• 미생물학회지
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• 제24권3호
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• pp.251-262
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• 1986

High-molecular-weight ${\beta}-glucosidase$ (EC 3.2.1.21) was purified from the culture filtrate of Trichoderma koningii through a four-step procedure including chromatography on Bio-Gel P-150, DEAE-Sephadex A-50 and SP-Sephadex C-50; and chromatofocusing on Polybuffer exchanger PBE 94. The molecular weight of the enzyme was determined to be about 101,000 by SDS-polyacrylamide gel electrophoreses, and the isoelectric point was estimated to be 4.96 by analytical isoelectric focusing. The temperature optimum for activity was about $55^{\circ}C$, and the pH optimumwas 3.5. The enzyme was considerably thermostable, for no loss of activity was observed when the enzyme was preincubated at $60^{\circ}C$ for 5h. Km values for cellobiose, gentiobiose, sophorose, salicin and $p-nitrophenyl-{\betha}-D-glucoside$ were 99.2, 14.7, 7.09, 3.15 and 0.70 mM, respectively, which indicates that the enzyme has much higher affinity towards $p-nitrophenyl-{\betha}-D-glucoside$ than towards the other substrates, especially cellobiose. Substrate inhibition by $p-nitrophenyl-{\betha}-D-glucoside$ and salicin was observed at the conecntrations exceeding 5mM. Gluconolactone was a powerful inhibitor against the action of the enzyme on $p-nitrophenyl-{\betha}-D-glucoside\;(K_i\;37.9\;{\mu}M)$, wherease glucose was much less effective ($K_i$ 1.95 mM). Inhibition was of the competitive type in each case. Transglucosylation activity was detected shen the readtion products formed from $p-nitrophenyl-{\betha}-D-glucoside$ by the enzyme were analysed using high-performance liquid chromatography.

• Journal of Microbiology and Biotechnology
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• 제16권1호
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• pp.145-148
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• 2006

A new functional lipolytic enzyme (AT4) has recently been found from Agrobacterium tumefaciens C58 Cereon using a genome-wide approach. The enzyme has some sequence similarity to E. coli acetyl hydrolase, Emericella nidulans lipase, Moraxella sp. lipase, Acinetobacter lwoffii esterase, and Streptomyces hygroscopicus acetyl hydrolase. However, the sequence similarities are very low (less than $25\%$), suggesting that it is a new lipase/esterase enzyme. ill the present study, intact cell of the A. tumefaciens strain was shown to have lipolytic activity on a tributyrin-LB plate. The AT4 gene was then expressed at a high level in E. coli BL21 (DE3) cells and the enzyme was purified simply by Ni-NTA column chromatography. The purified enzyme showed hydrolytic activity toward p-nitrophenyl caproate, but not toward olive oil, suggesting that the AT4 enzyme was a typical esterase rather than lipase. AT4 esterase had a maximum hydrolytic activity at $45^{\circ}C$ and pH 8.0, when p-nitrophenyl caproate was used as a substrate. It was relatively stable up to $40^{\circ}C$ and at pH 5.0-9.0. Calcium ion and EDT A did not affect the activity and thermal stability of the enzyme. As for substrate specificity, AT4 enzyme could rapidly hydrolyze acetyl and butyl groups from p-nitrophenyl esters and 1-naphthyl esters. In addition, it also released acetyl residues from acetylated glucose and xylose substrates. Therefore, this new esterase enzyme might be used as a biocatalyst in acetylation and deacetylation reactions performed in the fine chemical industry.

• Journal of Microbiology and Biotechnology
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• 제10권5호
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• pp.612-619
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• 2000

To investigate the biochemical properties of V. mimicus metalloprotease, whose gene was isolated previously from Vibrio mimicus ATCC33653, overexpression and purification were attempted. The 1.9 kb of open reading frame was amplified by PCR from pVMC193 plasmid which ligated the VMC gene with pUC19 and introduced into Escherichia coli BL21 (DE3) using the overexpression vector, pET22b (+). The overexpressed metalloprotease (VMC) was purified with Ni-NTA column chromatography and characterized with various protease inhibitors, pHs, temperatures, and substrates. The purified VMC showed the proteolytic activity against gelatin, soluble and insoluble collagens, and synthetic peptides. Unlike the observations made with all metalloproteases originated from other Vibrio sp., the VMC did not hydrolyze the casein. The proteolytic activity was critically decreased when the VMC was treated with metal chelating reagents, such as EDTA, 2,2-bipyridine, and 1, 10-phenanthroline. In particular, the 71 kDa VMC exhibited the hemagglutinating activity against human erythrocyte. As the purified VMC was treated with $CuCl_2$ and $NiCl_2$ for the chemical modification of metal binding, the proteolytic activity and hemagglutinating activity were profoundly influenced. The multialignment analysis made on the reported Vibrio metalloproteases showed the difference of amino acid sequence similarity between the two distinctive classes of Vibrio metalloproteases.

• The Plant Pathology Journal
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• 제19권3호
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• pp.129-132
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• 2003

Fusarium wilt of cucumber caused by Fusarium oxy-sporum f. sp. cucumerinum is a serious vascular disease worldwide. Biological control of Fusarium wilt in several crops has been accomplished by introducing non-pathogenic Fusarium sup. and other biocontrol agents in soil or in infection courts. In this study, quantitative models were used to determine the biocontrol efficacy of inundatively applied antagonist formulations and the length of their effectiveness in controlling Fusarium wilt of cucumber. Quantitative model of the form [Y=L (1${-exp}^{-kx}$)] best described the relationship between disease incidence (Y, %) and inoculum density (X) of isolates F51 and F55. Isolate F51 was selected as a more virulent isolate based on the extent of its effectiveness in causing the wilt disease. The degree of disease control (Xi/X) obtained with the density of the biocontrol agent (Z), was described by the model [Xi/X=A (1${-exp}^{-cz}$)]. The zeolite-based antagonist formulation amended with chitosan (ZAC) was better at lower rates of application and peaked at around 5 g/ kg of the potting medium, whereas the peat-based antagonist formulation (PA) peaked at around 10 g/kg of the potting medium. ZAC formulation provided significantly better suppression of Fusarium wilt as described by the curvilinear relationship of the type Y= a+bX+c$X^2$, where Y represents percent disease incidence and X represents sustaining effect of the biocontrol agent.

• 한국수산과학회지
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• 제39권spc1호
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• pp.189-197
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• 2006

To elucidate the organic matter sources in soft-bottom macrozoobenthos at Tongyeong, Korea, macrozoobenthos and their potential food sources were sampled in May 2004, and their C and N stable isotope compositions were analyzed. In total we collected 128 macrozoobenthic species, which ranged from 465 to 3,775 individuals/$m^2$(average 2,108 ind.$m^2$) in density and from 47.8 to 539.9 gwwt/$m^2$(average 366.0 gwwt/$m^2$) in biomass. Cluster and multi-dimensional scaling analyses indicated that the macrozoobenthic community was divided into two distinct groups. Coastal inner stations, where commercial fish cages were established, were dominated by Amphioplus ancistrotus, Scoletoma longifoila and Tharyx sp., whereas open sea stations were dominated by Chaetozone spinosa, Scoletoma longifolia and Capitella capitata. ${\delta}^{13}C$ values of sedimentary organic matter showed a distinct gradient in the range of -18.4 to $-15.2\%_{\circ}$, with a declining trend from the coastal inner stations to open sea stations. This probably reflects the settling rate of organic wastes such as feces and pellets from fish cages near the coastal inner stations. The macrozoobenthos showed a broad ${\delta}^{13}C$ range from -19.5 to $-8.9\%_{\circ}$ at the coastal inner stations, and a relatively narrow range from -21.3 to $-12.9\%_{\circ}$ at the open sea stations. ${\delta}^{13}C$ values of macrozoobenthos paralleled those of sedimentary organic matter, Our isotope results suggested that macrozoobenthos near the coastal inner stations used organic wastes derived from fish cages along with phytoplankton, whereas the macrozoobenthos near the open sea stations used organic wasters derived mainly from phytoplankton.

• 한국수산과학회지
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• 제39권spc1호
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• pp.132-141
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• 2006

Artificial structures have been designed as pilot structures to promote the creation and restoration of tidal flats. However, little information is available as to whether such artificial construction affects the macrobenthic community structure. We monitored the variation of the macrobenthic community structure and species composition near natural and artificial structures (seaweed and a timber fence) on the tidal flats near the Iwon Dike, Korea. In total, 137 macrobenthic species were recorded during this study, predominantly crustaceans (47%), polychaetes (18%), and molluscs (27%). Polychaetes comprised over 50% of the total density, followed by gastropods (38%) and crustaceans (11%). Macrobenthic species composition in the artificial and natural areas, was initially similar, but it differed after 7 months. The gastropod Umbonium thomasi, the most dominant species, was present at both sites in the first month after the start of the experiment, but disappeared at the artificial sites within 7 months, suggesting disturbance by the environmental factors. The number of species and diversity (H') varied significantly within sites at the beginning of the experiment, but no difference was observed after 7 months. Multivariate analysis (multidimensional scaling) revealed significant differences in community structure between the artificial and the natural areas from 7 months after the start of the experiment, except from 18 to 21 months. The community structures were mainly influenced by U. thomasi. Community structure at the artificial sites was affected by environmental variables, such as carbon, COD/IL sulfide, loss of ignition, kurtosis and silt, which changed over time. We observed no significant correlations between environmental variables and the dominant species, except in the case of Spio sp. and Macrophthalmus dilatatus, suggesting that the biological interactions and temporary disturbances such as typhoon, as well as the effects of artificial structures may also be important regulating factors in this system.

• Korean Journal of Acupuncture
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• 제21권2호
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• pp.177-192
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• 2004

Objectives : This study is designed to investigate the pathology, acupuncture and moxibustion therapy applicable of the dementia through literature of oriental medicine. Results : The findings of this study are as follows ; 1. The cause of dementia is internal damage of seven emotions(七情內傷), deficiency of the kidney essence(腎精不足), retention of dampness in the mental(痰迷心竅). 2. The acupuncture meridians used often for dementia are $Tok-maek(GV),\;Chok-t'aeyang\;Pang-gwang-Ky\check{o}ng(BL),\;Chok-yangm\check{o}yng\;Wi-Ky\check{o}ng(ST),\;Su-gwor\check{u}m\;Shimp'o-Ky\check{o}ng(PC),\;Su-so\check{u}m\;Shim-Ky\check{o}ng(HT)$. 3. The spots on the body suiTable for acupuncture used for therapy of the dementia are 98 points and extra points are 6 points, total therapeutic acupunture points are 104 points. And acupunture points are frequently used Shinmun(神門-HT7, 24), Paek'oe(百會-GV20, 19), Chok-samni(足三里-ST36, 18), Sugu(水溝-GV26, 17), P'ungnyung(豊隆-ST40, 16), Taech'u(大椎-GV14, 14), $Sam\check{u}mgyo(三陰交-SP6,\;14)$, Shimsu(心兪-BL15, 14), $Taen\check{u}ng(大陵-PC7,\;11)$, Nogung(勞宮-PC8, 11), Sosang(少商-LU11, 10), Kansa(間使-PC5, 10), Naegwan(內關-PC6, 10). 4. The acupoints used often on moxibustion are Paek'oe(百會-GV20), Chang-gang(長强-GV1), Taech'u(大椎-GV14), Kansa(間使-PC5). Conclusion : It is recommended that further study of many sided investigations in the future.

• 한국식물병리학회지
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• 제14권5호
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• pp.452-457
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• 1998

Thirty-eight isolates of Phytophthora sp. caused rots on roots and basal stems were collected from five flowering plants from 1992 to 1997 at eight cultivation areas in Korea. All the isolates were identified as P. nicotianae based on following characteristics. The fungus produced markedly papillate, not caducous and ovoid to spherical sporangia, abundant chlamydospores, and small oospores with amphigynous antheridia only when paired with either A1 or A2 mating type. All isolates grew well at 35$^{\circ}C$ and showed distinct arachnoid colony patterns on CMA and PDA. Sizes of sporangia and chlamydospores of five representative isolates from each plant averaged 43-52$\times$30-38 ${\mu}{\textrm}{m}$ and 28 ~34 ${\mu}{\textrm}{m}$. Mating type of the isolates was either A1 or A2, and oogonia and oospores were measured as 28~31 ${\mu}{\textrm}{m}$ and 21~25 ${\mu}{\textrm}{m}$. PCR-RFLP analysis of rDNA of the five isolates resulted that restriction band patterns of the small subunit and ITS regions were identical to a perilla isolate of P. nicotianae, but distinct from P. cactorum and P. capsici. Cross inoculation tests showed that the five isolates had pathogenicity to lily, christmas cactus, anthurium, baby's breath and carnation with different degrees. However, each isolate showed stronger pathogenicity to its corresponding original host than others. Among five lily cultivars Georgia and Quririna were more susceptible than Napoli and others. This is first report of Phytophthora root and stem rot of lily, Christmas cactus, anthurium, baby's breath and monochoria in Korea.

• The Journal of Korean Physical Therapy
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• 제21권2호
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• pp.109-116
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• 2009

Purpose: $17\beta$-estradiol is the most active endogenous estrogen, which is related to favorable changes in the plasma lipid profile, to relaxation of the coronary vessels, and to a decrease in platelet aggregation and vascular smooth muscle cell migration. However, although the beneficial effect of estrogens on plasma lipoproteins (ie, lowering low-density lipoprotein and increasing high-density lipoprotein cholesterol) contributes to cardiovascular protection, it does not fully account for the protective effect, particularly in the application of physical therapy, including low frequency electrical stimulation. Methods: The aim of this study was to demonstrate the inhibition of stressors, such as endothelin-1 (ET-1), serotonin (5-hydroxytryptamine, 5-HT), prostaglandin $F2\alpha$ ($PGF2\alpha$), and a protein kinase C (PKC) activator 12-deoxyphorbol 13-isobutyrate (DPB), induced isometric tension by $17\beta$-estradiol in vascular smooth muscle strips, respectively. In addition, the effects of low frequency electrical stimulation at the meridian points (CV-3, -4, Ki-12, SP-6, LR-3, BL-25, -28, -32, -52) on the indirect antihypertensive effect were examined by monitoring the changes in the serum $17\beta$-estradiol concentration in healthy volunteers. Results: Isometric tension analysis showed that the responses of inhibited tension by $17\beta$-estradiol were similar to the same stressors in rat aortic smooth muscle strips. Furthermore, although the continued amplitude modulation (AM) type of electrical stimulation was not increased significantly by electrical stimulation, the current of the frequency modulation (FM) type of low frequency electrical stimulation increased the serum $17\beta$-estradiol concentration in normal volunteers. Conclusion: These results, in part, suggest that $17\beta$-estradiol has the capacity to supress stressor-induced muscle tension, and electrical stimulation, particularly current of the FM type, has a modulatory effect on the sex steroid hormones, particularly $17\beta$-estradiol, in healthy volunteers.

• 한국어병학회지
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• 제22권1호
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• pp.15-21
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• 2009

White spot syndrome virus (WSSV) is a strong causative agent for high mortality in cultured and wild shrimps. From this study, the WSSV prevalence in marine organisms around shrimp farm as well as live feed-mediated transmission of WSSV to farmed shrimps were investigated. Based on nested-PCR method, WSSV was detected in wide array of marine organisms including Perinereis aibuhitensis (81.3% of prevalence rate, 13/16), Enedrias fangi (100%, 16/16), Ruditapes philippinarum (20%, 2/10), crab larvae (100%, 10/10), copepoda (30%, 3/10), Periophthalmus modestus (50%, 5/10), Pachygrapsus crassipes (10%, 1/10), Helice tridens (20%, 2/10) and Neomysis sp. (70%, 7/10). On the other hand, WSSV was not detected in Bullacta exarata, Uca arcuata, and Reishia clavigera. The percent prevalence of WSSV in wild shrimps, Fenneropenaeus chinensis was only 6%, but markedly increased up to 56% after a feeding trial using polychaete worms for one month, indicating that the live feed is one of significant carriers of WSSV to shrimps under practical farming conditions.