• Journal of Life Science
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• v.28 no.4
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• pp.389-397
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• 2018

The structure of glycan residues attached to glycoproteins can influence the biological activity, stability, and safety of pharmaceutical proteins delivered from transgenic pig milk. The production of therapeutic glycoprotein in transgenic livestock animals is limited, as the glycosylation of mammary gland cells and the production of glycoproteins with the desired homogeneous glycoform remain a challenge. The ${\beta}$-1,3-N-acetylglucosaminylatransferase1 (B3GNT1) gene is an important enzyme that attaches N-acetylglucosamine (GlcNAc) to galactose (Gal) residues for protein glycosylation; however, there is limited information about pig glycosyltransferases. Therefore, we cloned the pig B3GNT1 (pB3GNT1) and investigated its functional properties that could attach N-acetylglucosamine to galactose residue. Using several different primers, a partial pB3GNT1 mRNA sequence containing the full open reading frame (ORF) was isolated from liver tissue. The ORF of pB3GNT1 contained 1,248 nucleotides and encoded 415 amino acid residues. Organ-dependent expression of the pB3GNT1 gene was confirmed in various organs from adult and juvenile pigs. The pB3GNT1 mRNA expression level was high in the muscles of the heart and small intestine but was lower in the lungs. For functional characterization of pB3GNT1, we established a stable expression of the pB3GNT1 gene in the porcine kidney cell line (PK-15). As a result, it was suggested that the glycosylation pattern of pB3GNT1 expression in PK-15 cells did not affect the total sialic acid level but increased the poly N-acetyllactosamine level. The results of this study can be used to produce glycoproteins with improved properties and therapeutic potential for the generation of desired glycosylation using transgenic pigs as bioreactors.

• 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.

• Nuclear Medicine and Molecular Imaging
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• v.43 no.5
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• pp.487-494
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• 2009

Purpose: Small size of recombinant scFv antibody has many advantages such as rapid blood clearances and improved targeting antibodies to tumor region. On the other hand owing to small size, number of amino group is insufficient in conjugation with chelator and fluorescence labeling. This study is to introduce poly lysine tag to the C-terminal end of scFv lym-1 sequence for fluorescence chelator conjugation. Materials and Methods: Poly lysine scFv lym-1 gene, cloned into pET-22b (+) vector, was expressed in E. coli BL21 (DE3) strain. Antibody purification was performed with Ni-NTA column and then size exclusion column chromatography. Expression and purification levels of poly lysine tagged scFv lym-1 antibody were confirmed by western blot analysis. I-124, I-125, I-131 and Tc-99m were used for radiolabeling of purified poly lysine scFv lym-1. Flow cytometry analysis of FIT( conjugated poly lysine scFv lym-1 was performed for confirmation of immunoreactivity of human Burkitt's lymphoma cells. Results: Poly lysine scFv lym-1 antibody was purified through two steps and identified as molecular weight of 48 KDa. Radiolabeling yields of I-124, I-125, I-131 and Tc-99m into poly lysine scFv lym-1 were >99%, >99%, >95% and >99%, respectively. Flow cytometry analysis of poly lysine scFv and scFv lym-1 was showed similar immunoreactivity to human Burkitt's lymphoma cells. Conclusion: Poly lysine tag was useful for the sufficient number of amino groups to scFv lym-1 antibody for chelator conjugation with minimizing loss of immunoreactivity.

• Research in Community and Public Health Nursing
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• v.15 no.4
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• pp.587-599
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• 2004

Purpose: The purpose of this study is to compare abdomen-fat rate, life style and social-support between normal weight women and obese women. Method: 304 women objectives from their 30 to 59 years of age were selected living in Je-chon city, Chung-Buck province and their height and weight were measured from April 1st to June 30th, 2003. Data were classified into low-weight group ($18.5kg/m^2$), normal-weight group ($18.5{\sim}22.9kg/m^2$), over weight group ($23{\sim}24.9kg/m^2$), and obese group ($25kg/m^2$) following the Korean Conference of Obesity, 2001. in which 119 people in the normal weight group and 91 people in the obese group, i.e. total 210 people were analyzed in sequence. Using SPSS Win 10.1 Program, frequency and percentile, and by ANOVA, $X^2-test$ and t-test were treated. Results: The average age of obese women was 46.68 distributing 40.7% of forties and 39.6% of fifties while normal-weight women were average 41.73-year old distributing 53.8% of forties and 34.5% of thirties, which revealed aged in obese women. The body fat rate of obese women averaged $37.52{\pm}4.17%$, in which 98.9% of obese women and 21.0% of normal weight women with a more than 30% of body-fat rate resulted in a higher body-fat rate in obese women. The waists of obese women averaged $88.37{\pm}8.22cm$, in which more than 85cm showed in obese women of 68.2% and normal weight women of 7.6% indicating a higher waist-fat rate in obese women. The abdomen-fat rate of more than 0.85 of waist vs hip-fat showed 74.7% in obese women and 58.4% in normal weight women, indicating a higher abdomen-fat rate in obese women. Obese women and normal weight women showed significant differences in education level, number of children, religion, menstrual status, and mother's weight. Especially, obese women ate hotter or saltier food than normal weight women preferring meat. However, no significant differences appeared in marital status, social economic status. occupation. eating habits. smoking. drinking and physical exercise. Social support levels showed a lower rate in obese women than in normal weight women, indicating a statistically significant difference (p<.05). Observing areas of social support, obese women showed lower rates in attachment/intimacy, social integrity, opportunity of foster and confidence in value except help and instruction, which indicated a statistically significant difference (p<.05). Social support for obese women showed significant differences in age, education level, social hierarchy, religion and menstrual status. Obese women were more negative than normal weight women in health recognition, indicating a statistically significant difference (p<.01). Normal weight women showed higher health recognition when provided high social support and significantly low (p<.01) health recognition when provided low social support. However, there was no significant difference in health recognition in obese women whether high or low social support was given. The health recognition of obese women showed significant differences in age, education level, social hierarchy, number of children, menstrual status, physical exercise, eating habits, eating taste and preference of food. Conclusion: Obese women showed elder than normal-weight women, higher body-fat rate and abdomen-fat rate, lower social support, and a tendency to more negative health recognition. Therefore, providing weight-control programs for the treatment of obesity and prevention of recurrence for obese women to prevent progressing to adult disease and promote a healthy life, we suggest that better eating habits and the encouragement of regular physical exercise should be included, as well as total approachment on change of health recognition and social support would be needed.

• Restorative Dentistry and Endodontics
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• v.34 no.4
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• pp.333-339
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• 2009

This study was done to determine if there is any difference in microleakage between experimental composite resins, in which various proportions of three component photoinitiators (Camphoroquinone, OPPI, Amine) were included. Four kinds of experimental composite resin were made by mixing 3.2% silanated barium glass (78 wt.%, average size; 1 ${\mu}m$) with each monomer system including variously proportioned photoinitiator systems used for photoinitiating BisGMA/BisEMA/TEGDMA monomer blend (37.5:37.5:25 wt.%). The weight percentage of each component were as follows (in sequence Camphoroquinone, OPPI, Amine): Group A - 0.5%, 0%, 1% / Group B - 2%, 0.2%, 2% / Group C - 0.2%, 1%, 0.2% / Group D - 1%, 1%, 2%. Each composite resin was used as a filling material for round class V cavities (diameter: 2/3 of mesiodistal width; depth: 1.5 mm) made on extracted human premolars and they were polymerized using curing light unit (XL 2500, 3M ESPE) for 40 s with an intensity of 600 mW/$cm^2$. Teeth were thermocycled fivehundred times between $50^{\circ}C$and $550^{\circ}C$for 30s at each temperature. Electrical conductivity (${\mu}A$) was recorded two times (just after thermocycling and after three-month storage in saline solution) by electrochemical method. Microleakage scores of each group according to evaluation time were as follows [Group: at first record / at second record; unit (${\mu}A$)]: A: 3.80 (0.69) / 13.22 (4.48), B: 3.42 (1.33) / 18.84 (5.53), C: 4.18 (2.55) / 28.08 (7.75), D: 4.12 (1.86) / 7.41 (3.41). Just after thermocycling, there was no difference in microleakage between groups, however, group C showed the largest score after three-month storage. Although there seems to be no difference in microleakage between groups just after thermocycling, composite resin with highly concentrated initiation system or classical design (Camphoroquinone and Amine system) would be more desirable for minimizing microleakage after three-month storage.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.49 no.3
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• pp.193-201
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• 2023

Hydrogen peroxide (H₂O₂) is a type of active oxygen species (ROS) that causes oxidative stress in cells and affects cell growth, proliferation, senescence, and death. The purpose of this study is to find active peptides that attenuate cytotoxicity of H₂O₂. A positional scanning synthetic tetrapeptide combinatorial library was screened to predict the sequence of potentially active peptides. As a result of comparing the effect of peptide pools on H₂O₂-induced death of human keratinocytes (HaCaT cells), various active peptide sequences were predicted. Especially, peptides containing cysteine (C) residue were predicted to be active. In follow-up experiments, the cytotoxicity and activity of cysteine-containing peptides of different lengths, such as C-NH₂, CC-NH₂, CCC-NH₂, and CCCC-NH₂ were examined. C-NH₂ and CC-NH₂ showed no significant cytotoxicity up to 1.0 mM, but CCC-NH₂, and CCCC-NH₂ showed relatively strong cytotoxicity. C-NH₂ and CC-NH₂ alleviated H₂O₂-induced cytotoxicity. CC-NH₂ was more cytoprotective compared to C-NH₂, C, N-acetyl cysteine (NAC), and glutathione (GSH). When intracellular ROS was measured by flow cytometry, H₂O₂ increased ROS production, and CC-NH₂ suppressed ROS production more effectively than C-NH₂, and it was as effective as C, NAC, and GSH. This study suggests that CC-NH₂ of the cysteine-containing peptides of different lengths has an antioxidant property that safely and effectively alleviates H₂O₂-induced cytotoxicity and ROS production.

• Pediatric Infection and Vaccine
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• v.6 no.2
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• pp.219-233
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• 1999

Purpose : Respiratory syncytial virus(RSV) is the major cause of lower respiratory tract infection in infants and young children. This study was performed to analyze antigenic and genetic variation of G protein of subgroup A RSV. Methods : One hundred seventy-nine strains isolated at the Seoul National University Children's Hospital over 8 years-period from 1990 through 1998 were analysed for antigenic and genetic variability. Analysis was made by reactivity with monoclonal antibodies raised against RSV, and by restriction mapping and, for selected strains, nucleotide sequencing following amplification of full sequence of G gene by reverse transcription-polymerase chain reaction. Results : Restriction fragment analysis of the amplified G protein gene revealed 23 restriction patterns, 12 of which included more than 2 isolate, and the most frequent genetic type comprised 30% of the strains. Indirect immunofluorescent staining with monoclonal antibodies revealed 6 antigenic types with one predominant pattern accounting for 91% of the total strains. The most frequent antigenic type had 21 restriction patterns, and some viruses with same restiction pattern had different monoclonal antibody reaction pattern. Nucleotide sequence homology of subgroup A was 91~93% between reference(A2, Long) and Korean isolates, 93~99% among Korean isolates. Maximum-parsimony analysis demonstrated that Korean isolates were distinct from reference strains and subgroup A strains were clustered in 4 groups. Conclusion : The restriction analysis pattern of G protein gene identified greater diversity within subgroup A than was seen with the monoclonal analysis and a variety of antigenic and genetic types of RSV are circulating in Korea which are different from reference strains or strains isolated from other countries.

• Journal of the Mineralogical Society of Korea
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• v.7 no.1
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• pp.49-61
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• 1994

As the groundwater flows along the fractures of crystalline rocks, it will be in contact with the fracture walls mostly coated by secondary minerals which are quite different form those of host rocks. The presence of fracture-filling minerals in crystalline rocks is important on the view point of radioactive waste disposal because of their great surface reactivity. The Surichi drill hole of 200 m in depth in the Yugu area composed mainly of Precambrian gneiss was selected to study the formation process of clay minerals on the fracture wall of gneiss, and their relation with present groundwater. The water-rock interaction in fractures resulted in the formation of gibbsite and clay minerals. They are formed by two different processes : (1) Incongruent dissolution of feldspar by groundwater diffused from a fracture path into rock matrix produced smectite and illite in situ, (2) on the wall of fracture, gibbsite, kaolinite, smectite and illite are formed by precipitation of dissolved species in groundwater. They show the paragenetic sequence such as gibbsite${\leftrightarrow}$kaolinite${\leftrightarrow}$smectite or illite. The paragenetic sequence of fracture-filling minerals was controlled by increase of pH of groundwater, decrease of fracture permeability by precipitation of fillings, and immobility of alkali or alkaline earths in groundwater. The groundwater from the Surichi borehole is a $Na-HCO_{3}$ type with pH range of 8.6-9.2. The sodium and bicarbonate in groundwater would be supplied by the dissolution of albite and calcite, respectively. The saturation index of groundwater and surface water calculated by WATEQ4F indicates that gibbsite and kaolinite are under precipitation to equilibrium state, and that smectite and illite are under equilibrium to redissolution environment. The stability relation of clay minerals in the $Na_{2}O-Al_{2}O_{3}-SiO_{2}-H_{2}O$ system shows that kaolinite is stable for all waters.

• Journal of Genetic Medicine
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• v.5 no.1
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• pp.26-33
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• 2008

Purpose : Phenylketonuria(PKU) is an inborn error of metabolism and a genetic disorder resulting from a deficiency of phenylalanine hydroxylase(PAH) and decreased activity of tetrahydrobiopterin(BH4).In this study the correlation between the DNA mutation and clinical manifestations was investigated and PAH DNA mutations were compared bewteen Asian and Caucasian populations. Methods : DNA was isolated from peripheral leukocytes. The PAH gene was amplified by Polymerase Chain Reaction(PCR) and the sequence was analyzed with Multiplex Ligation-dependent Probe Amplification(MLPA). Results : We characterized the PAH gene of 102 independent Korean patients with PKU. PAH nucleotide sequence analysis revealed 44 different mutations, including 10 novel mutations comprising 9 missense mutations(N207D, K95del, A447P, G344D, P69S, S391I, A202T, G103S, and I306L) and 1 novel splice-site variant mutation(IVS10-3C>G). R243Q was the most prevalent mutation in this study. A259T has not previously been reported in Asian populations, but we found that this mutation had a frequency of 10.1% in our study. Furthermore, the genotypes of $BH_4$ responsive patients were analyzed and were divided into two groups: $BH_4$ medication-only group and $BH_4$ medication with diet therapy group. In the $BH_4$ medication-only group and $BH_4$ medication with diet therapy group, R241C was the most common mutation. Conclusion : Novel mutations in the PAH gene of PKU patients are still being discovered. Additional information as to the frequency of mutations in the tetrahydrobiopterine responsive gene is also accumulating. We anticipate that knowledge of these PKU gene mutations will assist the diagnosis, genetic counseling, and therapeutic treatment of PKU patients in future.

• The Journal of the Petrological Society of Korea
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• v.25 no.4
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• pp.373-388
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• 2016

To understand human activity in the past, the information about past environmental change including geomorphological and climatic conditions is essential and this can be traced by using age dating and geochemical analysis of sediments from the prehistoric sites. The sedimentary sequence of Seokgwan-dong Paleolithic Site located in Seoul was 5m long unconsolidated sediments and consists of lower part bedrock weathering sediments, slope deposits and upper-part fluvial deposits. In this study, upper part sediments were used to reconstruct past environmental change through age dating and various physical and chemical analyses including grain size, magnetic susceptibility and mineral and elements. The fluvial sediments can be divided into 4 units including three organic layers. Grain size analysis results showed that the sediments were very poorly sorted with fining upward features. Magnetic susceptibility was relatively high in the organic layers, indicating environmental changes causing mineral composition change at that times. The mineral and major element composition are similar to Jurassic biotite granite which mainly consists of quartz, K-feldspar, biotite and muscovite. The radiocarbon age of $14,240{\pm}80yr$ BP was obtained from the lower most organic layer of Unit III(O), suggesting that the fluvial sediments formed at least from the early stage of deglacial period after the end of Last Glacial Maximum. Subsequent wet and warm climates and resultant fluvial process including slope sedimentation during the Holocene may have been responsible for the sedimentary sequence in Seokgwan-dong paleolithic site and surrounding area. The observed organic layers suggests frequent wetland occurrence combined with natural levee changes in this area.