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Development of the feedback resistant pheAFBR from E. coli and studies on its biochemical characteristics (E. coli 유래 pheA 유전자의 되먹임제어 저항성 돌연변이의 구축과 그 단백질의 생화학적 특성 연구)

  • Cao, Thinh-Phat;Lee, Sang-Hyun;Hong, KwangWon;Lee, Sung Haeng
    • Korean Journal of Microbiology
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    • v.52 no.3
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    • pp.278-285
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    • 2016
  • The bifunctional PheA protein, having chorismate mutase and prephenate dehydratase (CMPD) activities, is one of the key regulatory enzymes in the aromatic amino acid biosynthesis in Escherichia coli, and is negatively regulated by an end-product, phenyalanine. Therefore, PheA protein has been thought as useful for protein engineering to utilize mass production of essential amino acid phenylalanine. To obtain feedback resistant PheA protein against phenylalanine, we mutated by using random mutagenesis, extensively screened, and obtained $pheA^{FBR}$ gene encoding a feedback resistant PheA protein. The mutant PheA protein contains substitution of Leu to Phe at the position of 118, displaying that higher affinity (about $290{\mu}M$) for prephenate in comparison with that (about $850{\mu}M$) of wild type PheA protein. Kinetic analysis showed that the saturation curve of $PheA^{FBR}$ against phenyalanine is hyperbolic rather than that of $PheA^{WT}$, which is sigmoidal, indicating that the L118F mutant enzyme has no cooperative effects in prephenate binding in the presence of phenylalanine. In vitro enzymatic assay showed that the mutant protein exhibited increased activity by above 3.5 folds compared to the wild type enzyme. Moreover, L118F mutant protein appeared insensitive to feedback inhibition with keeping 40% of enzymatic activity even in the presence of 10 mM phenylalanine at which the activity of wild type $PheA^{WT}$ was not observed. The substitution of Leu to Phe in CMPD may induce significant conformational change for this enzyme to acquire feedback resistance to end-product of the pathway by modulating kinetic properties.

Preoperative Radiotherapy of Maxillary Sinus Cancer (상악동암에서 수술 전 방사선 치료의 효과)

  • Kim, Jae-Chul;Park, In-Kyu
    • Radiation Oncology Journal
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    • v.16 no.3
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    • pp.259-264
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    • 1998
  • Purpose : This study was to evaluate the effectiveness of preoperative radiotherapy in maxillary sinus cancer. Materials and Methods : A retrospective analysis was done for 42 patients with maxillary sinus cancer who were treated with radiation with or without surgery from April 1986 to September 1996. There were 27 male and 15 female patients. Patients' age ranged from 24 to 75 years (median 56 years). Stage distribution showed 2 in T2, 19 in T3, and 21 in T4 lesions The histologic type was squamous cell carcinoma in 38, undifferentiated carcinoma in 1, transitional cell carcinoma in 1, and adenoid cystic carcinoma in 2 patients. All patients were treated with radiation initially with a dosage range of 50.4-70.2 Gy (median 70.2 Gy) before further evaluation of remnant disease. Eleven patients were given induction chemotherapy (2cycles of 5-fluorouracil and cisplatin) concurrently with radiotherapy. Six to eight weeks after radiotherapy with or without chemotherapy computerized tomography (CT) of paranasal sinus was taken to evaluate remnant disease. If the CT finding showed remnant disease, a Caldwell-Luc procedure was done to get the specimen of suspicious lesions. A radical maxillectomy was done if the specimen was proven to contain malignancy. In contrast periodic follow-up examination was done without any radical surgery if the tissue showed only granulation tissue. Follow-up period ranged from 3 to 92 months with a median 16 months. Results : Nine (21.4$\%$) patients showed complete response (CR) and 33 patients (78.6$\%$) showed persistent disease (PER) to initial radiotherapy. Among the 9 CR patients, 7 patients had no evidence of disease (NED), 1 patient had local failure, and 1 patient had regional failure. Among 33 PER patients, salvage total maxillectomy was done in 10 patients, and the surgery was not feasible or refused in 23 patients. Following the salvage radical surgery, 2 patients were NED and 8 patients were PER status. Overall and disease- free survival rate at 5 years was 23.1$\%$ and 16.7$\%$, respectively. The only factors associated with the overall survival rate was the response to radiotherapy (P<0.01). Conclusion : The only factors associated with the overall survival rate was the response to radiotherapy. We could omit a radical mutilating surgery by preoperative irradiation in 7 of 42 patients (21.4$\%$) so as to preserve their facial integrity.

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Selective Algicidal Effects of a Newly Developed GreenTD against Red Tide Harmful Alga (GreenTD 물질을 이용한 유해 적조 발생 종의 선택적 살조능 평가)

  • Lee, Minji;Shin, Juyong;Kim, Jin Ho;Lim, Young Kyun;Cho, Hoon;Baek, Seung Ho
    • Korean Journal of Environmental Biology
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    • v.36 no.3
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    • pp.359-369
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    • 2018
  • Harmful algal blooms (HABs) are a serious problem for public health and fisheries industries, thus there exists a need to investigate the possible ways for effective control of HABs. In the present study, we investigated the algicidal effects of a newly developed GreenTD against the HABs (Chattonella marina, Heterosigma akashiwo, Cochlodinium polykriokides, and Heterocapsa circularisquama) and non-HABs (Chaetoceros simplex, Skeletonema sp. and Tetraselmis sp.), which is focused on the different population density and concentration gradients of algicidal substances. The time series viability of target alga was assessed based on the activity of Chl. a photosynthetic efficiency in terms of $F_v/F_m$, and in vivo fluorescence (FSU). Effective control of Raphidophyta, C. marina and H. akashiwo was achieved at a GreenTD concentration of $0.5{\mu}gL^{-1}$ and $0.2{\mu}gL^{-1}$, respectively, and regrowth of both the species was not observed even after 14 days. The inhibitory ratio of the dinoflagellate, C. polykriokides was more than 80% at $0.2{\mu}gL^{-1}$ of GreenTD. H. circularisquama was constantly affected in the presence of $0.2{\mu}gL^{-1}$ of GreenTD in the high- and low-population density experimental groups. On the other hand, diatoms, C. simplex, and Skeletonema sp. were not significantly affected even in the presence of $0.2{\mu}gL^{-1}$ of GreenTD and exhibited re-growth activity with the passage of incubation time. In particular, green alga Tetraselmis sp. remained unaffected even in the presence of the highest concentration of GreenTD ($1.0{\mu}gL^{-1}$), implying that non-HABs were not greatly influenced by the algicidal substances. As a result, the algicidal activity of GreenTD on the harmful and nonharmful algae was as follows: raphidophyte>dinoflagellates>diatoms>green alga. Consequently, our results indicate that inoculation of GreenTD substances into natural blooms at a threshold concentration ($0.2{\mu}gL^{-1}$) can maximize the algicidal activity against HABs species. If we consider the dilution and diffusion rate in the field application, it is hypothesized that GreenTD will demonstrate economic efficiency, thus leading to effective control against the target HABs in the closed bay.

Availability of Slurry Composting and Biofiltration for Cultivation of Cherry Tomato (방울 토마토 재배 시 퇴비단 여과 액비의 이용가능성)

  • Kim, Eun-Young;Park, Bong-Ju;Oh, Myung-Min
    • Journal of Bio-Environment Control
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    • v.22 no.4
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    • pp.385-391
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    • 2013
  • In this study, the availability of slurry composting and biofiltration (SCB) solution as an alternative for synthetic nutrient solution was determined by monitoring the growth, fruit yield, and fruit quality of cherry tomato (Solanum lycopersicum L. 'Unicon'). Treatments for nutrient solution were consist of SCB 1/2N, 1N, 2N, and commercial nutrient solution 1N (CNS 1N) based on nitrogen concentration (218.32 $mg{\cdot}L^{-1}$) of cherry tomato nutrient solution (control 1N). All nutrient solution including SCB solution (440~520 mL per day) was supplied to rock wool medium using a timer. After 31 days of transplanting, fresh and dry weights of shoots, leaf area, plant height, stem diameter, SPAD value and number of node were measured. After measuring growth characteristics of tomato plants, total fruit yield, ratio of marketable fruit yield, fruit weight, total soluble solids content, total acidity, total phenolic concentration, and antioxidant capacity were determined once a week for 7 weeks. As a result, among the SCB treatments, SCB 1/2N was similar to control 1N and CNS 1N in terms of fresh and dry weights of shoots, leaf area, stem diameter, number of node, and SPAD value. Increased N concentration of SCB inhibited the growth of tomato plants. Total fruit yield of SCB 1/2N was 47% of that of control 1N which showed the best result. Percentage of marketable fruit yield in SCB 1/2N was about 58%. Soluble solids contents, total acidity, total phenolic concentration and antioxidant capacity was the highest in SCB 2N and the other treatments were not shown any difference. Blossom-end rot rarely occurred in control 1N and CNS 1N while SCB treatments without Ca induced the physiological disorder of 7~19%. In conclusion, SCB 1/2N was good for the vegetative growth of cherry tomato plants but reduced yield and quality of fruit compared with control 1N and CNS 1N. Thus, it is possible to apply SCB solution to grow cherry tomato plants hydroponically but in the consideration of fruits yield and quality additional supply of several minerals would be required.

A Study on Strategy for developing LBS Entertainment content based on local tourist information (지역 관광 정보를 활용한 LBS 엔터테인먼트 컨텐츠 개발 방안에 관한 연구)

  • Kim, Hyun-Jeong
    • Archives of design research
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    • v.20 no.3 s.71
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    • pp.151-162
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    • 2007
  • How can new media devices and networks provide an effective response to the world's growing sector of cultural and historically-minded travelers? This study emerged from the question of how mobile handsets can change the nature of cultural and historical tourism in ubiquitous city environments. As wireless network and mobile IT have rapidly developed, it becomes possible to deliver cultural and historical information on the site through mobile handset as a tour guidance system. The paper describes the development of a new type of mobile tourism platform for site-specific cultural and historical information. The central objective of the project was to organize this cultural and historical walking tour around the mobile handset and its unique advantages (i.e. portability, multi-media capacity, access to wireless internet, and location-awareness potential) and then integrate the tour with a historical story and role-playing game that would deepen the mobile user's interest in the sites being visited, and enhance his or her overall experience of the area. The project was based on twelve locations that were culturally and historically significant to Korean War era in Busan. After the mobile tour game prototype was developed for this route, it was evaluated at the 10th PIFF (Pusan International Film Festival). After use test, some new strategies for developing mobile "edutainment content" to deliver cultural historical contents of the location were discussed. Combining 'edutainment' with a cultural and historical mobile walking tour brings a new dimension to existing approaches of the tourism and mobile content industry.

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Mushroom growth and cultivation environment at cultivation house of vinyl bag cultivation Shiitake mushroom on high-temperature period (고온기 표고 톱밥재배용 재배사 내의 환경 제어시스템과 버섯생육 온도)

  • Jhune, Chang-Sung;Kong, Won Sik;Park, Hye-Sung;Cho, Jae-Han;Lee, Kang-Hyo
    • Journal of Mushroom
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    • v.12 no.4
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    • pp.263-269
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    • 2014
  • Although sawdust cultivation of shiitake (Lentinula edodes) is becoming more common, it is insufficiently competitive in spring and autumn, the best time to breed shiitake. Thus, it is urgently needed to develop a technique for all year round cultivation of shiitake using mushroom growing beds. In the present study, the temperature changes according to the location of shiitake cultivation facilities were investigated. We confirmed that a refrigerator, an air conditioner, triple membranes, shiitake cultivation beds, fog nozzles which were installed in the shiitake cultivation facilities play an important role in keeping the low temperature. Bag cultivation of shiitake was tested in temperature variation from $14^{\circ}C$ to $29^{\circ}C$ with a $3^{\circ}C$ interval to know its cultivating temperature range in hot summer season. In summary, the sawdust cultivation of shiitake is possible when the temperature difference between top and bottom is maintained below $1^{\circ}C$. And the temperature of the shiitake cultivation facilities should be maintained below $23^{\circ}C$ in the induction period for fruitbody formation.

Antihepatotoxic effect of ethanol extracts from steam-dried ginseng berry on ᴅ-galactosamine/lipopolysaccharide-sensitized mice (ᴅ-galactosamine/lipopolysaccharide로 감작된 급성간독성 마우스 모델에서 인삼열매추출물의 간독성 개선 효과)

  • Jang, Su Kil;Park, Jun Sub;Ahn, Jeong Won;Jo, Boram;Kim, Hyun Soo;Kim, JeongHoon;Kim, Sang Yun;Park, Jung Youl;Lee, Do Ik;Park, Hee Yong;Joo, Seong Soo
    • Korean Journal of Food Science and Technology
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    • v.49 no.6
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    • pp.676-684
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    • 2017
  • The present study aimed to examine the hepatoprotective effects of ethanol extracts from steam-dried ginseng berry (SGBE) in both $\text\tiny{D}$-Galactosamine/Lipopolysaccharide ($\text\tiny{D}$-GalN/LPS)-sensitized mice and in vitro models. Our results clearly demonstrated that SGBE significantly reduced the level of aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, and lactate dehydrogenase in blood, and $TNF{\alpha}$ was normalized in 8 h after the treatment with $\text\tiny{D}$-GalN/LPS. Coincidently, major organs remained unimpaired when compared to $\text\tiny{D}$-GalN/LPS control group. Moreover, p38, which stimulates expression of NAFLD-associated cytokines, was markedly inhibited when treated with SGBE. In vitro analysis revealed that the main components of SGBE, linoleic acid and ginsenoside Re/Rd, may play a role in protecting liver from $\text\tiny{D}$-GalN/LPS-induced toxicity. Finally, we concluded that SGBE may be a promising therapeutic agent for preventing damage to the liver.

Development of Evaluation Method for Jointed Concrete Pavement with FWD and Finite Element Analysis (FWD와 유한요소해석을 이용한 줄눈콘크리트포장 평가법 개발)

  • Yun, Kyong-Ku;Lee, Joo-Hyung;Choi, Seong-Yong
    • International Journal of Highway Engineering
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    • v.1 no.1
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    • pp.107-119
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    • 1999
  • The joints in the jointed concrete pavement provide a control against transverse or longitudinal cracking at slab, which may be caused by temperature or moisture variation during or after hydration. Without control of cracking, random cracks cause more serious distresses and result in structural or functional failure of pavement system. However, joints nay cause distresses due to its inherent weakness in structural integrity. Thus, the evaluation at joint is very important. and the joint-related distresses should be evaluated reasonably for economic rehabilitation. The purpose of this paper was to develop an evaluation system at joints of jointed concrete pavement using finite element analysis program, ILLI-SLAB, and nondestructive testing device. FWD. To develop an evaluation system for JCP, a sensitivity analysis was performed using ILLI-SLAB program with a selected variables which might affect fairly to on the performance of transverse joints. The most significant variables were selected from precise analysis. An evaluation charts were made for jointed concrete pavement by adopting the field FWD data. It was concluded that the variables which most significantly affect to pavement deflections are the modulus of subgrade reaction(K) and the modulus of dowel/concrete interaction(G), and limiting criteria on the performance of joints at JCP are 300pci. 500,000 lb/in. respectively. Using these variables and FWD test, a charts of load transfer ratio versus surface deflection at joints were made in order to evaluate the performance of JCP. Practically, Chungbu highway was evaluated by these evaluation charts and FWD field data for jointed concrete pavement. For Chungbu highway, only one joint showed smaller value than limiting criterion of the modulus of dowel/concrete interaction(G). The rest joints showed larger values than limiting criteria of the modulus of subgrade reaction(K) and the modulus of dowel/concrete interaction(G).

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Impacts of Three-dimensional Land Cover on Urban Air Temperatures (도시기온에 작용하는 입체적 토지피복의 영향)

  • Jo, Hyun-Kil;Ahn, Tae-Won
    • Journal of the Korean Institute of Landscape Architecture
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    • v.37 no.3
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    • pp.54-60
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    • 2009
  • The purpose of this study is to analyze the impacts of three-dimensional land cover on changing urban air temperatures and to explore some strategies of urban landscaping towards mitigation of heat build-up. This study located study spaces within a diameter of 300m around 24 Automatic Weather Stations(AWS) in Seoul, and collected data of diverse variables which could affect summer energy budgets and air temperatures. The study also selected reflecting study objectives 6 smaller-scale spaces with a diameter of 30m in Chuncheon, and measured summer air temperatures and three-dimensional land cover to compare their relationships with results from Seoul's AWS. Linear regression models derived from data of Seoul's AWS revealed that vegetation volume, greenspace area, building volume, building area, population density, and pavement area contributed to a statistically significant change in summer air temperatures. Of these variables, vegetation and building volume indicated the highest accountability for total variability of changes in the air temperatures. Multiple regression models derived from combinations of the significant variables also showed that both vegetation and building volume generated a model with the best fitness. Based on this multiple regression model, a 10% increase of vegetation volume decreased the air temperatures by approximately 0.14%, while a 10% increase of building volume raised them by 0.26%. Relationships between Chuncheon's summer air temperatures and land cover distribution for the smaller-scale spaces also disclosed that the air temperatures were negatively correlated to vegetation volume and greenspace area, while they were positively correlated to hardscape area. Similarly to the case of Seoul's AWS, the air temperatures for the smaller-scale spaces decreased by 0.32% ($0.08^{\circ}C$) as vegetation volume increased by 10%, based on the most appropriate linear model. Thus, urban landscaping for the reduction of summer air temperatures requires strategies to improve vegetation volume and simultaneously to decrease building volume. For Seoul's AWS, the impact of building volume on changing the air temperatures was about 2 times greater than that of vegetation volume. Wall and rooftop greening for shading and evapotranspiration is suggested to control atmospheric heating by three-dimensional building surfaces, enlarging vegetation volume through multilayered plantings on soil surfaces.

A Study on the Installation of Groyne using Critical Movement Velocity and Limiting Tractive Force (이동한계유속과 한계소류력을 활용한 수제 설치에 관한 연구)

  • Kim, Yeong Sik;Park, Shang Ho;An, Ik Tae;Choo, Yeon Moon
    • Journal of Wetlands Research
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    • v.22 no.3
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    • pp.194-199
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    • 2020
  • Unlike in the past, the world is facing water shortages due to climate change and difficulties in simultaneously managing the risks of flooding. The Four Major Rivers project was carried out with the aim of realizing a powerful nation of water by managing water resources and fostering the water industry, and the construction period was relatively short compared to the unprecedented scale. Therefore, the prediction and analysis of how the river environment changes after the Four Major Rivers Project is insufficient. Currently, part of the construction section of the Four Major Rivers Project is caused by repeated erosion and sedimentation due to the effects of sandification caused by large dredging and flood-time reservoirs, and the head erosion of the tributaries occurs. In order to solve these problems, the riverbed maintenance work was installed, but it resulted in erosion of both sides of the river and the development of new approaches and techniques to keep the river bed stable, such as erosion and excessive sedimentation, is required. The water agent plays a role of securing a certain depth of water for the main stream by concentrating the flow so much in the center and preventing levee erosion by controlling the flow direction and flow velocity. In addition, Groyne products provide various ecological environments by forming a natural form of riverbeds by inducing local erosion and deposition in addition to the protection functions of the river bank and embankment. Therefore, after reviewing the method of determining the shape of the Groyne structure currently in use by utilizing the mobile limit flow rate and marginal reflux force, a new Critical Movement Velocity(${\bar{U}}_d$) and a new resistance coefficient formula considering the mathematical factors applicable to the actual domestic stream were developed and the measures applicable to Groyne installation were proposed.