• 한국균학회소식:학술대회논문집
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• 2015.05a
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• pp.59-59
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• 2015

Cryptococcus neoformans causes life-threatening meningoencephalitis in humans, but the treatment of cryptococcosis remains challenging. To develop novel therapeutic targets and approaches, signaling cascades controlling pathogenicity of C. neoformans have been extensively studied but the underlying biological regulatory circuits remain elusive, particularly due to the presence of an evolutionarily divergent set of transcription factors (TFs) in this basidiomycetous fungus. In this study, we constructed a high-quality of 322 signature-tagged gene deletion strains for 155 putative TF genes, which were previously predicted using the DNA-binding domain TF database (http://www.transcriptionfactor.org/). We tested in vivo and in vitro phenotypic traits under 32 distinct growth conditions using 322 TF gene deletion strains. At least one phenotypic trait was exhibited by 145 out of 155 TF mutants (93%) and approximately 85% of the TFs (132/155) have been functionally characterized for the first time in this study. Through high-coverage phenome analysis, we discovered myriad novel TFs that play critical roles in growth, differentiation, virulence-factor (melanin, capsule, and urease) formation, stress responses, antifungal drug resistance, and virulence. Large-scale virulence and infectivity assays in insect (Galleria mellonella) and mouse host models identified 34 novel TFs that are critical for pathogenicity. The genotypic and phenotypic data for each TF are available in the C. neoformans TF phenome database (http://tf.cryptococcus.org). In conclusion, our phenome-based functional analysis of the C. neoformans TF mutant library provides key insights into transcriptional networks of basidiomycetous fungi and ubiquitous human fungal pathogens.

• Economic and Environmental Geology
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• v.28 no.6
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• pp.635-646
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• 1995

Mineralized zone in the Dongyang talc deposits occurs on the lowest dolomite member of the Hyangsanri Dolomite belonging to the Ogcheon Supergroup. Ore bodies are emplaced as pipe-like body along the axis of minor folds plunging $40^{\circ}$ to the west developed in these dolomite layers. Amphibolite and chlorite schist are found along the upper or lower contact of all ore bodies (Kim et al., 1963; Park and Kim, 1966). Following the recrystallization and silicification of dolomite, tremolite and tabular and leafy talc(I) of the earlier stage formed, and microcrystalline talc(II) formed in the later stage. Talc(l) and tremolite formed by the reaction between dolomite and the fluid. Whereas talc (II) formed by the reaction between dolomite and fluid, or by the reaction between early formed tremolite and fluid. During the early stage of mineralization, the fluid was the $H_2O-CO_2$ system dominant in $CO_2$, In the later stage, the composition of the fluid changed to $H_2O-NaCl-CO_2$system, and finally to the $H_2O-NaCl$ system. The pressure and temperature conditions of the formation of tremolite associated with talc(I) were 1,640~2,530 bar, and $440{\sim}480^{\circ}C$, respectively. The pressure and temperature condition of talc(II) ore formation was 1,400~2,200 bar, and $360{\sim}390^{\circ}C$, respectively. These conditions are much lower than the metamorphic pressure and temperature of the rocks from the Munjuri Formation located about 5 km to the noJ:th of Dongyang talc deposit ${\delta}^{13}C$ and ${\delta}^{18}O$ values of dolomite which is the host rock of the talc ore deposit are 2.9~5.7‰ (PDB), and -7.4~l6.8‰ (PDB), respectively. These values are little higher than those from the Cambro-Ordovician limestones of the Taebaeksan region, but belong to the range of the unaltered sedimentary dolomite. ${\delta}^{18}O$and ${\delta}D$ values of the talc from Dongyang deposit are 8.6~15.8‰ (vs SMOW), and -65~-90‰ (vs SMOW), respectively, belonging to the range of magmatic origin. These values are quite different from those measured in the metamorphic rocks of Munjuri and Kyemyungsan Formation. ${\delta}^{34}S$ value of anhydrite is 22.4‰ (CDT), which is much lower than ${\delta}^{34}S$ (30‰ vs COT) of sulfate of early Paleozoic period, and indicates the possibility of the addition of magmatic sulfur to the system. Talc ores show the textures of weak foliation and well developed crenulation cleavages. Talc ore deposit in the area is concluded as hydrothermal replacement deposit formed before the latest phase of the deformations that Ogcheon Belt has undergone.

• Horticultural Science & Technology
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• v.33 no.3
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• pp.409-419
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• 2015

This study was conducted to establish an efficient screening method for watermelon plants resistant to Fusarium wilt (FW), which is caused by Fusarium oxysporum f. sp. niveum (Fon). An HA isolate was prepared from a wilted watermelon plant in Haman-gun and identified as F. oxysporum f. sp. niveum based on morphological characteristics, molecular analyses of ITS (internal transcribed spacer) and TEF (translation elongation factor $1{\alpha}$) sequences, and host specificity on cucurbits including watermelon, melon, oriental melon, and cucumber. The assay for disease response of watermelon differentials indicated that the HA isolate was race 0. Among seven liquid media tested, the highest amount of Fon spores was produced from V8-juice broth, which was selected as a medium for mass production of Fon. The disease assay for 21 watermelon and 11 watermelon-rootstock cultivars demonstrated that 20 watermelon cultivars except for 'Soknoranggul' were susceptible; 'Soknoranggul' was moderately resistant. All the tested rootstock cultivars were highly resistant to the HA isolate. The evaluation of disease development depending on various conditions suggested that an efficient screening method for FW resistance in watermelon plants is to dip the roots of 10-day-old seedlings in spore suspension of $1.0{\times}10^5-1.0{\times}10^6conidia{\cdot}mL^{-1}$ for 30 min., to transplant the seedlings to plastic pots with a fertilized soil, and then to cultivate the plants at $25^{\circ}C$ for 3 weeks.

• Korean Journal of Microbiology
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• v.51 no.3
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• pp.256-262
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• 2015

In order to study the effect of the receptor protein (SeaR), which is isolated from Saccharopolyspora erythraea, we introduced the SeaR gene to Streptomyces virginiae as host strains. An effective transformation procedure for S. virginiae was established based on transconjugation by Escherichia coli ET12567/pUZ8002 with a ${\varphi}C31$-derived integration vector, pSET152, which contained int, oriT, attP, and $ermEp^{\ast}$ (erythromycin promotor). Therefore, the pEV615 was introduced into S. virginiae by conjugation and integrated at the attB locus in the chromosome of the recipients by the ${\varphi}C31$ integrase (int) function. Transformants of S. virginiae containing the SeaR gene were confirmed by PCR and transcriptional expression of the SeaR gene in the transformants was analyzed by RT-PCR, respectively. And, we examined the production time of virginiamycins in the culture media of both the transformants and the wild type. The production time of virginiamycins in the wild type and transformants was the same. When 100 ng/ml of synthetic $VB-C_6$ was added to the state of 6 or 8 hour cultivation of wild type and transformants, respectively, the virginiamycins production was induced, meaning that the virginiamycins production in the wild type was detected 2 h early than transformants. From these results, SeaR expression was also affected to virginiamycins production in transformants derived from S. virginiae. In this study, we showed that the SeaR protein worked as a repressor in transformants.

• Korean journal of applied entomology
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• v.47 no.4
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• pp.447-456
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• 2008

Five effective strains against tobacco cutworm, Spodoptera litura (Lepidoptera: Noctuidae), Steinernema carpocapsae (GSN1), Steinernema sp. (GSNUS-10), Steinernema sp. (GSNUS-14), Heterorhabditis bacteriophora Hamyang (HbH), and Heterorhabditis sp. (GSNUH-1) were selected among 14 isolates of Korean entomopathogenic nematode in laboratory tests. $LC_{50}$ values of above five strains against tobacco cutworm were various by different nematode strains and developmental stages of tobacco cutworm. $LC_{50}$ value of S. carpocapsae (GSN1) was the lowest by $4.0{\sim}8.3$ infective juveniles (Ijs) and 2nd instars of tobacco cutworm was most susceptible. Pathogenicity of five effective strains against tobacco cutworm depends on nematode strain, concentration, and application times. The most effective strain was determined as S. carpocapsae (GSN1). Two or three times of applications were effective regardless of nematode strain, or concentration. Efficacy of S. carpocapsae (GSN1), Steinernema (GSNUS-10), Steinernema (GSNUS-14), and Heterorhabditis (GSNUH-1) was variable depending on nematode strain, concentration, application times, and host variety. S. carpocapsae (GSN1) was the most effective and inoculation of 100,000 infective juveniles per m2 (720,000 Ijs/7.2 $m^2=1{\times}10^9$ Ijs/ha) resulted in higher efficacy. Three times of application of nematodes led to higher control efficacy than one or two applications. Efficacy of nematodes was higher on Chinese cabbage than cabbage or kale.

• Korean Journal of Environment and Ecology
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• v.28 no.5
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• pp.550-558
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• 2014

This study was conducted to investigate the distribution and the population dynamics of Hipparchia autonoe by using a line transect and Mark-Release-Recapture (MRR) at the Mt. Halla in Jeju Island. The results showed that H. autonoe was found from 1,500 m above the sea level. Total 1,493 H. autonoe with 978 males and 515 females were captured and released in the MRR study site. Among them, 518 individuals including 284 males and 234 females were recaptured. The average survival time was 2.31 days with 2.14 days for males and 3.47 days for females, indicating longer survival time in case of females than males. The daily population size of males estimated in the MRR study site was maintained about 1,000 individuals in July and gradually decreased less than 200 in August. The number of females showed peak at 335 individuals on July 24, and gradually decreased less than 120 in August. Thus, female population was 1/3 of males. The average travel distance of male and female H. autonoe were $116.8{\pm}191.9m$ and $118.4{\pm}161.5m$, respectively, indicating almost same between sexes. H. autonoe in the Mt. Halla formed single population group in the wide meadow around the Baekrokdam Lake. The highest population density of H. autonoe was occurred in the restored area from damages, where host plants such as the sheep's fescue or the food plant are abundant by artificial restoration efforts.

• Korean Journal of Plant Taxonomy
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• v.46 no.2
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• pp.256-266
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• 2016

A total of 13,000 individuals of Dendrobium moniliforme (L.) Sw. artificially propagated in laboratories and greenhouses were restored in their natural habitat of Bogildo Island, Wandogun, in the southern part of Korea in June of 2013. The growing conditions of the individuals were monitored for two years. The parental individuals for the restoration were obtained from a wild population in southern Korea, from which seeds were produced via artificial crossings. These seeds were germinated and cultivated in growing media and two-year-old plants were then grown in greenhouse beds. The genetic diversity among the propagated individuals was confirmed by examining DNA sequences of five regions of the chloroplast genome and the nuclear ITS region. The diversity values were as high as the average values of natural populations. All propagated individuals were transplanted into two different sites on Bogildo by research teams with local residents and national park rangers. After restoration, we counted and measured the surviving individuals, vegetative propagated stems, and growth rates in June of both 2014 and 2015. There was no human interference, and 97% of the individuals survived. The number of propagules increased by 227% in two years. In contrast, the average length of the stems decreased during the period. In addition, different survival and propagation rates were recorded depending on the host plants and the restored sites. The shaded sides of rock cliffs and the bark of Quercus salicina showed the best propagation rates, followed by the bark of Camellia japonica. A few individuals of D. moniliforme successfully flowered, pollinated, and fruited after restoration. Overall, our monitoring data over two years indicate that the restored individuals were well adapted and vigorously propagated at the restored sites. In order to prevent human disturbance of the restored sites, a CCTV monitoring system powered by a solar panel was installed after the restoration. In addition, a human surveillance system is operated by national park rangers with local residents.

• Applied Biological Chemistry
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• v.35 no.3
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• pp.179-185
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• 1992

Rhizobium meliloti populated in five Korean pasture soils were characterized by symbiotic effectiveness and intrinsic antibiotic resistance using whole-soil inoculum and 11 antibiotics, respectively. Most probable number (MPN) of naturalized rhizobia counted with alfalfa Vernal[Medicago sativa (L.)] as a host ranged $1.7{\times}10^2\;cells/g$. soil(Chunghyo, Kyeongiu)-$1.0{\times}10^5\;cells/g$. soil(Gampo, Kyeongiu) and ended to be positively associated with soil pH. On the whole, the effectiveness of population as compared to TAL mix inoculum (TAL 380+TAL 1372+TAL 1373) was very low. Nevertheless, there were two highly effective strains, YCK 539 and YCK 542, which were not inferior to TAL 1372, from Ogpo, Dalseong among the total of 30 of 6 isolates per each soil. As long as mean $N_2$ fixing ability of each soil isolate, the isolates from Hyeongog, Kyeonju were outstanding and the rest were in order of Ogpo, Dalseong>Chunghyo, Kyeongju>Hwaweon, Dalseong>Gampo, Kyeongiu. Isolates as a whole were resistant to erythromycin(67崙), nalidixic acid(77%), and streptomycin sulfate(8051), which had the concentration of $100\;{\mu}g/ml$, $160\;{\mu}g/ml$, and $10\;{\mu}g/ml$, respectively and divided into 14 patterns of resistance. Association between resistances in each soil was not clear. And there was no relationship of resistance pattern to effectiveness. The best effective strain YCKa 542 exclusively fell into No. X pattern having resistance to erythromycin, nalidixic acid, and neomycin sulfate.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.36 no.5
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• pp.366-374
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• 2010

Introduction: This study evaluated the capability of silk fibroin (SF) and recombinant human bone morphogenetic protein-2 loaded SF (SF-BMP) as a bone defect replacement matrix when grafted in a calvarial bone defect of rats in vivo. Materials and Methods: A total 70 calvarial critical size defects (5.0 mm in diameter) made on 35 adult female Sprague-Dawley rats were used in this study. The defects were transplanted with (1) rhBMP-2 loaded silk fibroin graft (SF-BMP: 0.8+$10\;{\mu}g$), (2) Silk fibroin (SF: $10\;{\mu}g$), and (3) no graft material (Raw). The samples were evaluated with soft x-rays, alkaline phosphatase activity, calcium/phosphate quantification, histological and histomorphometric analysis at postoperative 4 and 8 weeks. Results: The SF-BMP group ($48.86{\pm}14.92%$) had a significantly higher mean percentage bone area than the SF group ($24.96{\pm}11.01%$) at postoperative 4 weeks.(P<0.05) In addition, the SF-BMP group ($40.01{\pm}12.43%$) had a higher % bone area at postoperative 8 weeks than the SF group ($33.26{\pm}5.15%$). The mean ratio of gray scale levels to the host bone showed that the SF-BMP group ($0.67{\pm}0.08$) had a higher mean ratio level than the SF group ($0.61{\pm}0.09$) at postoperative 8 weeks. These differences were not statistically significant.(P=0.168 and P=0.243, respectively) The ratio of the calcium and phosphate contents of the SF-BMP ($0.93{\pm}0.22$) group was lower than that of the SF ($1.90{\pm}1.42$) group at postoperative 4 weeks. However, the SF-BMP group ($0.75{\pm}0.31$) had a higher Ca/$PO_4$ ratio than the SF ($0.68{\pm}0.04$) at postoperative 8 weeks. These differences were not statistically significant.(P=0.126 and P=0.627, respectively) For the bone-specific alkaline phosphatase (ALP) activity, which is recognized as a reliable indicator of the osteoblast function, the SF-BMP ($23.71{\pm}8.60\;U/L$) groups had a significantly higher value than the SF group ($12.65{\pm}6.47\;U/L$) at postoperative 4 weeks.(P<0.05) At postoperative 8 weeks, the SF-BMP ($21.65{\pm}10.02\;U/L$) group had a lower bone-specific ALP activity than the SF group ($16.72{\pm}7.35\;U/L$). This difference was not statistically significant.(P=0.263) For the histological evaluation, the SF-BMP group revealed less inflammation, lower foreign body reactions and higher bone healing than the SF group at postoperative 4 and 8 weeks. The SF group revealed more foreign body reactions at postoperative 4 weeks. However, this immunogenic reaction decreased and the remnant of grafted material was observed at postoperative 8 weeks. For histomorphometric analysis, the SF-BMP group had a significantly longer bone length to total length ratio than those of the SF group at postoperative 4 and 8 weeks.(P<0.05) Conclusion: The rhBMP-2 loaded silk fibroin graft revealed fewer immunoreactions and inflammation as well as more new bone formation than the pure silk fibroin graft. Therefore, silk fibroin may be a candidate scaffold for tissue engineered bone regeneration.

• Tuberculosis and Respiratory Diseases
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• v.48 no.2
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• pp.155-165
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• 2000

Background: The phagolysosomal function of alveolar macrophage against M. tuberculosis infection is influenced by Nramp1, which is encoded by the NRAMP1 gene. There are several genetic polymorphisms in NRAMP1, and these polymorphisms affect the innate host resistance through the defect in production and function of Nramp1. To investigate this relationship, the NRAMP1 genetic polymorphism in patients with primary tuberculous pleurisy was determined. Methods: Fifty-six primary tuberculous pleurisy patient, who were diagnosed by pleural biopsy, were designated to the pleurisy group and 45 healthy adults were designated to the healthy control group. Three genetic polymorphisms of NRAMP1, such as a single point mutation in intron 4(469+14G/C, INT4), a nonconservative single-base substitution at codon 543 that changes aspartic acid to asparagine(D543N) and a TGTG deletion in the 3' untranslated region(1729+55delI4, 3'UTR), were determined. Polymerase chain reaction(PCR) and polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP) were used. Results: The frequencies of mutant genotypes of INT4 and 3'UTR were significantly high in pleurisy group(p=0.001, p=0.023). But the frequencies of D543N were not significantly different between the two groups(p=0.079). The odds ratios, which are a comparison with wild genotype for determining mutant genotypes, were 8. 022(95% confidence interval=2.422-26.572) for INT4 and 5.733(95% confidence interval = 1.137~28.916) for 3'UTR ; these were statistically significant But the ratio for D543N was not significant In the combined analysis of the INT4 and 3'UTR polymorphisms, the odds ratios were 6.000(95% confidence interval = 1.461~24.640) for GC/++ genotype and 14.000(95% confidence interval=1.610~121.754) for GC/+del when compared with GG/++ homozygotes ; these were statistically significant. Conclusion: Among the NRAMP1 genetic polymorphisms, a single point mutation in intron 4(469+14G/C, INT4) and a TGTG deletion in the 3' untranslated region(1729+55del4, 3'UTR) were closely related to the primary tuberculous pleurisy.