Exploring the Role of FUS Mutants from Stress Granule Incorporation to Nucleopathy in Amyotrophic Lateral Sclerosis: A Dissertation

Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease characterized by preferential motor neuron death in the brain and spinal cord. The rapid disease progression results in death due to respiratory failure, typically within 3-5 years after disease onset. While ~90% of cases occur sporadically, remaining 10% of ALS cases show familial inheritance, and the number of genes linked to ALS has increased dramatically over the past decade. FUS/TLS (Fused in Sarcoma/ Translocated to liposarcoma) is a nucleic acid binding protein that may regulate several cellular functions, including RNA splicing, transcription, DNA damage repair and microRNA biogenesis. More than 50 mutations in the FUS gene are linked to 4% of familial ALS, and many of these may disrupt the nuclear localization signal, leading to variable amounts of FUS accumulation in the cytoplasm. However, the mechanism by which FUS mutants cause motor neuron death is still unknown. The studies presented in this dissertation focused on investigating the properties of FUS mutants in the absence and presence of stress conditions. We first examined how ALS-linked FUS mutants behaved in response to imposed stresses in both cell culture and zebrafish models of ALS. We found that FUS mutants were prone to accumulate in stress granules in proportion to their degree of cytoplasmic mislocalization under conditions of oxidative stress, ER stress, and heat shock. However, many FUS missense mutants are retained predominantly in the nucleus, and this suggested the possibility that these mutants might also perturb one or more nuclear functions. In a human cell line expressing FUS variants and in human fibroblasts from an ALS patient, mutant FUS expression was associated with enlarged promyelocytic leukemia nuclear bodies (PML-NBs) under basal condition. Upon oxidative insult with arsenic trioxide (ATO), PML-NBs in control cells increased acutely in size and were turned over within 12-24 h, as expected. However, PML-NBs in FUS mutant cells did not progress through the expected turnover but instead continued to enlarge over 24 h. We also observed a persistent accumulation of the transcriptional repressor Daxx and the 11S proteasome regulator in association with these enlarged PML-NBs. Furthermore, the peptidase activities of the 26S proteasome were decreased in FUS mutant cells without any changes in the expression of proteasome subunits. These results demonstrate that FUS mutant expression may alter cellular stress responses as manifested by (i) accumulation of mutant FUS into stress granules and (ii) inhibition of PML-NB dynamics. These findings suggest a novel nuclear pathology specific to mutant FUS expression that may perturb nuclear homeostasis and thereby contribute to ALS pathogenesis

Why do Family Firms Pay Cash Dividends in Emerging Markets? Corporate Control and Family Succession in Korea

Following the economic crisis in 1997, the Korean government introduced the enhanced corporate governance and reform policy, which drove family-controlled firms to search strategic reaction for control succession and wealth transfer. This paper explores alternative explanations for why Korean firms choose to pay cash dividends around this corporate reform period. What lead firms to pay cash dividends remains largely unexplained by the reducing agency cost, signaling, or life-cycle theories. This study focuses on relations between the ownership structure and cash dividends payout, seeking effects deriving from (i) controlling shareholder (CS) and (ii) their family members. The logit analysis result shows that firms with large control rights, especially higher ownership of other family members of CS are more likely to pay cash dividends. After adjusting for the characteristics that affect the degree of cash dividends, ownership variables are positively related to payout ratios and dividend yields. CS family members' ownership has a statistically stronger effect on payout ratios than CS's. These results provide the evidence of incentive for corporate control succession within the family with least costs carried by the family members of controlling shareholders who positively influence payout decisions and dividend ratios

Development of simple and rapid procedures for purification of egg yolk immuno-globulin and ovotransferrin from chicken egg and antimicrobial activity of ovo-transferrin against Escherichia coli O157:H7 and Listeria monocytogenes

The overall goal of this study was to develop practical, natural, and efficient antimicrobials from egg. To accomplish the goal, novel protocols for large-scale production of egg yolk immunoglobulin and ovotransferrin were developed. Also antibacterial activity of ovotransferrin against Escherichia coli O157:H7 and Listeria monocytogenes was investigated and the antimicrobial activity of ovotransferrin was improved by combinations with other antimicrobials. The developed protocol for large-scale production of egg yolk immunoglobulin (IgY) consisted of several steps such as using 0.01% charcoal to remove lipoproteins from IgY extract, ultrafiltration to concentrate IgY solution, and 40% ammonium sulfate to precipitate IgY following dialysis. Morevoer, apo-ovotransferrin was isolated from 2 time-diluted egg white by following steps: as iron saturation of egg white, extraction of iron-bound (holo) ovotransferrin using 43% ethanol, precipitation of holo-bound ovotransferrin by 59% ethanol, dissolving the precipitate with distilled water, and removal iron from holo-ovotransferrin using AG RTM1-X 2 ion exchange resin. After separating apo-ovotransferrin, the effects of citric acid, Zn- and Fe- binding, and sodium bicarbonate (NaHCO3) on the antibacterial activity of ovotransferrin against E. coli O157:H7 and L. monocytogenes in model systems using BHI broth were investigated. Synergistic effects of EDTA and lysozyme on the antimicrobial activities of ovotransferrin were investigated toward E. coli O157 and L. monocytogenes on model systems using BHI broth and pork chops and hams.;According to this study, IgY obtained by developed protocol was approximately 70--80% recovery and approximately 80% purity. The new procedure seems to be economical and practical for a large-scale purification of IgY from egg yolk since ammonium sulfate precipitation could accommodate of large volume of egg yolk. Also, the recovery rate and purity of ovotransferrin produced by the newly developed protocol was around 94% and 80% respectively. As the procedure is consisted of only a few steps, it is suitable to a large-scale production of ovotransferrin. Also becasuse ethanol and AG RTM1-X 2 ion exchange resins can be regenerated, it seems to be economical and efficient. In study related to antibacterial activity of ovotransferrin, 100 mM-NaHCO3 significantly increased the antibacterial activity of ovotransferrin against both E. coli O157:H7 and L. monocytogenes. 0.5% citric acid enhanced the antibacterial activities of ovotransferrin against E. coli O157:H7, Zn-bound ovotransferrin appeared to inhibit the growth of L. monocytogenes. So, this study suggests that the iron binding capacity of ovotransferrin was not the major mechanism of antimicrobial actions of ovotransferrin because acidic pH or Zn2+ promotes antibacterial activity of ovotransferrin toward E. coli O157:H7 or L. monocytogenes respectively. Also, 2 mg/ml EDTA improved antibacterial activity of ovotransferrin containing 100 mM-NaHCO3 against E. coli O157:H7. Also when lysozyme was combined with OS, the antibacterial activity of OS against L. monocytogenes increased slightly in vitro test. However, synergistic effect of lysozyme on antimicrobial activity of OS against L. monocytogenes did not show significantly in this study. Contrary to the model system results using BHI broth, ovotransferrin plus EDTA or/and lysozyme did not show any antimicrobial activities toward E. coli O157:H7 and L. monocytogenes in pork chops and hams.;Developing preservatives such as IgY and ovotransferrin are significant not only for the food industry but also for consumers because they are derived from natural sources and generally regarded as safe. Use of these products will significantly increase the value of eggs, which is important for egg industry. Furthermore, this study claims that ovotransferrins combined with NaHCO3, EDTA, or lysozyme have high potential as natural antimicrobial agents to control E. coli O157:H7 and L. monocytogenes. Also, further studies are required to elucidate the antibacterial mechanisms of ovotransferrin and to overcome the limitations of applying ovotransferrin in food products in the future

Asian Dance/Movement Therapy Educators’ Experiences of Teaching Dance/Movement Therapy in East Asia after Training in the US

This phenomenological study aimed to explore the lived learning and teaching experiences of native East Asian DMT educators who completed their DMT education in the US and then returned to their home countries. The researcher intended to learn how participants experienced their Western DMT education and how they adapted what they had learned to teaching DMT to students in East Asia. Six participants were successfully recruited from four East Asian countries (Hong Kong, Japan, Korea, and Taiwan). Three types of data were gathered via a researcher-designed demographic questionnaire, the Asian American Values Scale-Multidimensional, and individual interviews. The Nvivo10 software program was used to ensure efficient organization. Member checking and peer debriefing were used in different stages of data analysis to heighten internal validity. Three phases of coding processes were conducted. Approximately 100 pages of interview transcription yielded 638 meaningful segments during the first coding process, unrelated or unclear segment codes were deleted (559 codes), and similar or identical codes were then combined (516 codes). Finally, data analysis revealed 8 themes, 22 categories, and 72 subcategories. The eight themes were (1) emergence of new personal and professional identity, (2) DMT training experience in the US, (3) unwanted return and not welcomed home, (4) challenges as a DMT educator in one’s home country, (5) unfamiliar and different learning styles, (6) problems and needs, (7) efforts to develop the field of DMT as a DMT educator, and (8) giving advice and sharing meaningful moments. These findings support previous research on culturally competent teaching and contribute to creating a communicative and educational DMT bridge between the East and West, improving mutual understanding. More research is needed to enhance the understanding of culturally competent DMT teaching and indigenization in East Asia

TWENTIETH CENTURY VARIATIONS ON BORROWED THEMES

Mergers and acquisitions are words that are usually associated with the modern business world. Such joint efforts toward improvement, however, existed long before our time, in the form of musical partnerships. It was not unusual for composers to share in each other's works, borrowing themes and recreating them to generate new meanings; in the process, new masterpieces were often created. My performance project, Twentieth Century Variations on Borrowed Themes, explores the fruits of such labor. The main objective of this project is to demonstrate how certain composers of the twentieth century have taken famous themes and used them to create variations, imbuing their own creative ideas, musical styles and pianistic challenges. This objective was accomplished by performing three recorded public recitals. These programs consisted of early to late twentieth century pieces that are based on borrowed themes, either in theme and variations form, fantasia form, paraphrase form, or transformal variation form. I have selected the pieces based on their artistic merits and technical challenges, thus allowing me to grow as a pianist and artist. In addition, I wanted to choose some pieces that are rarely performed, as I believe the public delights in hearing unfamiliar gems. The first recital consisted of the music of two legendary pianists: Variations on a Theme of Chopin by Rachmaninoff and Goldberg Variations by BachIBusoni. The second program featured Grand Fantasy on Gershwin's Porgy and Bess by Earl Wild, Sonatina No. 6 (Fantasy on Bizet 's "Carmen") by Busoni, and Rhapsody on a Theme by Paganini, op.43 by Rachmaninoff. Some unusual and seldom-performed pieces, as well as a familiar favorite, were spotlighted in the third recital. The pieces performed on this program were John Rea's Twenty-one Transformal Variations on the "Kindersznen " by Robert Schumann (Las Meninas), Muczynski's Desperate Measures (Paganini Variations), Busoni's Elegie No. 3 (Turandot 's room), and Rhapsodie Espagnole by LisztIBuson. These composers artfully breathed new life into the material borrowed from others, and in the process, the "borrowed" themes became undoubtedly and uniquely their own music

Active site phosphoryl groups in the biphosphorylated phosphotransferase complex reveal dynamics in a millisecond time scale

AbstractThe N-terminal domain of Enzyme I (EIN) and phosphocarrier HPr can form a biphosphorylated complex when they are both phosphorylated by excess cellular phosphoenolpyruvate. Here we show that the electrostatic repulsion between the phosphoryl groups in the biphosphorylated complex results in characteristic dynamics at the active site in a millisecond time scale. The dynamics is localized to phospho-His15 and the stabilizing backbone amide groups of HPr, and does not impact on the phospho-His189 of EIN. The dynamics occurs with the kex of ∼500s−1 which compares to the phosphoryl transfer rate of ∼850s−1 between EIN and HPr. The conformational dynamics in HPr may be important for its phosphotransfer reactions with multiple partner proteins.Structured summary of protein interactionsEIN and HPr bind by nuclear magnetic resonance (View Interaction)

EDTA and Lysozyme Improves Antimicrobial Activities of Ovotransferrin against Escherichia coli O157:H7

The aim of this study was to evaluate the effect of ethylenediaminetetraacetic acid (EDTA) or/and lysozyme on the antibacterial activity of ovotransferrin against E. coli O157:H7. Ovotransferrin solution (20 mg/ml) containing 100 mM-NaHCO3 (OS) was added with EDTA (2.0 or 2.5 mg/ml), lysozyme (1.0, 1.5, or 2.0 mg/ml) or both were prepared. Antibacterial activities of OS (20 mg/ml ovotransferrin + 100 mM-NaHCO3), OSE (OS+ EDTA), or OSL (OS + lysozyme) against E. coli O157:H7 in model systems were investigated by turbidity and viability tests. Also, OSE, OSL or OSEL (OS + EDTA + lysozyme) was applied on irradiated pork chops and commercial hams to determine if the solutions have antibacterial activity on meat products. The effect of initial cell population on the antibacterial activity of ovotransferrin and EDTA or lysozyme combinations was also determined. EDTA at 2 mg/ml plus OS (OSE2) induced 3 ~ 4 log reduction in viable E. coli O157:H7 cells in brain heart infusion (BHI) broth media, and 1 mg/ml lysozyme plus OS (OSL1) resulted in 0.5 ~ 1.0 log reduction during 35 oC incubation for 36 hr. However, OSE or OSEL did not show significant antibacterial effect on pork chops and hams during storage at 10 oC. The initial cell number in media did not affect the antibacterial activity of OSE or OSEL against E. coli O157:H7. This study demonstrates that combinations of ovotransferrin, NaHCO3, and EDTA (OSE) have potential to control E. coli O157:H7

Pulsed Corona Discharge for Oxidation of Gaseous Elemental Mercury

Positive pulsed corona discharge has been applied for the oxidation of gaseous elemental mercury (Hg0) from a simulated flue gas. The oxidation of Hg0 to HgO and HgCl2 can significantly enhance the mercury removal from flue gas. At a gas condition of O2 (10%), H2O (3%), and N2 (balance), Hg0 oxidation efficiency of 84% was achieved at an input energy density of 45 J/l. The presence of NO, however, hinders Hg0 oxidation due to the preferential reaction of NO with O and O3. On the contrary, SO2 shows little effect on Hg0 oxidation due to its preferential reaction with OH. It has been also observed that the HCl in gas stream can be dissociated to Cl and Cl2 and can induce additional Hg0 oxidation to HgCl2