• Title/Summary/Keyword: Replication Control

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Development of Trans-Splicing Aptazyme Which Can Specifically Modify Hepatitis C Virus Genome (C형 간염바이러스(HCV) 유전체를 특이적으로 변형할 수 있는 Trans-Splicing Aptazyme 발굴)

  • Kim, Ju-Hyun;Lee, Chang-Ho;Jang, Sun-Young;Lee, Seong-Wook
    • Korean Journal of Microbiology
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    • v.44 no.3
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    • pp.186-192
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    • 2008
  • For the development of specific and effective basic genetic materials to inhibit replication of hepatitis C virus (HCV), HCV genome-targeting trans-splicing aptazyme, which activity is allosterically regulated by a specific ligand, was developed. The aptazyme was designed to be comprised of sequence of RNA aptamer to the ligand, communication module sequence which can transfer structural transition for inducing ribozyme activity upon binding the ligand to the aptamer, and trans-splicing ribozyme targeting +199 nt of HCV IRES. Especially, when the aptamer and the communication module was inserted at both P6 and P8 catalytic domain of the specific ribozyme, allosteric activity of the aptazyme was the most induced. The aptazyme was shown to induce activity of trans-splicing reaction specifically and efficiently only in the presence of the specific ligand, but neither in the absence of any ligand nor in the presence of control ligand. This aptazyme can be used as a specific and effective genetic agent against HCV, and a tool for the isolation of anti-HCV lead compounds.

Studies on the catalytic charcoaling (촉매적(觸媒的) 제탄(製炭)에 관(關)한 시험(試驗))

  • Park, Tae Sik;Park, Meung Gue
    • Journal of Korean Society of Forest Science
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    • v.3 no.1
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    • pp.18-22
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    • 1963
  • 1. Objects The experiments of catalytic aharcoaling were carried out for the fallowing purposes. (1) To determine the economically desirable amount of catalytic materials to be used when a catalytic charcoaling is practiced. (2) To observe the rate of carbonization of non-treated charcoal wood when the catalytic charcoaling is proceeded in the same charcoal pit. 2. Meterials (1) Small sample chips made of oak (Q. accutissima Carr.), measured by 0.5cm in width and thickness, respectively, and 1cm in length, were used as charcoal wood in each experiment. (2) Ammonium chloride was used as a catalytic material and electric kiln as a charcoaling apparatus. 3. Experiment (1) The sample chips were put into a electric oven for three hours at the temperature $60^{\circ}{\sim}70^{\circ}C$ in order to reduce some water contents. (2) Oven dried sample chips were then soaked for an hour in solution of ammonium chloride. Three kinds of solution were prepared, that is, 2.5%, 5%, and 10%, solution in which the amount of ammonium chloride used was weighed at the rate of 0.5%, 1.0%, and 2.0% to the total weight of the sample chips, resppectivelly. (3) Soaked sample chips were put in the air for 12 hours to reduce some water contents, and then were put into electric oven for 2 hours at the temperature $105^{\circ}{\sim}110^{\circ}C$. (4) Dried sample chips were kept in a desiccator with control sample chips which were treated excarly the same process as the treated sample chips except only not using the ammonium chloride in the process of soking. (5) Sample chips kept in the desiccator were used at random in each charcoaling experiment. (6) Charcoaling in the electric kiln were carried out by using small crucibles with complete cover to reduce the amount of ash. At each charcoaling experiment four crucibles filled with sample ships, weighed about 20gr, were put into electric kiln. The charcoaling was continued for an hour at the temperature $400^{\circ}{\sim}450^{\circ}C$. (7) In order to investigate the influence given by the gases produced during the catalytic charcoaling to the rate of carbonization of non-treated sample chips, the following experiment was done. (a) A crueible was divided into two parts by inserting a fine iron net at the middle of the crucible, and then non-treated sample chips, weighed about 10gr, were put in the upper part of the crucible and treated sample chips, weighed also about 10gr, were put in the under part. (b) The crucibles filled with two kinds of sample chips were put into a electric kiln for an hour at the temperature $400^{\circ}{\sim}450^{\circ}C$. 4. Results. Results for two replications (with four crucibles in one replication) for each experiment designed are as follows : (1) The rats of carbonization of the non treated sample chips, and that of the treated sample chips with ammonium chloride at the rate of 1.5%, 1.0%, and 2.0% to the total weight of the sample chips used were averaged at 19.85%, 22.63%, 24.14%, and 26.60%, respectively. (2) The rats of carbonization of the non-treated sample chips were averaged at (a) 20.04% (0.5% treatment), (b) 20.28% (1.0% treatment), and (c) 20.61% (2.0% treatment) when the treated sample chips were carbonized in the same crucible.

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Revised STandards for Reporting Interventions in Clinical Trials of Acupuncture(STRICTA) : Extending the CONSORT Statement (STRICTA(침 임상연구에서 중재 보고를 위한 표준) 개정판: CONSORT Statement의 확충안)

  • Lee, Hyang-Sook;Cha, Su-Jin;Park, Hi-Joon;Seo, Jung-Chul;Park, Jong-Bae J.;Lee, Hye-Jung
    • Korean Journal of Acupuncture
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    • v.27 no.3
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    • pp.1-23
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    • 2010
  • Objectives and methods : The Standards for Reporting Interventions in Clinical Trials of Acupuncture (STRICTA) were published in five journals in 2001 and 2002. These guidelines, in the form of a checklist and explanations for use by authors and journal editors, were designed to improve reporting of acupuncture trials, particularly the interventions, thereby facilitating their interpretation and replication. Subsequent reviews of the application and impact of STRICTA have highlighted the value of STRICTA as well as scope for improvements and revision. To manage the revision process a collaboration between the STRICTA Group, the CONSORT Group, and the Chinese Cochrane Centre was developed in 2008. An expert panel with 47 participants was convened that provided electronic feedback on a revised draft of the checklist. At a subsequent face-to-face meeting in Freiburg, a group of 21 participants further revised the STRICTA checklist and planned dissemination. Results : The new STRICTA checklist, which is an official extension of CONSORT, includes six items and 17 sub-items. These set out reporting guidelines for the acupuncture rationale, the details of needling, the treatment regimen, other components of treatment, the practitioner background, and the control or comparator interventions. In addition, and as part of this revision process, the explanations for each item have been elaborated, and examples of good reporting for each item are provided. In addition, the word "controlled" in STRICTA is replaced by "clinical", to indicate that STRICTA is applicable to a broad range of clinical evaluation designs, including uncontrolled outcome studies and case reports. Conclusions : It is intended that the revised STRICTA, in conjunction with both the main CONSORT Statement and extension for nonpharmacologic treatment, will raise the quality of reporting of clinical trials of acupuncture.

Effects of Regulate in Feed Intakes on Performance and Meat Quality in Old Laying Hens (산란성계에서 사료 급이량 조절이 생산성과 계육품질에 미치는 영향)

  • Kang, Hwan Ku;Kim, Chan Ho
    • Korean Journal of Poultry Science
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    • v.42 no.3
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    • pp.205-214
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    • 2015
  • This study aimed to investigate the effects of reducing feed intake on performance and meat quality in old laying hens. A total of 200 Hy-Line Brown laying hens (100 weeks old) were randomly allotted to five dietary treatments: control (100% daily feed intake), 90%, 60%, 50%, and 20% daily feed intake. Each treatment was replicated four times with 10 birds per replication and two birds per cage. Ten-bird units were arranged according to a randomized block design. The feeding trial lasted for 4 weeks under a 16L:8D lighting regimen. The results indicated that the daily feed intake correlated with hen-day egg production and feed conversion ratios (P<0.05). The carcass yields and partial ratios were also correlated with daily feed intake (P<0.05). The levels of leukocytes (without basophils) were higher in the 50% and 20% daily feed intake groups than in the other groups. The concentrations of dry matter, crude ash, crude fat, and crude protein, water holding capacity, cooking loss, and fatty acids in the breast meat did not decrease as the daily feed intake decreased. In conclusion, reducing daily feed intake decreased laying performance and carcass yield but had no effect on breast meat quality.

The Effect of Nonviolent Communication Program on Nurse's Interpersonal Competence, Job Stress and Spiritual Wellness (비폭력대화프로그램이 간호사의 대인관계능력, 직무스트레스, 정신적 웰니스에 미치는 효과)

  • Sim, Bong-Hee;Lee, Young-Sook;Ahn, Seong-Ah
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.17 no.9
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    • pp.375-386
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    • 2016
  • This study was conducted to investigate the effectiveness of a nonviolent communication program on nurse's interpersonal competence, job stress and spiritual wellness. This study was a inequality similar to the control group experimental design study of repeated measurements before and after. The data for this study were collected from June 1 to October 20, 2013 from 29 clinical nurses. The self-administered questionnaires used consisted of questions regarding general characteristics, interpersonal competence, job stress and spiritual wellness. The results were: 1. For interpersonal competence, the interaction between time and group was significant (F = 7.726, p = 0.002). 2. For job stress analysis, the interaction between time and group was not significant (F = 0.142, p = 0.851). 3. For spiritual wellness, the interaction between time and group was significant (F = 31.355, p < .001). These results suggest that the nonviolent communication program had a positive effect on rejected clinical nurse's interpersonal competence and spiritual wellness, but not on job stress. A follow-up replication study will be necessary.

Growth Suppression by Adenovirus-mediated Gene Transfer of p16/INK4a in Glioma Cell Lines (사람의 신경교종 세포주에서 아데노바이러스 벡터를 이용한 p16/INK4a 유전자 전달에 의한 종양성장 억제)

  • Kim, Mi-Suk;Kwon, Hee-Chung;Kang, Hee-Seog;Park, In-Chul;Rhee, Chang-Hun;Kim, Chang-Min;Lee, Choon-Taek;Hong, Seok-Il;Lee, Seung-Hoon
    • Journal of Korean Neurosurgical Society
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    • v.29 no.4
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    • pp.471-476
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    • 2000
  • Objective : p16/INK4a, a kind of tumor suppressor genes, encodes a specific inhibitor of the cyclin D-dependent kinases CDK4 and CDK6. This prevents the association of CDK4 with cyclin D1, and subsequently inhibits phosphorylation of retinoblastoma tumor suppressor protein(pRb), thus preventing exit from the G1 phase. According to previous reports, over 50% of glioma tissue and 80% of glioma cell lines have been demonstrated inactivation of p16/INK4a gene. The purpose of this study was to determine whether recombinant adenovirus-p16 virus is a suitable candidate for gene replacement therapy in cases of glioma. Methods : Three human glioma cell lines(U251MG, U87MG and U373MG) that express mutant p16 protein were used. Replication-deficient adenovirus was utilized as an expression vector to transfer exogenous p16 cDNA into the cells ; control cells were infected with the Ad-${\beta}$-gal expressing ${\beta}$-galactosidase. To monitor gene transfer and the expression of exogenous genes, we used Western Blotting analysis. Flow cytometry studies of cellular DNA content were performed to determine the cell cycle phenotype of the glioma cells before and after treatment. Results : We showed here that restoration of p16/INK4a expression in p16 negative U87MG, U251MG and partially deleted U373MG by Ad-CMV-p16 induced growth suppression in vitro. Flow cytometric study revealed that Ad-CMV-p16 infected U87MG cells were arrested during the G0-G1 phase of the cell cycle. Expression of p16 transferred by Ad-CMV-p16 in glioma cells was highly efficient and maintained for more than seven days. Conclusions : Our results suggest that Ad-CMV-p16 gene therapy strategy is potentially useful and warrants further clinical investigation for the treatment of gliomas.

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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.

Cotransformation of Saccharomyces cerevisiae with Heterogenous Plasmids (이종(異種) Plasmid에 의한 Saccharomyces cerevisiae의 동시형질(同時形質) 전환(轉換))

  • Kang, Byung Tae;Park, Jong Sung;Rhee, In Koo
    • Current Research on Agriculture and Life Sciences
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    • v.5
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    • pp.52-58
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    • 1987
  • The yeast S. cerevisiae DBY747 was transformed with E. C - S. C shuttle vector YIp5, YEp13 and YRp7 by the method of spheroplast. The transformation frequency of YEp13 and YRp7 in S. cerevisiae DBY747 was $1.2{\times}10^3$ and $1.0{\times}10^2$ per $10{\mu}g$ of DNA, respectively. The transformants with YIp5 plasmid incapable of autonomous replication in S. cerevisiae were not detected in the condition of this experiment, but YIpS plasmid expressed the gene carried on it when cotransformed with a helper plasmid such as YEp13 or YRp7 : autonomously replicating plasmid. When plasmids were used in covalently closed circular form, cotransformation frequency of Ylp5-YEpl3 and Ylp5-YRp7 was 210 and 95 per $10{\mu}g$ of DNA, respectively. In cotransformation of linear plasmids, transformation frequency of the same cohesive ends was similar to that of noncomplementary cohesive ends. Transformants by the cotransformation with circular plasmids have been shown much higher frequency than with linear plasmids in S. cerevisiae DBY 747. The mitotic segregation stability test suggested that the cotransformant of YIpS-YEp13 was more stable than that of YIpS-YRp7.

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Obesity-Associated Metabolic Signatures Correlate to Clinical and Inflammatory Profiles of Asthma: A Pilot Study

  • Liu, Ying;Zheng, Jing;Zhang, Hong Ping;Zhang, Xin;Wang, Lei;Wood, Lisa;Wang, Gang
    • Allergy, Asthma & Immunology Research
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    • v.10 no.6
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    • pp.628-647
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    • 2018
  • Purpose: Obesity is associated with metabolic dysregulation, but the underlying metabolic signatures involving clinical and inflammatory profiles of obese asthma are largely unexplored. We aimed at identifying the metabolic signatures of obese asthma. Methods: Eligible subjects with obese (n = 11) and lean (n = 22) asthma underwent body composition and clinical assessment, sputum induction, and blood sampling. Sputum supernatant was assessed for interleukin $(IL)-1{\beta}$, -4, -5, -6, -13, and tumor necrosis factor $(TNF)-{\alpha}$, and serum was detected for leptin, adiponectin and C-reactive protein. Untargeted gas chromatography time-of-flight mass spectrometry (GC-TOF-MS)-based metabolic profiles in sputum, serum and peripheral blood monocular cells (PBMCs) were analyzed by orthogonal projections to latent structures-discriminate analysis (OPLS-DA) and pathway topology enrichment analysis. The differential metabolites were further validated by correlation analysis with body composition, and clinical and inflammatory profiles. Results: Body composition, asthma control, and the levels of $IL-1{\beta}$, -4, -13, leptin and adiponectin in obese asthmatics were significantly different from those in lean asthmatics. OPLS-DA analysis revealed 28 differential metabolites that distinguished obese from lean asthmatic subjects. The validation analysis identified 18 potential metabolic signatures (11 in sputum, 4 in serum and 2 in PBMCs) of obese asthmatics. Pathway topology enrichment analysis revealed that cyanoamino acid metabolism, caffeine metabolism, alanine, aspartate and glutamate metabolism, phenylalanine, tyrosine and tryptophan biosynthesis, pentose phosphate pathway in sputum, and glyoxylate and dicarboxylate metabolism, glycerolipid metabolism and pentose phosphate pathway in serum are suggested to be significant pathways related to obese asthma. Conclusions: GC-TOF-MS-based metabolomics indicates obese asthma is characterized by a metabolic profile different from lean asthma. The potential metabolic signatures indicated novel immune-metabolic mechanisms in obese asthma with providing more phenotypic and therapeutic implications, which needs further replication and validation.

CDC6 mRNA Expression Is Associated with the Aggressiveness of Prostate Cancer

  • Kim, Ye-Hwan;Byun, Young Joon;Kim, Won Tae;Jeong, Pildu;Yan, Chunri;Kang, Ho Won;Kim, Yong-June;Lee, Sang-Cheol;Moon, Sung-Kwon;Choi, Yung-Hyun;Yun, Seok Joong;Kim, Wun-Jae
    • Journal of Korean Medical Science
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    • v.33 no.47
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    • pp.303.1-303.10
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    • 2018
  • Background: Cell division cycle 6 (CDC6) is an essential regulator of DNA replication and plays important roles in the activation and maintenance of the checkpoint mechanisms in the cell cycle. CDC6 has been associated with oncogenic activities in human cancers; however, the clinical significance of CDC6 in prostate cancer (PCa) remains unclear. Therefore, we investigated whether the CDC6 mRNA expression level is a diagnostic and prognostic marker in PCa. Methods: The study subjects included 121 PCa patients and 66 age-matched benign prostatic hyperplasia (BPH) patients. CDC6 expression was evaluated using real-time polymerase chain reaction and immunohistochemical (IH) staining, and then compared according to the clinicopathological characteristics of PCa. Results: CDC6 mRNA expression was significantly higher in PCa tissues than in BPH control tissues (P = 0.005). In addition, CDC6 expression was significantly higher in patients with elevated prostate-specific antigen (PSA) levels (> 20 ng/mL), a high Gleason score, and advanced stage than in those with low PSA levels, a low Gleason score, and earlier stage, respectively. Multivariate logistic regression analysis showed that high expression of CDC6 was significantly associated with advanced stage (${\geq}T3b$) (odds ratio [OR], 3.005; confidence interval [CI], 1.212-7.450; P = 0.018) and metastasis (OR, 4.192; CI, 1.079-16.286; P = 0.038). Intense IH staining for CDC6 was significantly associated with a high Gleason score and advanced tumor stage including lymph node metastasis stage (linear-by-linear association, P = 0.044 and P = 0.003, respectively). Conclusion: CDC6 expression is associated with aggressive clinicopathological characteristics in PCa. CDC6 may be a potential diagnostic and prognostic marker in PCa patients.