• Title/Summary/Keyword: Positive Column

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Immunological Characterization of Full and Truncated Recombinant Clones of ompH(D:4) Obtained from Pasteurella multocida (D:4) in Korea

  • Kim, Young-Hwan;Cheong, Ki-Young;Shin, Woo-Seok;Hong, Sung-Youl;Woo, Hee-Jong;Kwon, Moo-Sik
    • Journal of Microbiology and Biotechnology
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    • v.16 no.10
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    • pp.1529-1536
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    • 2006
  • We cloned a gene of ompH(D:4) from pigs infected with P. multocida D:4 in Korea [16]. The gene is composed of 1,026 nucleotides coding 342 amino acids (aa) with a signal peptide of 20 aa (GenBank accession number AY603962). In this study, we analyzed the ability of the ompH(D:4) to induce protective immunity against a wild-type challenge in mice. To determine appropriate epitope(s) of the gene, one full and three different types of truncated genes of the ompH(D:4) were constructed by PCR using pET32a or pRSET B as vectors. They were named ompH(D:4)-F (1,026 bp [1-1026] encoding 342 aa), ompH(D:4)-t1 (693 bp [55-747] encoding 231 aa), ompH(D:4)-t2 (561 bp [187-747] encoding 187 aa), and ompH(D:4)-t3 (540 bp [487-1026] encoding 180 aa), respectively. The genes were successfully expressed in Escherichia coli BL21(DE3). Their gene products, polypeptides, OmpH(D:4)-F, -t1, -t2, and -t3, were purified individually using nickel-nitrilotriacetic acid (Ni-NTA) affinity column chromatography. Their $M_rs$ were determined to be 54.6, 29, 24, and 23.2 kDa, respectively, using SDS-PAGE. Antisera against the four kinds of polypeptides were generated in mice for protective immunity analyses. Some $50{\mu}g$ of the four kinds of polypeptides were individually provided intraperitoneally with mice (n=20) as immunogens. The titer of post-immunized antiserum revealed that it grew remarkably compared with pre-antiserum. The lethal dose of the wild-type pathogen was determined at $10{\mu}l$ of live P. multocida D:4 through direct intraperitoneal (IP) injection, into post-immune mice (n=5, three times). Some thirty days later, the lethal dose ($10{\mu}l$) of live pathogen was challenged into the immunized mouse groups [OmpH(D:4)-F, -t1, -t2, and -t3; n=20 each, two times] as well as positive and negative control groups. As compared within samples, the OmpH(D:4)-F-immunized groups showed lower immune ability than the OmpH(D:4)-t1, -t2, and -t3. The results show that the truncated-OmpH(D:4)-t1, -t2, and -t3 can be used for an effective vaccine candidate against swine atrophic rhinitis caused by pathogenic P. multocida (D:4) isolated in Korea.

Characteristics and Pathways of the Somatosensory Evoked Field Potentials in the Rat (흰쥐에서 체감각유발장전위의 기록부위별 특성과 경로분석)

  • Shin, Hyun Chul;Park, Yong Gou;Lee, Bae Hwan;Ryou, Jae Wook;Zhao, Chun Zhi;Chung, Sang Sup
    • Journal of Korean Neurosurgical Society
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    • v.30 no.7
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    • pp.831-841
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    • 2001
  • Objective : Somatosensory evoked potentials(SSEPs) have been used widely both experimentally and clinically to monitor the function of central nervous system and peripheral nervous system. Studies of SSEPs have reported the various recording techniques and patterns of SSEP. The previous SSEP studies used scalp recording electrodes, showed mean vector potentials which included relatively constant brainstem potentials(far-field potentials) and unstable thalamocortical pathway potentials(near-field potentials). Even in invasive SSEP recording methods, thalamocortical potentials were variable according to the kinds, depths, and distance of two electrodes. So they were regarded improper method for monitoring of upper level of brainstem. The present study was conducted to investigate the characteristics of somatosensory evoked field potentials(SSEFPs) of the cerebral cortex that evoked by hindlimb stimulation using ball electrode and the pathways of SSEFP by recording the potentials simultaneously in the cortex, VPL nucleus of thalamus, and nucleus gracilis. Methods : In the first experiment, a specially designed recording electrode was inserted into the cerebral cortex perpendicular to the cortical surface in order to recording the constant cortical field potentials and SSEFPs mapped from different areas of somatosensory cortex were analyzed. In the second experiment, SSEPs were recorded in the ipsilateral nucleus gracilis, the contralateral ventroposterolateral thalamic nucleus(VPL), and the cerebral cortex along the conduction pathway of somatosensory information. Results : In the first experiment, we could constantly obtain the SSEFPs in cerebral cortex following the transcutaneous electrical stimulation of the hind limb, and it revealed that the first large positive and following negative waves were largest at the 2mm posterior and 2mm lateral to the bregma in the contralateral somatosensory cortex. The second experiment showed that the SSEPs were conducted by way of posterior column somatosensory pathway and thalamocortical pathway and that specific patterns of the SSEPs were recorded from the nucleus gracilis, VPL, and cerebral cortex. Conclusion : The specially designed recording electrode was found to be very useful in recording the localized SSEFPs and the transcutaneous electrical stimulation using ball electrode was effective in evoking SSEPs. The characteristic shapes, latencies, and conduction velocities of each potentials are expected to be used the fundamental data for the future study of brain functions, including the hydrocephalus model, middle cerebral artery ischemia model, and so forth.

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Marine Environmental Characteristics around the Test Phase of Offshore Wind Farm in the Southwestern Coast of Yellow Sea (서남해 해상풍력 실증단지 해역의 해양 환경 특성)

  • Seo, Jinsung;Maeng, Junho;Lim, Eunpyo;Jin, Seungjoo;Kim, Hyunmin;Kim, Taeyun
    • Journal of Environmental Impact Assessment
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    • v.28 no.5
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    • pp.457-470
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    • 2019
  • This study aims to present the characteristics of marine physics, water quality, and sediment quality around offshore wind farm near the Gochang and Buan sea areas through the analysis of monitoring data. The relationships between suspended solid and wave height as well as suspended solid and flow velocity were analyzed. We found that Correlation Coefficient values of 0.61~0.69 between wave height and suspended solid, and suspended solid concentration reaches 75 mg/L or higher when wave height of more than 1m occurred in the spring (1 month). The water quality index (WQI) was used to identify the status of the water quality in the study area. Most of the measured points were rated first grade (very good). The variation of chlorophyll-a was relatively hight compared to the other criteria, indicating that it is a major factor affecting the quality index. In the sediment column, all heavy metals were detected below the Threshold Effects Level(TEL), and ignition loss and grain size show a positive correlation.

Isolation and Identification of Antimicrobial Active Substances from Rhodiola sachlinensis (홍경천(Rhodiola sachlinensis)에서 항균성 물질의 분리 및 동정)

  • 심창주;이규희;정재홍;이상덕;김영호;오만진
    • Food Science and Preservation
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    • v.11 no.1
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    • pp.63-70
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    • 2004
  • The antimicrobial substances from Rhodiola sachalinensis were extncted, isolated and identified. The highest level of antimicrobial activity and its yield were obtained in methanol extract. The minimum inhibition concentrations of the methanol extract were 500 $\mu\textrm{g}$mL on agar plate and 100 $\mu\textrm{g}$mL in broth media for four gram positive and four gram negative microbials. The methanol extract was fractionated by n-hexane, chloroform, ethyl ether, ethyl acetate, and butanol, orderly. The separate was developed on the TLC plate with different solvent system ratio of chloroform and methanol. Nine substances were isolated from chloroform and methanol mixture(9:1, v/v). Among them, three isolates showed antimicrobial activity. Three substances separated by HPLC were identified by GC/MS(EI) spectrum and $^1$H, /sup13/C-NMR spectrum. They were gallic acid, (-)-epicatechin and kaempferol. The antimicrobial activities of each substances were shown gallic acid, (-)-epicatechin, kaempferol orderly.

Recombinant Production and Antimicrobial Activity of an Antimicrobial Model Peptide (Uu-ilys-CF) Derived from Spoon Worm Lysozyme, Uu-ilys (개불 라이소자임 유래 항균성 모델 펩타이드(Uu-ilys-CF)의 재조합 단백질 생산 및 항균 활성)

  • Oh, Hye Young;Go, Hye-Jin;Park, Nam Gyu
    • Journal of Life Science
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    • v.31 no.1
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    • pp.83-89
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    • 2021
  • Uu-ilys, an i-type lysozyme from spoon worm (Urechis unicinctus), is an innate immune factor that plays an important role in the defense against pathogens. It also possesses non-enzymatic antibacterial activity. Thus, there is a possibility to develop an antimicrobial model peptide from Uu-ilys. In this study, we report the design, production, and antibacterial activity of an Uu-ilys analog that exhibits antibacterial activity. The Uu-ilys structure was fragmented according to its secondary structures to predict the regions with antimicrobial activity using antimicrobial peptide (AMP) prediction tools from different AMP databases. A peptide containing the C-terminal fragment was predicted to exert antimicrobial activity. The chosen fragment was designated as an Uu-ilys analog containing the C-terminal fragment, Uu-ilys-CF. To examine the possibility of developing an AMP using the sequence of Uu-ilys-CF, recombinant fusion protein (TrxA-Uu-ilys-CF) was produced in an expression system that was heterologous. The produced fusion protein was cleaved after methionine leaving Uu-ilys-CF free from the fusion protein. This was then isolated through high performance liquid chromatography and reverse phase column, CapCell-Pak C18. The antibacterial activity of Uu-ilys-CF against different microbial strains (four gram-positive, six gram-negative, and one fungal strain) were assessed through the ultrasensitive radial diffusion assay (URDA). Among the bacterial strains tested, Salmonella enterica was the most susceptible. While the fungal strain tested was not susceptible to Uu-ilys-CF, broad spectrum antibacterial activity was observed.

Cephalometric study on head posture according to the Classification of Malocclusion (부정교합 분류에 따른 두경부 위치의 두부방사선 계측학적 연구)

  • Hwang, Chung-Ju;Kim, Suk-Hyun;Kil, Jae-Kyung
    • The korean journal of orthodontics
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    • v.27 no.2
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    • pp.221-230
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    • 1997
  • It has been known that head posture may influence directly and/or indirectly the growth and development of craniofacial morphology and can also be influenced by the funtional demand of physiologic activity. It was reported that facial morphology has close relationships with hyoid bone position and head posture. In many previous studies, Natural Head Posture(NHP) was guided, and also it was shown that NHP has high degree of reproducibility. Otherwise, There was few study about the relationship of head posture, with routine cephalometric film which is used for clinical orthodontic purpose. In this study, according to the Wits and ANB of initial cephalometric film which was taken with vertical pendulum as representative of true vertical reference line. We classified the subjects which is comprised of 60 adult female patients into Class I, II, III (Cl I, II, III)and we tried to find out the correlation of head posture and hyoid bone position according to the classification of malocclusion. As a result of our research, we found the followigs. 1. In comparison of vertical position of hyoid bone relative to the cranial base. the position of hyoid bone of Cl III was lower than that of Cl II. 2. In comparison of anteriorpostes or position of hyoid bone, relative to the cervical column. The position of hyoid bone of Cl III was more anterior than that of a II 3. in comparison of vertical position of hyoid bone relative to mandible. There was no significant correlation aumoug the groups of malocclusion. 4. ANB and Wits showed no significant correlation with hyoid bone position. 5. The relative extension of head, which was noted in Cl II, showed negative with Sum, ANB. 6. In Cl II and Cl III, Post to Ant facial height showed positive correlation with NSL/VER.

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Development of Simultaneous Analytical Method of Veterinary Antibiotics in Manure using Liquid Chromatography Coupled with Tandem Mass Spectrometry (LC-MS/MS를 이용한 퇴비 및 액비 중 항생제 동시 분석법 개발)

  • Chung, Hyung Suk;Lee, Young Jun;Lee, Han Sol;Rahman, Md. Musfiqur;Kabir, Md. Humayun;Park, Byung-Jun;Kim, Jang-Eok;Shim, Jae-Han
    • Korean Journal of Environmental Agriculture
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    • v.36 no.3
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    • pp.201-210
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    • 2017
  • BACKGROUND: The current study was to monitor of 9 veterinary antibiotics (ceftiofur, clopidol, florfenicol, sulfamethazine, sulfamethoxazole, sulfathiazole, tetracycline, tiamulin, and tylosin) in manure using liquid chromatography-tandem mass spectrometry (LC-MS/MS) in positive and negative electrospray ionization mode. METHODS AND RESULTS: Sample preparation was carried out using Mcllvaine buffer and citrate salts to adjust the pH of the sample followed by purification with dispersive solid phase extraction (d-SPE). Separation of analytes during LC-MS/MS analysis was conducted using an Eclipse Plus $C_{18}$ column and the mobile phase was in gradient mode with, 0.1% formic acid and 5 mM ammonium formate in methanol (A) and 0.1% formic acid and 5 mM ammonium formate in distilled water (B). The linearity of the matrix-matched calibrations of all tested antibiotics was good, with $R^2$ determination coefficients ${\geq}0.9920$. The limit of detection (LOD) and quantifications (LOQ) were $0.1-67.0{\mu}g/kg$ and $0.4-200.0{\mu}g/kg$, respectively. Analysis of 13 solid and liquid manure samples taken from the Republic of Korea revealed concentrations less than $0.7{\mu}g/kg$ for tiamulin, $1497.6{\mu}g/kg$ for sulfamethazine. CONCLUSION: To monitor 9 veterinary antibiotics from manure samples in 13 provincial areas throughout the Republic of Korea, an analytical method was developed. The developed method was fully validated and successfully applied for monitoring various veterinary antibiotics in manure samples.

Bioactivities and Isolation of Functional Compounds from Decay-Resistant Hardwood Species (고내후성 활엽수종의 추출성분을 이용한 신기능성 물질의 분리 및 생리활성)

  • 배영수;이상용;오덕환;최돈하;김영균
    • Journal of Korea Foresty Energy
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    • v.19 no.2
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    • pp.93-101
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    • 2000
  • Wood of Robinia pseudoacacia and bark of Populus alba$\times$P. glandulosa, Fraxinus rhynchophylla and Ulmus davidiana var. japonica were collected and extracted with acetone-water(7:3, v/v) in glass jar to examine whether its bioactive compounds exist. The concentrated extracts were fractionated with hexane, chloroform, ethylacetate and water, and then freeze-dried for column chromatography and bioactive tests. The isolated compounds were sakuranetin-5-O-$\beta$-D-glucopyranoside from Populus alba $\times$Pl glandulosa, 4--ethyoxy-(+)-leucorobinetinidin frm R. pseudoacacia and fraxetion from F. rhynchophylla and were characterized by $^1H$ and$^{13}C $ NMR and positive FAB-MS. Decay-resistant activity was expressed by weight loss ratio and hyphae growth inhibition in the wood dust agar medium inoculated wood rot fungi. R. pseudoacacia showed best anti-decaying property in both test and its methanol untreated samples, indicating higher activity than methanol treated samples in hyphae grwoth test. In antioxidative test, $\alpha$-tocopherol, one of natural antioxidants, and BHT, one of synthetic antioxidants, were used as references to cmpare with the antioxidant activities of the extacted fractions. Ethylacetate fraction of F. rhynchophylla bark indicated the hightest activity in this test and all fractions of R. pseudiacacia extractives also indicated higher activities compared with the other fractions. In the isolated compounds, aesculetin isolated from F. rhynchophylla bark showed best activity and followed by robonetinidin from R. pseudoacaica.

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Rice Growth and Grain Quality in No-till and Organic Farming Paddy Field as Affected by Different Rice Cultivars (무경운 및 유기농 논에서 품종에 따른 벼 생육 및 미질 특성 구명)

  • Lee, Young-Han
    • Korean Journal of Soil Science and Fertilizer
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    • v.43 no.2
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    • pp.209-216
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    • 2010
  • This study was to find out optimum rice cultivars for organic farming in no-tillage paddy. A field research was conducted the yield, yield components, and qualities of thirty two Korean rice cultivars, and two Japanese rice cultivars. The column length of early rice plant was shorter in medium maturing rice cultivars compared to early, and medium-late maturing rice cultivars. Planthopper population per20-plant was lower in Ilpumbyeo, Dongjinbyeo, Hojinbyeo, Donganbyeo, and Sobeebyeo in that order while was higherin rice cultivars of early maturity. The disease severity of sheath blight was higher in Saechucheongbyeo>Chucheongbyeo>Namwonbyeo>Sangmibyeo in that order, on the other hand, that was lowerinHwaseongbyeo>Junambyeo>Saesangjubyeo=Hitomebore>Ilpumbyeo>Hwayeongbyeo in that order, respectively. The grain yield of Korean rice cultivars was significantly higher in 2.95 Mg $ha^{-1}$ for Hwasinbyeo, 2.91 Mg $ha^{-1}$ for Ilpumbyeo, 2.86 Mg $ha^{-1}$ for Sobeebyeo, 2.73 Mg $ha^{-1}$ for Naepungbyeo, and 2.70 Mg $ha^{-1}$ for Saegyehwabyeo compared with 1.38 Mg $ha^{-1}$ for Hwabongbyeo, 1.62 Mg $ha^{-1}$ for Hwayeongbyeo, and 1.78 Mg $ha^{-1}$ for Haepyeongbyeo of medium maturing rice cultivars, respectively. Toyo taste value of medium maturing rice cultivars tended to higher than that of early, and medium-late maturing rice cultivars. Ripening rate of rice cultivars was significantly positive correlated with Toyo taste value while negative correlated with protein content. According to principal component analysis, these results show that Sangmibyeo, Sangsanbyeo, and Odaebyeo for early maturity, Naepungbyeo, and Sobeebyeo for medium maturity, and Hwasinbyeo, Chucheongbyeo, and Sindongjinbyeo for medium-late maturity were optimum rice cultivars for organic farming in no-tillage paddy.

Dynamics of Phytoplankton and Zooplankton of a Shallow Eutrophic Lake (lake llgam) (수심이 얕은 부영양 인공호(일감호)의 동 ${\cdot}$ 식물플랑크톤 동태학)

  • Kim, Ho-Sub;Park, Je-Chul;Hwang, Soon-Jin
    • Korean Journal of Ecology and Environment
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    • v.36 no.3 s.104
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    • pp.286-294
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    • 2003
  • This study was attempted to understand seasonal dynamics of phyto- and zooplankton communities in shallow, eutrophic Lake llgam and to compare them with the PEG (Plankton Ecology Group) model. Seasonal succession pattern of phytoplankton community was similar to PEG model as Chlorophyceae and Baciliphyceae increase during spring and autumn fellowed by increase of Cyanophyceae. However, based on the cell density and biomass, a dominant phytoplankton community differed with PEG model: Cyanophyceae had been a dominant community throughout a year, except for ice-cover period during which Chlorophyceae was a dominant group. In spring, when ice melted and dissolved nutrients in water column increased, the increase of Chlorophyceae occurred: when nutrients (DIN and DIP) rapidly decreased, Cyanophyceae increase occurred. Microcystis, Oscillatoria, Lyngbya, Merismopedia were maior dominant species of Cyanophyceae and their cell density and/or biomass was the highest in October 2000 (12.9${\pm}$5.8${\times}10^5$ cells/ml, 3.5${\pm}$0.9${\times}10^3{\mu}gC/l$). Cyanophyceae biomass showed positive relationship with chlorophyll a ($r^2$ = 0.71,P< 0.001) and TP concentration ($r^2$ = 0.62, P< 0.001). Small-sized rotifers such as Keratella cochlearis, increased between March and May when Chlorophyceae increased. Both high standing crop of copepods and cladocerans, such as Diaphanosoma brachyrum and Bosmina longirostris occurred between June and September accompanied with the increase of Dinophyceae and Bacillariophyceae. There was no evidence that clear-water phase was caused by zooplankton grazing. The diversity and evenness index of phyto- and/or zooplankton increased with chlorophyll a concentration. These results suggest zooplankton grazing and limiting nutrient deficiency could lead to change of phytoplankton biomass, but not the phytoplankton community in Lake llgam.