RBM20 and Cardiomyopathy

RBM20 is a vertebrate-specific RNA-binding protein with two zinc finger (ZnF) domains, one RNA-recognition motif (RRM)-type RNA-binding domain and an arginine/serine (RS)-rich region. RBM20 has initially been identified as one of dilated cardiomyopathy (DCM)-linked genes. RBM20 is a regulator of heart-specific alternative splicing and Rbm20ΔRRM mice lacking the RRM domain are defective in the splicing regulation. The Rbm20ΔRRM mice, however, do not exhibit a characteristic DCM-like phenotype such as dilatation of left ventricles or systolic dysfunction. Considering that most of the RBM20 mutations identified in familial DCM cases were heterozygous missense mutations in an arginine-serine-arginine-serine-proline (RSRSP) stretch whose phosphorylation is crucial for nuclear localization of RBM20, characterization of a knock-in animal model is awaited. One of the major targets for RBM20 is the TTN gene, which is comprised of the largest number of exons in mammals. Alternative splicing of the TTN gene is exceptionally complicated and RBM20 represses >160 of its consecutive exons, yet detailed mechanisms for such extraordinary regulation are to be elucidated. The TTN gene encodes the largest known protein titin, a multi-functional sarcomeric structural protein specific to striated muscles. As titin is the most important factor for passive tension of cardiomyocytes, extensive heart-specific and developmentally regulated alternative splicing of the TTN pre-mRNA by RBM20 plays a critical role in passive stiffness and diastolic function of the heart. In disease models with diastolic dysfunctions, the phenotypes were rescued by increasing titin compliance through manipulation of the Ttn pre-mRNA splicing, raising RBM20 as a potential therapeutic target

Alternative Splicing Regulator RBM20 and Cardiomyopathy

RBM20 is a vertebrate-specific RNA-binding protein with two zinc finger (ZnF) domains, one RNA-recognition motif (RRM)-type RNA-binding domain and an arginine/serine (RS)-rich region. RBM20 has initially been identified as one of dilated cardiomyopathy (DCM)-linked genes. RBM20 is a regulator of heart-specific alternative splicing and Rbm20ΔRRM mice lacking the RRM domain are defective in the splicing regulation. The Rbm20ΔRRM mice, however, do not exhibit a characteristic DCM-like phenotype such as dilatation of left ventricles or systolic dysfunction. Considering that most of the RBM20 mutations identified in familial DCM cases were heterozygous missense mutations in an arginine-serine-arginine-serine-proline (RSRSP) stretch whose phosphorylation is crucial for nuclear localization of RBM20, characterization of a knock-in animal model is awaited. One of the major targets for RBM20 is the TTN gene, which is comprised of the largest number of exons in mammals. Alternative splicing of the TTN gene is exceptionally complicated and RBM20 represses >160 of its consecutive exons, yet detailed mechanisms for such extraordinary regulation are to be elucidated. The TTN gene encodes the largest known protein titin, a multi-functional sarcomeric structural protein specific to striated muscles. As titin is the most important factor for passive tension of cardiomyocytes, extensive heart-specific and developmentally regulated alternative splicing of the TTN pre-mRNA by RBM20 plays a critical role in passive stiffness and diastolic function of the heart. In disease models with diastolic dysfunctions, the phenotypes were rescued by increasing titin compliance through manipulation of the Ttn pre-mRNA splicing, raising RBM20 as a potential therapeutic target

Development of a Si/CdTe semiconductor Compton telescope

We are developing a Compton telescope based on high resolution Si and CdTe imaging devices in order to obtain a high sensitivity astrophysical observation in sub-MeV gamma-ray region. In this paper, recent results from the prototype Si/CdTe semiconductor Compton telescope are reported. The Compton telescope consists of a double-sided Si strip detector (DSSD) and CdTe pixel detectors, combined with low noise analog LSI, VA32TA. With this detector, we obtained Compton reconstructed images and spectra from line gamma-rays ranging from 81 keV up to 356 keV. The energy resolution is 3.8 keV and 7.9 keV at 122 keV and 356 keV, respectively, and the angular resolution is 9.9 degrees and 5.7 degrees at 122 keV and 356 keV, respectively.Comment: 12 pages, 14 figures, submitted to SPIE conference proceedings vol. 5501, "High-Energy Detectors in Astronomy", Glasgow UK, 6/21-6/24 200

Low-molecular weight fractions of Japanese soy sauce act as a RAGE antagonist via inhibition of RAGE trafficking to lipid rafts

Advanced glycation end-products (AGE) have been implicated in aging and the pathogenesis of diabetic complications, inflammation, Alzheimer\u27s disease, and cancer. AGE engage the cell surface receptor for AGE (RAGE), which in turn elicits intracellular signaling, leading to activation of NF-κB to cause deterioration of tissue homeostasis. AGE are not only formed within our bodies but are also derived from foods, endowing them with flavor. In the present study, we assessed the agonistic/antagonistic effects of food-derived AGE on RAGE signaling in a reporter assay system and found that low-molecular weight AGE can antagonize the action of AGE-BSA. Foods tested were Japanese soy sauce, coffee, cola, and red wine, all of which showed fluorescence characteristics of AGE. Soy sauce and coffee contained Nε-carboxymethyl-lysine (CML). Soy sauce, coffee, and red wine inhibited the RAGE ligand-induced activation of NF-κB, whereas cola had no effect on the ligand induction of NF-κB. The liquids were then fractionated into high-molecular weight (HMW) fractions and low-molecular weight (LMW) fractions. Soy sauce-, coffee-, and red wine-derived LMW fractions consistently inhibited the RAGE ligand induction of NF-κB, whereas the HMW fractions of these foods activated RAGE signaling. Using the LMW fraction of soy sauce as a model food-derived RAGE antagonist, we performed a plate-binding assay and found that the soy sauce LMW fractions competitively inhibited AGE-RAGE association. Further, this fraction significantly reduced AGE-dependent monocyte chemoattractant protein-1 (MCP-1) secretion from murine peritoneal macrophages. The LMF from soy sauce suppressed the AGE-induced RAGE trafficking to lipid rafts. These results indicate that small components in some, if not all, foods antagonize RAGE signaling and could exhibit beneficial effects on RAGE-related diseases. © 2013 The Royal Society of Chemistry

Genome-wide association study of lung adenocarcinoma in East Asia and comparison with a European population

Lung adenocarcinoma is the most common type of lung cancer. Known risk variants explain only a small fraction of lung adenocarcinoma heritability. Here, we conducted a two-stage genome-wide association study of lung adenocarcinoma of East Asian ancestry (21,658 cases and 150,676 controls; 54.5% never-smokers) and identified 12 novel susceptibility variants, bringing the total number to 28 at 25 independent loci. Transcriptome-wide association analyses together with colocalization studies using a Taiwanese lung expression quantitative trait loci dataset (n = 115) identified novel candidate genes, including FADS1 at 11q12 and ELF5 at 11p13. In a multi-ancestry meta-analysis of East Asian and European studies, four loci were identified at 2p11, 4q32, 16q23, and 18q12. At the same time, most of our findings in East Asian populations showed no evidence of association in European populations. In our studies drawn from East Asian populations, a polygenic risk score based on the 25 loci had a stronger association in never-smokers vs. individuals with a history of smoking (P interaction  = 0.0058). These findings provide new insights into the etiology of lung adenocarcinoma in individuals from East Asian populations, which could be important in developing translational applications

Genome-wide association study of lung adenocarcinoma in East Asia and comparison with a European population.

Lung adenocarcinoma is the most common type of lung cancer. Known risk variants explain only a small fraction of lung adenocarcinoma heritability. Here, we conducted a two-stage genome-wide association study of lung adenocarcinoma of East Asian ancestry (21,658 cases and 150,676 controls; 54.5% never-smokers) and identified 12 novel susceptibility variants, bringing the total number to 28 at 25 independent loci. Transcriptome-wide association analyses together with colocalization studies using a Taiwanese lung expression quantitative trait loci dataset (n = 115) identified novel candidate genes, including FADS1 at 11q12 and ELF5 at 11p13. In a multi-ancestry meta-analysis of East Asian and European studies, four loci were identified at 2p11, 4q32, 16q23, and 18q12. At the same time, most of our findings in East Asian populations showed no evidence of association in European populations. In our studies drawn from East Asian populations, a polygenic risk score based on the 25 loci had a stronger association in never-smokers vs. individuals with a history of smoking (Pinteraction = 0.0058). These findings provide new insights into the etiology of lung adenocarcinoma in individuals from East Asian populations, which could be important in developing translational applications

CELF family RNA-binding protein UNC-75 regulates two sets of mutually exclusive exons of the unc-32 gene in neuron-specific manners in Caenorhabditis elegans.

An enormous number of alternative pre-mRNA splicing patterns in multicellular organisms are coordinately defined by a limited number of regulatory proteins and cis elements. Mutually exclusive alternative splicing should be strictly regulated and is a challenging model for elucidating regulation mechanisms. Here we provide models of the regulation of two sets of mutually exclusive exons, 4a-4c and 7a-7b, of the Caenorhabditis elegans uncoordinated (unc)-32 gene, encoding the a subunit of V0 complex of vacuolar-type H(+)-ATPases. We visualize selection patterns of exon 4 and exon 7 in vivo by utilizing a trio and a pair of symmetric fluorescence splicing reporter minigenes, respectively, to demonstrate that they are regulated in tissue-specific manners. Genetic analyses reveal that RBFOX family RNA-binding proteins ASD-1 and FOX-1 and a UGCAUG stretch in intron 7b are involved in the neuron-specific selection of exon 7a. Through further forward genetic screening, we identify UNC-75, a neuron-specific CELF family RNA-binding protein of unknown function, as an essential regulator for the exon 7a selection. Electrophoretic mobility shift assays specify a short fragment in intron 7a as the recognition site for UNC-75 and demonstrate that UNC-75 specifically binds via its three RNA recognition motifs to the element including a UUGUUGUGUUGU stretch. The UUGUUGUGUUGU stretch in the reporter minigenes is actually required for the selection of exon 7a in the nervous system. We compare the amounts of partially spliced RNAs in the wild-type and unc-75 mutant backgrounds and raise a model for the mutually exclusive selection of unc-32 exon 7 by the RBFOX family and UNC-75. The neuron-specific selection of unc-32 exon 4b is also regulated by UNC-75 and the unc-75 mutation suppresses the Unc phenotype of the exon-4b-specific allele of unc-32 mutants. Taken together, UNC-75 is the neuron-specific splicing factor and regulates both sets of the mutually exclusive exons of the unc-32 gene

Through-hole drilling characteristics of alumina ceramics using diamond-coated carbide drill

The drilling characteristics of alumina ceramics during through-hole drilling with a diamond-coated carbide drill were investigated experimentally. The drilling characteristics were evaluated by cutting edge behavior, cutting force (thrust force and torque), and chipping state at the exit side of the drilled hole in relation to the number of drilled holes. Additionally, the effects of the diamond-coating thickness and drill feed rate were investigated. The drills had a typical twist shape and were made of tungsten carbide base material with diamond coatings at 10 μm and 20 μm thick. The drilling machine was a standard machining center with a simple unidirectional drill feed without vibration. In through-hole drilling, as in blind-hole drilling, the coating on the rake face flaked during the initial stage of drilling, resulting in a sharp cutting edge on the diamond-coating ridge remaining on the flank face. However, flaking of the diamond coating on the rake face was observed earlier during through-hole drilling than in blind-hole drilling. Although the chipping area increased with increasing drill feed rate, coating flaking on the rake face significantly suppressed it at all feed rates. There was a clear correlation between the chipping area and cutting force since the chipping area decreased with decreasing thrust force. Before the theoretical hole penetration, hat-shaped chipping was observed with two crack propagation directions, accompanied by a sharp decrease in thrust force. Moreover, even after observing a sharp decrease in thrust force, a gradual decrease in thrust force was observed in the case of a good chipping state. The chipping area was significantly improved by reducing the coating thickness, and the thrust force was reduced

The UGCAUG stretch and the RBFOX family proteins are involved in the neuron-specific selection of exon 7a from the <i>unc-32</i> exon 7 reporter.

<p>(<i>A</i>) Fluorescence images of the exon 7 reporter worms <i>ybIs1622 [rgef-1::unc-32E7a-EGFP rgef-1::unc-32E7b-mRFP]</i> with dual-bandpass (Dual), green (GFP2) and red (DSR) filters and a bright field image (BF). Note that individual neurons differentially express E7a-EGFP and E7b-mRFP in a cell-type-specific pattern. (<i>B</i>) Nucleotide sequence alignment of intron 7b from <i>C. briggsae</i>, <i>C. elegans</i> and <i>C. remanei</i>. Residues shaded in black and gray are conserved among three and two species, respectively. Conserved stretches are boxed in red and the sequences of the <i>M1</i> and <i>M2</i> mutant pairs of the reporter minigenes are indicated. (<i>C</i>) Schematic illustrations of the mutated pairs of the exon 7 reporter minigenes. Red arrows indicate the positions of the modification. (<i>D</i>) A fluorescence image of the <i>M1</i> mutant reporter worms with a dual-bandpass filter. (<i>E</i>) Fluorescence images of the <i>ybIs1622</i> worms in the <i>asd-1 (yb978)</i> (top), <i>fox-1 (e2643)</i> (middle) and <i>asd-1; fox-1</i> (bottom) backgrounds with a dual-bandpass filter. Scale bars, 200 µm in (<i>A</i>), (<i>D</i>) and (<i>E</i>). (<i>F</i>) Top, radiolabelled wild-type (WT) and mutant (Mut) intron 7b probes. The substituted bases are underlined. Lowercase indicates the sequence derived from the T7 promoter. Bottom left, Neutral PAGE and CBB staining of the recombinant FLAG-tagged ASD-1 and FOX-1 proteins. Bottom right, EMSA using the WT and Mut probes without (−) or with 4-fold dilution series of FLAG-ASD-1 and -FOX-1.</p