• Journal of Microbiology and Biotechnology
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• 제27권9호
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• pp.1593-1601
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• 2017

Vibrio species are generally recognized as pathogens predominant in seafood along coastal areas. The food industry has sought to develop efficient microbial detection methods. Owing to the limits of conventional methods, this study aimed to establish a rapid identification method for Vibrio isolated from Korea, based on matrix-assisted laser-desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Four different preparation procedures were compared to determine the appropriate means to pretreat Vibrio species, using 17 isolates and five reference strains. Extended direct transfer and full formic acid extraction methods using bacterial colonies on agar plates revealed very low identification rates. Formic acid and trifluoroacetic acid (TFA) extractions using bacterial broth cultures were also performed. All Vibrio isolates and reference strains prepared by TFA extraction were successfully identified to the species level (17/22, 77.3%) and to the genus level (5/22, 22.7%). Thus, TFA extraction was considered the most appropriate method to pretreat Vibrio species for MALDI-TOF MS. The remaining 33 isolates and two reference strains were prepared by TFA extraction and analyzed by MALDI-TOF MS. Overall, 50 isolates were identified to the species level (40/50, 80%) and to the genus level (10/50, 20%). All isolates were identified as 43 V. alginolyticus, six V. parahaemolyticus, and one V. vulnificus species. V. alginolyticus and V. parahaemolyticus were isolated from fish offal (87.5% and 12.5%, respectively), seawater (91.3%, 8.7%), and shellfish (62.5%, 37.5%), whereas V. alginolyticus and V. vulnificus were isolated from sediment (90.9% and 9.1%, respectively). This study established a reliable system of MALDI-TOF MS preparation and analysis for Vibrio identification.

• Reproductive and Developmental Biology
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• 제29권2호
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• pp.83-91
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• 2005

Somatic cell nuclear transfer in cattle has limited efficiency in terms of production of live offspring due to high incidence of fetal failure after embryo transfer to recipients. Such low efficiency of cloning could possibly arise from abnormal and poorly developed placenta. In the present study the placental proteome in late pregnancy established from in vitro fertilization (IVF) and nuclear transfer (NT) was analysed. Proteome alternation was tested using two-dimensional polyacrylamide gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI- TOF). Comparing placenta from NT embryos to those from IVF counterparts, significant changes in expression level were found in 18 proteins. Of these proteins 12 were not expressed in NT placenta but expressed in IVF counterpart, whereas the expression of the other 6 proteins was limited only in NT placenta. Among these proteins, cytokeratin 8 and vimentin are considered to be involved in regulation of post-implantation development. In particular, cytokeratin 8 and vimentin may be used as makers for placental development during pregnancy because their expression levels changed considerably in NT placental tissue compared with its IVF counterpart. Data from 2-DE suggest that protein expression was disorientated in late pregnancy from NT, but this distortion was eliminated with progression of pregnancy. These findings demonstrate abnormal placental development during late pregnancy from NT and suggest that alterations of specific placental protein expression may be involved in abnormal function of placenta.

• 한국대기환경학회지
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• 제33권4호
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• pp.411-423
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• 2017

The spacial potential source contribution function (PSCF) method was utilized by considering topography and height of back trajectories based on the measurement of organic typo matter (OM), $NO_3{^-}$, $SO{_4}^{2-}$, and $NH_4{^+}$ at the Baegryungdo Super Site ($37^{\circ}57^{\prime}N$, $124^{\circ}37^{\prime}E$, 135 m a.s.l. (above sea level)) for three selected periods (i.e., January~April, May~August, and September~December) in 2013. The PSCF were calculated on the contributions of trans-boundary transport to the hourly mean concentrations using a high-resolution time-of-flight aerosol mass spectrometer (HR-ToF-AMS). The cluster analysis using back trajectories was performed to identify the major airflows to the sampling site. The upper atmosphere in the Tianjin area of China and the lower atmosphere in the western coast area of Korea can be the major source of trans-boundary pollution to the sampling site during January~April resulted from PSCF. The area in Lianyungang-city and Liaoning-sheng, China can be responsibile for the nitrogen related secondary compounds during May~August, and Shandong Peninsula in China is the major source area during September~December. In addition, relationships between the cluster analysis of back trajectories and PSCF were investigated for the statistically significance level for the source areas.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2003년도 생물공학의 동향(XII)
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• pp.40-44
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• 2003

Soil-borne infectious disease including Pythium aphanidermatum and Rhizoctonia solani causes severe damage to plants, such as cucumber. This soil-borne infectious disease was not controlled effectively by chemical pesticide. Since these diseases spread through the soil, chemical agents are usually ineffective. Instead, biological control, including antagonistic microbe can be used as a preferred control method. An efficient method was developed to select an antagonistic strain to be used as a biological control agent strain. In this new method, surface tension reduction potential of an isolate was included in the ‘decision factor’ in addition to the other factors, such as growth rate, and pathogen inhibition rate. Considering these 3 decision factors by a statistical method, an isolate from soil was selected and was identified as Bacillus sp. GB16. In the pot test, this strain showed the best performance among the isolated strains. The lowest disease incidence rate and fastest seed growth was observed when Bacillus sp. GB16 was used. Therefore this strain was considered as plant growth promoting rhizobacteria (PGPR). The action of surface tension reducing component was deduced as the enhancement of wetting, spreading, and residing of antagonistic strain in the rhizosphere. This result showed that new selection method was significantly effective in selecting the best antagonistic strain for biological control of soil-borne infectious plant pathogen. The antifungal substances against P. aphanidermatum and R. solani were partially purified from the culture filtrates of Bacillus sp. GB16. In this study, lipopeptide possessing antifungal activity was isolated from Bacillus sp. GB16 cultures by various purification procedures and was identified as a surfactin-like lipopeptide based on the Fourier transform infrared spectroscopy (FT-IR), nuclear magnetic resonance (NMR), high performance liquid chromatography mass spectroscopy (HPLC-MS), and quadrupole time-of-flight (Q-TOF) ESI-MS/MS data. The lipopeptide, named GB16-BS, completely inhibited the growth of Pythium aphanidermatum, Rhizoctonia solani, Penicillium sp., and Botrytis cineria at concentrations of 10 and 50 mg/L, respectively. A novel method to prevent the foaming and to provide oxygen was developed. During the production of surface active agent, such as lipopeptide (surfactin), large amount of foam was produced by aeration. This resulted in the carryover of cells to the outside of the fermentor, which leads to the significant loss of cells. Instead of using cell-toxic antifoaming agents, low amount of hydrogen peroxide was added. Catalase produced by cells converted hydrogen peroxide into oxygen and water. Also addition of corn oil as an oxygen vector as well as antifoaming agent was attempted. In addition, Ca-stearate, a metal soap, was added to enhance the antifoam activity of com oil. These methods could prevent the foaming significantly and maintained high dissolved oxygen in spite of lower aeration and agitation. Using these methods, high cell density, could be achieved with increased lipopeptide productivity. In conclusion to produce an effective biological control agent for soil-borne infectious disease, following strategies were attempted i) effective screening of antagonist by including surface tension as an important decision factor ii) identification of antifungal compound produced from the isolated strain iii) novel oxygenation by $H_2O_2-catalase$ with vegetable oil for antifungal lipopeptide production.

• 한국초지조사료학회지
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• 제31권2호
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• pp.99-106
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• 2011

프로테오믹스 기법을 이용하여 벼 고온 스트레스 관련 단백질을 분리 동정하기 위하여 $42^{\circ}C$에서 고온처리한 벼의 줄기로부터 단백질을 분리하였다. 분리한 단백질로부터 Rubisco 단백질을 제거하기 위해 15% PEG fractionation을 실시한 후 상등액 분획의 단백질을 이차원전기 영동한 후, CBB 염색을 통해 차별적 발현을 보이는 단백질을 분석하였다. 총 46개의 단백질 spot이 발현양에 변화를 보였으며, 그 중 24개의 단백질이 고온 스트레스에 의해 발현이 증가되었으며, 22개의 단백질이 감소하는 발현 양상을 나타내었다. 이들 단백질을 MALDI-TOF MS와 database를 통해 동정한 결과 에너지 대사관련 단백질, 산화 환원 관련 단백질 및 저분자량 small HSP 등, 10개의 단백질이 동정되었다. 이들 동정된 단백질들은 식물의 고온 스트레스에 대한 적응기작을 이해하는데 중요한 단서를 제공할 것이며, 특히 미토콘드리아 small HSP는 프로테옴 분석법에 의해 최초로 동정되었으며, 금후 내하고성 목초 분자육종에 활용될 수 있는 좋은 유전자로 판단된다.

• 원예과학기술지
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• 제34권6호
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• pp.924-939
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• 2016

The great economic losses caused by Cucumber mosaic virus (CMV) infection of peppers has led to the development of genetically modified (GM) CMV-resistant peppers. We developed virus-resistant pepper plants using Agrobacterium tumefaciens -mediated transformation. The expressed recombinant protein was purified using nickel-nitrilotriacetic acid resin and immunoaffinity chromatography, and purity was assessed by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Immunoblot analysis revealed the purified CMV coat protein (CMV-CP) had a molecular mass of 25 kDa. After in-gel digestion and desalting, the internal peptide fragments of CMV-CP were sequenced by matrix-assisted laser desorption/ionization-time of flight. Most GM pepper and Escherichia coli BL21 internal peptides had identical peptide sequences and contained 137 of 183 whole peptides in CMV-CP. A quantitative enzyme-linked immunosorbent assay was performed to detect CMV-resistant GM peppers. We also provide basic information about the expressed protein in GM peppers for further safety assessment. The contents of soluble protein and CMV-CP were measured in GM and control peppers cultivated in three different areas of Korea. Statistical significance in terms of cultivation areas, harvest times, generations, and plant tissue origin were determined based on a P value of 0.05. The highest amount of CMV-CP was detected at the seedling stage from plant grown in each region. T3 and T5 showed significantly different levels of CMV-CP from T4 in leaves in the whorl stage. No statistical differences were observed among GM peppers at different stages of maturity in any cultivation area. The results from this study contribute to the safety evaluation of newly designed CMV-resistant GM peppers and provide a standard against which to compare other virus-resistant GM peppers.

• 대한인간공학회지
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• 제28권1호
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• pp.37-51
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• 2009

The nuclear power industry in the world has recognized the importance of integrating non-technical and team skills training with the technical training given to its control room operators to reduce human errors since the Three Mile Island and Chernobyl accidents. The Nuclear power plant (NPP) industry in Korea has been also making efforts to reduce the human errors which largely have contributed to 120 nuclear reactor trips from the year 2001 to 2006. The Crew Resource Management (CRM) training was one of the efforts to reduce the human errors in the nuclear power industry. The CRM was developed as a response to new insights into the causes of aircraft accidents which followed from the introduction of flight recorders and cockpit voice recorders into modern jet aircraft. The CRM first became widely used in the commercial airline industry, but military aviation, shipboard crews, medical and surgical teams, offshore oil crews, and other high-consequence, high-risk, time-critical industry teams soon followed. This study aims to develop a CRM training program that helps to improve plant performance by reducing the number of reactor trips caused by the operators' errors in Korean NPP. The program is; firstly, based on the work we conducted to develop a human factors training from the applications to the Nuclear Power Plant; secondly, based on a number of guidelines from the current practicable literature; thirdly, focused on team skills, such as leadership, situational awareness, teamwork, and communication, which have been widely known to be critical for improving the operational performance and reducing human errors in Korean NPPs; lastly, similar to the event-based training approach that many researchers have applied in other domains: aircraft, medical operations, railroads, and offshore oilrigs. We conducted an experiment to test effectiveness of the CRM training program in a condition of simulated control room also. We found that the program made the operators' attitudes and behaviors be improved positively from the experimental results. The more implications of the finding were discussed further in detail.

• 한국식품저장유통학회지
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• 제30권2호
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• pp.235-246
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• 2023

Yellowball (Citrus hybrid cv. Yellowball ) is a new citrus hybrid between Haruka (C. tamurana × natsudaidai ) and Kiyomi (C. unshiu × sinensis) and is known to possess strong antioxidant activity. However, detailed information on the antioxidant components of its peel has not yet been reported. This study evaluated the antioxidant activity of the peel and identified the antioxidant components by fractionating a methanolic extract of Yellowball peels using liquid-liquid extraction with n-hexane, ethyl ether (ether), ethyl acetate (EA), butanol, and water. The phenolic contents and antioxidant activities of the n-hexane, ether, and EA fractions were higher than those of the other fractions, and these fractions were further separated by semi-preparative high-performance liquid chromatography (HPLC). Four antioxidant peaks, EA1, EA2, EA3, and He1, were isolated and analyzed using ultra-performance liquid chromatography-quadrupole-time- of-flight mass spectrometry (UPLC-Q-TOF MS). Sinapoyl glucoside and hesperidin were identified in EA2 and EA3, respectively, and a polymethoxylated flavone (PMF) complex (5-hydroxy-3,6,7,8,3',4'-hexamethoxyflavone, natsudaidain, tetrameth- oxyflavone, and tangeretin) was identified in He1. A compound in EA1 with m/z 223.0246 [M-H] could not be identified and was named unknown2. The antioxidant activity of unknown2 (IC₅₀=69.17 ㎍/mL) was similar to that of Trolox, which was noted as a major antioxidant in Yellowball peel. Further studies on the antioxidant capacity of Yellowball peel are required; however, these results provide a foundation for using Yellowball peel as an antioxidant.

• 한국가금학회:학술대회논문집
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• 한국가금학회 2005년도 제22차 정기총회 및 학술발표회
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• pp.66-67
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• 2005

배자 생식세포 발달에 관련된 메카니즘을 밝혀내기 위해서, 닭 배자 생식기에서 추출한 원시 생식세포의 단백질체 지도를 만들었다. 총 500 배자를 6일간 배양하여 배자 생식기를 획득했고, 7-10일 배양 후, 배양된 원시생식세포는 2차원 젤 전기 영동법에 의해 분할되어 졌다. 유의적 발현 수준을 나타낸 많은 단백질 스팟 들은 MALDI-TOP 와 LC-MS/MS에 의해 확인되었으며, 89개의 단백질 스팟 중에 50개의 mass spectra 들이 데이터베이스에서 조류 단백질과 일치함을 확인하였다. 본 실험에서 행한 단백질체 지도는 형질전환 연구와 생식세포 생물학 분야에 중요한 참고 문헌으로 가치를 가질수 있을 것이다.

• Journal of Korean Neurosurgical Society
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• 제62권4호
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• pp.398-404
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• 2019

Objective : Recently, three-dimensional (3D) printed models of the intracranial vascular have served as useful tools in simulation and training for cerebral aneurysm clipping surgery. Precise and realistic 3D printed aneurysm models may improve patients' understanding of the 3D cerebral aneurysm structure. Therefore, we created patient-specific 3D printed aneurysm models as an educational and clinical tool for patients undergoing aneurysm clipping surgery. Herein, we describe how these 3D models can be created and the effects of applying them for patient education purpose. Methods : Twenty patients with unruptured intracranial aneurysm were randomly divided into two groups. We explained and received informed consent from patients in whom 3D printed models-(group I) or computed tomography angiography-(group II) was used to explain aneurysm clipping surgery. The 3D printed intracranial aneurysm models were created based on time-of-flight magnetic resonance angiography using a 3D printer with acrylonitrile-butadiene-styrene resin as the model material. After describing the model to the patients, they completed a questionnaire about their understanding and satisfaction with aneurysm clipping surgery. Results : The 3D printed models were successfully made, and they precisely replicated the actual intracranial aneurysm structure of the corresponding patients. The use of the 3D model was associated with a higher understanding and satisfaction of preoperative patient education and consultation. On a 5-point Likert scale, the average level of understanding was scored as 4.7 (range, 3.0-5.0) in group I. In group II, the average response was 2.5 (range, 2.0-3.0). Conclusion : The 3D printed models were accurate and useful for understanding the intracranial aneurysm structure. In this study, 3D printed intracranial aneurysm models were proven to be helpful in preoperative patient consultation.