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Molecular Dynamics Simulation Study for Ionic Strength Dependence of RNA-host factor Interaction in Staphylococcus aureus Hfq

  • Lazar, Prettina;Lee, Yun-O;Kim, Song-Mi;Chandrasekaran, Meganathan;Lee, Keun-Woo
    • Bulletin of the Korean Chemical Society
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    • v.31 no.6
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    • pp.1519-1526
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    • 2010
  • The behavior of peptide or protein solutes in saline aqueous solution is a fundamental topic in physical chemistry. Addition of ions can strongly alter the thermodynamic and physical properties of peptide molecules in solution. In order to study the effects of added ionic salts on protein conformation and dynamics, we have used the molecular dynamics (MD) simulations to investigate the behavior of Staphylococcus aureus Hfq protein under two different ionic concentrations: 0.1 M NaCl and 1.0 M NaCl in presence and absence of RNA (a hepta-oligoribonucleotide AU5G). Hfq, a global regulator of gene expression is highly conserved and abundant RNA-binding protein. It is already reported that in vivo the increase of ionic strength results in a drastic reduction of Hfq affinity for $Q{\beta}$ RNA and reduces the tendency of aggregation of Escherichia coli host factor hexamers. Our results revealed the crucial role of 0.1 M NaCl Hfq system on the bases with strong hydrogen bonding interactions and by stabilizing the aromatic stacking of Tyr42 residue of the adjacent subunits/monomers with the adenine and uridine nucleobases. An increase in RNA pore diameter and weakened compactness of the Hfq-RNA complex was clearly observed in 1.0 M NaCl Hfq system with bound RNA. Aggregation of monomers in Hfq and the interaction of Hfq with RNA are greatly affected due to the presence of high ionic strength. Higher the ionic concentration, weaker is the aggregation and interaction. Our results were compatible with the experimental data and this is the first theoretical report for the experimental study done in 1980 by Uhlenbeck group for the present system.

Effect of Glycine Betaine on Follicle-Stimulating Hormone Production by Chinese Hamster Ovary Cells at Low Culture Temperature (CHO 세포의 저온배양에서 Glycine Betaine이 재조합 FSH의 생산에 미치는 영향)

  • Yoon, Sung-Kwan;Ahn, Yong-Ho
    • KSBB Journal
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    • v.22 no.2
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    • pp.109-113
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    • 2007
  • Suspension culture of recombinant Chinese hamster ovary (CHO) cells producing follicle-stimulating hormone was performed to investigate the effect of glycine betaine on cell growth and FSH production at low culture temperature. At 28$^{\circ}C$, cell growth was suppressed, but cell viability remained high for a longer culture period. When the culture temperature was lowered from 37$^{\circ}C$ to 28$^{\circ}C$, more than 14-fold increase in the maximum FSH titer was achieved. In batch culture at 28$^{\circ}C$, the use of 15 mM glycine betaine (GB) to culture medium resulted in the enhancement of maximum cell density and FSH titer by 11% and 17%, respectively, compared to the culture without GB. In pseudo-perfusion culture at 28$^{\circ}C$ with the exchange of fresh medium containing 15 mM GB, a final FSH of $2,058{\mu}g$ which is approximately 1.4-fold higher as compared to the culture without GB was obtained. This enhanced FSH production with 15 mM GB was not just because of enhanced specific FSH productivity (qFSH), but mainly because of the extended culture longevity. Taken together, this result demonstrates that the application of GB at low culture temperature is feasible to enhance the production of recombinant proteins in rCHO cells.

Chemical transformation and target preparation of saponins in stems and leaves of Panax notoginseng

  • Wang, Ru-Feng;Li, Juan;Hu, Hai-Jun;Li, Jia;Yang, Ying-Bo;Yang, Li;Wang, Zheng-Tao
    • Journal of Ginseng Research
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    • v.42 no.3
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    • pp.270-276
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    • 2018
  • Background: Notoginsenoside Ft1 is a promising potential candidate for cardiovascular and cancer disease therapy owing to its positive pharmacological activities. However, the yield of Ft1 is ultralow utilizing reported methods. Herein, an acid hydrolyzing strategy was implemented in the acquirement of rare notoginsenoside Ft1. Methods: Chemical profiles were identified by ultraperformance liquid chromatography coupled with quadruple-time-of-flight and electrospray ionization mass spectrometry (UPLC-Q/TOF-ESI-MS). The acid hydrolyzing dynamic changes of chemical compositions and the possible transformation pathways of saponins were monitored by ultrahigh-performance LC coupled with tandem MS (UHPLC-MS/ MS). Results and conclusion: Notoginsenoside Ft1 was epimerized from notoginsenoside ST4, which was generated through cleaving the carbohydrate side chains at C-20 of notoginsenosides Fa and Fc, and vinaginsenoside R7, and further converted to other compounds via hydroxylation at C-25 or hydrolysis of the carbohydrate side chains at C-3 under the acid conditions. High temperature contributed to the hydroxylation reaction at C-25 and 25% acetic acid concentration was conducive to the preparation of notoginsenoside Ft1. C-20 epimers of notoginsenoside Ft1 and ST4 were successfully separated utilizing solvent method of acetic acid solution. The theoretical preparation yield rate of notoginsenoside Ft1 was about 1.8%, which would be beneficial to further study on its bioactivities and clinical application.

Factors Predicting the Interface Pressure Related to Pressure Injury in Intensive Care Unit Patients (중환자실 환자의 욕창 관련 경계압력 예측요인)

  • Shine, Ji Seon;Kim, Soo Jin;Lee, Ji Hyun;Yu, Mi
    • Journal of Korean Academy of Nursing
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    • v.47 no.6
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    • pp.794-805
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    • 2017
  • Purpose: Interface pressure is a factor that contributes to the occurrence of pressure injuries. This study aimed to investigate interface pressure at common sites of pressure injury (occipital, gluteal and peritrochanteric areas), to explore the relationships among risk factors, skin condition and interface pressure, and to identify risk factors influencing interface pressure. Methods: A total of 100 patients admitted to the intensive care unit were enrolled at a tertiary teaching hospital in Korea. Interface pressure was recorded by a scanning aid device (PalmQ). Patient data regarding age, pulmonary disease, Braden Scale score, body mass index, serum albumin, hemoglobin, mean blood pressure, body temperature, and oxygen saturation were included as risk factors. Data collected from July to September 2016 were analyzed using binary logistic regression. Results: The mean interface pressure of the occipital, gluteal, and right and left peritrochanteric areas were 37.96 (${\pm}14.90$), 41.15 (${\pm}16.04$), 53.44(${\pm}24.67$), and 54.33 (${\pm}22.80$) mmHg, respectively. Predictive factors for pressure injuries in the occipital area were age ${\geq}70$ years (OR 3.45, 95% confidence interval [CI]: 1.19~9.98), serum albumin deficit (OR 2.88, 95% CI: 1.00~8.26) and body temperature ${\geq}36.5^{\circ}C$ (OR 3.12, 95% CI: 1.17~8.17); age ${\geq}70$ years (OR 2.81, 95% CI: 1.10~7.15) in the right peritrochanteric area; and body temperature ${\geq}36.5^{\circ}C$ (OR 2.86, 95% CI: 1.17~6.98) in the left peritrochanteric area. Conclusion: Our findings suggest that old age, hypoalbuminemia, and high body temperature may be contributory factors to increasing interface pressure; therefore, careful assessment and nursing care of these patients are needed to prevent pressure injury. Further studies are needed to establish cutoff values of interface pressure for patients with pressure ulcers.

Molecular Isolation and Characterization of the 2CysPrx Gene from Salicornia herbacea (퉁퉁마디로부터 2CysPrx 유전자 분리 및 특성 분석)

  • Kim, Suk-Kyu;Chung, Sang Ok;Na, Gil-Jong
    • Korean Journal of Environment and Ecology
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    • v.30 no.5
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    • pp.810-820
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    • 2016
  • This study is focused on the investigation of the genes which are induced by various stresses of the halophyte Salicornia herbacea. One of the factors influencing in the germination of Salicornia herbacea is salt stress. The highest germination rate was found in the condition without NaCl, and the upper limit of the NaCl concentration for the germination of Salicornia herbacea was 7%. The optimal temperature of $20^{\circ}C$showed a germination rate of 98%. Among genes induced by stress the 2CysPrx gene was cloned and analyzed for this study. The 2CysPrx gene has two cysteine conserved residues and is composed of 275 amino acids with molecular weight of 30.1kDa. The 2CysPrx gene appeared to be one copy in the genome and consists of 6 introns and 7 exons. Quantitative real-time PCR revealed that the highest transcription rate induced by NaCl and $H_2O_2$ appeared to be at the concentration of 3.5% NaCl and 40mM $H_2O_2$, respectively. The amount of transcript induced by high temperature($40^{\circ}C$) and $75{\mu}M$ of ABA was respectively highest. The gene at low temperature ($4^{\circ}C$) appeared not to be expressed. We are conducting to clone other peroxyredoxin genes induced by various environmental stresses.

Effects of Composition on Soft Magnetic Properties and Microstructures of Fe-Hf-O Thin Films (Fe - Hf - O계 박막에서 조성이 미세구조 및 연자기 특성에 미치는 효과)

  • 박진영;김종열;김광윤;한석희;김희중
    • Journal of the Korean Magnetics Society
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    • v.7 no.5
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    • pp.237-242
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    • 1997
  • The microstructure and soft magnetic properties of as-deposited Fe-Hf-O thin film alloys, which are produced at $P_{O2}=10%$ by rf magnetron sputtering method in $Ar+O_2$ mixed gas atmosphere, is investigated. Newly developed $Fe_{82}Hf_{3.4}O_{14.6}$ film exhibits good soft magnetic properties with $4{\pi}M_s=17.7$ kG, $H_c=0.7$ Oe and ${\mu}_{eff}$(0.5~100MHz)=2,500, respectively. The Fe-Hf-O films are composed of $\alpha$-Fe nanograins and amorphous phase with larger amounts of Hf and O elements which chemically combine each other. With increasing Hf area fraction, Hf and O contents increased proportionally. It was considered that O content in films was determined by Hf contents, because O was chemically combined with Hf. It results from decreasing the $\alpha$-Fe grain size by precipitates (Hf and O), high electrical resistivity. The $Fe_{82}Hf_{3.4}O_{14.6}$ film exhibits the quality factor (Q=$\mu$'/$\mu$") of 25 at 20 MHz. These good frequency characteristics are considered to be superior to other films already reported.o other films already reported.

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Analysis on the actual condition of consciousness for related regulations with revised Water Leisure Safety Regulation (수상레저안전법의 개정에 따른 관련규제에 관한 의식실태분석)

  • Kim Jun-Hoon;Gug Seung-Gi
    • Proceedings of the Korean Institute of Navigation and Port Research Conference
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    • 2005.10a
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    • pp.189-194
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    • 2005
  • The interest of people is rising on water leisure and the population of water sports is increasing with expanded nationwide roads and 5-day work system for income elevation. However, there are many discussions about the way to prevent the accident related to water leisure activities. Water Leisure Safety Regulation was enacted in 1999 as a part of the way and revised in 2005. Therefore it needs to grasp the present state of water leisure activities and analyze the actual condition of consciousness. For the analysis. the question survey was conducted on water leisurers in the focus with regulation of the revised so that the degree of relevance and correlation between mutual variables were analyzed by Frequency and Crosstabs. From this study we found that more than $60\%$ of water leisurers thought registration, safety instruction, insurance, and safety inspection necessary and there was a characteristic common inclination to all for necessity to impose legal controls on water leisure activities as the level of dissatisfaction for water leisure apparatus and cognition for necessity of insurance was high.

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Research on Oral Status of Hearing Impaired Youth by Using QLF-D (QLF-D를 이용한 청각장애 청소년의 구강상태에 관한 조사)

  • Kim, Chang-Suk
    • The Journal of the Korea Contents Association
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    • v.13 no.9
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    • pp.305-311
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    • 2013
  • This study analyzed the oral status after recording the images by using QLF-D with targets of 38 youth people with hearing impairment and hearing language impairment. In order to investigate the state of oral hygiene, plaque index (O'Leary index) and contents of investigation of the state of the teeth included the number of sound teeth, the number of caries teeth, dental caries experience and the number of filling teeth. The following results were obtained. First, women lacked the management on plaque and had more caries teeth compared with men. In terms of impairment classification, subjects with both hearing and language impairment lacked the management on plaque and had more caries teeth. Second, subjects who did not get an oral exam for one year had more caries teeth. Oral hygiene score was the highest with the brushing time for 3-4 minutes. The number of sound teeth was increased as the brushing time was increased. In addition, the oral hygiene management time was the highest when cleaning the teeth, gums and tongue at the same time. Third, it was shown that the satisfaction of oral health education by using the new equipment was high. As a result of this study, in order to improve the oral health level of impaired students, they shall be trained to manage their teeth by themselves and educated to increase their motivation and practice. Thus, it is thought that various approaches which are differentiated from existing methods are required to be tried.

Detection of Novel Mutations in the FABP3 Promoter Region and Association Analysis with Intramuscular Fat Content in Pigs (돼지 FABP3 Promoter 부위 내 신규 돌연변이 탐색과 근내지방도와의 연관성 분석)

  • Kim, J.H.;Park, E.W.;Park, J.J.;Choi, B.W.;Kim, T.H.;Seo, B.Y.;Cheong, I.C.;Lim, H.T.;Oh, S.J.;Lee, J.G.;Jeon, J.T.
    • Journal of Animal Science and Technology
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    • v.47 no.1
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    • pp.1-10
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    • 2005
  • Intramuscular fat content(lMF) is considered as one of major economic traits in the pig breeding and industry. In general, high IMF results in better meat quality. Several approaches to detect quantitative trait 10ci( QTL) for IMF indicated a strong possibility of the existence of a QTL related to IMF between the microsatellite marker SW71 and SW1881 on SSC6q. Porcine FABP3 has been considered as a candidate gene affecting IMF due to its physiological roles and position on the pig genome. Two novel mutations, g.-114T> C and g.-158T>G were detected by duplicate sequencing of the porcine FABP3 promoter region. These two mutations were identified as absolute linkage disequilibrium. The g.-158T> G mutation was used for investigating relationships with growth and fat deposition traits. The GG genotype of the g.-158T> G polymorphism showed highly negative effects(P< 0.01) on body weights at 3 and 12 weeks of age, and a positive effect(P< 0.05) on IMF. However, backfat thickness(BF) and carcass fat(CF) content were not significantly associated with the genotype. The result indicates that the novel mutations, identified in this study, could be utilized as possible genetic markers to improve IMF, independent with BF.

Genome-wide association study for the interaction between BMR and BMI in obese Korean women including overweight

  • Lee, Myoungsook;Kwon, Dae Young;Kim, Myung-Sunny;Choi, Chong Ran;Park, Mi-Young;Kim, Ae-jung
    • Nutrition Research and Practice
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    • v.10 no.1
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    • pp.115-124
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    • 2016
  • BACKGROUND/OBJECTIVES: This is the first study to identify common genetic factors associated with the basal metabolic rate (BMR) and body mass index (BMI) in obese Korean women including overweight. This will be a basic study for future research of obese gene-BMR interaction. SUBJECTS/METHODS: The experimental design was 2 by 2 with variables of BMR and BMI. A genome-wide association study (GWAS) of single nucleotide polymorphisms (SNPs) was conducted in the overweight and obesity (BMI > $23kg/m^2$) compared to the normality, and in women with low BMR (< 1426.3 kcal/day) compared to high BMR. A total of 140 SNPs reached formal genome-wide statistical significance in this study (P < $1{\times}10^{-4}$). Surveys to estimate energy intake using 24-h recall method for three days and questionnaires for family history, a medical examination, and physical activities were conducted. RESULTS: We found that two NRG3 gene SNPs in the 10q23.1 chromosomal region were highly associated with BMR (rs10786764; $P=8.0{\times}10^{-7}$, rs1040675; $2.3{\times}10^{-6}$) and BMI (rs10786764; $P=2.5{\times}10^{-5}$, rs10786764; $6.57{\times}10^{-5}$). The other genes related to BMI (HSD52, TMA16, MARCH1, NRG1, NRXN3, and STK4) yielded P < $10{\times}10^{-4}$. Five new loci associated with BMR and BMI, including NRG3, OR8U8, BCL2L2-PABPN1, PABPN1, and SLC22A17 were identified in obese Korean women (P < $1{\times}10^{-4}$). In the questionnaire investigation, significant differences were found in the number of starvation periods per week, family history of stomach cancer, coffee intake, and trial of weight control in each group. CONCLUSION: We discovered several common BMR- and BMI-related genes using GWAS. Although most of these newly established loci were not previously associated with obesity, they may provide new insights into body weight regulation. Our findings of five common genes associated with BMR and BMI in Koreans will serve as a reference for replication and validation of future studies on the metabolic rate.