• Title/Summary/Keyword: 전기영동법

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The relation between angiotensin converting enzyme (ACE) gene polymorphism and neonatal hyperbilirubinemia in Korea (한국인 신생아 황달과 안지오텐신 전환효소 유전자의 다형성)

  • Kim, Mi Yeoun;Lee, Jae Myoung;Kim, Ji Sook;Kim, Eun Ryoung;Lee, Hee Jae;Yoon, Seo Hyun;Chung, Joo Ho
    • Clinical and Experimental Pediatrics
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    • v.50 no.1
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    • pp.28-32
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    • 2007
  • Purpose : Human angiotensin converting enzyme (ACE) gene shows an insertion/deletion polymorphism in 16 intron, and three genotypes are determined by whether a 287 bp fragment of the DNA is present or not; II, ID and DD genotype. DD genotype has been suggested as a risk factor of chronic nephrotic disease such as IgA nephropathy and diabetic nephropathy, various cardiovascular diseases and several other diseases. ACE activity increases in acute hepatitis, chronic persistent hepatitis, chronic active hepatitis and cirrhosis. On the other hand, patients with fatty livers have normal ACE activity. This study was designed to find out the relation between polymorphsims of the ACE genes and neonatal hyperbilirubinemia in Koreans. Methods : The genomic DNA was isolated from 110 full-term Korean neonates who had hyperbilirubinemia with no obvious causes (serum bilirubin$${\geq_-}12mg/dL$$) and 164 neonates of a control population (serum bilirubin <12 mg/dL). We performed polymerase chain reaction (PCR) to see the allele of the ACE gene. Electrophoresis was done in the PCR products in 1.5 percent agarose gel, and then DNA patterns were directly visualized under ethidium bromide staining. Results : ACE genotypes in the hyperbilirubinemia group are as follows; 26.36 percent for II, 53.64 percent for ID, 20.00 percent for DD, 0.532 for I allele and 0.468 for D allele. These distributions were not significantly different from those in the control group; 24.39 percent for II, 51.83 percent for DI, 23.78 percent for DD, 0.503 for I allele and 0.497 for D allele. Conclusion : In this study, ACE gene polymorphism was detected in the neonatal hyperbilirubinemia and control group. The most frequent genotype was ID. Our results indicate that the ACE gene polymorphism is not associated with the prevalence of neonatal hyperbilirubinemia in Koreans.

Germination and Protein Pattern of Phragmites communis Population according to the Soil Salt Contents of Habitats (생육지의 토양염분농도에 따른 갈대( Phragmites communis)개체군의 발아 및 단백질 패턴)

  • Lee, Ho-Joon;Hyo-Sik Yang;Sung-Jong Kim
    • The Korean Journal of Ecology
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    • v.16 no.2
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    • pp.135-149
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    • 1993
  • In order to study and ecotypic variation of Phragmites communis Trin., we investigated germination rates, velocities, and protein band patte군 of seeds of three population of salt marsh, estuary and fresh water areas of Muan Peninsular in southwestern coast of Korea from March, 1990 to October, 1992. The highest germination rates of seeds were observed at $25^{\circ}C$; those of population of the estuary and fresh water are were 100% and that of the salt marsh was 95%. Similar germination rates were observed from the populations of estuary and fresh water areas at $30^{\circ}C$ and $20^{\circ}C$, but they decreased at $15^{\circ}C$. The onset of germination of seeds of three population was earlier $^{\circ}C$, but they decreased at $15^{\circ}C$. The onset of germination of seeds of th three populations was earlier at both $25^{\circ}C$, which was higher than those of any other areas, while that of fresh water areas was the lowest. were different; those of salt marsh and estuary decreased to 30% and 2.5%, respectively, at 3.0% of salt content, but seeds of the fresh water area did not germinate at all at the same salt content. The onset of germination was delayed in the order of the salt marsh, esturay and fresh water areas as salt content of culture solution increased. Germination of seeds from the population of salt marsh was found to begin earliest. The highest germination velocity of three populations was observed in the culture containing no salt. The germination velocity constant decreased as salt content of culture solution increased from 0.5% to 3.0%: those of the populations of the salt marsh, estuary, and fresh water areas were 9.50, 0.75 and 0.00, respectively, at the salt concentration of 3.0%. Soluble protein patterns of seedings from the three populations were analyzed by SDS-PAGE method. The results showed that protein patterns of the three populations were distinctly different qualitatively and quantitatively. The present study suggests that populations of Phragmites communis Trin. in the coast of Korea had taken ecotypic variations of habitats, i.e., fresh water, estuary, and salt marsh types, according to the salt content.

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The effects of high-dose intravenous immunoglobulin on plasma protein and lipid levels in the patients with Kawasaki disease (가와사끼병에서 고용량 정맥용 면역글로불린이 혈장 단백 및 지질에 미치는 영향(면역글로불린 투여 후 단백질 농도 변화))

  • Lee, Keun Young;Kim, Dong-Un;Lee, Hyun Seung;Jang, Pil Sang;Kim, Young-Hoon;Kim, Jin Tack;Kim, Hyun Hee;Lee, Kyung-Yil;Lee, Joon-Sung
    • Clinical and Experimental Pediatrics
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    • v.49 no.12
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    • pp.1348-1353
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    • 2006
  • Purpose : The reticuloendothelial system is composed of sinusoidal capillaries, through which even large protein molecules are freely movable between plasma and interstitial space, including the lymphatic system. Therefore, high-dose intravenous immunoglobulin (IVIG) would cause a redistribution of proteins between two compartments. To investigate this hypothesis, we measured plasma protein and lipid levels in patients with Kawasaki disease before and after high-dose IVIG treatment. Methods : Thirty four children with Kawasaki disease who had complete responses to high-dose IVIG treatment (1 g/kg/day for two consecutive days), were analyzed. Before and after the administration of IVIG, serum analyses were performed for such parameters as total protein, albumin, ${\gamma}$-globulins (IgG, IgM, IgA), ${\alpha}1-$, ${\alpha}2-$, and ${\beta}-$ globulin fractions, and lipid profiles (total cholesterol, HDL-cholesterol, LDL-cholesterol and triglyceride). Results : The levels of ${\gamma}$-globulins including IgG, IgM, IgA were significantly increased, and IgG was increased by $1,779{\pm}304mg/dL$ after two-dose of IVIG infusion. The levels of albumin, ${\alpha}1-$, ${\alpha}2-$, and ${\beta}$ globulin fractions were significantly decreased by 18 percent, 24 percent, 19 percent and 12 percent, respectively. HDL-cholesterol level was significantly decreased by 20 percent, while LDL-cholesterol and triglyceride levels were significantly increased by 21 percent and 50 percent, respectively. The total cholesterol level was not changed. Conclusion : High-dose IVIG treatment decreased the levels of a variety of proteins except immunoglobulins, and the increase of IgG after IVIG treatment was lower than expected. Our results suggest that a part of infused IVIG and plasma proteins, including etiologic proteins for Kawasaki disease, may be distributed to the extravascular compartments. The rapid improvement of symptoms induced by IVIG in Kawasaki disease might be explained by this mode of action of IVIG.

A Comparison of Gene Extraction Methods for the Identification of Raw Materials from Processed Meat Products (식육추출가공품의 사용원료 확인을 위한 유전자추출 방법의 비교 및 검토)

  • Park, Yong-Chjun;Kim, Mi-Ra;Lim, Ji-Young;Park, Young-Eun;Shin, Jun-Ho;Hwang, Cho-Rong;Lim, Jan-Di;Kim, Kyu-Heon;Lee, Jae-Hwang;Cho, Tae-Yong;Lee, Hwa-Jung;Han, Sang-Bae
    • Journal of Food Hygiene and Safety
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    • v.27 no.2
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    • pp.146-151
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    • 2012
  • In this study, effective gene extraction methods were compared to identify raw materials of processed meat products through molecular biological methods. Species specific primers were used to identify ingredients of processed foods and, as a sample, 13 kinds of processed meat products including beef, pork and chicken. According to the type of sample, 13 kinds of samples were classified into liquid type, source type and powder type. The samples were pre-treated (centrifugation) and (or) performed Whole Gene Amplification (WGA) kit for amplification of the extracted DNA. As a result, it was possible to identify the raw material of products through the centrifugation of sample 1 ml for liquid type of processed meat products. For source type of products after gene extraction, it was required to perform WGA for the identification of ingredients. For powder type products did not required any further pre-treatment and WGA. In this study, it was an opportunity to confirm the possibility of identification of raw material from the gene extraction of processed meat products and this method could be used to examine the authenticity of raw material of products.

Allele Distribution and Frequency of Human Surfactant Protein-A2 in Korean Neonates (한국 신생아의 폐 표면 활성제 단백-A2(Human Surfactant Protein-A2) 유전자 대립형질의 분포와 빈도)

  • Kim, Nyeon Cheon;Yoon, Hee Chul;Suk, Jung Su;Ko, Jung Ho;Yoo, Ook Joon;Lee, In Kyu;Oh, Myung Ho;Bae, Chong Woo
    • Clinical and Experimental Pediatrics
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    • v.46 no.4
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    • pp.340-344
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    • 2003
  • Purpose : We evaluated allele frequencies and distribution of surfactant protein A2(SP-A2) in Korean neonates in order to estimate the prevalence of RDS, to find out new SP-A alleles, and to establish new steroid therapy. Methods : Genomic DNA was extracted from 71 neonates and served as a template in PCR for genotype analysis. SP-A gene-specific amplications and gene-specific allele determinations were performed using PCR-cRFLP methods. Results : The distribution for the alleles of the SP-A2 gene in the study population was 1A, $1A^0$, $1A^1$, $1A^2$, $1A^3$, $1A^5$, $1A^6$, $1A^7$, $1A^8$, $1A^9$, $1A^{11}$, $1A^{12}$. The specific frequencies for the alleles of the SP-A2 gene in the study population were : 1A=11.3%, $1A^0=38%$, $1A^1=12.7%$, $1A^2=9.2%$, $1A^5=15.5%$, $1A^7=2.9%$, $1A^8=4.9%$, $1A^9=2.2%$, others=3.3%. Conclusion : The frequency of $1A^0$ was higher than the other SP-A2 alleles in Korean neonates. This finding suggests that the prevalence of RDS in Korea may be low compared with other countries. However, this finding also suggests that Korean neonates have a high risk of infection.

Loss of Heterozygosity on the Long Arm of Chromosome 21 in Non-Small Cell Lung Cancer (비소세포폐암에서 21q 이형체 소실)

  • Chai, Po-Hee;Bae, Nack-Cheon;Lee, Eung-Bae;Park, Jae-Yong;Kang, Kyung-Hee;Kim, Kyung-Rok;Bae, Moon-Seob;Cha, Seung-Ik;Chae, Sang-Chul;Kim, Chang-Ho;Jung, Tae-Hoon
    • Tuberculosis and Respiratory Diseases
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    • v.50 no.6
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    • pp.668-675
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    • 2001
  • Background : Non-smalll lung cancer(NSCLC) develops as a result of the accumulation of multiple genetic abnormalities. Loss of heterozygosity(LOH) is one of the most frequent genetic alterations that is found in NSCLC, and the chromosomal regions that display a high rate of LOH are thought to harbor tumor suppressor genes(TSGs). This study was done to determine the frequency of LOH in 21q with the aim of identifying potential TSG loci. Method : Thirty-nine surgically resected NSCLCs were analysed. Patients peripheral lymphocytes were used as the source of the normal DNA. Five microsatellite Inarkers of 21q were used to study LOH : 21q21.1(D21S1432, and D21S1994); 21q21.2-21.3(D21S1442) ; 21q22.1(21S1445) ; and 21q22.2-22.3(D21S266). The fractional allelic loss(FAL) in a tumor was calculated as the ratio of the number of markers showing LOH to the number of informative markers. Result : LOH for at least one locus was detected in 21 of 39 tumors(53.8%). Among the 21 tumors with LOH, 5(21.8%) showed LOH at almost all informative loci. Although statistically not significant, LOH was found more frequently in squamous cell carcinomas(15 of 23, 65.2%) than in adenocarcinomas(6 of 16, 37.5%). In the squamous cell carcinomas the frequency of LOH was higher in stage II-III (80.0%) than in stage I (53.8%). The FAL value in squamous cell carcinomas($0.431{\pm}0.375$) was significantly higher than that found in adenocarcinomas($0.l92{\pm}0.276$). Conclusion : These results suggest that LOH on 21q may be involved in the development of NSCLC, and that TSG(s) that contribute to the pathogenesis of NSCLC may exist on 21q.

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Effects of Extraction Method on Anserine, Protein, and Iron Contents of Salmon (Oncorhynchus keta) Extracts (연어(Oncorhynchus keta) 추출물 중의 Anserine, 단백질 및 철분 함량에 미치는 추출방법의 영향)

  • Min, Hye-Ok;Park, In-Myoung;Song, Ho-Su
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.2
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    • pp.220-228
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    • 2017
  • Effects of extraction methods on reducing concentrations of pro-oxidants (total iron and protein) of salmon was determined. For development of the extraction process, the effectiveness of several extraction methods was determined and compared, including heat treatment (60, 80, and $100^{\circ}C$), ion exchange and carboxymethyl (CM)-cellulose column chromatography, and ultrafiltration (UF). Protein, total iron, and anserine contents of salmon extracts were 23.64 mg/mL, $16.20{\mu}g/mL$, and 5.47 mg/mL in non-heated extracts, 7.40 mg/mL, $2.32{\mu}g/mL$, and 5.20 mg/mL in heated extracts at $60^{\circ}C$, 7.64 mg/mL, $1.20{\mu}g/mL$, and 5.21 mg/mL at $80^{\circ}C$, and 7.04 mg/mL, $0.68{\mu}g/mL$, and 4.04 mg/mL at $100^{\circ}C$, respectively. Heating and UF decreased contents of protein and total iron, whereas only UF slightly decreased anserine content. Application of the primary ion exchange method increased the content of anserine up to 16%. Protein and total iron contents by the primary ion exchange method decreased by 70 and 98%, respectively. Secondary ion exchange (CM-cellulose) treatment after primary ion exchange and UF resulted in lower anserine content than the primary ion exchange method. However, the content of impurities (protein, total iron) was lower than in all other salmon extracts. Therefore, primary ion exchange, UF, and secondary ion exchange method were the best extraction processes in this study.

Recombination and Expression of VP1 Gene of Infectious Pancreatic Necrosis Virus DRT Strain in a Baculovirus, Hyphantria cunea Nuclear Polyhedrosis Virus (전염성 췌장괴저바이러스 DRT Strain VP1유전자의 Baculovirus Hyphantria cunea Nuclear Polyhedrosis Virus에 재조합과 발현)

  • Lee, Hyung-Hoan;Chang, Jae-Hyeok;Chung, Hye-Kyung;Cha, Sung-Chul
    • The Journal of Korean Society of Virology
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    • v.27 no.2
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    • pp.239-255
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    • 1997
  • Expression of the cDNA of the VP1 gene on the genome RNA B segment of infectious pancreatic necrosis virus (IPNV) DRT strain in E. coli and a recombinant baculovirus were carried out. The VP1 gene in the pMal-pol clone (Lee et al. 1995) was cleaved with XbaI and transferred into baculovirus transfer vector, pBacPAK9 and it was named pBacVP1 clone. The VP1 gene in the pBacVP1 clone was double-digested with SacI and PstI and then inserted just behind T5 phage promoter and the $6{\times}His$ region of the pQE-3D expression vector, and it was called pQEVPl. Again, the $6{\times}$His-tagged VP1 DNA fragment in the pQEVP1 was cleaved with EcoRI and transferred into the VP1 site of the pBacVP1, resulting pBacHis-VP1 recombinant. The pBacHis-VP1 DNA was cotransfected with LacZ-Hyphantria cunea nuclear polyhedrosis virus (LacZ-HcNPV) DNA digested with Bsu361 onto S. frugiperda cells to make a recombinant virus. One VP1-gene inserted recombinant virus was selected by plaque assay. The recombinant virus was named VP1-HcNPV-1. The $6{\times}$His-tagged VP1 protein produced by the pQEVP1 was purified with Ni-NTA resin chromatography and analyzed by SDS-PAGE and Western blot analysis. The molecular weight of the VP1 protein was 94 kDa. The recombinant virus, VP1-HcNPV-1 did not form polyhedral inclusion bodies and expressed VP1 protein with 95 kDa in the infected S. frugiperda cells, which was detected by Western blot. The titer of the VP1-HcNPV-1 in the first infected cells was $2.0{\times}10^5\;pfu/ml$ at 7 days postinfection.

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In Situ Surfactant Flushing of Contaminated Site (계면 활성제를 이용한 In Situ 토양 세척)

  • 염익태;안규홍
    • Journal of Korea Soil Environment Society
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    • v.2 no.2
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    • pp.9-24
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    • 1997
  • Surfactant-aided in situ soil flushing has been proposed as an alternative for the expensive and time consuming 'pump and treat' technology in remediation of contaminated soil and groundwater Injected surfactants can effectively solubilize contaminants sorbed to the soil matrix or nonaqueous phase liquids(NAPLs) in residual saturation. The contaminants solubilized in groundwater are recovered and treated further. The theoretical background of the technology and the results of the field operations, mostly in the US. were summarized. In addition, the factors crucial to the successful application of the technology were discussed. Cost analyses and technical limitations in current applications were also discussed. In conclusion, it is likely that in situ surfactant flushing become a viable option for soil remediation in limited cases. Currently, further advances with respect to operation cost and to treatment efficiency are required for more extensive application of the technology. However, the current trends in soil remediation, specially the growing emphasis on risk based corrective action and natural attenuation, will increase the competitiveness of the technology. For example, removal of easily washable contaminants by short term soil flushing followed by long term monitoring and natural attenuation can greatly reduce the operation cost and time.

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Gene Expression of Metalloproteinases, Tissue Inhibitors of Metalloproteinases and Cytokines in Adriamycin-induced Cardiomyopathy (아드리아마이신으로 유도된 심근증에서 Metalloproteinase, Metalloproteinase 조직억제자, Cytokine 유전자 발현에 대한 연구)

  • Hong, Young Mi
    • Clinical and Experimental Pediatrics
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    • v.48 no.2
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    • pp.197-203
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    • 2005
  • Purpose : Changes in metalloproteinases(MMP) activity have been demonstrated in several disease states, including rheumatoid arthritis and tumor metastasis. More importantly, increased myocardial MMP activity has been reported to occur in both clinical and experimental forms of dilated cardiomyopathy. There was no report about MMP in adriamycin(ADR)-induced cardiomyopathy. The purpose of this study was to investigate gene expression of MMP and tissue inhibitor of metalloproteinases(TIMP) in ADR-induced cardiomyopathy and clarify the relationship between MMP and cytokines. Methods : Male Sprague-Dawley rats were divided into two groups. The first group was control. The second group was given intraperitoneal injections of ADR(5 mg/kg) twice a week over two weeks. Serum concentrations of MMP, TIMP, interleukin(IL)-6 and tumor necrosis factor(TNF)-${\alpha}$ were measured. RNA extraction was performed from frozen rat hearts. Reverse transcription polymerase chain reaction(RT-PCR) was employed. cDNA Microarray analysis was performed by using a set of 5,184 sequence-verified rat cDNA clones. Results : Serum MMP and TIMP levels were not significantly different between the two groups. IL-6 was $36.8{\pm}2.8pg/mL$ and TNF-${\alpha}$ $2.2{\pm}2.7pg/mL$ in the ADR group. They were significantly higher than in the control group. Serum MMP correlated significantly with TNF-${\alpha}$(r=0.41, P<0.05). There was no gene expression of MMP, IL-6 or TNF-${\alpha}$ in the hearts of both groups. Gene expression of TIMP was significantly depressed in the hearts of the ADR group. Conclusion : These results suggested a potential role for TNF-${\alpha}$ in the regulation of extracellular matrix remodeling in ADR induced cardiomyopathy. Rapid screening of multiple decreased gene expression by DNA chip may be a useful diagnostic test to detect early cardiac injury before developing ADR induced cardiomyopathy.