• Clinical and Experimental Pediatrics
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• v.48 no.10
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• pp.1068-1075
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• 2005

Purpose : The role of ghrelin, which promotes the secretion of growth hormone, was not well known until now. Recently it was found that the mutation of ghrelin gene is related to obesity and diabetes. This study is to find the screening method that can easily and effectively detect the polymorphism of Leu72Met in ghrelin gene of obesity patients and apply it to clinical usage. Methods : We compared PCR-RFLP, PCR-SSCP and ARMS methodologies for analyzing of the polymorphism of Leu72Met in ghrelin gene of obesity children, and also studied the merits and demerits of these methodologies. Results : In this study, we were able to find out the band of peculiar allele of Leu72Met in ghrelin gene using PCR-RFLP, PCR-SSCP and ARMS analyses. The polymorphism of Leu72Met in ghrelin gene determined by all above methodologies was in complete agreement. Compared to the PCR-RFLP and PCR-SSCP, ARMS analysis is simple, inexpensive and also consume less time. It is very sensitive to analyze the polymorphism and easy to understand the results of test. Conclusion : Though PCR-RFLP, PCR-SSCP and ARMS analyses were sensitive to analyze the polymorphism of Leu72Met in ghrelin gene, ARMS analysis appears to be more efficient than PCR-RFLP and PCR-SSCP. Therefore, we conclude that ARMS analysis is suitable to analyze the polymorphism of Leu72Met in ghrelin gene for large quantity of specimens.

• Journal of Korean Society of Environmental Engineers
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• v.32 no.7
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• pp.656-664
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• 2010

We investigated the levels and distribution patterns of 16 priority polycyclic aromatic hydrocarbons (PAHs), 12 Coplanar polychlorinated biphenyls (Co-PCBs) and organochlorine pesticides (OCPs) in the surface sediments from the Nakdong river basin, Korea. The levels of 16 PAHs and 12 Co-PCBs in the river sediment samples ranged from not detected (N.D.)~969.3 ng/g-dry, 4.2~7716.5 pg/g-dry (0.0~10.1 pg-TEQ/g-dry), respectively. Also, the levels of 16 PAHs were from 5.8~2987.2 ng/g-dry and 4.3~461.1 pg/g-dry (0.0~0.6 pg-TEQ/g-dry) for 12 Co-PCBs in the lake sediment samples. Only dichloro-diphenyl-trichloroethane (DDT) were detected among target OCPs and the concentration ranged from N.D.~1.5 ng/g-dry in the river sediment samples. These contamination levels were far below the guideline values suggested for environmental quality assessment. Low molecular weight PAHs were dominant in the river sediment samples, while middle and high molecular weight PAHs were dominant among 16 PAHs in the lake sediment samples. PCB-118 and PCB-105 were predominant congeners in the sediment, which were similar to the results obtained from previous studies. With these results, the assessment of potential sources of PAHs and Co-PCBs contamination in the sediments of the Nakdong river basin was performed. The pyrogenic-PAHs originated from combustion of fossil fuel and biomass were related with the PAHs contribution in most of the sediment samples, and Co-PCBs in the sediment samples were related with commercial PCB products.

• 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.

• Research in Plant Disease
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• v.12 no.3
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• pp.298-302
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• 2006

An isolate of Cucumber mosaic virus(CMV), called as Can-CMV, was originally isolated from Canna generalis showing typical streak mosaic foliar symptoms, and its properties were investigated in this study. Whereas all known isolates of CMV could induce symptoms on their systemic hosts(four kinds of Nicotiana spp and a zucchini squash), Can-CMV induced no symptoms on its systemic hosts tested. Replication and movement of the virus on upper leaves as well as inoculated leaves-were confirmed by RT-PCR suggesting that Can-CMV could only infect systemically on N. benthamiana and N. glutinosa. Size of local lesions on the Can-CMV-inoculated leaves of Chenopodium amaranticolor was much smaller than that of Fny-CMV. Whereas Fny-CMV and LS-CMV could induce distinct necrotic local lesions on Vigna unguiculata 2 to 3 days postinoculation(dpi), chlorotic spots symptom was expressed by Can-CMV 4 to 5 dpi. Virus-specific 4 kinds of dsRNAs were isolated from leaves of N. benthamiana infected with Can-CMV, and these dsRNAs corresponded to the viral genomic RNAs and subgenomic RNAs and their patterns were indistinguishable to those of Fny-CMV and LS-CMV. By restriction mapping analysis of 950 bp of RT-PCR amplified products of coat protein gene of the virus as well as by serological analysis of gel diffusion test, Can-CMV belongs to a typical member of CMV subgroup IA. These results suggest that the Can-CMV isolated from C. generalis possesses unique pathological properties to understand further insight into the various interactions between virus and host.

• Korean Journal of Environmental Agriculture
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• v.30 no.3
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• pp.288-294
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• 2011

BACKGROUND: Methylmercury is analyzed by HPLC-ICP/MS because of the simplicity for sample preparation and interference. However, most of the pre-treatment methods for methylmercury need a further pH adjustment of the extracted solution and removal of organic matter for HPLC. The purpose of this study was to establish a rapid and accurate analytical method for determination of methylmercury in fish by using HPLC-ICP/MS. METHOD AND RESULTS: We conducted an experiment for pre-treatment and instrument conditions and analytical method verification. Pre-treatment condition was established with aqueous 1% L-cysteine HCl and heated at $60^{\circ}C$ in microwave for 20 min. Methylmercury in $50{\mu}L$ of filtered extract was separated by a C18 column and aqueous 0.1% L-cysteine HCl + 0.1% L-cysteine mobile phase at $25^{\circ}C$. The presence of cysteine in mobile phase and sample solution was essential to eliminate adsorption, peak tailing and memory effect problems. Correlation coefficient($r^2$) for the linearity was 0.9998. The limits of detection and quantitation for this method were 0.15 and $0.45{\mu}g/kg$ respectively. CONCLUSION: Result for analytical method verification, accuracy and repeatability of the analytes were in good agreement with the certified reference materials values of methylmercury at a 95% confidence level. The advantage of the established method is that the extracted solution can be directly injected into the HPLC column without additional processes and the memory effect of mercury in the ICP-MS can be eliminated.

• Korean Journal of Clinical Laboratory Science
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• v.38 no.2
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• pp.99-104
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• 2006

Human Papilloma viruses (HPVs) are etiological agents for cervical cancer and are classified into low- and high-risk categories. The aim of this study was to determine the frequency of the HPV genotype in the HPV screening test of Korean women using PCR-direct sequencing. Consensus primers of L1 legion were used for the amplification of HPV DNA and the PCR products (450 bps) obtained were analyzed by automatic sequencing. Sequences were compared with those in GenBank by using the BLAST program. Cervical swab samples of 3,978 women (20-73 years) were tested and the average age was 37.6 years. In this study, 1,174 samples were HPV positive out of 3,978 cervical swab samples screened (29.5%) and 136 samples (11.6%) showed a double infection. A total of 1,310 HPV genotypes were analyzed. The HPV positive rate was the lowest in the 20 years group (69.5%) and most of the samples of the > 60 years group were found HPV positive. Among thirty seven different HPV types identified by sequencing, 21 were HPV high risk types and 16 HPV low risk types were 69.8% (914/1,310) and 26.0% (340/1,310), respectively. In HPV high-risk types, 16 (13.21%), was the most frequently found. HPV 53 (9.62%) and 58 (9.24%) were also frequently found. This group was followed by HPV types 70 (5.50%), 33 (4.73%), 66 (4.20%), 18 (4.05%), 52 (4.05%), 31 (3.97%) and 56 (3.51%) in descending order of frequency. Among HPV low-risk types, 62 (4.20%), 6 (3.59%), 81 (3.59%), 84 (3.51%), and 11 (2.6%) were frequently found. In conclusion, PCR-direct sequencing could be used for quick and reliable typing of known and novel HPVs from clinical specimens. This data could be useful for epidemiological study of HPV and it also allows type-specific follow-up of women who have been treated for cervical intraepithelial neoplasia.

• The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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• v.26 no.1
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• pp.37-48
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• 2021

A new high speed rotation type-passive sampling device (HSR-PSD), which can rotate seawater at high speed and absorb easily and quickly the freely dissolved hydrophobic organic contaminants from seawater, was developed and then applied around the Korean Peninsula. Freely dissolved concentrations (Cfree) of polycyclic aromatic hydrocarbons (PAHs) were determined using the HSR-PSD with low density polyethylene (LDPE) sheets as a passive sampler. Furthermore, dissolved concentrations (Cdissolved) of PAHs in seawater were also obtained from high volume water sampling as a conventional method to account for actual bioavailability. When the LDPE sheets were rotated in the HSR-PSD at 900 rpm, PAHs with log KOW 3.4 ~ 5.2 were equilibrated between the LDPE and water in 5 hours. Although the high molecular weight PAHs with log KOW 5.6 ~ 6.8 was expected to be 2 to 30 days to reach the equilibrium, the Cfree of the PAHs at equilibrium could be corrected using performance reference compounds in 5 hours. Meanwhile, the total Cfree of PAHs were from 0.32 to 1.2 ng/L, which were higher than reported values in other oceans, but lower than in coastal water such as estuary, harbor, or shore. A bioavailability from the detected PAHs was highest at the sampling line near the dumping site of the Yellow Sea. Predicted residual concentrations in biota were relatively higher in offshore including the dumping site than in coastal regions.

• Korean Journal of Poultry Science
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• v.42 no.1
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• pp.15-26
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• 2015

In this study, genotyping was executed by using 27 microsatellite markers for genetic diversity of 469 Korean Native Chickens [20 population, each population is 24 samples but Hanhyup A line is 13 samples). in total 469 samples were collected from National Institute of Animal Science (Korean Native Chicken (NR, NY, NG, NL and NW), Ogye (NO), Leghorn F,K (NF and NK), Black and Brown cormish (NH and NS), Rhode Island Red C, D (NC and ND), Total is 12 populations] and Hanhyup [H line (HH), F line (HF), G line (HG), V line (HV), S line (HS), W line (HW), Y line (HY), A line (HA), total is 8 populations]. [The allele number were observed 5 (ADL0268) to 20 (MCW0127) each markers. Observed heterozygostiy ($H_{obs}$), expected heterozygosity ($H_{exp}$), polymorphism Information Content (PIC) were observed 0.359 to 0.677, 0.668 to 0.881 and 0.646 to 0.869, respectively. Using these markers, the calculated the heterozygote deficit within chicken line ($F_{is}$) value each population from mean 0.117. Phylogenetic tree showing the genetic relationship among 20 population using standard genetic distance calculated from 27 microsatellite markers. genetic distances revealed the closest (0.175) between NC and ND. on the other hand, Farthest genetic distances (0.710) revealed between NF and HV. STRUCTURE analysis and Principal Components Analysis (PCA) showed that results of similar phylogenetic tree. The expected probability of identity values on random individuals (Total population and only Hanhyup line) was estimated at $8.80{\times}10^{-83}$ and $3.87{\times}10^{-117}$, respectively. In conclusion, This study shows the useful data that be utilized as a basic data of Korean Native Chicken breeding and development for commercial chicken industry to meet the consumer's demand.

• Journal of the Korean Society of Food Science and Nutrition
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• v.12 no.1
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• pp.46-50
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• 1983

In order to find out the possibility of utilizing red pepper seed as food resources of fats and proteins, a series of studies were conducted. The red pepper seed contained 27.6% of crude fat and 22.2% of crude protein. The lipid fractions obtained by silicic acid column chromatography were mainly composed of 95.4% neutral lipid, where as compound lipid were 4.6%. Among the neutral lipid separated by thin layer chromatography, triglyceride was 85.6%, sterol ester 4.9%, free fatty acids 3.4%, diglyceride 2.5%, sterol 2.2% and monoglyceride 1.1%, respectively. The predominant fatty acids of red pepper seed oil were linoleic acid (57.1-75.4%), palmitic acid (13.9-21.3%) and oleic acid (8.0-15.1%), especially glycolipid contained 1.7% of linolenic acid and small amount of myristic acid and arachidic acid. The salt soluble protein of red pepper seed was highly dispersible in 0.02M sodium phosphate buffer containing 1.0M $MgSO_4$, and the extractability of seed protein was about 25.0%. Glutamic acid and arginine were major amino acids of red pepper seed protein. The electrophoretic analysis showed 6 bands in seed protein, and the collection rate of the main protein fraction purified by sephadex G-100 and G-200 was about 62.2%. Glutamic acid (19.9%) was major amino acid of the main protein, followed by glycine and alanine. The molecular weight of the main protein was estimated to be 93,000.

• Journal of Life Science
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• v.22 no.6
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• pp.703-712
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• 2012

Adeno-associated virus (AAV) has been considered to be a very safe and efficient gene delivery system. However, the major obstacles to therapeutic usage of AAV have been to achieve highly efficient and reproducible production processes, and also to develop a reliable quantifying method of various serotypes with a simple protocol. We compared the efficiency of the conventional production protocol of AAV2 and adenovirus (Ad) co-infection to that of a new method containing AAV2 infection followed by pHelper transfection. We tested HEK293 and 293T, and further examined the time-dependent changes of AAV2 production. The new method of AAV2 and pHelper DNA gave about ten times higher production efficiency than that of the conventional protocol. The highest production efficiency in 293T was achieved as $1.61{\times}10^5$ virus genomes (v.g.)/cell by the new method of 10 MOI of AAV2 infection and 5 days post-infection. This protocol of the highest efficiency was then applied to produce various AAV serotypes and showed the efficiencies higher than $10^5$ v.g./cell. Next, we designed the universal PCR primers of highly conserved regions for various AAV serotypes to develop a simple and reliable titration method. The universal primers could amplify all the tested AAV serotypes with similar sensitivities by ten molecular copies. Therefore, this pair of universal primers can be further utilized to detect AAV contaminants in therapeutic adenoviral vectors.