Nicotinic acetylcholine receptors (nAChRs) play important roles in nervous system functions and are involved in a variety of diseases. We previously demonstrated that ginsenosides, the active ingredients of Panax ginseng, inhibit subsets of nAChR channel currents, but not ${\alpha}7$, expressed in Xenopus laevis oocytes. Mutation of the highly conserved Leu247 to Thr247 in the transmembrane domain 2 (TM2) channel pore region of ${\alpha}7$ nAChR induces alterations in channel gating properties and converts ${\alpha}7$ nAChR antagonists into agonists. In the present study, we assessed how point mutations in the Leu247 residue leading to various amino acids affect 20(S)-ginsenoside $Rg_3$ ($Rg_3$) activity against the ${\alpha}7$ nAChR. Mutation of L247 to L247A, L247D, L247E, L247I, L247S, and L247T, but not L247K, rendered mutant receptors sensitive to $Rg_3$. We further characterized $Rg_3$ regulation of L247T receptors. We found that $Rg_3$ inhibition of mutant ${\alpha}7$ nAChR channel currents was reversible and concentration-dependent. $Rg_3$ inhibition was strongly voltage-dependent and noncompetitive manner. These results indicate that the interaction between $Rg_3$ and mutant receptors might differ from its interaction with the wild-type receptor. To identify differences in $Rg_3$ interactions between wild-type and L247T receptors, we utilized docked modeling. This modeling revealed that $Rg_3$ forms hydrogen bonds with amino acids, such as Ser240 of subunit I and Thr244 of subunit II and V at the channel pore, whereas $Rg_3$ localizes at the interface of the two wild-type receptor subunits. These results indicate that mutation of Leu247 to Thr247 induces conformational changes in the wild-type receptor and provides a binding pocket for $Rg_3$ at the channel pore.
Kim, Chang-Yung;Cho, Gyu-Taek;Baek, Hyung-Jin;Lee, Sok-Young;Lee, Myung-Chul;Lee, Young-Yi;Choi, Yu-Mi
Korean Journal of Plant Resources
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v.26
no.2
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pp.328-335
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2013
The National Agrobiodiversity Center is the nodal agency assigned as the National Agricultural Genebank of the Rural Development Administration. Its main role is to collect, conserve, evaluate and distribute plant genetic resources. As of 2010, NAC has distributed a total of 380,981 accessions in the last 20 years (1991-2010) or an average of about 19,000 accessions per year. To meet customers' demands for germplasm and derive quality improvements, a mail survey in 2011 was conducted among the genetic resource users in 2010. Most of the clients obtained information on the germplasm conserved in the national genebank from the NAC website or NAC staff, and they sought specific traits in the samples. Most users received the materials within 15 days, and wanted useful data together with genetic resource. Korean landrace was the most frequently requested accessions. According to the survey results, it is supposed that useful genetic resources should be preferentially collected and their characterization/evaluation should be strengthened to enhance the utilization of genetic resources.
Journal of the Korean Association of Oral and Maxillofacial Surgeons
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v.36
no.5
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pp.346-352
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2010
Introduction: Heat shock protein70 (HSP70) is a highly conserved family of proteins produced after a variety of stresses. Many studies reported that the overexpression of HSP70 can improve the prognosis of the patients with sepsis through a reduction of the nitric oxide concentration. However, these results only revealed the effect of HSP70 and nitric oxide. No studies have examined the relationship between HSP70 and nitric oxide. The aim of this study was to evaluate the effect of the overexpression of HSP70 on the expression of inducible nitric oxide synthase and the nitric oxide concentration. In addition, the mechanism of the relationship of HSP70 and inducible nitric oxide synthase (iNOS) in sepsis was examined. Materials and Methods: The experiments were performed on male sprague-dawley rats. Sepsis was induced by a cecal ligation and puncture (CLP). Glutamine (GLN) or saline was administered 1 hour after the initiation of sepsis. Serum and lung tissues were acquired from the rats 12 hours or 24 hours after the initiation of sepsis. The nitric oxide concentration, the expression of HSP70 in lung, and the gene expression of iNOS in lung were analyzed. The three groups, sham operation, CLP and CLP+GLN, were compared. Results: Compared to the other groups, in CLP+GLN, GLN administered after the initiation of sepsis enhanced the expression of HSP70 in the lung at 12 hours ($47.19{\pm}10.04$ vs. $33.22{\pm}8.28$, P=0.025) and 24 hours ($47.06{\pm}10.60$ vs. $31.90{\pm}4.83$, P=0.004). In CLP+GLN, GLN attenuated the expression of iNOS messenger RNA (mRNA) in the lung at 12 hours ($5,513.73{\pm}1,051.60$ vs. $4,167.17{\pm}951.59$, P=0.025) and 24 hours ($18,740.27{\pm}8,241.20$ vs. $9,437.65{\pm}2,521.07$, P=0.016), and reduced the concentration of nitric oxide in the serum at 12 hours ($0.86{\pm}0.48$ vs. $3.82{\pm}2.53$, P=0.016) and 24 hours ($0.39{\pm}0.25$ vs. $1.85{\pm}1.70$, P=0.025). Conclusion: The overexpression of HSP70 induced by the administration of GLN in sepsis attenuates the expression of the iNOS gene but reduces the nitric oxide concentration.
The present study was conducted to analyze the quality of the habitats of Abies koreana WILS. by using the InVEST model based on the analytic hierarchy process (AHP) technique and to evaluate the economic value by estimating the carbon fixation. Abies koreana WILS., an original biological species of South Korea, may be an essential element in establishing the national biological sovereignty in the future. The subjects of the present study were the national parks in Mt. Halla, Mt. Jiri, and Mt. Sobaek, which are the habitats of Abies koreana WILS. As suggested by previous studies as a limitation of the InVEST model, the utilization of the data from relevant international publications as the input data, due to the lack of the domestic input data, may decrease the accuracy of the modeling. Therefore, the AHP technique was applied for the input data. The modeling was performed with reference to the years of 1980, 1990, and 2000 for the scenario analysis. The result of the modeling showed that the habitat quality was changed most in the national park in Mt. Halla, as the habitat quality score was decreased from 0.96 in 1980 to 0.97 in 1990 and 0.94 in 2000. In the national part of Mt. Sobeak, the habitat quality was changed most in the sub-alpine zone, as the habitat quality score was decreased from 0.98 in 1980 and 0.98 in 1990 to 0.97 in 2000. The habitat quality was best conserved in the national part in Mt. Jiri, as the habitat quality score was 0.98 in 1980, 0.99 in 1990, and 0.99 in 2000. The estimated economic loss by the change of the habitat quality was 19,280,000 USD for Mt. Halla and 8,030,000 USD for Mt. Sobeak. In the present study, the habitat quality of the Abies koreana WILS, the original species of South Korea, was evaluated and the economic value of the ecological services provided by the habitats was estimated quantitatively. The result showed that the ecosystem service model may be used to qualitatively analyze the quality of a habitat located in a specific region and to estimate the economic value quantitatively. The objective evaluation of ecosystem services demonstrated in the present study may be applied to promote sustainable utilization of natural resources and conservation of the ecosystem by predicting the changes that may be caused by external factors including the development of preservation areas.
The evaluation of soybean germplasm has mainly been carried out by morphological characters at Genetic Resources Division, Rural Development Administration (RDA). However, this information has been limited serving a diverse information for user and effectively managing the soybean germplasm. To resolve this problem, soybean collection conserved at RDA gene bank was profiled using nine soybean SSR (Simple Sequence Repeat) markers. Soybean SSR allele was confirmed using genescan and genotyper softwares of automatic sequencer for accurate genotyping of each accession and continuous accumulation of data. SSR profiling of soybean germplasm has been carried out from 2,855 (Satt458) to 4,368 (Satt197) accessions by locus. The number of allele revealed 267 with an average of 29.6 in total accession, and varied from a low of 21 (Satt532 and Satt141) to a high of 58 (Sat_074). Although the number of accessions of wild soybean is less than that of soybean landraces, Korean wild soybean is more variable than other soybean landraces populations in total number of alleles. However, Korean soybean landraces were more variable than Korean wild soybeans in 5 loci. In the allele frequency, wild soybean accessions showed an even distribution in all alleles and higher distribution in low ladder than in high ladder. Also, Korean soybean landraces revealed a high condensed frequency in Satt286 (202 bp, 232 bp), Chinese soybean landraces in Satt197 (171 bp) and Satt458 (173 bp), and Japanese soybean landraces in Sat_074 (244 bp) and Satt458 (170 bp). These SSR profile information will be provided as indications of redundancies or omissions of accessions and can aid in managing soybean collection held at RDA gene bank. The information on diversity analysis could help to enlarge the genetic diversity of materials in breeding program, and could be used to develop a core collection of soybean germplasm.
Lee, Seungbum;Jang, Ik;Hyun, Do Yoon;Lee, Jung-Ro;Kim, Seong-Hoon;Yoo, Eunae;Lee, Sookyeong;Cho, Gyu-Taek;Lee, Kyung Jun
KOREAN JOURNAL OF CROP SCIENCE
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v.65
no.4
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pp.485-495
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2020
Melon (Cucumis melo L.), one of the most important fruit crop species, is cultivated worldwide. In this study, a total of 206 melon accessions conserved at the National Agrobiodiversity Center (NAC) in RDA were characterized for nine morphological characteristics according to the NAC descriptor list. In addition, to confirm the genetic composition of each melon accession, genetic profiling was performed using 20 SSR markers. Among the 206 melon accessions, 159 (77.2%) were collected from Asia. The color of fruit flesh and skin were mostly 'white' (56.0%) and 'green' (49%), respectively. Days to female flowering (FD) and maturity (MD) of the accessions ranged from 58 to 72 and 17 to 63, respectively. The fruit length and width of the accessions ranged from 6.0 to 29.3 and 3.6 to 17.2 cm, respectively. The sugar content (SU) ranged from 2.5% to 13.2% with an average of 7.0%. In correlation analysis, SU showed positive and negative correlations with MD and FD, respectively. The accessions were classified into four clusters by cluster analysis. From the results of genetic profiling using 20 SSR markers, three accessions (K189118, K100486, and K190292) were expected to be inbred lines among 206 melon accessions. These results could expand the knowledge of the melon germplasm, providing valuable material for the development of new melon varieties to suit consumer tastes.
The purpose of this study was to characterize equine heat-shock protein (Hsp) genes and analyze their expression pattern in various horse tissues and blood leukocytes after exercise. In a previous study, RNA sequencing of blood and skeletal muscles of thoroughbreds before and after exercise was performed using differently expressed gene (DEG) analysis. Three Hsp genes (HspH1, Hsp90${\alpha}$ and Hsp70) were selected by DEG analysis and were found to be differentially expressed in either blood or muscle. To validate and extend previous observations on these genes, we performed RT-PCR analyses of horse tissue as well as real-time qPCR analyses of blood leukocytes after exercise. mRNA expression of these Hsp genes was found to be ubiquitous in the analyzed tissues (including thyroid, colon, skeletal muscle, cecum, kidney, spinal cord, heart, and lung). In addition, Hsp mRNA expression of these genes in extracted whole blood increased after 120 minutes of exercise compared to the baseline condition. These results are in agreement with the results of human and other experimental animals, suggesting that regulatory mechanisms that are responsible for upregulation of Hsp gene transcription may be conserved among species. Further investigations to correlate Hsp gene expression patterns with athletic performance or recovery processes after exercise are warranted.
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.
Park, Hye-Mi;Yoo, Seon-Hee;Lee, Seon-Ho;Kim, Nyun-Ok
The Korean Journal of Nuclear Medicine Technology
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v.20
no.2
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pp.75-79
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2016
Purpose The ASAN Medical Center, Nuclear Medicine performs TSH (Thyroid stimulating hormone) and FT4 (Free Thyroxine) tests 8 times per day. Accordingly, 70 ~ 80 kit tubes are consumed every day for the measurements and the time consumed for reagent dispensing averages over 170 seconds, where the TAT (turnaround time) may be effected when the number of test samples is larger than expected. Therefore, the following test was conducted with the purpose to reduce the number of kit tubes consumed, and reduce the time for reagent dispensing. Materials and Methods The test is based on applying the same reagent for tests where the number of samples is 30 or less. The test for TSH was conducted 9 times from July $1^{st}$ 2015 to July $10^{th}$ 2015. The test for FT4 was conducted 4 times from June $18^{th}$ 2015 to June $22^{nd}$, 2015. Standard Solution No.2 (0.153 uU/mL) and No.5 (4.96 uU/mL) was selected as the two-point standards for the TSH test, and Standard Solution No.3 (0.777 ng/dL) and No.4 (2.044 ng/dL) was selected as the two-point standards for the FT4 test. 38 test samples were subject to correlation analysis. Results For TSH, the result of the normal test shows ranges of 0.20 ~ 0.37 uU/mL for Control1, 0.53 ~ 0.71 uU/mL for Control2, and 6.77 ~ 7.94uU/mL for Control3, while the result of two-point calibration curve test shows ranges of 0.18 ~ 0.27 uU/mL for Control1, 0.53 ~ 0.71 uU/mL for Control2, and 7.30 ~ 8.52 uU/mL for Control3. For FT4, the result of the normal test shows ranges of 0.85 ~ 0.94 ng/dL for Control1 and 4.23 ~ 4.57 ng/dL for Control2, while the result of two-point calibration curve test shows ranges of 0.61 ~ 0.75 ng/dL for Control1 and 3.88 ~ 5.71 ng/dL for Control2. For TSH, the CV% of the normal test for Control1, Control2 and Control3 are 10.5, 3.3 and 3.6 respectively, while the CV% of the two-point calibration curve test for Control1 and Control1 are 12.4, 8.2 and 5.1 respectively. The result shows an outstanding correlation of TSH: y = 0.9985x - 0.0459 $R^2=0.9986$. For FT4, the CV% of the normal test for Control1 and Control2 are 0.70 and 0.71 respectively, while the CV% of the two-point calibration curve test for Control1 and Control1 are 8.7 and 16.2 respectively. The result shows an outstanding correlation of FT4: y = 1.2674x - 0.1133 $R^2=0.9824$. Conclusion The two-point calibration curve can be efficiently applied for TSH in cases where the number of test samples is not large, since the number of samples to be re-tested increases when the result is abnormal from the calibration curve. The two-point calibration curve test should not be applied for FT4 where the results do not consistently comply with the quality assessment range. Depending on how the two-point calibration curve is applied, up to 5 test tubes can be conserved per test, and the reduced time for reagent dispensing is anticipated to have a positive effect on the TAT (turnaround time).
Kim, Woon-Su;Lee, Se-Young;Park, Hye-Sun;Baik, Woon-Kee;Lee, Jun-Heon;Seo, Seong-Won
Korean Journal of Poultry Science
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v.37
no.3
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pp.275-282
/
2010
Chicken is an important livestock as a valuable biomedical model as well as food for human, and there is a strong rationale for improving our understanding on metabolism and physiology of this organism. The first draft of chicken genome assembly was released in 2004, which enables elaboration on the linkage between genetic and metabolic traits of chicken. The objectives of this study were thus to reconstruct metabolic pathway of the chicken genome and to construct a chicken specific pathway genome database (PGDB). We developed a comprehensive genome database for chicken by integrating all the known annotations for chicken genes and proteins using a pipeline written in Perl. Based on the comprehensive genome annotations, metabolic pathways of the chicken genome were reconstructed using the PathoLogic algorithm in Pathway Tools software. We identified a total of 212 metabolic pathways, 2,709 enzymes, 71 transporters, 1,698 enzymatic reactions, 8 transport reactions, and 1,360 compounds in the current chicken genome build, Gallus_gallus-2.1. Comparative metabolic analysis with the human, mouse and cattle genomes revealed that core metabolic pathways are highly conserved in the chicken genome. It was indicated the quality of assembly and annotations of the chicken genome need to be improved and more researches are required for improving our understanding on function of genes and metabolic pathways of avian species. We conclude that the chicken PGDB is useful for studies on avian and chicken metabolism and provides a platform for comparative genomic and metabolic analysis of animal biology and biomedicine.
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