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cDNA Cloning and Stage-Dependant Expression of Arylphorin Gene from Chinese Oak Silkworm, Antheraea pernyi (작잠(Antheraea pernyi) 아릴포린(Arylphorin) 유전자의 cDNA 클로닝 및 아릴포린 유전자의 발육시기 의존성 발현양상)

  • Lee, Sang-Mong;Hwang, Jae-Sam;Park, Nam-Sook;Kim, Yong-Gyun;Kim, Keun-Ki;Son, Hong-Joo;Park, Hyun-Chul;Jin, Byung-Rae
    • Journal of Life Science
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    • v.20 no.8
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    • pp.1193-1200
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    • 2010
  • The cDNA cloning and developmental profiles of the mRNA for A. pernyi arylphorin was determined. The complete A. pernyi arylphorin cDNA sequence comprised 2,234 bp (without the poly $A^+$ tail), including an open reading frame of 2,112 bp beginning with a methionine ATG at bp34. The A. pernyi arylphorin contained 704 amino acids which are highly enriched in aromatic amino acids, phenylalanine and tyrosine. The calculated molecular mass of the A. pernyi arylphorin from the ORF was 83,439 Da. The deduced amino acid sequence of A. pernyi arylphorin showed 78, 71, 62 and 64% identity with those of H. cecropia, M. sexta $\alpha$ subunit, M. sexta $\beta$ subunit and B. mori storage protein. In Northern blot analysis, the A. pernyi arylphorin mRNA only in the fat body of the 5th instar larvae was responsible for gene expression of the protein, and the synthetic activity of the mRNA was detected strongly in the early larvae, but not in the middle or late-stage larvae. In addition, a very weak signal in mRNA activity was detected in pupal stages, but this was considered to be inactive mRNA after reviewing the results of the labeling experiment of this protein.

Genome-Wide Analysis of DNA Methylation before- and after Exercise in the Thoroughbred Horse with MeDIP-Seq

  • Gim, Jeong-An;Hong, Chang Pyo;Kim, Dae-Soo;Moon, Jae-Woo;Choi, Yuri;Eo, Jungwoo;Kwon, Yun-Jeong;Lee, Ja-Rang;Jung, Yi-Deun;Bae, Jin-Han;Choi, Bong-Hwan;Ko, Junsu;Song, Sanghoon;Ahn, Kung;Ha, Hong-Seok;Yang, Young Mok;Lee, Hak-Kyo;Park, Kyung-Do;Do, Kyoung-Tag;Han, Kyudong;Yi, Joo Mi;Cha, Hee-Jae;Ayarpadikannan, Selvam;Cho, Byung-Wook;Bhak, Jong;Kim, Heui-Soo
    • Molecules and Cells
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    • v.38 no.3
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    • pp.210-220
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    • 2015
  • Athletic performance is an important criteria used for the selection of superior horses. However, little is known about exercise-related epigenetic processes in the horse. DNA methylation is a key mechanism for regulating gene expression in response to environmental changes. We carried out comparative genomic analysis of genome-wide DNA methylation profiles in the blood samples of two different thoroughbred horses before and after exercise by methylated-DNA immunoprecipitation sequencing (MeDIP-Seq). Differentially methylated regions (DMRs) in the pre-and post-exercise blood samples of superior and inferior horses were identified. Exercise altered the methylation patterns. After 30 min of exercise, 596 genes were hypomethy-lated and 715 genes were hypermethylated in the superior horse, whereas in the inferior horse, 868 genes were hypomethylated and 794 genes were hypermethylated. These genes were analyzed based on gene ontology (GO) annotations and the exercise-related pathway patterns in the two horses were compared. After exercise, gene regions related to cell division and adhesion were hypermethylated in the superior horse, whereas regions related to cell signaling and transport were hypermethylated in the inferior horse. Analysis of the distribution of methylated CpG islands confirmed the hypomethylation in the gene-body methylation regions after exercise. The methylation patterns of transposable elements also changed after exercise. Long interspersed nuclear elements (LINEs) showed abundance of DMRs. Collectively, our results serve as a basis to study exercise-based reprogramming of epigenetic traits.

Simplistic QA for an Enhanced Dynamic Wedge using the Reversed Wedge Pair Method (역방향 조사방식을 통한 동적쐐기의 품질관리)

  • Lee Jeong Woo;Hong Semie;Suh Tae Suk
    • Progress in Medical Physics
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    • v.15 no.3
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    • pp.161-166
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    • 2004
  • A simplistic quality assurance (QA) method was designed for a Linac built-in enhanced dynamic wedge (EDW), which can be utilized to make wedged beam distributions. For the purpose of implementing the EDW symmetry QA, a film dosimetry system, low speedy dosimetry film, film densitometer and 3D RTP system were used, and the films irradiated by means of a 60$^{\circ}$ Reversed wedge pair (REWP) method. The profiles were then analyzed in terms of their symmetries, including partial treatment, which is the case of stopping it abruptly during EDW irradiation, and the measured and calculated values compared using the Cad Plan Golden Segmented Treatment Table (Golden STT). The result of this experiment was in good agreement, within 1 %, of the 'reversed wedge pair counterbalance effect'. For the QA of the effective wedge factor (EWF), the authors measured EWFs in relation to the 10$^{\circ}$, 15$^{\circ}$, 20$^{\circ}$, 25$^{\circ}$, 30$^{\circ}$, 45$^{\circ}$ and 60$^{\circ}$ EDW, which were compared with the calculated values using the correction factor derived from the Golden STT and the log files produced automatically during the process of EDW irradiation. By means of this method it was capable of check up the safety of effective wedge factor without any other dosimetry system. The EDW QA was able to be completed within 1 hour from irradiation to analysis as a consequence of the simplified QA procedure, with maximized effectiveness. Unlike the metal wedge system, the EDW system was heavily dependent on the dose rates and jaw movements; therefore, its features could potentially cause inaccuracy. The frequent simplistic QA for the EDW is essential, and could secure against the flaw of dynamic treatment that uses the EDW.

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Characteristics of Kawasaki Disease Patients who are Unresponsive to High-dose Intravenous Immunoglobulin Therapy (가와사끼병에서 정맥용 면역글로불린 불응군의 특성)

  • Kook, Dong-Hyuk;Ko, Yong-Min;Lee, Keun-Young;Kim, Dong-Un;Lee, Kyung-Yil;Lee, Joon-Sung
    • Pediatric Infection and Vaccine
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    • v.15 no.2
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    • pp.180-187
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    • 2008
  • Purpose : We wanted to determine the characteristics of patients with Kawasaki disease (KD) who were unresponsive to intravenous immunoglobulin (IVIG). Methods : The patients with KD were divided into two groups: the IVIG responsive group (25 cases) and the IVIG unresponsive group (14 cases). We analyzed various parameters before and after the administration of IVIG, including the complete blood cell count with the differential count (%), the erythrocyte segmentation rate (ESR), the C-reactive protein (CRP) level and the protein and lipid profiles. Results : The IVIG unresponsive group had a prolonged duration of fever and a higher incidence of CAL compared to the IVIG responsive group (P<0.001, respectively). Before IVIG infusion, the neutrophil differential, the ESR and the CRP values were higher (P<0.001), and the total protein and albumin values were lower in the IVIG unresponsive group (P=0.01) compared to the IVIG responsive group. After IVIG infusion, there were no significant changes in the WBC count and CRP levels in the IVIG unresponsive group. The reduction of the HDL-cholesterol levels by IVIG was more significant in the unresponsive group (P=0.02). Conclusion : A more severe and prolonged inflammatory response occurred in the IVIG unresponsive group at an early stage, and this finding can be detected by such inflammatory parameters as the neutrophil count and the CRP and HDL-cholesterol levels after IVIG infusion.

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Correlation of Hepatic $^{18}F-Fluorodeoxyglucose$ Uptake with Fatty Liver (간의 $^{18}F-Fluorodeoxyglucose$ 섭취 양상과 지방간의 관계)

  • An, Young-Sil;Yoon, Joon-Kee;Hong, Seon-Pyo;Joh, Chul-Woo;Yoon, Seok-Nam
    • Nuclear Medicine and Molecular Imaging
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    • v.40 no.5
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    • pp.243-248
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    • 2006
  • Purpose: Liver demonstrates heterogeneous FDG uptake and sometimes it shows abnormally increased uptake even though there is no malignant tissue. However, there was no previous study to correlate these various pattern of hepatic FDG uptake with benign liver disease. Therefore, we evaluated the significance of hepatic FDG uptake associated with various clinical factors including fatty liver, liver function tests and lipid profiles. Materials and Methods: We reviewed a total of 188 patients (male/female: 120/68, mean age: $50{\pm}9$) who underwent PET/CT for screening of malignancy. Patients with DM, impaired glucose tolerance, previous severe hepatic disease or long-term medication history were excluded. The FDG uptake in liver was analyzed semi-quantitatively using ROI on transaxial images (segment 8) and we compared mean standardized uptake value (SUV) between fatty liver and non-fatty liver group. We also evaluated the correlation between hepatic FDG uptake and various clinical factors including serum liver function test (ALT, AST), ${\gamma}-GT$, total cholesterol and triglyceride concentration. The effect of alcoholic history and body mass index on hepatic FDG uptake was analyzed within the fatty liver patients. Results: The hepatic FDG uptake of fatty liver group was significantly higher than that of non-fatty liver group. Serum total cholesterol and triglyceride concentration showed significant correlation with hepatic FDG uptake. However, there was no significant correlation between other factors (ALT, AST, and ${\gamma}-GT$) and FDG uptake. Also there was no difference of mean SUV between normal and abnormal groups on the basis of alcoholic history and body mass Index within fatty liver patients. Fatty liver and high serum triglyceride concentration were the independent factors affecting hepatic FDG uptake according to multivariate analysis. Conclusion: In conclusion, hepatic FDG uptake was strongly correlated with fatty liver and serum triglyceride concentration.

Environmental Impact Assessment by Marine Cage Fish Farms: II. Estimation of Hydrogen Sulfide Oxidation Rate at $O_2$-H$_2$S Interface and Sulfate Reduction Rate in Anoxic Sediment Layer (해상 어류가두리양식장의 환경영향 평가: II. 가두리 양식장 퇴적물의 산소-황화수소 경계면에서 황화수소의 산화율 및 무산소 퇴적층에서 황산염 환원율 추정)

  • Lee, Jae-Seong;Kim, Kee-Hyun;Yu, Jun;Lee, Pil-Yong;Jung, Rae-Hong;Lee, Wong-Chan;Han, Jung-Jee;Lee, Yong-Hwa
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.9 no.2
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    • pp.64-72
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    • 2004
  • We measured the vertical profiles of $O_2$, H$_2$S, and pH in sediment pore water beneath marine cage fish farms using a microsensor with a 25 ${\mu}{\textrm}{m}$ sensor tip size. The sediments are characterized by high organic material load. The oxygen consumption, hydrogen sulfide oxidation, and sulfate reduction rates in the microzonations (derived from the vertical distribution of chemical species concentration) were estimated by adapting a simple one-dimensional diffusion-reaction model. The oxygen penetration depth was 0.75 mm. The oxic microzonations were divided into upper and lower layers. Due to hydrogen sulfide oxidation within the oxic zone, the oxygen consumption rate was higher in the lower layer. The total oxygen consumption rate integrated with reaction zone depth was estimated to be 0.092 $\mu$mol $O_2$cm$^{-2}$ hr$^{-1}$ . The total hydrogen sulfide oxidation rate occurring within 0.7 mm thickness was estimated to be 0.030 $\mu$mo1 H$_2$S cm$^{-2}$ hr$^{-1}$ , and its turnover time in the oxic sediment layer was estimated to be about 2 minutes. This suggests that hydrogen sulfide was oxidized by both chemical and microbial processes in this zone. The molar consumption ratio, calculated to be 0.84, indicates that either other electron accepters exit on hydrogen sulfide oxidation, or elemental sulfur precipitation occurs near the $O_2$- H$_2$S interface. Total sulfate reduction flux was estimated to be 0.029 $\mu$mol cm$^{-2}$ hr$^{-1}$ , which accounted for more than 60% of total $O_2$ consumption flux. This result implied that the degradation of organic matter in the anoxic layer was larger than in the oxic layer.

Differences of Garlic Powder Ingestion and Exercise Training on Blood Lipids, MDA and SOD in Rats (운동훈련과 마늘가루 섭취에 의한 흰쥐의 혈중 지질 및 MDA, SOD 활성 차이)

  • Jeon, Byung-Duk;Kim, Jin-Ha;Ryu, Sung-Pil
    • Journal of Life Science
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    • v.19 no.10
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    • pp.1337-1345
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    • 2009
  • This study investigated the effect of exercise training and garlic powder ingestion on blood lipids and antioxidants activity in rats. Twenty-eight male Sprague Dawley rats were fed a high-fat diet with or without garlic powder (500 mg/kg) for four weeks as grouped in control (CON), exercise (EXE), garlic (GAR), and garlic + exercise training (GAREXE), respectively. EXE and GAREXE were trained on the treadmill for the same periods. Weight of fats (mesentery, perirenal, and epididymal) were weighed and blood glucose, triglycerides (TG), total cholesterol (TC), and high density lipoprotein- cholesterol (HDL-C) were analyzed and low density lipoprotein-cholesterol (LDL-C) was calculated. Malondialdehyde (MDA) and superoxide dismutase (SOD) for lipid peroxidation were analyzed in liver tissue. Body weight in GAREXE was significantly lower in the statistics than that in other groups (p<0.05), and the volume of fat in GAR and GAREXE was also much lower (p<0.05). Blood glucose was significantly lower in EXE and GAR (p<0.05), however, there was no effect of exercise training. Blood TG was lower in GAR and GAREXE (p<0.05), however, there was no effect of exercise training. HDL-C was significantly improved in EXE and GAR compared to CON (p<0.05), and GAREXE was higher than EXE (p<0.05). MDA content was considerably lower in GAREXE compared to EXE (p<0.05), and SOD activity was much higher in other groups compared to CON (p<0.05). In addition, GAREXE was significantly higher than EXE and GAR, thus there was significant increase when a garlic diet was carried out together with exercise (p<0.05). These results suggested that garlic powder ingestion during the training periods had a beneficial effect of lowering glucose and enhancing blood lipids profiles. Moreover, it also has antioxidant effects, which means that it could possibly suppress aging. It is necessary to inspect various effects of garlic with a variety of research methods regarding sampling process, production process, intake method, etc.

Effect of Onion Peel Extracts on Blood Lipid Profile and Blood Coagulation in High Fat Fed SD Rats (고지방 섭취 흰쥐에서 양파 껍질 추출물의 보충 섭취가 혈중 지질농도와 혈행 개선에 미치는 효과)

  • Chung, Hye-Kyung;Shin, Min-Jeong;Cha, Yong-Jun;Lee, Kyung-Hea
    • The Korean Journal of Food And Nutrition
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    • v.24 no.3
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    • pp.442-450
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    • 2011
  • Numerous studies have suggested that dietary flavonoids contribute to prevent cardiovascular disease. Onion contains many functional phytochemicals such as quercetin. The aim of this study was to examine whether onion peel extracts supplementation affect blood lipid profiles and blood coagulation in animal model. Total 48 Sprague-Dawley male rats at 5 weeks old were divided into 6 groups with different diets(C: control, HF: high fat diet, HFOE 0.01%: high fat+onion peel extract 0.01% diet, HFOE 0.02%, HFOE 0.05%, HFOE 0.1%) for 8 weeks. Onion peel extract supplementation significantly decreased serum levels of LDL-cholesterol and increased HDL-cholesterol, while total cholesterol and triglyceride levels were not affected. Hematological parameters(hematocrit, white blood cell, red blood cell, and platelet count) and blood coagulation parameters(prothrombin time, activated partial thromboplastin time, thrombin time, and fibrinogen) were not significantly different among 6 groups. However, activated partial thromboplastin time of HFOE 0.05% group was significantly longer than that of HF group. These results indicate that onion peel extract supplementation displays hypocholestrolemic effects but does not seem to have anti-coagulation effects in high fat fed SD rats.

Membrane Protein Alterations Associated with Anticancer Drug Resistance in Mouse Lymphoblastic Leukemia L1210 Cells (항암제에 내성인 생쥐의 백혈병세포 L1210의 세포막 단백질의 변환)

  • Kim, Seong-Yong;Son, Sung-Kweon;Kim, Jae-Ryong;Kim, Jung-Hye
    • Journal of Yeungnam Medical Science
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    • v.10 no.2
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    • pp.432-444
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    • 1993
  • Multidrug resistance(MDR) phenotype is frequently observed in animal and human cancer cell lines selected for in vitro resistance to a single chemotherapeutic agent. It is characterized by the diminished drug accumulation and is related to the drug efflux mechanism in resistant cells. In the present study, adriamycin resistant cells(L1210-$AdR_6$ : $10^{-6}M$ adriamycin, $-AdR_5$ : $10^{-5}M$) and vincristine resistant cells (L1210-$VcR_7$ : $10^{-7}M$ vincristine, $-VcR_6$ : $10^{-6}M$) were produced from mouse lymphoblastic leukemia cell line L1210. Growth profiles of survived cells were observed for 5 days with MTT(thiazolyl blue) assay and resistance was compared with $IC_{50}$(drug concentration of 50% survival reduction in absorbance). Resistant cells proliferated more slowly than sensitive cell. Doubling times were 29.7hr in L1210, 68.7hr in L1210-$AdR_5$ and 58.2hr in $-VcR_6$. MDRs expressed as resistance factor were as follows, L1210-$AdR_5$ was 76.4 times for vincristine, L1210-$VcR_6$ was 96.4 times for adriamycin. The cell membrane proteins with three different M.W. were recognized to be related resistance, 220, 158, and 88 kd in L1210-$AdR_5$, 158, 140 and 88 kd in L1210-$VcR_6$ by SDS-PAG electrophoresis. Cell surface membrane proteins were identified by radio-iodination and autoradiogram, their molecular weights were 158, 72.8, and 42.4 Kd in L1210-$VcR_6$.

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Effect of 5-aza-2'-deoxycytidine on Cell Proliferation of Non-small Cell Lung Cancer Cell Line A549 Cells and Expression of the TFPI-2 Gene

  • Dong, Yong-Qiang;Liang, Jiang-Shui;Zhu, Shui-Bo;Zhang, Xiao-Ming;Ji, Tao;Xu, Jia-Hang;Yin, Gui-Lin
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.7
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    • pp.4421-4426
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    • 2013
  • Objective: The present study employed 5-aza-2'-deoxycytidine (5-Aza-CdR) to treat non-small cell lung cancer (NSCLC) cell line A549 to investigate the effects on proliferation and expression of the TFPI-2 gene. Methods: Proliferation was assessed by MTT assay after A549 cells were treated with 0, 1, 5, 10 ${\mu}mol/L$ 5-Aza-CdR, a specific demethylating agent, for 24, 48 and 72h. At the last time point cells were also analyzed by flow cytometry (FCM) to identify any change in their cell cycle profiles. Methylation-specific polymerase chain reaction (MSPCR), real time polymerase chain reaction(real-time PCR) and western blotting were carried out to determine TFPI-2 gene methylation status, mRNA expression and protein expression. Results: MTT assay showed that the growth of A549 cells which were treated with 5-Aza-CdR was significantly suppressed as compared with the control group (0 ${\mu}mol/L$ 5-Aza-CdR). After treatment with 0, 1, 5, 10 ${\mu}mol/L$ 5-Aza-CdR for 72h, FCM showed their proportion in G0/G1 was $69.7{\pm}0.99%$, $76.1{\pm}0.83%$, $83.8{\pm}0.35%$, $95.5{\pm}0.55%$ respectively (P<0.05), and the proportion in S was $29.8{\pm}0.43%$, $23.7{\pm}0.96%$, $15.7{\pm}0.75%$, $1.73{\pm}0.45%$, respectively (P<0.05), suggesting 5-Aza-CdR treatment induced G0/G1 phase arrest. MSPCR showed that hypermethylation in the promoter region of TFPI-2 gene was detected in control group (0 ${\mu}mol/L$ 5-Aza-CdR), and demethylation appeared after treatment with 1, 5, 10 ${\mu}mol/L$ 5-Aza-CdR for 72h. Real-time PCR showed that the expression levels of TFPI-2 gene mRNA were $1{\pm}0$, $1.49{\pm}0.14$, $1.86{\pm}0.09$ and $5.80{\pm}0.15$ (P<0.05) respectively. Western blotting analysis showed the relative expression levels of TFPI-2 protein were $0.12{\pm}0.01$, $0.23{\pm}0.02$, $0.31{\pm}0.02$, $0.62{\pm}0.03$ (P<0.05). TFPI-2 protein expression in A549 cells was gradually increased significantly with increase in the 5-Aza-CdR concentration. Conclusions: TFPI-2 gene promoter methylation results in the loss of TFPI-2 mRNA and protein expression in the non-small cell lung cancer cell line A549, and 5-Aza-CdR treatment could induce the demethylation of TFPI-2 gene promoter and restore TFPI-2 gene expression. These findings provide theoretic evidence for clinical treatment of advanced non-small cell lung cancer with the demethylation agent 5-Aza-CdR. TFPI-2 may be one molecular marker for effective treatment of advanced non-small cell lung cancer with 5-Aza-CdR.