• 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.

• Korean Journal of Clinical Laboratory Science
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• v.48 no.3
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• pp.169-175
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• 2016

Saccharin (o-benzoic sulfimide) is the first artificial and non-caloric sweetener that was first synthesized in 1879. In this study, we examined the biological activity of saccharin against various human cancer cell lines and human bone marrow-derived mesenchymal stem cells. A viability assay based on the conversion of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was performed to test for the cytotoxicity of saccharin about the four human cancer cell lines (H460, H157, A549 and SKOV3), one murine cancer cellline (Raw264.7), and MSCs. In order to find the differentially expressed gene in saccharin-treated MSCs against untreated MSCs, we performed annealing control primer (ACP)-based differential display reverse transcriptionp-olymerase chain reaction (DDRT-PCR). All tested cells were treated with saccharin at various concentrations (0.0, 4.8, 7.2, 9.6, 12.0, 14.4 mg/mL) for 48 hr. The number of metabolically active cancer cells decreased when treated with the saccharin at various concentrations for 48 hr as compared with the untreated cells. The decrease in cell survival was more evident with increasing concentrations of saccharin. Moreover, novel candidate genes, which were differentially expressed in MSCs in response to saccharin, were identified in 16 bands on 2% agarose gel. This revealed 16-7 up-regulated and 9 down-regulated-differentially expressed genes indicated by arrows. One of these candidate genes was a FK506-binding protein gene. The functional roles of FK506 binding proteins, with respect to the activities of stem cell proliferation, were not characterized. Further studies are required to get a better understanding of FK506-binding proteins in its roles in increasing stem cell proliferative activities from using saccharin.

• Korean Journal of Agricultural Science
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• v.25 no.1
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• pp.79-88
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• 1998

This study was performed in order to apply to effective breeding of Korean native cattle on the molecular genetic level obtained from PCR and nucleotide sequencing analysis of BoLA DRB3 exon2 that has important roles in host immune defence. Genomic DNA used in this study was prepared from the blood of Korean native cattle in Korean Native Cattle Improvement Center of National Livestock Cooperation. The results obtained from this study are summarized as follows: 1. Genomic DNA extracted from the blood of Korean native cattle was subjected to electrophoresis on 1.5% agarose gel. Major band was bigger than 12.2kb, indicating that genomic DNA was well prepared for PCR. Amplified products of 284bp fragments was obtained the amplification of BoLA DRB3 exon2 gene by PCR. 2. Cloning of BoLA DRB3 exon2 of Korean native cattle with pCR2.1 vector was conformed by 300bp fragment from recombinent plasmid that restricted with enzyme digestion of EcoRI. 3. Homology of BoLA DRB3 exon2 alleles of parent was 82.0% between sire's alleles and 90.1% between dam's alleles. 4. In pedigree analysis using BoLA DRB3 exon2 gene, sequencing result of BoLA DRB3 exon2 genes showed inheritance by Mendelian mode through the parents to their offspring. 5. Taking together those experimental results, pedigree was confirmed on the basis of sequencing for the alleles of parents and offspring. This knowledge by the molecular biological approach could be served for the improvement of Korean native cattle.

• Investigative Magnetic Resonance Imaging
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• v.12 no.2
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• pp.131-141
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• 2008

Purpose : To investigate the feasibility and accuracy of Proton Resonance Frequency (PRF) shift based magnetic resonance (MR) temperature mapping utilizing the self-developed center array-sequencing phase unwrapping (PU) method for non-invasive temperature monitoring. Materials and Methods : The computer simulation was done on the PU algorithm for performance evaluation before further application to MR thermometry. The MR experiments were conducted in two approaches namely PU experiment, and temperature mapping experiment based on the PU technique with all the image postprocessing implemented in MATLAB. A 1.5T MR scanner employing a knee coil with $T2^*$ GRE (Gradient Recalled Echo) pulse sequence were used throughout the experiments. Various subjects such as water phantom, orange, and agarose gel phantom were used for the assessment of the self-developed PU algorithm. The MR temperature mapping experiment was initially attempted on the agarose gel phantom only with the application of a custom-made thermoregulating water pump as the heating source. Heat was generated to the phantom via hot water circulation whilst temperature variation was observed with T-type thermocouple. The PU program was implemented on the reconstructed wrapped phase images prior to map the temperature distribution of subjects. As the temperature change is directly proportional to the phase difference map, the absolute temperature could be estimated from the summation of the computed temperature difference with the measured ambient temperature of subjects. Results : The PU technique successfully recovered and removed the phase wrapping artifacts on MR phase images with various subjects by producing a smooth and continuous phase map thus producing a more reliable temperature map. Conclusion : This work presented a rapid, and robust self-developed center array-sequencing PU algorithm feasible for the application of MR temperature mapping according to the PRF phase shift property.

• Journal of Sericultural and Entomological Science
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• v.23 no.2
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• pp.11-36
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• 1982

Forty-nine plant species as additives to silkworm artificial diet and 5 species as cellulose sources for artificial diet were screened for their economic values as feed-resources for the silkworm. Feeding response to artificial diet was tested on 82 silkworm strains. The effect of rearing conditions on feeding response and enzyme activities in the silkworm was investigated. The results were summarized as follows. 1. Seven species out of 49, Vigna sinensis ENDL, Ipomoea vatatas Lamarck, Cyperus anuricus Var. Laxus, Alnus japonica Stendel, Trifolium repens L, Prunus serrulata Lindley. Var, Glycine max L increased feeding response, compared with the basic formula of artificial diet. 2. The economic values of Vigna sinensis ENDL, Ipomoea vatatas Lamarck, Cyperus anuricus Var. Laxus, Ainus japonica Stendel, Cassia tera L, Erigeron canedensis L as feed-resources for artificiale diet were recognized, through feeding experiment during the entire larval stage. 3. Mulberry cellulose showed the best results in rearing and cocoon characteristics. 4. The extent of feeding response varied according to strains and varieties. Varieties in japanese strains showed higher feeding response than those in chinese and european varieties, with considerable variations among a varieties in strains. 5. The begining of 4th instar seems to be a proper time to convert from mulberry to artificial diet, or artificial diet to mulberry, however the middle of 3rd instar seems acceptable. 6. The optimum temperature for artificial diet rearing is 30$^{\circ}C$ during the period of 1st-3rd instar and 28$^{\circ}C$ for 4th-5th instar. 7. Electrophoretic isozyme patterns of esterase and acid phosphatase on agarose gel, as affected by strain. rearing temperature and feed-resources, were observed as follow. (1) Isozyme patterns of mid-gut esterase varied, depending on instar. One or two more isozyme bands were observed in the larvae than feed on the mulberry fed for the artificial diet. (2) A strain, chinese-15 with a higher feeding response, had 1∼2 more bands than chinese-60 with a lower feeding response. (3) Five bands of mid-gut esterase in 3rd and 4th instar larvae reared at 28$^{\circ}C$. and 4 for 3rd instar and 6∼7 for 4th instar larvae at 35$^{\circ}C$ were observed. (4) No similar esterase bands could be found among mid-gut, blood and silkgland. There are five esterase bands in the midgut, one in blood and three in silkgland. (5) There was rather small digerence in acid phosphatase types of mid-gut and blood according to varieties and rearing temperature. No active band was shown in silkgland. In midgut, there was one acid phosphatase band at 3rd instar, two at 4th instar and three at 5th instar. In blood, one active band at 3rd or 4th instar and three bands at 5th inster wire detected.

• Korean Journal of Clinical Laboratory Science
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• v.39 no.2
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• pp.113-121
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• 2007

Hearing loss is a common congenital disorder that is frequently associated with mutations in the Cx26 gene (GJB2). Recently, the mutation analysis of GJB2 has been used in a newborn screening test for the detection of hearing impairment. Population-based studies should be performed before the application of genetic testing for the identification of deaf newborns. In this study, 8 positions of GJB2 mutations-including 35delG, 167delT, 235delC, V27I, V37I, M34T, E114G, and I203T-were analyzed using PCR-direct sequencing in a total of 437 healthy Korean neonates. DNAs from dried blood spots were extracted using a commercial DNA extraction kit. The PCR-amplified products (783 bps) of the GJB2 gene were detected using 2% agarose gel electrophoresis and subjected to direct sequencing. The sequences were compared with those in the GenBank database by using the BLAST program. In this study, 5 GJB2 mutations -including V27I (79G>A), V37I (109G>A), E114G (341A>G), I203T (608T>C), and 235delC- were found. Of the 437 neonate samples, 301 subjects showed GJB2 mutations (68.9%, 301/437). The V27I mutation was found in 271 subjects and was the most frequent (62.0%, 271/437). The E114G, I203T and V37I mutations were shown in 146, 17 and 14 subjects, respectively. The 235delC mutation was found in 1 subject. The E114G mutation was frequently accompanied by the V27I mutation. V27I/E114G (97.2%, 143/147) was the most common double mutation and 3 subjects had the double mutation V27I/I203T. A triple mutation, V27I/E114G/I203T, was found in 1 subject. In conclusion, PCR-direct sequencing is a convenient tool for the rapid detection of GJB2 mutations and this data might provide information for the genetic counseling of the GJB2 gene.

• Korean Journal of Food Science and Technology
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• v.30 no.5
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• pp.1214-1221
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• 1998

The study was carried out to investigate the antioxidant effects of ethanol, methanol, and water extracts or fractions of ethanol extract of Ligularia fischeri on low density lipoprotein(LDL) and ethanol extract was further fractionated. The methanol extract and ethyl acetate fraction showed strong antioxidant effect with 71.7% (13.36 nmol/mg) and 95% (1.35 nmol/mg) inhibition in the presence of $15\;{\mu}g/mL$, respectively. In the value of malondialdehyde(MDA) with oxidation time, ethanol, methanol and water extract in the presence of $25\;{\mu}g/mL$ inhibited the oxidation up to 4h incubation and ethyl acetate fraction showed strong antioxidant effect up to 8h incubation. Also, the ethanol, methanol and water extract showed antioxidant effects in the agarose gel electrophoresis test. The conjugated diene formation by lipid oxidation with addition of $Cu^{2+}$ in the Ligularia fischeri extracts and their fractions was decreased approximately 1.1 to 2.8 times and 2.2 to 3.2 times, respectively compared to control. In the degradation of apo B-100 by oxidation using SDS-PAGE, ethanol, methanol and water extract showed similar degradation band pattern compared to that of native LDL band. Apo B-100 contents using densitometer were 77.8, 92.5% and 82.3% in the ethanol, methanol and water extract, respectively, compared to 100% of native LDL. In the meanwhile, apo B-100 contents in hexane, ethyl acetate and water fraction were 38.8, 94.5 and 65.5% respectively. This results indicated that ethyl acetate fraction showed the strongest antioxidant effect on MDA value or apo B-100 contents of LDL.

• Radiation Oncology Journal
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• v.16 no.2
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• pp.91-98
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• 1998

Purpose : The relationship between environmental PH on the radiation induced-apoptosis in SCK mammary adenocarcinoma cells and cell cycle dependence was investigated. Material and Methods : Mammary adenocarcinoma cells of A/J mice(SCK cells) in exponential growth phase were irradiated with a $l37^Cs$ irradiator at room temperature. The cells were irradiated 1 hour after the media was replaced with fresh media at a different pHs. After incubation at $37^{\circ}C$ for 0-48 h, the extent of apoptosis was determined using agarose gel electrophoresis and flow cytometry. The progression of cells through the cell cycle after irradiation in different pHs was also determined with flow cytometry. Bssults : The induction of apoptosis by irradiation in pH 6.6 medium was markedly less than that in pH 7.5 medium. When the cells were irradiated and maintained in pH 7.5 medium, the percentage of cells in $G_2/M$ phase rapidly increased to about $70\%$ at 12 h after an exposure to 120y and returned to control level by 36 h. The percentage of cells in G1 phase decreased as the percentage of cells in $G_2/M$ increased. On the other hand, in pH 6.6 medium the percentage of cells in G2/M phases gradually increased to about $45\%$ at 24 h after 12Gy irradiation and then slowly recessed and consequently, as much as $30-35\%$ of the cells were still in the Ga/M phase 48 h after irradiation. The percentage of cells in G1 phase then increased as the Ga/M arrest began to recede. The radiation-induced Ga/M arrest in PH 0.0 medium lasted markedly longer than that in pH 7.5 medium. Conclusion : Radiation-induced apoptosis in SCK tumor cells are reversely suppressed in an acidic environment. Radiation-induced Ga/M arrest is prolonged in an acidic environment indicating that the suppression of radiation-induced apoptosis and prolongation of radiation-induced Ga/M arrest in an acidic environment are related.

• Proceedings of the Korean Society for Food Science of Animal Resources Conference
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• 2000.11a
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• pp.59-75
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• 2000

This study was carried out to develop a DNA marker for identifying between Korean cattle (Hanwoo) and other breeds. First experiment was performed to isolate Hanwoo specific DNA marker at sequence characterized amplified regions (SCARs). Five breeds of cattle including Hanwoo, Holstein, Hereford, Angus and Charolais were represented with the from 8 to 20 individuals. Fourteen primers of 300 arbitrary primers of 10 nucleotides showed reproducible polymorphism across the breeds. An amplified band of 0.9 kb in the primer MG-3 showed the specificity to Holstein breed. And MG-6 and MG-12 detected the Hereford and Hanwoo specific markers at the size of 2.0 kb and 1.0 kb, respectively. A 1.0 kb band of MG-12 was cloned and sequenced. A SCAR primer was designed based on the obtained sequences. It was possible to identify the Hanwoo from Holstein breed. Second experiment was carried out to observe the genotype frequencies of MC1R in 1,044 samples of imported beef and eight different cattle breeds including Hanwoo, Holstein, Angus, Brown-Swiss, Charolais, Limousin, Simmental and Hereford. The primers for the amplification of bovine MC1R gene were designed based on a bovine MC1R gene sequence (GenBank accession no.Y19103). A size of 350 bp was amplified by polymerase chain reaction(PCR), digested with two different restriction enzyme, BsrFI and MspA II, and electrophoresed in 2.5% Metaphore agarose gel for determination of genotypes. Genotype frequencies of Hanwoo were 0.10 in E+e and 0.90 in ee. Allele ED was shown in all of Holstein and Angus breeds tested which have black coat color phenotypes. We suggested that SCAR marker and the bovine MC1R gene could be used as a DNA marker for distinguishing beef between Hanwoo and Holstein.

• Journal of Life Science
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• v.19 no.11
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• pp.1581-1590
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• 2009

Hizikia fusiforme is a kind of brown edible seaweed that mainly grows in the temperate seaside areas of the northwest pacific, including Korea, Japan and China, and has been widely used as a health food for hundreds of years. Recently, H. fusiforme has been known to exert pharmacological activities including antioxidant, antimutagenic and anticoagulant activities. However, the molecular mechanisms of H. fusiforme in malignant cells have not been clearly elucidated yet. In this study, the effects of ethyl alcohol extract of H. fusiforme (EAHF) on the anti-proliferative effects of MDA-MB-231 and MCF-7 human breast cancer cells were investigated. EAHF treatment resulted in a concentration-dependent growth inhibition by including apoptosis in MDA-MB-231 cells and G1 phase arrest in MCF-7 cells, which could be proved by MTT assay, DAPI staining, agarose gel electrophoresis and flow cytometry analysis. In MDA-MB-231 cells, the increase in apoptosis induced by EAHF treatment correlated with up-regulation of pro-apoptotic Bax expression. EAHF treatment induced the proteolytic activation of caspase-3 and caspase-9, and a concomitant inhibition of poly (ADP-ribose) polymerase, $\beta$-catenin, phospholipase-${\gamma}1$ protein and DNA fragmentation factor 45/inhibitor of caspase-activated DNase. Taken together, these findings provide important new insights into the possible molecular mechanisms of the anti-cancer activity of H. fusiforme.