• Research in Plant Disease
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• v.22 no.2
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• pp.65-71
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• 2016

Cymbidium mosaic virus (CymMV) is a major virus infecting orchid plants and causing economic loss. In this study, the incidence of viral infection in Calanthe spp. at the Korean Institute of Calanthe was investigated using reverse transcription polymerase chain reaction. The CymMV infection rate was 42%, and the two viruses Odontoglossum ringspot virus and Cucumber mosaic virus had frequencies of 8% and 2%, respectively. Additionally, we characterized an isolate of CymMV, CymMV-GW, using biological tests and examined the nucleotide sequence properties of its complete genome. CymMV-GW induced chlorotic ringspots and chlorotic spot symptoms in inoculated leaves of Chenopodium amaranticolor and Nicotiana benthamiana, respectively. In this study, we have for the first complete genome sequence of CymMV-GW in Korea. The CymMV-GW genome was 6,225 nucleotides in length, excluding the poly-(A) tail, and showed whole-genome nucleotide and amino acid sequence identities of 97.7% and 100%, respectively, with the NJ-1 isolate of CymMV. Here, we report the complete genome sequence of the CymMV-GW isolate and viral infection rates for Calanthe spp. in Korea.

• Radiation Oncology Journal
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• v.33 no.2
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• pp.149-154
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• 2015

Purpose: We performed exome sequencing in a breast cancer family without BRCA mutations. Materials and Methods: A family that three sisters have a history of breast cancer was selected for analysis. There were no family members with breast cancer in the previous generation. Genetic testing for BRCA mutation was negative, even by the multiplex ligation-dependent probe amplification method. Two sisters with breast cancer were selected as affected members, while the mother of the sisters was a non-affected member. Whole exome sequencing was performed on the HiSeq 2000 platform with paired-end reads of 101 bp in the three members. Results: We identified 19,436, 19,468, and 19,345 single-nucleotide polymorphisms (SNPs) in the coding regions. Among them, 8,759, 8,789, and 8,772 were non-synonymous SNPs, respectively. After filtering out 12,843 synonymous variations and 12,105 known variations with indels found in the dbSNP135 or 1000 Genomes Project database, we selected 73 variations in the samples from the affected sisters that did not occur in the sample from the unaffected mother. Using the Sorting Intolerant From Tolerant (SIFT), PolyPhen-2, and MutationTaster algorithms to predict amino acid substitutions, the XCR1, DLL1, TH, ACCS, SPPL3, CCNF, and SRL genes were risky among all three algorithms, while definite candidate genes could not be conclusively determined. Conclusion: Using exome sequencing, we found 7 variants for a breast cancer family without BRCA mutations. Genetic evidence of disease association should be confirmed by future studies.

• BMB Reports
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• v.33 no.2
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• pp.184-187
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• 2000

Prostaglandin $E_2$ ($PGE_2$) plays an important role in the regulation of various gastric functions, and the growth-inhibitory activities on tumor cells are studied in vitro and in vivo. Although the mechanisms have attracted many researchers in the past decade, the molecular mechanisms of cell cycle arrest, or induction of apoptosis by $PGE_2$, is unclear. We investigated the effects of $PGE_2$ on the growth of the human gastric carcinoma cell line SNU1 and genes that are regulated by $PGE_2$ and isolated them using differential display RT-PCR (DD RT-PCR). FACS analysis suggested that SNU1 cells were arrested at the G1 phase by $PGE_2$ treatment. This growth inhibitory effect was in a time- and dose-dependent manner. Treatment of SNU1 cells with $10\;{\mu}g/ml$ $PGE_2$, followed by DD RT-PCR analysis, revealed differently expressed bands patterns from the control. Among the differently expressed clones, we found an unidentified cDNA clone (HGP-27) overexpressed in $PGE_2$-treated cells. The full-length cDNA of HGP-27 was isolated using RACE, which consisted of a 30-nt 5'-noncoding region, a 891-nt ORF encoding the 296 amino acid protein, and a 738-nt 3'-noncoding region including a poly(a) signal. This gene was localized on the short arm of chromosome number 11. Using the Motif Finder program, a myb-DNA binding repeat signature was detected on the ORF region. The COOH-terminal half was shown to have similarity with the $NH_3$-terminal domain of thioredoxin (Trx). This relation between HGP-27 and Trx implied a potential role for HGP-27 in modulating the DNA binding function of a transcription factor, myb.

• BMB Reports
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• v.44 no.4
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• pp.238-243
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• 2011

We generated 16,993 expressed sequence tags (ESTs) from two libraries containing full-length cDNAs from the brain and liver of the Korean Jindo dog. An additional 365,909 ESTs from other dog breeds were identified from the NCBI dbEST database, and all ESTs were clustered into 28,514 consensus sequences using StackPack. We selected the 7,305 consensus sequences that could be assembled from at least five ESTs and estimated that 12,533 high-quality single nucleotide polymorphisms (SNPs) were present in 97,835 putative SNPs from the 7,305 consensus sequences. We identified 58 Jindo dog-specific SNPs in comparison to other breeds and predicted seven synonymous SNPs and ten non-synonymous SNPs. Using PolyPhen, a program that predicts changes in protein structure and potential effects on protein function caused by amino acid substitutions, three of the non-synonymous SNPs were predicted to result in changes in protein function for proteins expressed by three different genes (TUSC3, ITIH2, and NAT2).

• Analytical Science and Technology
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• v.30 no.2
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• pp.75-81
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• 2017

5-methylthioninhydrin (5-MTN) is an amino acid sensitive reagent used for the development of latent fingermarks deposited on porous surfaces such as paper and wood. The present study demonstrates that the 5-MTN can be used as a latent footwear impression enhancement reagent, by reacting with trace multivalent metal ions, which are the main components of the latent footwear impression. 5-MTN and L-alanine complex (MTN-ALA) used for the latent footwear impression development was prepared, by mixing $4.5{\times}10^{-3}M$ 5-MTN (in methanol) and $4.5{\times}10^{-3}M$ L-alanine (in methanol) in 1:1 ratio, and keeping undisturbed at room temperature for 24 h. The latent footwear impressions were deposited on white and black non-porous surfaces (glass plate, polyethylene panel, polypropylene panel, acryl panel, polyvinyl chloride (PVC) panel, poly(methyl methacrylate) (PMMA) panel, acrylonitrile-butadiene-styrene (ABS) panel, tile), and a semi-porous surfaces (painted wood). The latent footwear impressions on these surfaces were treated with MTN-ALA complex by spraying. The fluorescence of footwear impressions (occurred due to the reaction between MTN-ALA and metal complexes) was observed under a 505 nm forensic light source and an orange barrier filter. The enhancement of latent footwear impression was achieved from black surfaces without any blurring. However, the fluorescence (enhancement) of footwear impression was not observed on the white PVC, PMMA, and ABS surfaces, because the incident light interfered and reflected on the surface. The sensitivity of MTN-ALA was superior to 2,2'-dipyridil, which is a representative non-fluorescing footwear impression enhancement reagent, and similar to 8-hydroxyquinoline, which is a representative fluorescing footwear impression enhancement reagent.

• The Korea Journal of Herbology
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• v.31 no.3
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• pp.49-57
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• 2016

Objectives： Polygonati Rhizoma (PR) has containing the bioactive compounds such as poly sccharide A,B,C, oligosaccharide, amino acid, it has reported to anti-diabetes and hypertension, atherosclerosis. In this study, we were evaluates antioxidant and anti-physical fatigue effects of PR and steamed PR.Methods : The sample was divided into 5 groups-PR0 (PR without steaming process), PR1 (PR with once steaming process), PR3 (PR with third steaming process), PR6 (PR with sixth steaming process), PR9 (PR with ninth steaming process). We measured anti-oxidant activity through contents of polyphenol, flavonoid and DPPH, ABTS free radical scavenging capacity. And, anti-physical fatigue effect was evaluated using the swimming test, and the AMPK protein expressions in soleus muscle.Results ： As a result, polyphenol, flavonoid, DPPH, ABTS free radical scavenging capacity of PR were increased as steaming times. Anti-physical fatigue effects by swimming test, PR0 have significantly increased, but steamed PR groups were decreased. The AMPK protein expressions of PR0 and PR1 groups were increased comparing with PR3, PR6 and PR9. All groups had effects on decreasing TG, creatine in blood serum, but had no effects on TC in blood serum.Conclusions ： In conclusion, PR with 9 steaming process was more excellent than not-processed PR in anti-oxidant effect such as DPPH, ABTS radical scavenging activity and contents of polyphenol, flavonoid, but, not-processed PR increased swimming times than processed PR. These results suggest that processed PR has anti-oxidant effect as steaming times, and not-processed PR may be a novel potential anti-physical fatigue agents than processed PR.

• Biomedical Science Letters
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• v.12 no.3
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• pp.131-137
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• 2006

RGS is a negative regulator of G-protein signaling and can be identified by the presence of a conserved $120{sim}125$ amino acid motif, which is referred to as the RGS box. A number of RGSs are induced in response to a wide variety of stimuli. Increased levels of RGSs lead to significant decreases in GPCR responsiveness. To obtain further evidence of a role of RGS proteins in rat C6 astrocytoma cells, we first determined the expression profile of RGS-specific mRNA in C6 cells using reverse transcription-polymerase chain reaction (RT-PCR) with a poly dT18 primer and transcript-specific primers. We found that RGS2, RGS3, RGS6, RGS9, RGS10, RGS12, and RGS16 were differentially expressed in C6 astrocytoma cells. The highest expression rate was found for RGS3, followed by RGS16, RGS10 and RGS9, whereas the expression level for RGS2 was barely detectable. We next assessed whether forskolin regulated the expression of RGSs expressed in C6 astrocytoma cells. The present study found that forskolin dose-dependently stimulated the expression of RGS2 transcripts. This up-regulation of RGS2 gene was abrogated by H-89, potent and broad-spectrum protein kinase A (PKA) inhibitors. Actinomycin D completely inhibited the up-regulation of RGS2 gene induced by forskolin $(10{\mu}M)$, indicating that the regulation of RGS2 gene is controlled at the transcriptional level. In addition, forskolin did significantly activate transcriptional cAMP response element (CRE) in either HEK 293 cells or C6 cells and did not modulate the $NF-{\kappa}B$ and AP-l activity as measured by luciferase reporter gene assay. Finally, forskolin induced the expression of RGS2 mRNA in C6 astrocytoma cells, which depend on the PKA pathway and CRE transcriptional pathways.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.6
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• pp.892-897
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• 2005

Macrophage colony-stimulating factor (M-CSF) is a growth factor required for growth and differentiation of mononuclear phagocyte lineage. Total and 16 poly (A) mRNA of bovine M-CSF were isolated from healthy bovine peripheral mononuclear cells stimulated by phobol 12-myristste 13-acetate (TPA). The more compatible cultured mononuclear cells were 5${\times}$10/ml for RNA isolation. TPA-activated mononuclear cells increased the level of M-CSF-mRNA more than concanavalin A (Con A) and lipopolysaccharide (LPS). The optimal analysis of reverse transcriptase-polymerase chain reaction (RT-PCR) for14 Macrophage colonystimulating factor (M-CSF) as a growth factor required for bovine M-CSF was denaturation at 94$^{\circ}C$ for 1 minute, annealing at 57$^{\circ}C$ for 1 minute, extension at 72$^{\circ}C$ for 1 minute for 30 cycles. The size of cDNA of bovine M-CSF by RT-PCR was 774 base pairs. A 774 base pairs cDNA encoding bovine M-CSF was synthesized by reverse transcriptase polymerase chain reaction (RT-PCR). Ligated cDNA was transformed to competent cells and then plasmid isolation and digestion was performed. Molecular cloning and sequencing were performed for cDNA of bovine M-CSF. The size of cloned cDNA of bovine M-CSF was 774base pairs. The homology of base sequence and amino acid sequence was 88% and 86% compared with known human M-CSF, respectively. From a high degree of sequence similarity, the obtained cDNA of bovine M-CSF is thought be a specific gene of bovine M-CSF.

• Research in Plant Disease
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• v.19 no.2
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• pp.77-83
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• 2013

Bean yellow mosaic virus (BYMV; genus Potyvirus, family Potyviridae) causes severe losses to various legume species and a number of non-legume species, particularly freesia plants. In a survey of virus diseases in Gyeonggi province, Korea, BYMV isolates were identified from many cultivated freesia species. Here, we determined the complete nucleotide sequences of a BYMV freesia isolate (BYMV-Fr; accession number FJ492961). BYMV-Fr genome consists of 9,545 nucleotides (nt) excluding the poly (A) tail and encodes 3,057 amino acid (aa), with an AUG start and UAG stop codon, containing one open reading frame typical of a potyvirus polyprotein. The polyprotein of BYMV-Fr was divided to ten proteins and the cleavage sites of each protein were determined. The coat protein (CP) and polyprotein of BYMV-Fr were compared at the aa level with those of the previously reported 4 BYMV isolates. BYMV-Fr shared 90.1 to 97.1 and 91.0 to 92.5% at the CP and polyprotein homology. Interestingly, BYMV-Fr showed identities of a lower level at the nt level of 5' noncoding region (61.4 to 67.6%) and at the aa level of P1 (71.4 to 72.8%), comparing with four BYMV isolates. Based on the aa sequence diversity of CP and polyprotein, phylogenetic analysis with the four BYMV isolates showed two distinct groups and BYMV-Fr and most BYMV isolates were most closely related to the clover yellow vein virus among 52 potyviruses. To our knowledge, this is the first report of the complete genome sequence of BYMV freesia strain.

• Macromolecular Research
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• v.12 no.4
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• pp.367-373
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• 2004

We have developed a rigorous heat treatment method to improve the biocompatibility of chitosan as a tissue-engineered scaffold. The chitosan scaffold was prepared by the controlled freezing and lyophilizing method using dilute acetic acid and then it was heat-treated at 110$^{\circ}C$ in vacuo for 1-3 days. To explore changes in the physicochemical properties of the heat-treated scaffold, we analyzed the degree of deacetylation by colloid titration with poly(vinyl potassium sulfate) and the structural changes were analyzed by scanning electron microscopy, Fourier transform infrared (FT-IR) spectroscopy, wide-angle X-ray diffractometry (WAXD), and lysozyme susceptibility. The degree of deacetylation of chitosan scaffolds decreased significantly from 85 to 30% as the heat treatment time increased. FT-IR spectroscopic and WAXD data indicated the formation of amide bonds between the amino groups of chitosan and acetic acids carbonyl group, and of interchain hydrogen bonding between the carbonyl groups in the C-6 residues of chitosan and the N-acetyl groups. Our rigorous heat treatment method causes the scaffold to become more susceptible to lysozyme treatment. We performed further examinations of the changes in the biocompatibility of the chitosan scaffold after rigorous heat treatment by measuring the initial cell binding capacity and cell growth rate. Human dermal fibroblasts (HDFs) adhere and spread more effectively to the heat-treated chitosan than to the untreated sample. When the cell growth of the HDFs on the film or the scaffold was analyzed by an MTT assay, we found that rigorous heat treatment stimulated cell growth by 1.5∼1.95-fold relative to that of the untreated chitosan. We conclude that the rigorous dry heat treatment process increases the biocompatibility of the chitosan scaffold by decreasing the degree of deacetylation and by increasing cell attachment and growth.