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Use of Human Serum Albumin Fusion Tags for Recombinant Protein Secretory Expression in the Methylotrophic Yeast Hansenula polymorpha (메탄올 자화효모 Hansenula polymorpha에서의 재조합 단백질 분비발현을 위한 인체 혈청 알부민 융합단편의 활용)

  • Song, Ji-Hye;Hwang, Dong Hyeon;Oh, Doo-Byoung;Rhee, Sang Ki;Kwon, Ohsuk
    • Microbiology and Biotechnology Letters
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    • v.41 no.1
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    • pp.17-25
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    • 2013
  • The thermotolerant methylotrophic yeast Hansenula polymorpha is an attractive model organism for various fundamental studies, such as the genetic control of enzymes involved in methanol metabolism, peroxisome biogenesis, nitrate assimilation, and resistance to heavy metals and oxidative stresses. In addition, H. polymorpha has been highlighted as a promising recombinant protein expression host, especially due to the availability of strong and tightly regulatable promoters. In this study, we investigated the possibility of employing human serum albumin (HSA) as the fusion tag for the secretory expression of heterologous proteins in H. polymorpha. A set of four expression cassettes, which contained the methanol oxidase (MOX) promoter, translational HSA fusion tag, and the terminator of MOX, were constructed. The expression cassettes were also designed to contain sequences for accessory elements including His8-tag, $2{\times}(Gly_4Ser_1)$ linkers, tobacco etch virus protease recognition sites (Tev), multi-cloning sites, and strep-tags. To determine the effects of the size of the HSA fusion tag on the secretory expression of the target protein, each cassette contained the HSA gene fragment truncated at a specific position based on its domain structure. By using the Green fluorescence protein gene as the reporter, the properties of each expression cassette were compared in various conditions. Our results suggest that the translational HSA fusion tag is an efficient tool for the secretory expression of recombinant proteins in H. polymorpha.

Residue analysis of penicillines in livestock and marine products (국내 유통 축·수산물 중 페니실린계 동물용의약품에 대한 잔류실태조사)

  • Song, Ji-Young;Hu, Soo-Jung;Joo, Hyun-Jin;Kim, Mi-Ok;Hwang, Joung-Boon;Han, Yoon-Jung;Kwon, Yu-Jihn;Kang, Shin-Jung;Cho, Dae-Hyun
    • Analytical Science and Technology
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    • v.25 no.4
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    • pp.257-264
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    • 2012
  • Penicillins belong to the ${\beta}$-lactam class of antibiotics, and are frequently used in human and veterinary medicine. Despite the positive effects of these drugs, improper use of penicillins poses a potential health risk to consumers. This study has been undertaken to determinate multi-residues of penicillins, including amoxicillin, ampicillin, oxacillin, bezylpenicillin, cloxacillin, dicloxacillin, and nafcillin, using liquid chromatographic tandem mass spectrometer (LC-MS/MS). The developed method was validated for specificity, precision, recovery, and linearity in livestock and marine products. The analytes were extracted with 80% acetonitrile and clean-up by a single reversed-phase solid-phase extraction step. Six penicillins presented recoveries higher than 76% with the exception of Amoxicillin. Relative standard deviations (RSDs) were not more than 10%. The method was applied to 225 real samples. Benzylpenicillin was detected in 12 livestock products and 7 marine products. Amoxicillin, ampicillin, cloxacilllin, dicloxacillin, nafcillin and oxacillin were not detected. The detected levels were 0.001~0.009 mg/kg in livestock products excluding eggs and milk. In marine products, the detected levels were under 0.03 mg/kg. They were under the MRL levels. As monitoring results, it is identified to be safe but it is considered that safety management of antibiotics should continue by monitoring.

Modified QuEChERS Multi-Residue Analysis Method for 61 pesticides in Fruits using with HPLC and GC-ECD/NPD (HPLC 및 GC-ECD/NPD를 이용한 과일 중 61종 농약의 QuEChERS 전처리 다성분 분석법 개발)

  • Lee, Ju-Young;Hong, Su-Myeong;Kim, Taek-Kyum;Min, Zaw Win;Kim, Yang-Hyeon;Song, Kyung-Ae;Kwon, Hye-Yong;Lee, Hee-Dong;Im, Geon-Jae;Kim, Doo-Ho;Kim, Jang-Eok
    • The Korean Journal of Pesticide Science
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    • v.16 no.3
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    • pp.242-256
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    • 2012
  • QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) method has been a lot of research for pesticide analysis, because it is very simple and fast. However, this method requires high sensitivity instrument such as LC-MS/MS because of the use of small sample volume and many impurities compared to the conventional method. So, QuEChERS method needs to be modified for using with HPLC and GC-ECD/NPD. The aim of this work was to study the application of the QuEChERS method as well as its modification for the extraction and preconcentration of 5 groups of 61 pesticides from 4 fruits prior to their determination by HPLC-PDA, GC-ECD/NPD, and LC-MS/MS. The method was validated using spiking levels at 0.1 mg/kg (or 0.01 mg/kg) in apple, grapes, pear and persimmon. The average recovery by QuEChERS AOAC Official 2007. 01 version using the LC-MS/MS varied from 71.1127.4% for 61 pesticides. The average recovery rates using modified QuEChERS varied from 70.9~126% for 61 pesticides by HPLC-PDA and GC-ECD/NPD. The results satisfied the criteria of multiple pesticide residue analysis, setting 70~130% for recovery rates and below 30% for CV.

A Study on the Applicability of Soilremediation Technology for Contaminated Sediment in Agro-livestock Reservoir (농축산저수지 오염퇴적토의 토양정화기술에 대한 적용성 연구)

  • Jung, Jaeyun;Chang, Yoonyoung
    • Journal of Environmental Impact Assessment
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    • v.29 no.3
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    • pp.157-181
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    • 2020
  • Sediments from rivers, lakes and marine ports serve as end points for pollutants discharged into the water, and at the same time serve as sources of pollutants that are continuously released into the water. Until now, the contaminated sediments have been landfilled or dumped at sea. Landfilling, however, was expensive and dumping at sea was completely banned due to the London Convention. Therefore, this study applied contaminated sedimentation soil of 'Royal Palace Livestock Complex' as soil purification method. Soil remediation methods were applied to pretreatment, composting, soil washing, electrokinetics, and thermal desorption by selecting overseas application cases and domestically applicable application technologies. As a result of surveying the site for pollutant characteristics, Disolved Oxigen (DO), Suspended Solid (SS), Chemical Oxygen Demand (COD), Total Nitrogen (TN), and Total Phosphorus (TP) exceeded the discharged water quality standard, and especially SS, COD, TN, and TP exceeded the standard several tens to several hundred times. Soil showed high concentrations of copper and zinc, which promote the growth of pig feed, and cadmium exceeded 1 standard of Soil Environment Conservation Act. In the pretreatment technology, hydrocyclone was used for particle size separation, and the fine soil was separated by more than 80%. Composting was performed on organic and Total Petroleum Hydrocarbon (TPH) contaminated soils. TPH was treated within the standard of concern, and E. coli was analyzed to be high in organic matter, and the fertilizer specification was satisfied by applying the optimum composting conditions at 70℃, but the organic matter content was lower than the fertilizer specification. As a result of continuous washing test, Cd has 5 levels of residual material in fine soil. Cu and Zn were mostly composed of ion exchange properties (stage 1), carbonates (stage 2), and iron / manganese oxides (stage 3), which facilitate easy separation of contamination. As a result of applying acid dissolution and multi-stage washing step by step, hydrochloric acid, 1.0M, 1: 3, 200rpm, 60min was analyzed as the optimal washing factor. Most of the contaminated sediments were found to satisfy the Soil Environmental Conservation Act's standards. Therefore, as a result of the applicability test of this study, soil with high heavy metal contamination was used as aggregate by applying soil cleaning after pre-treatment. It was possible to verify that it was efficient to use organic and oil-contaminated soil as compost Maturity after exterminating contaminants and E. coli by applying composting.

Red Tide Detection through Image Fusion of GOCI and Landsat OLI (GOCI와 Landsat OLI 영상 융합을 통한 적조 탐지)

  • Shin, Jisun;Kim, Keunyong;Min, Jee-Eun;Ryu, Joo-Hyung
    • Korean Journal of Remote Sensing
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    • v.34 no.2_2
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    • pp.377-391
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    • 2018
  • In order to efficiently monitor red tide over a wide range, the need for red tide detection using remote sensing is increasing. However, the previous studies focus on the development of red tide detection algorithm for ocean colour sensor. In this study, we propose the use of multi-sensor to improve the inaccuracy for red tide detection and remote sensing data in coastal areas with high turbidity, which are pointed out as limitations of satellite-based red tide monitoring. The study area were selected based on the red tide information provided by National Institute of Fisheries Science, and spatial fusion and spectral-based fusion were attempted using GOCI image as ocean colour sensor and Landsat OLI image as terrestrial sensor. Through spatial fusion of the two images, both the red tide of the coastal area and the outer sea areas, where the quality of Landsat OLI image was low, which were impossible to observe in GOCI images, showed improved detection results. As a result of spectral-based fusion performed by feature-level and rawdata-level, there was no significant difference in red tide distribution patterns derived from the two methods. However, in the feature-level method, the red tide area tends to overestimated as spatial resolution of the image low. As a result of pixel segmentation by linear spectral unmixing method, the difference in the red tide area was found to increase as the number of pixels with low red tide ratio increased. For rawdata-level, Gram-Schmidt sharpening method estimated a somewhat larger area than PC spectral sharpening method, but no significant difference was observed. In this study, it is shown that coastal red tide with high turbidity as well as outer sea areas can be detected through spatial fusion of ocean colour and terrestrial sensor. Also, by presenting various spectral-based fusion methods, more accurate red tide area estimation method is suggested. It is expected that this result will provide more precise detection of red tide around the Korean peninsula and accurate red tide area information needed to determine countermeasure to effectively control red tide.

Effect of Implant Types and Bone Resorption on the Fatigue Life and Fracture Characteristics of Dental Implants (임플란트 형태와 골흡수가 임플란트 피로 수명 및 파절 특성에 미치는 효과에 관한 연구)

  • Won, Ho-Yeon;Choi, Yu-Sung;Cho, In-Ho
    • Journal of Dental Rehabilitation and Applied Science
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    • v.26 no.2
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    • pp.121-143
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    • 2010
  • To investigate the effect of implant types and bone resorption on the fracture characteristics. 4 types of Osstem$^{(R)}$Implant were chosen and classified into external parallel, internal parallel, external taper, internal taper groups. Finite elements analysis was conducted with ANSYS Multi Physics software. Fatigue fracture test was performed by connecting the mold to the dynamic load fatigue testing machine with maximum load of 600N and minimum load of 60N. The entire fatigue test was performed with frequency of 14Hz and fractured specimens were observed with Hitachi S-3000 H scanning electron microscope. The results were as follows: 1. In the fatigue test of 2 mm exposed implants group, Tapered type and external connected type had higher fatigue life. 2. In the fatigue test of 4 mm exposed implants group, Parallel type and external connected types had higher fatigue life. 3. The fracture patterns of all 4 mm exposed implant system appeared transversely near the dead space of the fixture. With a exposing level of 2 mm, all internally connected implant systems were fractured transversely at the platform of fixture facing the abutment. but externally connected ones were fractured at the fillet of abutment body and hexa of fixture or near the dead space of the fixture. 4. Many fatigue striations were observed near the crack initiation and propagation sites. The cleavage with facet or dimple fractures appeared at the final fracture sites. 5. Effective stress of buccal site with compressive stress is higher than that of lingual site with tensile stress, and effective stress acting on the fixture is higher than that of the abutment screw. Also, maximum effective stress acting on the parallel type fixtures is higher. It is careful to use the internal type implant system in posterior area.

Targeted Therapies and Radiation for the Treatment of Head and Neck Cancer (두경부 암의 표적 지향적 방사선 치료)

  • Kim, Gwi-Eon
    • Radiation Oncology Journal
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    • v.22 no.2
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    • pp.77-90
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    • 2004
  • Purpose: The purpose of this review Is to provide an update on novel radiation treatments for head and neck cancer Recent Findings: Despite the remarkable advances In chemotherapy and radiotherapy techniques, the management of advanced head and neck cancer remains challenging. Epidermal growth factor receptor (EGFR) Is an appealing target for novel therapies In head and neck cancer because not only EGFR activation stimulates many important signaling pathways associated with cancer development and progression, and importantly, resistance to radiation. Furthermore, EGFR overexpression Is known to be portended for a worse outcome in patients with advanced head and neck cancer. Two categories of compounds designed to abrogate EGFR signaling, such as monoclonal antibodies (Cetuxlmab) and tyrosine kinase inhibitors (ZD1839 and 051-774) have been assessed and have been most extensively studied In preclinical models and clinical trials. Additional TKIs In clinical trials include a reversible agent, Cl-1033, which blocks activation of all erbB receptors. Encouraging preclinical data for head and neck cancers resulted In rapid translation Into the clinic. Results from Initial clinical trials show rather surprisingly that only minority of patients benefited from EGFR inhibition as monotherapy or In combination with chemotherapy. In this review, we begin with a brief summary of erbB- mediated signal transduction. Subsequently, we present data on prognostic-predictive value of erbB receptor expression in HNC followed by preclinlcal and clinical data on the role of EGFR antagonists alone or in combination with radiation In the treatment of HNC. Finally, we discuss the emerging thoughts on resistance to EGFR biockade and efforts In the development of multiple-targeted therapy for combination with chemotherapy or radiation. Current challenges for investigators are to determine (1 ) who will benefit from targeted agents and which agents are most appropriate to combine with radiation and/or chemotherapy, (2) how to sequence these agents with radiation and/or cytotoxlc compounds, (3) reliable markers for patient selection and verification of effective blockade of signaling in vivo, and (4) mechanisms behind intrinsic or acquired resistance to targeted agents to facilitate rational development of multi-targeted therapy, Other molecuiar-targeted approaches In head and neck cancer were briefly described, Including angloenesis Inhibitors, farnesyl transferase inhibitors, cell cycle regulators, and gene therapy Summary: Novel targeted theraples are highly appealing in advanced head and neck cancer, and the most premising strategy to use them Is a matter of intense Investigation.

A Study on the Distributional Characteristics of Unminable Manganese Nodule Area from the Investigation of Seafloor Photographs (해저면 영상 관찰을 통한 망간단괴 채광 장애지역 분포 특성 연구)

  • Kim, Hyun-Sub;Jung, Mee-Sook;Park, Cheong-Kee;Ko, Young-Tak
    • Geophysics and Geophysical Exploration
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    • v.10 no.3
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    • pp.173-182
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    • 2007
  • It is well known that manganese nodules enriched with valuable metals are abundantly distributed in the abyssal plain area in the Clarion-Clipperton (C-C) fracture zone of the northeast Pacific. Previous studies using deep-sea camera (DSC) system reported different observations about the relation of seafloor topographic change and nodule abundance, and they were sometimes contradictory. Moreover, proper foundation on the estimation of DSC underwater position, was not introduced clearly. The variability of the mining condition of manganese nodule according to seafloor topography was examined in the Korea Deep Ocean Study (KODOS) area, located in the C-C zone. In this paper, it is suggested that the utilization of deep towing system such as DSC is very useful approach to whom are interested in analysing the distributional characteristics of manganese nodule filed and in selecting promising minable area. To this purpose, nodule abundance and detailed bathymetry were acquired using deep-sea camera system and multi-beam echo sounder, respectively on the seamount free abyssal hill area of southern part ($132^{\circ}10'W$, $9^{\circ}45'N$) in KODOS regime. Some reasonable assumptions were introduced to enhance the accuracy of estimated DSC sampling position. The accuracy in the result of estimated underwater position was verified indirectly through the comparison of measured abundances on the crossing point of neighboring DSC tracks. From the recorded seafloor images, not only nodules and sediments but cracks and cliffs could be also found frequently. The positions of these probable unminable area were calculated by use of the recorded time being encountered with them from the seafloor images of DSC. The results suggest that the unminable areas are mostly distributed on the slope sides and hill tops, where nodule collector can not travel over.

A Review of Statistical Methods in the Korean Journal of Orthodontics and the American Journal of Orthodontics and Dentofacial Orthopedics (대한치과교정학회지(KJO)와 미국교정학회지(AJODO)에서 사용된 통계기법의 비교분석 및 고찰(1999-2003))

  • Lim, Hoi-Jeong
    • The korean journal of orthodontics
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    • v.34 no.5 s.106
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    • pp.371-379
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    • 2004
  • The purpose of this study was to investigate the changes and types of statistical methods used in the Korean Journal of Orthodontics (KJO) and the American Journal of Orthodontics and Dentofacial Orthopedics (AJODO) from )999 to 2003. The frequency of use, transitions, assumption check of statistical methods and types of advanced statistical methods were examined from each journal. The study consisted of 247 articles published in the KJO and randomly chosen 50 articles per year which were original articles and used statistical methods T-test, analysis of variance(ANOVA), correlation analysis, nonparametric analysis. regression analysis chi-square test. factor analysis, were the order of statistical methods most frequently used in the KJO, while t-test. ANOVA, nonparametric analysis, correlation analysis, regression analysis, chi-square test. factor analysis. were the order of statistical methods used in the AJODO The changes of statistical methods observed in the KJO were not significant $(X^2=17.4\;p=0.5881)$ but the changes observed in the AJODO was seen to be significant $(x^2=42.4,\;p=0.0397)$ Some of the studies examined had overlooked the assumptions of the statistical methods employed. Data investigation such as outlier should be performed before analysis and alternative statistical approaches are applied for a small sample size. Types of advanced statistical methods were factor analysis and discriminant analysis in the KJO and Intention-To-Treat (ITT) analysis in clinical trials through multi-center, survival analysis and Generalized Estimating Equations (GEE) in the AJODO. Appropriate analysis approaches and interpretations should be applied for the correlated and repeated measurements of the orthodontic data set.

Management Plan for Humanistic and Ecological Characteristics of Suweol Village Forest in Tongyoung (통영 수월숲의 인문학적 특성 및 생태적 특성을 고려한 관리방안)

  • Lim, Eui-Jea;Lee, Soo-Dong;Kim, Mi-Jeong
    • Korean Journal of Environment and Ecology
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    • v.27 no.1
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    • pp.85-98
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    • 2013
  • In order to propose effective conservation management plan, this study verified ecological characteristics, humanities and Social characteristics. The research site is private property which is owned belonging to the Kim's of Gimhae that have long history. The study site is more than a thousand years old and was created for protecting from typhoon. There held the religious ritual what is called Dongsinje until 1960s. There have been protected and managed by the villagers. As the results of analysis, the area of windbreak are $12,392.69m^2$. The windbreak is dominated old years deciduous broad-leaved tree such as Zelkova serrata, Celtis sinensis, Aphananthe aspera. Around there were farmlands(52.1%), urbanized area(26.3%), forest area(16.6%). The vegetation communities of windbreak were classified by considering the dominant species and current status of forest. The forest types are following as; A. aspera community(I) which is using less pressure, Platycarya strobilacea-Carpinus coreana-Z. serrata community(II). Z. serrata community(III) which is using high pressure, Z. serrata-A. aspera community(IV), Z. serrata community(V) which is damaged under canopy trees. The windbreak was in good condition whereas, there were concerns the some wrong status was being undermined such as the wrong forest restoration projects in the past, the trails that is penetrating inside the forest, building up education facilities. Therefore, in order to restore the value of windbreak what is so called Suwol forest, we should improve the problems of forest ecosystem such as wrong management, forest fragmentation by facilities and decline in forest by lack of growing the next generation trees. In addition, we should remove excessive resting facilities and lead to passive use of forest. to improve the way of wrong management, moreover, we should close off he trails that is penetrating inside the forest for improving fragmentation. We should restore vegetation restoration and fostering the next generation trees for forest ecosystem. In order to restore unique of histo-cultural and ecological forest landscape, the next generation trees should be grown up that is the dominant species in Suwol forest. Moreover, as a results of comparing the between good vegetation communities and damaged vegetation communities, it is necessary to complementary planting for demeged vegetation communities, therefore there needs to 10.8 under canopy trees, 79.7 shrubs.