• Title/Summary/Keyword: Rad4

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Transcriptional Regulation of a DNA Repair Gene in Saccharomyces cerevisiae

  • Jang, Yeon-Kyu;Sancar, Gwen-B.;Park, Sang-Dai
    • Proceedings of the Zoological Society Korea Conference
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    • 1998.10b
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    • pp.113-113
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    • 1998
  • In Saccharomyces cerevisiae UV irradiation and a variety of chemical DNA -damaging agents induce the transcription of specific genes, including several involved in DNA repair. One of the best characterized of DNA -damage inducible genes is PHRI, which encodes the apoenzyme for DNA photolyase. Basal-level and damage-induced expression of PHRI require an upstream activation sequence, UASPHRI. Here we report the identification of the UlvIE6 gene of S. cerevisiae as a regulator of UASPHRl activity. Surprisingly, the effect of deletion of UME6 is growth phase dependent. In wild-type cells PHRI is induced in late exponential phase, concomitant with the initiation of glycogen accumulation that precedes the diauxic shift. Deletion of UNIE6 abolishes this induction, decreases the steady-state concentration of photolyase molecules and PHRI mRNA, and increases the UV sensitivity of a rad2 mutant. The results suggest that UM E6 contributes to the regulated expression of a subset of damage-responsive genes in yeast. Furthermore, the upstream repression sequence, URSPHRI, is required for repression and damage-induced expression of PHRl. Here we show identification of YER169W and YDR096W as putative regulators acting through $URS_{PHRI}$. These open reading frames were designated as RPHI (YERl69W) and RPH2 (YDR096W) indicating regulator of PHRI. Simultaneous disruption of both genes showed a synergistic effect, producing a four-fold increase in basal level expression and a similar decrease m the induction ratio following treatment of methyl methanesulfonate(MMS). Mutation of the sequence ($AG_4$) bound by Rphlp rendered the promoter of PHRI insensitive to changes in RPHI or RPH2 status. The data suggest that RPHI and RPH2 act as damage-responsive negative regulators of PHRI. Surprisingly, the sequence bound by Rphlp in vitro is found to be $AG_4$ which is identical to the consensus binding site for the regulators Msn2p and Msn4p involved in stress-induced expression. Deletion of MSN2 and MSN4 has little effect on the induction$.$ ratio following DNA damage. However, all deletions led to a significant decrease in basal-level and induced expression of PHRI. These results imply that MSN2 and MSN4 are positive regulators of P HRI but are not required for DNA damage repression. [Supported by grant from NIH]om NIH]

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Residual, Unresectable and Recurrent Rectal Cancer : Role of External Radiation Therapy in 46 Patients (국소 재발성 또는 진행된 직장암의 방사선 치료 -46예의 치료 성적 분석-)

  • Gil, Hack-Joon;Oh, Yoon-Kyeong;Yoon, Sei-Chul;Shinn, Kyung-Sub;Bahk, Yong-Whee
    • Radiation Oncology Journal
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    • v.6 no.1
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    • pp.55-61
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    • 1988
  • Fifty patients with residual, unresectable or recurrent rectal cancer were treated with external irradiation using a 6-MV linear accelerator at the Division of Therapeutic Radiology, Department of Radiology, Kangnam St. Mary's Hospital, Catholic University Medical College during the period of April 1983 to December 1987. This paper describes the results of a retrospective analysis of the results of external irradiation for the residual, unresectable and recurrent rectal cancer in 46 patients. Four patients were lost to follow-up. Of the 46 patients, $18 (39\%)$ presented with unresectable primary lesions and $28 (61\%)$ with residual or recurrent rectal cancer. In $93\%$, the pathologic diagnosis was adenocarcinoma. Resonse to irradiation was observed in $22 (73\%)$ out of 30 patients who were treated for pain, $12 (86\%)$ out of 14 patients who were treated for mass, and $17 (77\%)$ out of 22 patients who were treated for bloody discharge. The actuarial postoperative 2-year and 3-year survival rates in recurrent and unresectable patients were $43\%$ and $22\%$, respectively. However, the post-RT 2-year survival rate was $13\% (6/46)$.

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Measurement of the Radiolysis Gases Generated in Several Waste Forms by External Irradiation (${\gamma}$-조사에 의한 방사성폐기물의 방사분해가스 발생량 평가)

  • Kwak, Kyung-Kil;Ryue, Young-Gerl;Kim, Ki-Hong;Je, Whan-Gyeong;Kim, Dong-Ho
    • Journal of Nuclear Fuel Cycle and Waste Technology(JNFCWT)
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    • v.4 no.4
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    • pp.345-352
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    • 2006
  • The cemented and paraffin wastes form which are incorporated the concentrated wastes, the cemented waste form which is incorporated the spent ion-exchange resins, and the miscellaneous waste(decontamination paper) were irradiated up to $10^{+8}$ rads at $5.43{\times}10^{+5}$ rads/hr with Co-60(72,023.9 Ci) as an external irradiation source. As a result, the radiolysis gases such as $H_2,\;CH_4,\;N_2,\;C_2H_6,\;O_2,\;CO\;and\;CO_2$, were measured in all the wastes. The major gas which was generated in all the wastes was hydrogen($H_2$). The volume of the generated gases showed a difference from $0.029{\sim}0.788\;cm^3.atm/1.1g$ according to the type of wastes, and more was generated in the cemented waste form incorporated a spent ion-exchange resin than in the other wastes. More hydrogen($H_2$) gas was generated in the decontamination paper waste than in the other wastes, and the G($H_2$) value was 0.12.

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Radioprotective Effect of Methylene Blue: 2. Electron Microscopy of the Effect of Methylene Blue on the Liver and Heart of Rats following Gamma-Irradiation (Methylene Blue의 방사선방어결과 2. Methylene Blue가 $\gamma$선에 조사한 흰줘의 간 및 심장조직에 미치는 전자현미경적 연구)

  • Sang Yul Nam;Seung Han Chang
    • The Korean Journal of Zoology
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    • v.12 no.4
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    • pp.114-122
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    • 1969
  • Electron microscopic examination of the liver and heart tissues of methylene blue-treated rats before gamma-irradiation was observed in this study. 1. It was observed severe alteration and degeneration of organelles: accumulation of glycogen particles, severe swollen mitochondria, and broken endoplasmic reticulum in liver tissue of saline-treated rat(control) opposed by emthylene blue-treated rat at 64 and 212 hours following gamma-irradiation. 2. Heart muscles of both methylene blue-treated and saline-treated rats showed no significant alterations, but it was observed that slightly elongated mitochondria with broken cristae and some of vacuoles as well as increased glycogen particles in sarcoplasmic reticulum at 212 hours following gamma-irradiation. 3. It may be considered that methylene blue greatly reduces the sensitivities of rats to gamma-irradiation.

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Development and Assessment Individual Maximum Permissible Dose Method of I-131 Therapy in High Risk Patients with Differentiated Papillary Thyroid Cancer (물리학 선량법을 이용한 갑상선암의 개인별 최대안전용량 I-131 치료법 개발과 유용성 평가)

  • Kim, Jeong-Chul;Yoon, Jung-Han;Bom, Hee-Seung;JaeGal, Young-Jong;Song, Ho-Chun;Min, Jung-Joon;Jeong, Heong;Kim, Seong-Min;Heo, Young-Jun;Li, Ming-Hao;Park, Young-Kyu;Chung, June-Key
    • The Korean Journal of Nuclear Medicine
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    • v.37 no.2
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    • pp.110-119
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    • 2003
  • Purpose: Radioiodine (I-131) therapy is an effective modality to reduce both recurrence and mortality rates in differentiated thyroid cancer. Whether higher doses shows higher therapeutic responses was still debatable. The purpose of this study was to validate curve-fitting (CF) method measuring maximum permissible dose (MPD) by a biological dosimetry using metaphase analysis of peripheral blood lymphocytes. Materials and Methods: Therapeutic effects of MPD was evaluated in 58 patients (49 females and 9 males, mean age $50{\pm}11$ years) of papillary thyroid cancer. Among them 43 patients were treated with ${\Leq}7.4GBq$, while 15 patients with ${\geq}9.25GBq$. The former was defined as low-dose group, and the latter high-dose group. Therapeutic response was defined as complete response when complete disappearance of lesions on follow-up I-131 scan and undetectable serum thyroglobulin levels were found. Statistical comparison between groups were done using chi-square test. P value less than 0.05 was regarded as statistically significant. Results: MPD measured by CF method using tracer and therapeutic doses were $13.3{\pm}1.9\;and\;13.8{\pm}2.1GBq$, respectively (p=0.20). They showed a significant correlation (r=0.8, p<0.0001). Exposed doses to blood measured by CF and biological methods were $1.54{\pm}0.03\;and\;1.78{\pm}0.03Gy$ (p=0.01). They also showed a significant correlation (r=0.86, p=0.01). High-dose group showed a significantly higher rate of complete response (12/15, 80%) as compared to the low-dose group (22/43, 51.2%) (p=0.05). While occurrence of side effects was not different between two groups (40% vs. 30.2%, p=0.46). Conclusion: Measurement of MPD using CF method is reliable, and the high-dose I-131 therapy using MPD gains significantly higher therapeutic effects as compared with low-dose therapy.

Evaluation of IH-1000 for Automated ABO-Rh Typing and Irregular Antibody Screening (ABO 및 RhD 혈액형 검사와 비예기항체 선별검사를 위한 자동화장비 IH-1000의 평가)

  • Park, Youngchun;Lim, Jinsook;Ko, Younghuyn;Kwon, Kyechul;Koo, Sunhoe;Kim, Jimyung
    • The Korean Journal of Blood Transfusion
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    • v.23 no.2
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    • pp.127-135
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    • 2012
  • Background: Despite modern advances in laboratory automated medicine, work-process in the blood bank is still handled manually. Several automated immunohematological instruments have been developed and are available in the market. The IH-1000 (Bio-Rad Laboratories, Hercules, CA, USA), a fully automated instrument for immunohematology, was recently introduced. In this study, we evaluated the performance of the IH-1000 for ABO/Rh typing and irregular antibody screening. Methods: In October 2011, a total of 373 blood samples for ABO/Rh typing and 303 cases for unexpected antibody screening were collected. The IH-1000 was compared to the manual tube and slide methods for ABO/Rh typing and to the microcolumn agglutination method (DiaMed-ID system) for antibody screening. Results: For ABO/Rh typing, concordance rate was 100%. For unexpected antibody screening, positive results for both column agglutination and IH-1000 were observed in 10 cases (four cases of anti-E and c, three of anti-E, one of anti-D, one of anti-M, and one of anti-Xg) and negative results for both were observed in 289 cases. The concordance rate between IH-1000 and column agglutination was 98.7%. Sensitivity and specificity were 90.9% and 99.3%, respectively. Conclusion: The automated IH-1000 showed good correlation with the manual tube and slide methods and the microcolumn agglutination method for ABO-RhD typing and irregular antibody screening. The IH-1000 can be used for routine pre-transfusion testing in the blood bank.

Screening of Wood-Rot Fungi Based on RBBR Decolorization and Its Laccase Activity (RBBR 탈색능을 이용한 목재부후균의 선발 및 이들 균의 Laccase 효소활성)

  • Choi, Yun-Jeong;Shin, Yoo-Su;Cho, Nam-Seok
    • Journal of the Korean Wood Science and Technology
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    • v.34 no.4
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    • pp.46-53
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    • 2006
  • This study was to screen white-rot fungi possesing strong lignin degrading enzymes, glucose-1 oxidase (GOD), laccase (LAC) and Mn-peroxidase (MnP), based on their decolorization activity of Remazol Brilliant Blue R (RBBR). In the midst of 20 tested fungi, 9 isolates were shown 4 kinds of activities such as RBBR decolorization, GOD, LAC and MnP. Relatively high active strains were identified as Phlebia radiata, Trametes versicolor, Abortiporus biennis, Gleophyllum odoratum and Cerrena unicolor. In particular, T. versicolor, G. odoratum, and C. unicolor, which have high activities of LAC, were used to confirm the optimal temperature and pH and to evaluate the effect of inducer, 2,5-xylidine on their LAC activity. The optimum temperatures for mycelial growth were $28^{\circ}C$ for T. versicolor and G. odoratum, and $25^{\circ}C$ for C. unicolor. The optimum pH for mycelial growth was 5.5. Three strains showed the increase of LAC enzyme activity by the addition of 2,5-xylidine. T. versicolor had the highest LAC activity of $22,700nkat/{\ell}$, corresponding to 11.3 times, G. odoratum $15,400nkat/{\ell}$, 9 times and C. unicolor $17,330nkat/{\ell}$, 5.5 times higher than those of the control.

Two-dimensional gel Electrophoresis of Helicobacter pylori for Proteomic Analysis

  • Jung, Tae-Sung;Kang, Seung-Chul;Choi, Yeo-Jeong;Jeon, Beong-Sam;Park, Jeong-Won;Jung, Sun-Ae;Song, Jae-Young;Choi, Sang-Haeng;Park, Seong-Gyu;Choe, Mi-Young;Lee, Byung-Sang;Byun, Eun-Young;Baik, Seung-Chul
    • The Journal of the Korean Society for Microbiology
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    • v.35 no.2
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    • pp.97-108
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    • 2000
  • Two-dimensional gel electrophoresis (2-DE) is an essential tool of proteomics to analyse the entire set of proteins of an organism and its variation between organisms. Helicobacter pylori was tried to identify differences between strains. As the first step, whole H. pylori was lysed using high concentration urea contained lysis buffer [9.5 M Urea, 4% CHAPS, 35 mM Tris, 65 mM DTT, 0.01% SDS and 0.5% Ampholite (Bio-Rad, pH 3-10)]. The extract ($10\;{\mu}g$) was rehydrated to commercially available immobilised pH gradient (IPG) strips, then the proteins were separated according to their charges as the first dimensional separation. The IPG strips were placed on Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE) to separate according to molecular mass of the proteins as the second dimension. The separated protein spots were visualised by silver staining in order to compare different expression of proteins between strains. Approximately 120 spots were identified in each mini-protein electrophoresised gel, furthermore about 65 to 75 spots were regarded as identical proteins in terms of pI value and molecular weight between strains used. In addition, distinct differences were found between strains, such as 219-1, Y7 and Y14, CH150. Two representative strains were examined using strips which had pH range from 4 to 7. This strips showed a number of isoforms which were considered large spots on pH range 3-10. Furthermore, the rest of spots on pH 4-7 IPG strips appeared very distinctive compared to broad range IPG strips. 2-DE seems to be an excellent tool for analysing and identifying variations between H. pylori strains.

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Comparative Analysis of Muscle Proteome from Porcine White and Red Muscles by Two-dimensional Electrophoresis (이차원전기영동법을 이용한 white muscle과 red muscle간의 단백질 발현양상의 비교분석)

  • Kim, N.K.;Joh, J.H.;Chu, K.S.;Park, H.R.;Park, B.Y.;Kim, O.H.;Lee, C.S.
    • Journal of Animal Science and Technology
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    • v.45 no.5
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    • pp.731-738
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    • 2003
  • The technique known as proteomics is useful for characterizing the protein expression pattern of a particular tissue or cell type as well as quantitatively identifying differences in the levels of individual proteins. In present study, we carried out the comparative expression patterns of white and red muscles. We used the two-dimensional electrophoresis(2-DE) for analyzing the protein expression. Proteins isolated from porcine white and red muscles were separated by 12% poly-acrylamide gel and then were detected by coomassie blue and silver staining. More than 600 protein spots were detected on each 2-DE gel. By visual analysis of the stained gel, five proteins were identified to be differentially expressed in the white vs red muscle. By database searching based on the molecular weights and pI(isoelectric point) of the five proteins, three of them were found to be most close to troponin I, T and myoglobin. However, further researche is needed for identification and functional analysis of the unidentified proteins. In conclusion, we found five proteins, which are differentially expressed in the white vs red muscle. The functional analysis of the differentially expressed proteins will provide valuable information on biochemical characteristics of the muscle type.

Rheological and Pasting Properties of Potato Flour Dispersions (감자가루 분산액의 유변학적 및 페이스팅 특성 분석)

  • Heo, Hyemi;Won, Chuin;Jin, Yong-Ik;Chang, Dong-Chil;Kim, Misook;Lee, Youngseung;Chang, Yoon Hyuk
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.9
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    • pp.1407-1413
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    • 2015
  • The objective of this study was to investigate the rheological and pasting properties of potato flour dispersions at different concentrations (3, 4, 5, 6, and 7%, w/w). A potato cultivar 'Goun', used in this study, was developed by Highland Agriculture Research Center, RDA. Potato flour dispersions showed shear-thinning behaviors (n=0.44~0.51) at $25^{\circ}C$. Apparent viscosity (${\eta}_{a,100}$), consistency index (K), and yield stress (${\sigma}_{oc}$) significantly increased with an increase in potato flour concentration. Storage modulus (G') and loss modulus (G") significantly increased, whereas complex viscosity (${\eta}^*$) was significantly reduced with increasing frequency (${\omega}$) from 0.63 to 63.8 rad/s. Magnitudes of G' and G" were significantly increased with elevation of potato flour concentration. G' values were considerably greater than G" over the entire range of frequency (${\omega}$) with a high dependence on ${\omega}$. Cox-Merz rule was not applicable to potato flour dispersions. Rapid Visco Analyzer data showed that peak viscosity, trough viscosity, breakdown viscosity, final viscosity, and set back viscosity of samples significantly increased with an increase in potato flour concentration.