. Here, we identified nuclear factor I-A (NFIA) as a transcriptional regulator of brown fat by a genome-wide open chromatin analysis of murine brown and white fat followed by motif analysis of brown-fat-specific open chromatin regions. NFIA and the master transcriptional regulator of adipogenesis, PPARγ, co-localize at the brown-fat-specific enhancers. Moreover, the binding of NFIA precedes and facilitates the binding of PPARγ, leading to increased chromatin accessibility and active transcription. Introduction of NFIA into myoblasts results in brown adipocyte differentiation. Conversely, the brown fat of NFIA-knockout mice displays impaired expression of the brown-fat-specific genes and reciprocal elevation of muscle genes. Finally, expression of NFIA and the brown-fat-specific genes is positively correlated in human brown fat. These results indicate that NFIA activates the cell-type-specific enhancers and facilitates the binding of PPARγ to control the brown fat gene program.

We have established a new transgenesis protocol based on CRISPR-Cas9, “New and Easy XenopusTransgenesis (NEXTrans),” and identified a novel safe harbor site in African clawed frogs, Xenopus laevis. We describe steps in detail for the construction of NEXTransplasmid and guide RNA, CRISPR-Cas9-mediated NEXTransplasmid integration into the locus, and its validation by genomic PCR. This improved strategy allows us to simply generate transgenic animals that stably express the transgene.

A surface marker that distinctly identifies cardiac progenitors (CPs) is essential for the robust isolation of these cells, circumventing the necessity of genetic modification. Here, we demonstrate that a Glycosylphosphatidylinositol-anchor containing neurotrophic factor receptor, Glial cell line-derived neurotrophic factor receptor alpha 2 (Gfra2), specifically marks CPs. GFRA2 expression facilitates the isolation of CPs by fluorescence activated cell sorting from differentiating mouse and human pluripotent stem cells. Gfra2mutants reveal an important role for GFRA2 in cardiomyocyte differentiation and development both in vitro and in vivo. Mechanistically, the cardiac GFRA2 signaling pathway is distinct from the canonical pathway dependent on the RET tyrosine kinase and its established ligands. Collectively, our findings establish a platform for investigating the biology of CPs as a foundation for future development of CP transplantation for treating heart failure.

Programmable nucleases enable engineering of the genome by utilizing endogenous DNA double-strand break (DSB) repair pathways. Although homologous recombination (HR)-mediated gene knock-in is well established, it cannot necessarily be applied in every cell type and organism because of variable HR frequencies. We recently reported an alternative method of gene knock-in, named the PITCh (Precise Integration into Target Chromosome) system, assisted by microhomology-mediated end-joining (MMEJ). MMEJ harnesses independent machinery from HR, and it requires an extremely short homologous sequence (5–25 bp) for DSB repair, resulting in precise gene knock-in with a more easily constructed donor vector. Here we describe a streamlined protocol for PITCh knock-in, including the design and construction of the PITCh vectors, and their delivery to either human cell lines by transfection or to frog embryos by microinjection. The construction of the PITCh vectors requires only a few days, and the entire process takes ∼1.5 months to establish knocked-in cells or ∼1 week from injection to early genotyping in frog embryos.

The 2007 to 2009 financial crises also known as 'global financial crisis' has shown the importance of contagion and interconnectedness among financial nodes. This trickles down the crisis effect to small nodes and whole system becomes fragile. We analysed selected EU members banking system network in terms of their pre- and post-crisis status. By using the matrix of bilateral obligations and their capital levels, conducted stress test simulations for both periods. Different levels of shocks entail most aspects of stress tests in terms of identifying networks strength and interconnectedness. Short-terms to long-term clearing scenarios are used to identify strong and weak nodes within system during both time frames. Default probabilities for different levels of shocks shows realistic picture of the banking systems, as they received huge bailouts and financing from governments and regulators. Default probabilities are used to identify the level of difficulty faced by particular institution or whole system.

While the concept of Cloud Radio Access Networks (C-RANs) presents a promising solution to provide required quality of service (QoS) for the future network environment, i.e., more than 10 Gbps capacity, less than 1 ms latency, and connectivity for numerous devices, it is still susceptible to quality of experience (QoE) problems. Until now, only a few researchers considered the design and operation of C-RANs based on QoE. In this article we describe our envisioned C-RAN based on passive optical networks (PONs) exploiting power over fiber (PoF), which can be installed with low installation cost and is capable of providing communication services without external power supply for remote radio head (RRH), and describe QoE requirement on the envisioned network. For all users in the envisioned network to satisfy their QoE, effective network design and operation approaches are then presented. Our proposed design and operation approaches demonstrate how to construct the envisioned network, i.e. the numbers of RRHs and optical line terminals (OLTs), and sleep scheduling of RRHs for an energy-efficient optical power transmission.

Fiber Wireless (FiWi) access networks is widely used to connect users to the core network. The FiWi access networks consist of the edge router, the Passive Optical Networks (PONs), and the wireless networks. In this environment, the edge router, which is the terminal to the core network, can have heavy congestion. To cope with this problem, we propose a Multi Point Control Protocol (MPCP)-based centralized rate control method to centrally control the traffic rate at each Home GateWay (HGW) so as to avoid congestion at the edge router. In the proposed scheme, the transmission delay due to rate control overhead can be mitigated. The performance of the proposed scheme is verified by numerical analysis and computer simulations.

The local thermal deformation of the chips mounted by area-arrayed fine bumps has increased drastically because of the decrease of the flexural rigidity of the thinned chips. In this paper, the dominant structural factors of the local residual stress in a silicon chip are investigated quantitatively based on the measurement of the local residual stress in a chip using stress sensor chips. The piezoresistive strain gauges were embedded in the sensor chips. The length of each gauge was 2 µm and a unit cell consisted of four gauges with different crystallographic directions. This alignment of strain gauges enables to measure the tensor component of three-dimensional stress fields separately. Test flip chip substrates were made by silicon chip on which the area-arrayed tin/copper bumps were electroplated. The width of a bump was fixed at 200 µm and the bump pitch was varied from 400 µm to 1,000 µm. The measured amplitude of the residual stress increased from about 30 MPa to 250 MPa. It was confirmed that both the material constant of underfill and the alignment structure of fine bumps are the dominant factors of the local deformation and stress of a silicon chip mounted on area-arrayed metallic bumps.

The relationship between the electrical properties and crystallographic quality (crystallinity) of electroplated copper thin-film interconnections was investigated. The crystallinity of the grains and grain boundaries of the interconnections was evaluated on the basis of the image quality (IQ) value obtained by electron back-scatter diffraction (EBSD) analysis. The electrical properties of the interconnections vary markedly depending on their crystallinity. The crystallinity also changed markedly as functions of electroplating conditions and the annealing temperature after electroplating. Although the electro migration (EM) resistance of the annealed interconnection was improved, stress-induced migration (SM) was activated by a high residual stress after annealing. To improve electrical reliability without heat treatment after electroplating, the effects of the seed layer under the interconnections on the crystallinity were investigated. As a result, the crystallinity was improved by changing the seed layer from Cu to Ru. In addition, the decrease in current density during electroplating also improved the crystallinity. Therefore, both introducing the Ru seed layer and decreasing the current density during electroplating are effective for developing highly reliable copper interconnections.

Smart Fiber Wireless (SFiWi) networks have attracted great attention due to their ability to become the next generation broadband access networks. FiWi networks integrate optical networks and wireless networks. Access networks consume much energy due to the increase of network participants and the volume of content. To conserve energy consumption in Passive Optical Network (PON), Optical Network Unit (ONU) sleep mode was proposed. The basic idea of ONU sleep mode is to enter sleep state, for an ONU sleep state period while the ONU is in idle situation. Traditional research on ONU sleep showed that long ONU sleep state period results only in increased latency. In this research we prove that excessively long ONU sleep state periods not only cause long latency but also result in more energy consumption. Furthermore, we derive the optimal sleep state period to minimize energy consumption. Also, we show that our derived sleep state period achieves better latency compared to large ONU sleep state period.

Four isolates tentatively identified as Pseudaegerita matsushimaeon the basis of the morphology of bulbil-like propagules were collected from substrates submerged in water in Thailand and Japan. In culture studies the two Thai isolates were found to produce phialoconidia on conidiogenous cells and phialoconidiophores whose morphology was similar to that of Trichoderma. Phylogenetic analysis based on D1/D2 regions of LSU rDNA sequences showed that the four isolates were nested in Hypocrea/Trichoderma(Hypocreales) while P. corticalis, the type species of Pseudaegerita, belongs to Hyaloscypha(Helotiales). Preliminary analysis by ISTH Web tools based on 5.8S-ITS rDNA and phylogenetic analysis based on rpb2and tef1-int4genes showed that the isolates have specific sequences of Trichoderma(anchors 1–5) and belong to the Hamatum clade but they grouped apart from any known species of Trichoderma. The sequences of the tef1-int4gene, which were amplified from the authentic specimen of P. matsushimae(IMI 266915), also showed that it belongs to the Hamatum clade closely clustering with T. yunnanensebut separate from our four isolates. The morphology of P. matsushimae(IMI 266915), especially the sizes of phialides and phialoconidia, were different from T. yunnanense. Thus, we conclude that IMI 266915 and our isolates are to be assigned to two different species in the Hamatum clade of Trichoderma, although both species have similar morphology of bulbils and phialoconidia. Morphology and molecular data revealed that P. matsushimaeshould be assigned to the genus Trichoderma as T. matsushimaeand the Thai and Japanese isolates are placed in T. aeroaquaticumsp. nov. This finding supports the interpretation that aero-aquatic fungi have evolved from terrestrial fungi. We assume that these fungi probably were derived from typically soil-inhabiting species of Trichoderma; an adaptation to aquatic environments is shown by formation of bulbil-like propagules floating on water.

Recent evidence suggests that cell therapy such as the injection of bone marrow–derived mononuclear cells (BMMNCs) can exert protective effects in various conditions associated with ischemia-reperfusion injury. Here, we investigate the effects of BMMNCs on the organ injurydysfunction induced by hemorrhagic shock (HS). Thirty-seven anesthetized male Wistar rats were subjected to hemorrhage by reducing mean arterial pressure to 35 ± 5 mmHg for 90 min, followed by resuscitation with 20 mLkg Ringer’s lactate administered over 10 min and 50 of the shed blood over 50 min. Rats were killed 4 h after the onset of resuscitation. Bone marrow–derived mononuclear cells were freshly isolated from rat tibias and femurs using Percoll density gradient centrifugation, and BMMNCs (1 × 107cells per rat in 1 mLkg phosphate-buffered saline, i.v.) were administered on resuscitation. Hemorrhagic shock resulted in significant organ injurydysfunction (renal, hepatic, neuromuscular) and inflammation (hepatic, lung). In rats subjected to HS, administration of BMMNCs significantly attenuated (i) organ injurydysfunction (renal, hepatic, neuromuscular) and inflammation (hepatic, lung), (ii) increased the phosphorylation of Akt and glycogen synthase kinase-3, (iii) attenuated the activation of nuclear factor-B, (iv) attenuated the increase in extracellular signal–regulated kinase 12 phosphorylation, and (v) attenuated the increase in expression of intercellular adhesion molecule-1. Our findings suggest that administration of BMMNCs protects against the induction of early organ injurydysfunction caused by severe HS by a mechanism that may involve activation of Akt and the inhibition of glycogen synthase kinase-3 and nuclear factor-B.

We fabricated wavelength tunable laser diodes consisting of ring resonator wavelength filters with Si-wire waveguides. More than 45 nm wavelength tuning operation was obtained by about 100 mW heating power. We also measured spectral linewidth of the tunable laser, and observed about 200 kHz of spectral linewidth.

Two isolates, AC04Tand AC05, were isolated from the flowers of red ginger collected in Chiang Mai, Thailand. In phylogenetic trees based on 16S rRNA gene sequences, the two isolates were included within a lineage comprised of the genera Acidomonas, Gluconacetobacter, Asaia, Kozakia, Swaminathania, Neoasaia, Granulibacter, and Tanticharoenia, and they formed an independent cluster along with the type strain of Tanticharoenia sakaeratensis. The calculated pair-wise sequence similarities of isolate AC04Twere 97.8–92.5% to the type strains of the type species of the 11 genera of acetic acid bacteria. The DNA base composition was 66.0–66.1 mol % G+C with a range of 0.1 mol %. A single-stranded, labeled DNA from isolate AC04Tpresented levels of DNA-DNA hybridization of 100, 85, 4, and 3% respectively to DNAs from isolates AC04Tand AC05 and the type strains of Tanticharoenia sakaeratensisand Gluconacetobacter liquefaciens. The two isolates were unique morphologically in polar flagellation and physiologically in intense acetate oxidation to carbon dioxide and water and weak lactate oxidation. The intensity in acetate oxidation almost equaled that of the type strain of Acetobacter aceti. The two isolates had Q-10. Isolate AC04Twas discriminated from the type strains of the type species of the 11 genera by 16S rRNA gene restriction analysis using restriction endonucleases TaqI and Hin6I. The unique phylogenetic, genetic, morphological, physiological, and biochemical characteristics obtained indicate that the two isolates can be classified into a separate genus, and Ameyamaea chiangmaiensisgen. nov., sp. nov. is proposed. The type strain is isolate AC04T(=BCC 15744T, =NBRC 103196T), which has a DNA G+C content of 66.0 mol %.

The mechanism of the directional coarsening of ' phases (rafting) of Ni-base superalloy under an uni-axial strain was analyzed by molecular dynamics (MD) analysis. The stress-induced anisotropic diffusion of Al atoms perpendicular to the interface was observed clearly in a Ni(001)/Ni3Al(001) interface structure, The reduction of the diffusion of Al atoms perpendicular to the interface is thus, effective for improving the creep and fatigue resistance of the alloy. It was also found that the dopant elements in the superalloy also affected the strain-induced diffusion of Al atoms. Pd was one of the most effective elements which restrain Al atoms from moving around the interface.

Electrical conductivity of 10 mol% yttria-stabilized zirconia (10YSZ) and YBa2Cu3O7-d (YBCO) under stress were measured in the temperature range from 600oC to 800oC for 10YSZ and at 600oC for YBCO. It was observed that conductivity under stress slightly decreased for 10YSZ and increased for YBCO. In the case of 10YSZ, the conductivity change was caused by the change in mobility. In the case of YBCO, the conductivity change was caused by the change in mobility and change in carrier concentration.

A new hybrid phosphine was designed. The phosphine combines two common structural characteristics found among the effective phosphine ligands reported recently, namely, three tert-alkyl substituents binding to the phosphorus and an aryl group at an appropriate position. A hybrid phospine/palladium system is versatile and effective for the coupling reaction of various aryl halides with primary and secondary amines including carbazole.

Tanticharoenia sakaeratensisgen. nov., sp. nov. is proposed for three strains isolated from soil collected in Thailand. The three strains, AC37T, AC38, and AC39, were included within a lineage comprising the genera Asaia, Kozakia, Swaminathania, Neoasaia, Acetobacter, Gluconobacter, and Saccharibacterin a phylogenetic tree based on 16S rRNA gene sequences, but formed a quite different, independent cluster. Pair-wise sequence similarities of strain AC37Twere 96.5–92.1% to the type strains of Acetobacter aceti, Gluconobacter oxydans, Acidomonas methanolica, Gluconacetobacter liquefaciens, Asaia bogorensis, Kozakia baliensis, Swaminathania salitolerans, Saccharibacter floricola, Neoasaia chiangmaiensis, and Granulibacter bethesdensis. The three strains had DNA base compositions comprising respectively 65.6, 64.5, and 65.6 mol % G+C with a range of 1.1 mol %, and formed a single species. Phenotypically, the three strains did not oxidize acetate or lactate, but grew on 30% D-glucose (w/v). Chemotaxonomically, they had Q-10. The type strain is AC37T(= BCC 15772T= NBRC 103193T).

No Abstract

The removal properties of diesel exhaust particles (DEP) were investigated using an engine exhaust particle size spectrometer (EEPS), field emission-type scanning electron microscopy (FE-SEM) and time-of-flight secondary ion mass spectrometry (TOF-SIMS). DEP were treated using a dielectric barrier discharge (DBD) reactor installed in the tail pipe of a diesel engine, and a model DBD reactor fed with DEP in the mixture of N2and O2. When changing the experimental parameters of both the plasma conditions and the engine load conditions, we obtained characteristic information of DEP treated with plasma discharges from the particle diameter and the composition. In evaluating the model DBD reactor, it became clear that there were two types of plasma processes (reactions with active oxygen species to yield CO2and reactions with active nitrogen species to yield nitrogen containing compounds). Moreover, from the result of a TOF-SIMS analysis, the characteristic secondary ions, such as C2H6N+, C4H12N+, and C10H20N2+, were strongly detected from the DEP surfaces during the plasma discharges. This indicates that the nitrogen contained hydrocarbons were generated by plasma reactions.

Hydrogenated nitrile rubber (HNBR) and a ternary copolymer of vinylidene fluoride, tetrafluoropropylene, and perfluoromethyl vinyl ether (FKM) were blended and vulcanized rubbers that have the excellent low temperature characteristics and oil resistance were successfully obtained. The results of dynamic mechanical analysis demonstrated the great improvement of low temperature characteristics for one of the polymer blend materials (FKM/HNBR = 30/70). Subsequently, the structures of the polymer blends were analyzed by using atomic force microscopy (AFM) and transmission electron microscopy (TEM). The results of AFM and TEM showed that the polymer blend materials have phase‐separated structures with the sea‐island morphology. The sea regions were HNBR and the island regions were FKM for the samples containing less than 50% of FKM. On the other hand, the sea region was FKM for the sample containing 70% of FKM. Even in the stretched polymer blend, no gap or crazing was observed at the interface of FKM and HNBR. Therefore, some kind of strong bonding was presumed to exist between the two types of phases. © 2004 Wiley Periodicals, Inc. J Appl Polym Sci 95: 149–156, 2005

A novel therapeutic strategy for phenylketonuria (PKU) has been initiated in Japan. A total of 12 patients who met the criteria for tetrahydrobiopterin (BH4)-responsive hyperphenylalaninemia (HPA) with a mutant phenylalanine hydroxylase (PAH) (EC 1.14.16.1) gene were recruited at 12 medical centers in Japan between June 1995 and July 2001. Therapeutic efficacy of BH4was evaluated in single-dose, four-dose, and 1-wk BH4loading tests followed by long-term BH4treatment, and also examined in relation to the PAH gene mutations. The endpoints were determined as the percentage decline in serum phenylalanine from initial values after single-dose (>20%), four-dose (>30%), and 1-wk BH4(>50%) loading tests. Patients with mild PKU exhibiting decreases in blood phenylalanine concentrations of >20% in the single-dose test also demonstrated decreases of >30% in the four-dose test. The 1-wk test elicited BH4responsiveness even in patients with poor responses in the shorter tests. Patients with mild HPA, many of whom carry the R241C allele, responded to BH4administration. No clear correlation was noted between the degree of decrease in serum phenylalanine concentrations in the single- or four-dose tests and specific PAH mutations. The 1-wk test (20 mg/kg of BH4per day) is the most sensitive test for the diagnosis of BH4-responsive PAH deficiency. Responsiveness apparently depends on mutations in the PAH gene causing mild PKU, such as R241C. BH4proved to be an effective therapy that may be able to replace or liberalize the phenylalanine-restricted diets for a considerable number of patients with mild PKU.

Anuran larvae transform their epidermis to the adult counterpart during metamorphosis. The major event of this process is the proliferation of larval epidermal basal cells and their differentiation into adult ones. The present study isolated novel type I keratin cDNA dubbed xak-c (Xenopus adult keratin-c) that was exclusively expressed in adult epidermal basal cells. The gene started its expression in the larval epidermis at the onset of metamorphosis. Thyroid hormone (TH) induced the precocious expression of the gene in the epidermis of premetamorphic tadpoles. To study the transcriptional regulation of this gene in relation to epidermal metamorphosis, a 2.8 kb 5'-flanking region of xak-c was cloned and its promoter activity was investigated. Gene constructs were made so as to contain the xak-c promoter region and gene of EGFP or luciferase as a reporter gene and were transfected into various types of cells, which revealed that the 5'-flanking region had an epidermal cell-specific transcriptional activity in both anurans and mammals. Larval skin tissues of Xenopus were transfected with the constructs and cultured in the presence and absence of TH, which showed that the promoter region is responsive to TH, although the region did not contain the consensus TH response element-like sequence. In sharp contrast, the promoter region did not respond to TH in the adult skin, clearly indicating that the cloned region contains specific sequences that respond to metamorphosis-dependent transcription factor(s). © 2002 Wiley-Liss, Inc.

Rheological properties and processability of thermoforming were studied for high-density polyethylene (HDPE) and a blend of HDPE with crosslinked HDPE (xHDPE). Blending the xHDPE, which enhances melt strength and strain hardening in elongational viscosity of HDPE, helps the sheet avoid sagging in thermoforming. Moreover, the product of the blend obtained by vacuum forming has uniform wall thickness. Melt strength and strain hardening of the blend were, however, depressed by a processing history in a single-screw extruder, whereas reprocessing by a two-roll mill enhanced the melt strength again. It is considered that the processing history by a single-screw extruder, in which shear-dominant flow takes place, depresses the trapped entanglements between network chain of xHDPE and linear HDPE molecules, and results in low level of melt strength. © 2002 Wiley Periodicals, Inc. J Appl Polym Sci 86: 79–83, 2002

Blending a crosslinked high-density polyethylene (xHDPE) enhances melt strength and strain hardening behavior in elongational viscosity of high-density polyethylene (HDPE) to a great degree. Gel fraction of xHDPE has a stronger effect on the strain hardening than sol fraction, although sol fraction also enhances the strain hardening to some degree. Further, the xHDPE crosslinked by peroxide in a compression mold exhibits more pronounced effect than xHDPE by radiation, which is attributed to the difference in the amount of the gel fraction. The xHDPE, which enhances the strain hardening, has sparse crosslink points in the network. Moreover, it was found from linear viscoelastic measurements, such as oscillatory modulus and relaxation modulus, that the xHDPE is characterized as a critical gel, which was also supported by the result of tensile testing. © 2002 Wiley Periodicals, Inc. J Appl Polym Sci 86: 73–78, 2002

Heart transplantation has been extensively used in animal models, including studies on gene therapy for myocardial preservation. We investigated the feasibility of in situ left coronary artery (LCA) ligation as a physiological system for the examination of strategies to modulate myocardial tolerance against ischemia-reperfusion injury, such as gene therapy, using heterotopically transplanted rat hearts.

Objective:Skeletal myoblast transplantation is a promising alternative to treat heart failure. A single fiber, the minimal functional unit of skeletal muscle, retains skeletal myoblasts beneath the basal lamina. When surrounding muscle is injured, myoblasts migrate from the fiber into the damaged area to regenerate muscle. We hypothesized that such isolated fibers could be used as an efficient vehicle to deliver myoblasts into damaged myocardium, resulting in improved cardiac function. Methods:Living single fibers of rat skeletal muscle were isolated, and their behavior was characterized in vitro. Single fibers were injected into the myocardium (at 4 sites, each receiving a single fiber) of rats in 2 models of heart failure induced either by means of doxorubicin administration or left coronary artery occlusion. Results:Skeletal myoblasts dissociated from an isolated single fiber, proliferated, and differentiated into multinucleated myotubes in vitro. Within 3 days after grafting in vivo, original fibers provided putative myoblasts and disappeared. At 4 weeks, discrete loci consisting of several multinucleated myotubes were observed. Furthermore, single-fiber transplantation significantly improved cardiac function compared with the control treatment in either doxorubicin-treated hearts (maximum dP/dt, 4013.9 ± 96.1 vs 3603.1 ± 102.3 mm Hg/s; minimum dP/dt, −2313.7 ± 75.1 vs −2057.1 ± 52.4 mm Hg/s) or ischemic hearts (maximum dP/dt, 3905.6 ± 103.0 vs 3572.6 ± 109.7 mm Hg/s; minimum dP/dt, −2336.1 ± 69.7 vs −2106.4 ± 74.2 mm Hg/s). Conclusion:Single-fiber transplantation acts as a vehicle for delivering putative skeletal myoblasts that appear to differentiate into myotubes within the myocardium. This was associated with improved function of failing hearts, suggesting its efficacy as a novel graft for cellular cardiomyoplasty.

Helicobacter pyloriinduces severe neutrophilic infiltration in the lamina propria of the stomach, which leads to gastritis in humans. The possible involvement of a paracellular route for bacterial nutrients and etiologic agents that may play an important role in colonization of the bacteria and cause gastritis has been suggested. To study the functions of the paracellular barrier of gastric surface epithelium, SS1, a strain ofH. pyloriadapted to the murine stomach, was inoculated into the stomachs of C57BL/6 mice. At 4 months after inoculation, SS1 had achieved a high level of colonization (106–107colony-forming units/g tissue) associated with neutrophilic infiltration in the lamina propria of the junctional zone. Disruption of the paracellular barrier was observed in the SS1-infected stomachs, as revealed by the invasion of a lanthanum tracer into the paracellular space of the surface epithelium. Only 2% of junctions were permeable in control stomachs, whereas 72% of the paracellular barrier was disrupted in the SS1-infected gastric epithelia. Furthermore, distribution of tight junction-related molecules such as 7H6 antigen, occludin, and cortical actin was affected in the surface epithelium by SS1 infection. The linear expression pattern of occludin was disrupted and became irregular or punctuated. The 7H6 antigen accumulated as aggregates in the apical portion of the surface epithelium and cortical actin became irregular and punctuated. Taken together, these results indicate that infection by SS1 directly or indirectly caused an increase in paracellular permeability and altered the localization of tight junction-related molecules of the gastric surface epithelium. This observation suggests that the paracellular pathway may play a significant role in establishingH. pylori-induced gastritis in the clinical setting.

We examined the biochemical properties of YM‐57029 ({4‐[4‐(4‐Carbamimidoylphenyl)‐3‐oxopiperazin‐1‐yl]piperidino}acetic acid monohydrochloride trihydrate) and the pharmacodynamics and pharmacokinetics of its prodrug, YM128 (Ethyl (Z)‐(4‐{4‐[4‐(N2‐hydroxycarbamimidoyl)phenyl]‐3‐oxopiperazin‐1‐yl}piperidino)acetate), an orally‐active glycoprotein IIb/IIIa (GPIIb/IIIa) antagonist. YM‐57029 strongly inhibited aggregation of human platelets induced by various agonists, with IC50 values ranging from 3.6 to 51 nM. YM‐57029 specifically inhibited fibrinogen binding to purified GPIIb/IIIa about 1,000‐fold more potently than Arg‐Gly‐Asp‐Ser (RGDS). Moreover, YM‐57029 effectively inhibited an Arg‐Gly‐Asp (RGD) peptide binding to platelets, suggesting that YM‐57029 competed with the RGD sequence of ligand. YM‐57029 or YM128 dose‐dependently inhibited ex vivo platelet aggregation after iv bolus injection or oral administration to beagle dogs and cynomolgus monkeys. However, YM128 exerted more potent and prolonged inhibitory effects on platelet aggregation than YM‐57029 after oral administration to cynomolgus monkeys. Furthermore, YM‐57029 prolonged template bleeding time at a dose that inhibited ex vivo platelet aggregation during cumulative iv infusion to cynomolgus monkeys. Metabolic and pharmacokinetic studies showed that YM128 effectively converted into YM‐57029 in liver microsomes from humans as well as dogs and monkeys, and that bioavailabilities of YM128 in dogs and monkeys were 32.3 and 22.2%, respectively. These results suggest that YM128, a prodrug of YM‐57029, may be a valuable GPIIb/IIIa antagonist with good bioavailability in humans. Drug Dev. Res. 55:149–161, 2002. © 2002 Wiley‐Liss, Inc.