17 research outputs found

    Ultrasonic tomographic imaging of defects in industrial materials

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    Ultrasonic tomography has been fairly widely applied for imaging of inhomogeneities in isotropic materials, particularly in the medical field, however, little success has been made in its application to industrial materials. This is largely due to the complex nature of ultrasonic wave propagation in these anisotropic materials. The three dimensional characteristics of ultrasonic wave propagation in anisotropic materials have been thoroughly studied for single crystals and also studied recently for different composites [1,2,3]. Understanding these characteristics provides the theoretical background for developing appropriate ultrasonic tomographic imaging methods for industrial materials

    Rpb1 Sumoylation in Response to UV Radiation or Transcriptional Impairment in Yeast

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    Covalent modifications of proteins by ubiquitin and the Small Ubiquitin-like MOdifier (SUMO) have been revealed to be involved in a plethora of cellular processes, including transcription, DNA repair and DNA damage responses. It has been well known that in response to DNA damage that blocks transcription elongation, Rpb1, the largest subunit of RNA polymerase II (Pol II), is ubiquitylated and subsequently degraded in mammalian and yeast cells. However, it is still an enigma regarding how Pol II responds to damaged DNA and conveys signal(s) for DNA damage-related cellular processes. We found that Rpb1 is also sumoylated in yeast cells upon UV radiation or impairment of transcription elongation, and this modification is independent of DNA damage checkpoint activation. Ubc9, an E2 SUMO conjugase, and Siz1, an E3 SUMO ligase, play important roles in Rpb1 sumoylation. K1487, which is located in the acidic linker region between the C-terminal domain and the globular domain of Rpb1, is the major sumoylation site. Rpb1 sumoylation is not affected by its ubiquitylation, and vice versa, indicating that the two processes do not crosstalk. Abolishment of Rpb1 sumoylation at K1487 does not affect transcription elongation or transcription coupled repair (TCR) of UV-induced DNA damage. However, deficiency in TCR enhances UV-induced Rpb1 sumoylation, presumably due to the persistence of transcription-blocking DNA lesions in the transcribed strand of a gene. Remarkably, abolishment of Rpb1 sumoylation at K1487 causes enhanced and prolonged UV-induced phosphorylation of Rad53, especially in TCR-deficient cells, suggesting that the sumoylation plays a role in restraining the DNA damage checkpoint response caused by transcription-blocking lesions. Our results demonstrate a novel covalent modification of Rpb1 in response to UV induced DNA damage or transcriptional impairment, and unravel an important link between the modification and the DNA damage checkpoint response

    Systematic Two-Hybrid and Comparative Proteomic Analyses Reveal Novel Yeast Pre-mRNA Splicing Factors Connected to Prp19

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    Prp19 is the founding member of the NineTeen Complex, or NTC, which is a spliceosomal subcomplex essential for spliceosome activation. To define Prp19 connectivity and dynamic protein interactions within the spliceosome, we systematically queried the Saccharomyces cerevisiae proteome for Prp19 WD40 domain interaction partners by two-hybrid analysis. We report that in addition to S. cerevisiae Cwc2, the splicing factor Prp17 binds directly to the Prp19 WD40 domain in a 1∢1 ratio. Prp17 binds simultaneously with Cwc2 indicating that it is part of the core NTC complex. We also find that the previously uncharacterized protein Urn1 (Dre4 in Schizosaccharomyces pombe) directly interacts with Prp19, and that Dre4 is conditionally required for pre-mRNA splicing in S. pombe. S. pombe Dre4 and S. cerevisiae Urn1 co-purify U2, U5, and U6 snRNAs and multiple splicing factors, and dre4Ξ” and urn1Ξ” strains display numerous negative genetic interactions with known splicing mutants. The S. pombe Prp19-containing Dre4 complex co-purifies three previously uncharacterized proteins that participate in pre-mRNA splicing, likely before spliceosome activation. Our multi-faceted approach has revealed new low abundance splicing factors connected to NTC function, provides evidence for distinct Prp19 containing complexes, and underscores the role of the Prp19 WD40 domain as a splicing scaffold

    Gene Expression in a Drosophila Model of Mitochondrial Disease

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    Background A point mutation in the Drosophila gene technical knockout (tko), encoding mitoribosomal protein S12, was previously shown to cause a phenotype of respiratory chain deficiency, developmental delay, and neurological abnormalities similar to those presented in many human mitochondrial disorders, as well as defective courtship behavior. Methodology/Principal Findings Here, we describe a transcriptome-wide analysis of gene expression in tko25t mutant flies that revealed systematic and compensatory changes in the expression of genes connected with metabolism, including up-regulation of lactate dehydrogenase and of many genes involved in the catabolism of fats and proteins, and various anaplerotic pathways. Gut-specific enzymes involved in the primary mobilization of dietary fats and proteins, as well as a number of transport functions, were also strongly up-regulated, consistent with the idea that oxidative phosphorylation OXPHOS dysfunction is perceived physiologically as a starvation for particular biomolecules. In addition, many stress-response genes were induced. Other changes may reflect a signature of developmental delay, notably a down-regulation of genes connected with reproduction, including gametogenesis, as well as courtship behavior in males; logically this represents a programmed response to a mitochondrially generated starvation signal. The underlying signalling pathway, if conserved, could influence many physiological processes in response to nutritional stress, although any such pathway involved remains unidentified. Conclusions/Significance These studies indicate that general and organ-specific metabolism is transformed in response to mitochondrial dysfunction, including digestive and absorptive functions, and give important clues as to how novel therapeutic strategies for mitochondrial disorders might be developed.Public Library of Scienc

    Religion and forgiveness from a Hong Kong Chinese perspective

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    This study investigated the relationship between religion and forgiveness in a sample of Hong Kong Chinese teachers (n=230) and students (n=714). Findings indicated some influence from Chinese cultural values in the conceptualization of forgiveness. Religious affiliation was the strongest predictor of concepts of forgiveness, whereas religious practice predicted attitudes toward "forgivingness" and the practice of forgiveness. No significant difference in forgiveness between believers and non-believers in real life situations was reported. Implications for future research on forgiveness are discussed. Β© Springer Science+Business Media, Inc. 2006.link_to_subscribed_fulltex

    Isolation, characterization and expression analysis of a hypoxia-responsive glucose transporter gene from the grass carp, Ctenopharyngodon idellus

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    Glucose transporters (GLUTs) have been implicated in adaptive and survival responses to hypoxic stress in mammals. In fish, the expression and regulation of GLUT in relation to hypoxia remains unexplored. Here we describe the identification of a hypoxia-responsive glucose transporter gene (gcGLUT) and the corresponding full-length cDNA from the grass carp. The gene spans β‰ˆ 11 kb of genomic sequence and consists of 12 exons and 11 introns, and an open reading frame (ORF) of 1599 bp encoding a polypeptide of 533 amino acids, with a predicted molecular mass of β‰ˆ 57 kDa and a pI of 8.34. BLASTX analysis showed that the ORF shared high sequence identity with the GLUT1 (57-59%), GLUT3 (59-60%) and GLUT4 (55-59%) proteins from different vertebrates. Comparative analysis of GLUT genomic structures showed that the arrangement of exons and position of split codons are highly conserved amongst members of the class I GLUTs suggesting that these genes share a common ancestor. Phylogenetic analysis indicated that gcGLUT is most closely related to the GLUT3 proteins. Northern blot analysis showed that the 3.1-kb gcGLUT transcript was most abundantly expressed and responsive to hypoxia in kidney. Up-regulated expression by hypoxia was also evident in eye and gill, but differential patterns of expression were observed. Low expression levels detected in brain, heart, liver and muscle were not responsive to hypoxic stress.link_to_subscribed_fulltex

    Colorimetry

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