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The effects of enamel matrix derivatives on the proliferation and gene expression of PDL fibroblast, $SaOs_2$ cells and Cementum derived cells (법랑기질유도체가 치주인대세포, 불멸화 조골세포, 백악질 유래세포의 증식과 유전자 발현에 미치는 영향)

  • Jeong, Yoo-Jee;Kim, Kyoung-Hwa;Kim, Tae-Il;Seol, Yang-Jo;Ku, Young;Lee, Hae-Jun;Rhyu, In-Chul;Chung, Chong-Pyoung;Han, Soo-Boo;Lee, Yong-Moo
    • Journal of Periodontal and Implant Science
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    • v.35 no.2
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    • pp.321-333
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    • 2005
  • 1. 목적 in vitro 상에서 법랑기질유도체가 치주인대섬유아세포, 불멸화 조골세포와 백악질 유래세포의 증식과 유전자 발현에 미치는 영향을 알아보고자 하였다. 2. 연구방법 및 재료 <세포증식 연구> 교정을 목적으로 발거한 치아에서 분리, 배양한 치주인대섬유아세포와 백악질유래세포, 그리고 $SaOs_2$ 세포를 이용하였다. 법랑기질유도체가 세포 증식에 미치는 영향을 알아보기 위해, 35 mm Petri dish에 dish 당 $5{\times}10^3$ 개의 세포를 접종하였다. 대조군은 1% 항생제와 10% FBS를 포함한 DMEM 배지를 이용했고, 5mM 초산을 첨가한 군과 첨가하지 않은 두 개의 대조군이 이용되었다. 실험군은 100 ${\mu}g/ml$의 법랑기질유도제를 첨가한 군과 100 ${\mu}g/ml$의 법랑기질유도체와 5 mM의 초산을 첨가한 2개의 실험군이 이용되었다. 각 군은 세 개의 배양접시에 행해졌고, 1, 3, 8일에 세포의 수를 각각 측정하였다. 결과는 repeated measures ANOVA로 통계 처리하였다. <유전자 발현 연구> 각 세포의 형질 특성을 알아보기 위해 RT-PCR을 실시하여 조골세포 분화 표식자와 연관된 Human collagen type I(COL I), human osteopontin(OP), human osteocalcin(OC), human alkaline phosphatase(ALP)와 human bone sialoprotein(BSP)의 mRNA 발현을 실험 1, 3, 8일에 걸쳐, 세 군의 차이를 비교 관찰하였다. 3. 결과 <세포증식 연구> 치주인대세포와 백악질유래세포, 그리고 $SaOs_2$ 세포의 증식은 법랑기질유도체에 의해 영향을 받지 않았다. 대조군과 초산이 포함된 대조군 그리고 법랑기질유도체와 초산이 포함된 실험군에서 유의할 만한 세포 수의 차이가 실험 기간 1, 3, 8일에 걸쳐 나타나지 않았다(p<0.05). <유전자 발현 연구> ALP와 COL I은 세 군의 세포에서 모두 발현되었고, 발현 정도는 EMD에 영향을 받지 않았다. OC은 세 군에서 모두 비교적 약하게 발현되었고, 특히 $SaOs_2$ cell과 백악질유래세포에서 약하게 발현되었다. EMD는 OC의 발현정도를 약하게 하였다. OP은 백악질유래세포에서 1, 3, 8일에 걸쳐 EMD 유무에 관련 없이 발현되지 않았다. 그러나 치주인대세포와 $SaOs_2$ cell에서는 강하게 발현되었다. BSP는 치주인대세포와 $SaOs_2$ cell에서 1, 3, 8일에 걸쳐 비교적 고르게 발현되었다. EMD 배지에서 배양된 백악질유래세포는 8일에는 BSP가 발현되지 않았다. 4 결론 이번 실험 결과에 의하면 법랑기질유도체는 치주인대세포, 불멸화 조골세포와 백악질 유래세포의 증식에 있어 유의성 있는 효과를 나타내지 않았다. 그러나, 유전자 발현에 있어서는, 치주인대세포와 백악질유래세포, 그리고 $SaOs_2$ 세포 모두에서 OC mRNA의 발현을 억제하는 효과를 나타내었다. EMD는 세포의 증식에는 영향을 미치지 않지만, 유전자 발현에 있어 일부 영향을 미치는 것으로 보인다. 법랑기질유도체가 세포의 증식과 유전자 발현에 미치는 영향은 배양된 세포의 형질특성, 배양환경, 배양일수 등에 따라 달라질 수 있다. 그러므로 법랑기질유도체가 in vitro 상에서 세포에 미치는 영향은 보다 정량화된 연구가 필요하다.

Control of Trophoblast Gene Expression and Cell Differentiation

  • Cheon, Jong-Yun
    • 대한생식의학회:학술대회논문집
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    • 2001.03a
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    • pp.195-205
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    • 2001
  • 태반 영양배엽 (trophoblast)은 포유동물의 발생과정 중 가장 먼저 분화되는 세포로서, 자궁환경내에서 배아가 착상, 발생, 및 분화하기 위해서 반드시 필요한 태반을 형성하는 색심적인 세포이다. 영양배엽 세포의 분화과정중의 결함은 배아의 사산이나 임신질환 등의 치명적 결과를 초래한다. 하지만, 영양배엽 세포의 분화를 조절하는 분자생물학적인 메카니즘은 아직 규명되지 않고 있다. 영양배엽 세포의 분화를 조절하는 경로를 규경하기 위한 선결과제는 분화된 영양배엽 세포에서만 발현하는 많은 유전자들이 밝혀져야만 한다. 본 연구팀은 최근에 분화된 영양배엽 세포에서만 발현하는 두 종류의 새로운 유전자들을 찾았다. 한 종류는 homeobox를 보유하고 있는 조절 유전자 Psx이고, 다른 한 종류는 임신호르몬인 태반 프로락틴 라이크 단백질 유전자 PLP-C${\beta}$이다. 본 연구과제의 목표는 이들 유전자의 기능과 조절 메카니즘을 규명함으로써, 영양배엽 세포의 분화를 조절하는 조절경로를 밝히는 것이다. 이를 위하여 다음과 같은 일련의 연구를 수행할 것이다. 1) Psx 유전자가 분화된 영양배엽 세포에서만 발현케 하는 조절 메카니즘을 규명하기 위해 functional assays, in vitro footprinting, gel mobility shift assays, 생쥐형질전화, UV crosslinking, Southwestern blot 등의 방법을 통해 Psx 유전자의 cis-acting 요인과 trans-acting factor를 밝혀 분석한다. 2) 영양배엽 세포의 분화조절 경로를 규명하기 위해 random oligonuclotide library screening, DD-PCR, subtractive screening 등의 방법을 이용하여 Psx 유전자에 의해 조절되는 하부유전자를 밝힌다. 3) Psx 유전자를 knock-out시켜 영양배엽 세포가 발달 및 분화하는데 미치는 역할을 밝힌다. 4) Yeast two-hybrid screening방법을 이용하여 태반 프로락틴 유전자의 수용체를 찾아 이들의 신호전달 기전을 밝힌다. 제1차년 연구결과로서, mouse와 rat으로부터 각각 Psx 유전자의 genomic DNA를 클로닝하여, 유전자 구조를 비교한 결과, mouse Psx (mPsx2)는 4개의 exons으로 이루어져 있는 반면에, rat Psx (Psx3)는 3개의 exons으로 구성되어 있었다. 즉, rPsx3는 mPsx2의 exon1이 없었다. Notrhern blot과 in situ hybridization 분석에 의해 mouse와 rat에서 Psx 유전자가 다르게 발현 조절되는 현상을 밝혔다. 실제로 mPsx2와 rPsx3의 5'-flanking지역을 클로닝하여 염기서열 분석 결과 전혀 homology를 찾을 수 없었다. 또한, 이들 각각 promoter의 activity를 luciferase reporter를 이용하여 조사한 결과 Rcho-1 trophoblast cells에서 각기 다른 activity를 보여 주는 것을 발견하였다. Psx 유전자의 transcription start sites는 Primer extension에 의해 밝혔다. 또한 Psx2 유전자를 knock-out 시키기 위해 targeting vector를 Osdupde1에 제작하였다. 본 과제를 시작할 때 새로운 프로락틴 유전자 하나를 클로닝하여 이 유전자를 PLP-I라고 이름을 붙였다. 이 후 이 유전자 (PLP-I)는 PLP-C${\beta}$라고 이름을 붙이게 되었다. Mouse PLP-C${\beta}$ 유전자의 counterpart를 rat에서 찾아 염기서열을 비교한 결과 mouse와 rat에서 PLP-C${\beta}$유전자의 homology는 약 79% (amino acid level)였다. 본 연구과정을 통해 또 하나의 새로운 PLP-C subfamily member를 mouse로부터 클로닝 하였고, 이 유전자를 PLP-C${\gamma}$라 하였다. PLP-C${\beta}$와 PLP-C${\gamma}$의 발현 유형은 Northern blot과 in 냐셔 hybridization 분석에 의해 태반의 제한된 spongitrophoblast와 trophoblast giant cells에서만 발현하는 것을 밝혔다. 놀랍게도 이들 두 새로운 유전자는 alternative splicing에 의해 두 종류의 isoform이 있음을 밝혔다. PLP family member 유전자로서 splicing에 의한 isoforms을 보여 주는 유전자로는 PLP-C${\beta}$와 PLP-C${\gamma}$가 최초이다. 이들 isoform mRNAs의 발현 유형은 RT-PCR 방법을 이용하여 규명하였다. 또 하나의 새로운 발견은 PLP-C${\beta}$와 PLP-C${\gamma}$가 독특한 유전자 구조를 갖고 있었다. 즉, PLP-C${\beta}$는 exon3의 alternative splicing에 의해 5개 혹은 6개의 exons을 갖는 two isoforms이 생긴다. 반면에 PLP-C${\gamma}$는 exon2가 alternative splcing이 되면서 7개의 exons을 갖거나 6개의 exons을 갖는 isoforms을 만든다. 그리고, PLP-C${\gamma}$의 promoter activity를 trophoblast Rcho-l${\gamma}$ 세포주를 이용하여 PLP-C${\gamma}$ 의 1.5 kb 5'-flanking 지역이 trophoblast-specific promoter activity를 갖고 있음을 밝혔다. PLP-C${\gamma}$ 유전자의 transcription start site는 Primer extension에 의해 밝혔다. 제 1차 년도의 연구결과를 토대로, 2차년에서는 다음단계의 연구를 수행하고자 한다. 즉, 1) mPsx2와 rPsx3의 promoter를 비교분석 함으로서 mouse와 rat에서 Psx 유전자가 다르게 조절되는 메카니즘 규명, 2) Psx와 PLP-C 유전자의 promoter에 있는 cis-acting elements 탐색, 3) Psx2와 Psx3의 단백질을 이용하여 이들이 binding하는 target sequence 규명, 4) 제작한 Psx2 targeting vector를 이용하여 ES cells에서 Psx2 유전자 knock-out, 5) Psx 유전자를 과발현시키는 세포주를 만들고 Psx에 의해 조절되는 유전자 탐색, 6) 새로 밝히 PLP-C members 유전자들의 조절기전을 Rcho-1 세포주를 이용하여 여러 거지 성장인자와 다른 호르몬에 대한 반응을 탐색, 7) Psx와 PLP-C${\gamma}$ 유전자의 chromosomal mapping 등을 밝힐 것이다.

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Cardiomyogenic Potential of Human Adipose Tissue and Umbilical Cord Derived-Mesenchymal Like Stem Cells (사람의 지방 및 제대에서 유래된 유사중간엽 줄기세포로부터 심근세포로의 분화 유도)

  • Park, Se-Ah;Kang, Hyeon-Mi;Kim, Eun-Su;Kim, Jin-Young;Kim, Hae-Kwon
    • Clinical and Experimental Reproductive Medicine
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    • v.34 no.4
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    • pp.239-252
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    • 2007
  • Objectives: In the present study, we examined the differentiation potential of human adipose-(HAD) and human umbilical cord-derived mesenchymal like stem cells (HUC) into cardiomyocytes. Methods: Cells were initially exposed to 5-azacytidine for 24h cells and then were cultivated in the presence or absence of activin A, TGF-$\beta$1, or Wnt inhibitor with various combinations of BMP and FGF. Assessment of cardiomyogenic differentiation was made upon the expression of cardiomyocyte-specific genes using RT-PCR. Results: HAD that cultivated in control medium for 4 weeks after 5-azacytidine expose showed new expression of TnT gene and increased expression of Cmlc1 and kv4.3 genes. However, HAD cultivated in the presence of combinations of BMP-4/FGF-4 (B4/F4) and BMP-4/FGF-8 (B4/F8) showed new expression of $\beta$-MHC gene and more increased expression of Cmlc1, TnT, TnI, Kv4.3 genes. Significantly enhanced expression of Cmlc1, TnT, and Kv4.3 genes were also observed compared to that cultivated in the control medium. Treatment of HUC with either 5-azacytidine or combinations of BMP and FGF did not affect the expression profile of these genes. However, when activin A or TGF-$\beta$1 was present in addition to the BMP-2/FGF-8 (B2/F8) after 5-azacytidine exposure, HUC exhibited new expression of $\beta$-MHC gene and increased expression of $\alpha$-CA, TnT and Kv4.3 genes. When Wnt inhibitor was present in addition to BMP and FGF, HUC showed new expression of Cmlc1 gene and increased expression of $\alpha$-CA, TnT, TnI and Kv4.3 genes. Conclusions: Based on these observations, it is suggested that HAD and HUC could differentiate into cardiomyocytes which might be used as therapeutic cells for the heart diseases.

Effect of extract temperature and duration on antioxidant activity and sensory characteristics of Ulmus pumila bark extract (추출온도 및 시간에 따른 유백피 추출물의 항산화 활성과 음료의 관능적 특성)

  • Cho, Myoung Lae;Oh, Yu-Na;Ma, Jin-Gyeong;Lee, Su-Jin;Choi, Young-Hee;Son, Dong-Hwa;Jang, Eun Hee;Kim, Jong-Yea
    • Food Science and Preservation
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    • v.23 no.7
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    • pp.995-1003
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    • 2016
  • Ulmus pumila L. bark underwent distilled water extraction under three temperature condition ($4^{\circ}C$, room temperature, or $80^{\circ}C$) and two extraction times (1, or 5 min) in order to develop a functional beverage products. Changes in yield, pH, color, total phenolic (TP) content, tannin content and antioxidant activity of the aqueous extracts were evaluated for each extraction temperature and duration. Extraction conditions did not affect yield or pH value of the extracts; however CIE $b^*$ values were high in extracts prepared under high extraction temperature ($80^{\circ}C$) and long extraction duration (5 min) conditions. Both extraction temperature and duration affected the TP and tannin contents of the extracts; however, all extraction conditions resulted in ${\geq}450\;mg\;GAE/g$ TP content and ${\geq}80\;mg\;CE/g$ tannin content. All extracts exhibited ABTS and DPPH radical scavenging ability similar to that of vitamin C. Nitric oxide inhibition activity was lower in the 5 min duration sample than in the 1 min sample. The $4^{\circ}C$ extraction temperature produced an extract with the highest reducing power and hydrogen peroxide values. Extraction temperature also affected sensory evaluation results with the $80^{\circ}C$ extraction temperature producing significantly higher flavor, bitterness, and color score, than those obtained under $4^{\circ}C$ and room temperature extraction conditions.

Method Validation and Quantification of Lutein and Zeaxanthin from Green Leafy Vegetables using the UPLC System (UPLC를 이용한 lutein과 zeaxanthin의 분석법 검증 및 엽채류에서의 정량적 평가)

  • Kim, Suna;Kim, Ji-Sun
    • Korean Journal of Food Science and Technology
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    • v.44 no.6
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    • pp.686-691
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    • 2012
  • The objective of this research is to present method development and validation for the simultaneous determination of lutein and zeaxanthin using ultra performance liquid chromatography (UPLC). Also, rapid quantification was performed on six green leafy vegetables (Allium tuberosum, Aster scaber, Hemerocallis fulva, Pimpinella brachycarpa, Sedum sarmentosum and Spinacia oleracea) that are commonly consumed in Korea. Separation and quantification were successfully achieved with a Waters Acquity BEH C18 ($50{\times}2.1mm$, $1.7{\mu}m$) column by 85% methanol within 5 min. Two compounds showed good linearity ($r^2$ > 0.9968) in $1-150{\mu}g/mL$. Limit of detection (LOD) and quantification (LOQ) for lutein and zeaxanthin were 1.7 and 5.1 g/mL and 2.1 and 6.3 g/mL, respectively. The RSD for intra- and inter-day precision of each compound was less than 10.69%. The recovery of each compound was in the range of 91.75-105.13%. Aster scaber and Spinacia oleracea contained significantly higher amounts of lutein ($4.06{\pm}0.24$ and $3.97{\pm}0.10mg$/100 g of fresh weight), respectively.

Characterizations of Kefir Grains in Fermented Whey and Their Effects on Inflammatory Cytokine Modulation in Human Mast Cell-1 (HMC-1) (Kefir grain에 의한 유청발효액의 특성과 human mast cell-1 (HMC-1)에서 염증 cytokine 조절에 미치는 영향)

  • Son, Ji Yoon;Park, Young W.;Renchinkhand, Gereltuya;Han, Jung Pil;Bum, Jin Woo;Paik, Seung-Hee;Lee, Jo Yoon;Nam, Myoung Soo
    • Journal of Life Science
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    • v.26 no.6
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    • pp.689-697
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    • 2016
  • Kefir is an acidic-alcoholic fermented milk product originating from the Caucasian mountains. Kefir has long been known for its probiotic health benefits, including its immunomodulatory effects. The objectives of this study were to investigate the properties of a fermented whey product and to examine the effects of kefir grains on the in vitro immune-modulation of human mast cell-1 (HMC-1). The results showed that the whey fermented by kefir grains contained the maximum lactic acid bacteria and yeast for 16 hr by 1.83×108 and 6.5×105 CFU/ml, respectively, and lactose and whey proteins were partially hydrolyzed. The experimental whey fermented by kefir grains exhibited an in vitro anti-inflammatory effect on the HMC-1 line for 8, 16, and 24 hr, and this effect induced the expression of interleukin (IL)-4 as a pro-inflammatory cytokine, but not for 48 hr by RT-PCR in HMC-1 cells. In addition, the same phenomenon was observed for the expression of IL-8 as a pro-inflammatory cytokine by the kefir-fermented whey during the same periods of 8-48 hr under the same conditions. These cytokines resulted in the production of IL-4 at 20-25 ng in HMC-1 cells for 8, 16, and 24 hr, whereas 5 ng was produced for 48 hr by the fermented whey. In contrast, IL-8 was produced at 15-20 ng in HMC-1 cells during 4, 8, 16, and 24 hr, while 7 ng was produced at 48 hr. It was concluded that the whey fermented by kefir grains possesses a potential anti-inflammatory function, which could be used for an industrial application as an ingredient of functional foods and pharmaceutical products.

Expression of Anion Exchanger and CFTR in the Hepatocyte and Cholangiocytes in Bile Duct-Ligated Rat (담관 결찰 쥐의 간세포와 담관세포의 anion exchanger와 CFTR 발현)

  • Lee, Jae-Dong;Wang, Joon-Ho;Ki, Seung-Seog;Choe, Won-Hyeok;Park, Jae-Seung;Cho, Won-Kyoo;Park, Jung-Jun;Kim, Hong-Su
    • Journal of Life Science
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    • v.21 no.12
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    • pp.1772-1777
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    • 2011
  • Cystic fibrosis transmembrane conductance regulator (CFTR) gene and sodium-independent $Cl^-/HCO_3^-$ anion exchanger (AE) genes are expressed in a wide variety of mammalian tissues including cholangiocytes. They play an important role in the regulation of intracellular pH (pHi) as well as in transepithelial acid/base transport necessary for biliary bicarbonate secretion. The aim of this study was to examine the expression level of CFTR gene and AE genes (AE1, AE2 and AE3) in the cholangiocytes and the hepatocytes, and also measure AE2 gene expression level after bile duct ligation (BDL). As we previously described, isolated hepatocytes and cholangiocytes from the liver of normal and BDL rats were prepared and gene expression levels were measured by using RT-PCR. We found that AE1, AE2, and AE3 genes were expressed in both hepatocytes and cholangiocytes, but CFTR was only in cholangiocytes. AE2 gene expression level was higher in the BDL hepatocytes than normal hepatocytes, which was significantly different between two groups. AE2 gene expression level was lower in the BDL cholangiocytes than normal cholangiocytes. However, AE2 gene expression level in both hepatocytes and cholangiocytes were not changed with a longer duration of BDL. These results suggest that CFTR and AE2 may play an important role in the pathogenetic mechanism of biliary cholestatic liver disease.

A Study about the Factors Affecting Hearing loss in Adolescent's use of Personal Cassette Players(PCPs) (휴대용 카세트 사용 청소년의 청력관련 요인)

  • Lim, Kyung-Hee;Park, Kyung-Min;Park, Myung-Hwa
    • Research in Community and Public Health Nursing
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    • v.12 no.1
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    • pp.125-141
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    • 2001
  • The purpose of this study was to investigate the factors affecting hearing loss in adolescent's use of PCPs, and to give the basic data for adolescent's hearing conservation program development and prevention education against their hearing loss. This study was a descriptive research about three factors affecting hearing loss; the knowledge and attitude about noise, the perception of hearing loss and the hearing threshold. The subjects of this study were 383 students in two general high schools and two vocational high schools in Teagu. They have been using PCPs but with no current or past ear disease. This study was carried out from Sep. 1. 2000 to Oct. 24, 2000. The instrument used for the knowledge and attitude about noise was a questionnaire developed by Rhee. Kyung Yong and Yi. Kwan Hyung(1996). The instrument used for the perception of hearing loss was a Smith Hearing Screening Questionnaire. A Belton Model 112 Audiometer. air-conduction hearing test instrument. was used for the hearing threshold. Data was analysed by a SPSS/Win 10.0 program with frequency. percentage, t-test. ANOVA and pearson correlation. The results of this study were as follows; 1. The average of concern about hearing scored the highest $3.66{\pm}0.70$. The average of perceived susceptibility scored $2.64{\pm}0.85$ and the average of knowledge about noise scored $2.13{\pm}0.56$. The average of total knowledge and attitude about noise scored $2.82{\pm}0.46$. The average of discomfort of hearing loss($2.51{\pm}0.81$) scored higher than that fear of hearing loss($1.35{\pm}0.53$). The average of total perception of hearing loss scored $1.93{\pm}0.59$. The hearing threshold of the subjects scored the highest at 500Hz(Lt. $23.21{\pm}6.62$, Rt. $23.39{\pm}7.02$) and scored higher in order of 1000Hz, 2000Hz, 4000Hz and 8000Hz. 2. The knowledge and attitude about noise and the perception of hearing loss were both affected only by one important characteristic, which was general and vocational high schools. The knowledge and attitude about noise raked (t=5.258, p=0.000), and perception of hearing loss raked(t=2.241. p=0.026). However. several other important characteristics also impacted significantly on the knowledge and attitudes about noise. They included grade (t = 1. 987. p=0.048), father's education(F=2.745. p=0.043), marks(F=3.157, p=0.044), drinking(t=2.307, p=0.022) and smoking(t=2.587, p=0.010). The left hearing threshold differed significantly by sex at 1000Hz(t=5.175, p<0.001) and 8000Hz (t=3.334, p<0.01). According to general and vocational high schools (p<0.001), at 500Hz (t=-5.056), 1000Hz (t=-5.253), 2000Hz (t=-4.905), 4000Hz (t=-4.704) and 8000Hz (t=-5.204) significant differences were also shown. Marks were significant at 1000Hz (F=3.824, p<0.05) and drinking was found to be significant at 500Hz(t=2.203, p<0.05). The right hearing threshold differed significantly by sex at l000Hz(t=5.557. p<0.001). 4000Hz(t=2.234. p<0.05) and 8000Hz (t=2.730. p<0.01). According to general and vocational high schools(p<0.001) at 500Hz (t=-4.730), 1000Hz(t=-6.271). 2000Hz (t=-4.573). 4000Hz(t=-3.554) and 8000Hz (t=-3.405) significant differences were also shown. Grades impacted at 500Hz(t=2.201. p<0.05) and 4000Hz(t=2.511. p<0.05), while marks were significant at l000Hz(F=4.1l5. p<0.05) and drinking was significant at 500Hz(t=2.333. p<0.05). 3. The left hearing threshold in accordance with use of PCPs differed significantly at 2000Hz(F=2.996. p=0.03l) according to volume level and at 8000Hz(F=2.197. p=0.022) according to duration${\times}$hours per day. The right hearing threshold differed significantly at l000Hz(F=3.075. p=0.028) according to volume level and at 8000Hz(F=2.925. p=0.034) according to duration. 4. The knowledge and attitudes about noise showed a light positive correlation with the perception of hearing loss. A positive correlation was shown. as stated previously in all Hz, between the left hearing threshold and the right hearing threshold, especially the highest correlation at 2000Hz(r=0.761. p=0.000). This study has shown that the factors related to adolescent's use of PCPs are important as they impact significantly an adolescent's hearing. These results then indicate that in future, when designing a hearing conservation program and prevention education this data should be considered.

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Effect of Multidrug Resistance Gene-1 (mdr1) Overexpression on In-Vitro Uptake of $^{99m}Tc$-sestaMIBl in Murine L1210 Leukemia Cells (백혈병 세포에서 Multidrug Resistance Gene-1 (mdr1)의 과발현이 $^{99m}Tc$-sestaMIBl 섭취에 미치는 영향)

  • Chun, Kyung-Ah;Lee, Jae-Tae;Lee, Sang-Woo;Kang, Do-Young;Sohn, Sang-Kyun;Lee, Jong-Kee;Chung, June-Key;Jun, Soo-Han;Lee, Kyu-Bo
    • The Korean Journal of Nuclear Medicine
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    • v.33 no.2
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    • pp.152-162
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    • 1999
  • Purpose: To determine whether $^{99m}Tc$-MIBI is recognized by the multidrug resistant P-glycoprotein (Pgp), we have measured quantitatively $^{99m}Tc$-MIBI uptake in cancer cells. The effects of various Pgp reversing agents on cellular $^{99m}Tc$-MIBI uptake were also investigated in the presence of multidrug resistance gene-1 (mdr1 gene) overexpression. Materials and Methods: We measured percentage uptake of $^{99m}Tc$-MIBI at different incubation temperatures both in mdr1 positive and negative cells. The effects of verapamil, cyclosporin, and dipyridamole on cellular uptake of $^{99m}Tc$-MIBI were also evaluated with or without overex-pression of mdr1 gene in cultured murine leukemia Ll210 cells. Results: The mdr1 gene expressing cell lines were effectively induced in in vitro with continuous application of low-dose adriamycin or vincristine. Cellular uptake of $^{99m}Tc$-MIBI was higher in mdr1 negative Ll210 cells than those of mdr1 positive cells, and higher when incubated in $37^{\circ}C$ than $4^{\circ}C$. In the presence of verapamil, cyclosporin or dipyridamole, $^{99m}Tc$-MIBI uptake was increased upto 604% in mdr1 positive cells. Conclusion: Cellular uptake of $^{99m}Tc$-MIBI is lower in leukemia cells over-expressing mdr1 gene, and MBR-reversing agents increase cellular uptake. These results suggest that $^{99m}Tc$-MIBI can be used for characterizing Pgp expression and developing MDR-reversing agents in vitro.

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A Study on the Creep Fracture Life of Al 7075 alloy( I ) (Al 7075 합금의 크리이프 파단수명에 관한 연구( I ))

  • 강대민
    • Journal of the Korean Society of Safety
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    • v.8 no.4
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    • pp.27-40
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    • 1993
  • High temperature tensile tests, steady state creep tests, Internal stress tests and creep rupture tests using A17075 alloy( $T_{6}$ ) were performed over the temperature range of 9$0^{\circ}C$~50$0^{\circ}C$ (0.4 $T_{m}$ ~0.85 $T_{m}$ ) and stress range of 0.64~17.2(kgf/$\textrm{mm}^2$). The main results obtained in this paper were as follows. (1) The activation energies for yielding at the temperature of 0.4 $T_{m}$ ~0.75 $T_{m}$ were calculated to be 25.7~36.5kcal/mol, which were nearly equal to the activation energies for creep. (2) At around the temperature of 9$0^{\circ}C$~12$0^{\circ}C$ and under the stress level of 10~17.2(kgf/$\textrm{mm}^2$), and at around the temperature of 200~41$0^{\circ}C$ and under the stress level of 1.53~9.55(kgf/$\textrm{mm}^2$) and again at around the temperature of 470~50$0^{\circ}C$ and under the stress level of 0.62~l.02(kgf/$\textrm{mm}^2$), the applied stress dependence of steady state creep rate $n_{measu}$ measured were, respectively, 3.15, 6.62 and 1.1, which were in good agreement the calculated stress dependence $n_{ealeu}$ obtained by the difference of the applied stress dependence of the Internal stress and the ratio of the internal stress to the applied stress. (3) At the temperature range of 0.4~0.43 $T_{m}$ , and at the temperature range of 0.52~0.75 $T_{m}$ and again at the temperature range of 0.82~0.85 $T_{m}$ , the activation energies $Q_{measu}$ obtained by steady state creep rate, respective, 26. 16, 34.9, 36.2 and 36.1kcal/mol, which were in good agreement with those obtained with the activation energies under constant effective stress and the temperature dependence of Internal stress. (4) At the temperature range of the 0.52~0.73 $T_{m}$ and under the stress level of 1.53~9.55(kgf/$\textrm{mm}^2$), the stress dependence of rupture life(n’) measured was 6.3~6.6, which was in good agreement with the stress dependence of steady state creep rate(n). And at the same condition the activation energy for rupture( $Q_{f}$ ) measured was 32.0~36.9kca1/mol, which was also in good agreement with the activation energy obtained by steady state creep rate ( $Q_{c}$ ). (5) The rupture life( $t_{f}$ ) might be represented by athermal process attributed to the difference of the applied stress dependence of the internal stress and the ratio of the internal stress to the applied stress, and the thermal activated process attributied to the temperature dependence of the internal stress as $t_{f}$ = A'$\sigma$$_{a}$ {n(1-d $\sigma$$_{i}$ /d $\sigma$$_{a}$ )/(1-$\sigma$$_{i}$ / $\sigma$$_{a}$ )}.exp[{ $Q_{c}$ $^{*}$-( $n_{o}$ R $T^2$/ $E_{(T)}$) (d $E_{(T)}$/dT) - ( $n_{0}$ R $T^2$/ $\sigma$$_{a}$ - $\sigma$$_{i}$ ) (d $\sigma$$_{i}$ /dT)}/RT]. (6) The relationship betwween Larson-Miller rupture parameter and logarithmic stress was linearly decreased, so creep rupture life of Al 7075 alloy seemed to be predicted exactly with Larson-Miller parameter.meter.

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