포도와 오이즙액을 이용한 동충하초(Paecilomyces japonica) 균사체 배양액의 미백효과

이영훈,박상상,이상원,이성호,박기훈,최영주,갈상완,Lee, Young-Hoon,Park, Sang-Sang,Lee, Sang-Won,Lee, Sung-Ho,Park, Ki-Hoon,Choi, Young-Ju,Gal, Sang-Wan 한국생명과학회 2006 생명과학회지 Vol.16 No.5

포도와 오이즙액에 동충하초 균사체 배양액의 미백화장품 원료로서의 이용가능성을 조사하였다. B16BL6 mouse melanoma cell에 동충하초 균사체 배양액을 배양기간 및 농도별로 처리했을 때, 15일 동안의 배양조건과 $50\;{\mu}l/ml$ (5%)의 농도로 처리하였을 때 멜라닌 생성억제 효과가 가장 높은 것을 확인하였다. In vitro tyrosinase 활성억제효과에서는 포도와 오이즙액에 동충하초 균사체 배양액을 10% 농도로 첨가시 50%의 멜라닌 생성 억제율을 보였으며, 30% 농도에서는 100% 억제를 확인하였다. 결과적으로 포도와 오이즙액에 동충하초 균사체 배양액은 멜라닌 생성의 주 효소인 tyrosinase의 작용을 억제함으로써 미백 효과를 나타내는 것을 확인하였다. 또한 세포독성실험에서 포도, 오이즙액과 그 즙액에 동충하초 균사체 배양액을 각각의 농도로 B16BL6 mouse melanoma cell에 처리 한 결과 포도, 오이즙액의 5% 처리 시에는 50% 사멸, 9% 이상에서는 100% 세포가 사멸하는 것을 확인 할 수 있었다. 그러나 포도, 오이즙액에 동충하초 균사체를 배양한 배양액은 20% 까지 처리 시에도 세포독성이 나타나지 않는 것을 확인 하였다. 따라서 포도와 오이즙액에 동충하초 균사체를 배양한 배양액은 세포 독성이 없으며, 미백 효과가 뛰어난 것으로 미백화장품 원료로서 사용가능함을 확인하였다. This study was carried out to investigate the whitening effect of mycelial culture broth of P. japonica in the mixture of cucumber and grape extracts. In the inhibition test of melanin biosynthesis of melanoma cell, B16BL6, the culture broth of P. japonica more then $50\;{\mu}l/ml$ (5%) concentration inhibited the melanin biosynthesis of the cell entirely without cytotoxicity. More then 10 days incubation of P. japonica in the mixture was required to have the inhibition activity. In vitro inhibition test of melanin biosynthesis of the culture broth of P. japonica was investigated in the concentration dependent manner of 10% to 50%. 30% concentration of the culture broth inhibited completely tyrosinase activity. In the cytotoxicity test, cucumber and grape extract itself has a strong cytotoxicity to the melanoma cell, B16Bl6. The value of $IC_{50}$ of the cucumber and grape extracts against the melanoma cells was 5% concentration. However, the culture broth of P. japonica incubated in the cucumber and grape extracts did not show the cytotoxcity up to 20% against melanoma cell, B16BL6. Therefore, we concluded that the culture broth of P. japonica in the mixture of cucumber and grape extracts can be used as a whitening cosmetic resource.

흑두로 제조한 청국에서 분리된 Bacillus subtillus BB-1으로 부터 혈전용해효소 유전자 크로닝 및 특성규명

이영훈,이성호,전주미,김홍출,조용운,박기훈,최영주,갈상완,Lee Young-Hoon,Lee Sung-Ho,Jeon Ju-Mi,Kim Hong-Chul,Cho Yong-Un,Park Ki-Hoon,Choi Young-Ju,Gal Sang-Wan 한국생명과학회 2005 생명과학회지 Vol.15 No.4

흑두로 제조한 청국으로부터 혈전용해력이 우수한 균을 선발하여 동정하였으며, 그를 Bacillus subtilis BB-1로 명명하였다. 이 균은 혈전용해효소 isozyme을 적어도 5개이상 생성하는 균주로 확인되었다. 이 균으로부터 크로모좀을 분리하여 shot gun법으로 혈전용해효소 유전자를 크로닝하였으며, 이 유전자를 BSF-1이라 명명하였다. 이 유전자는 714개의 아미노산을 암호화하고 있으며 기존에 밝혀진 혈전용해효소 유전자와 상동성은 검출되지 않은 새로운 혈전용해효소 유전자였다. 혈전용해효소활성 최적 pH 및 온도는 5.0과 $35^{\circ}C$였다. 기질특이성은 적혈구 배지 또는 skim milk, gelatin등에 전혀 분해활성이 없었다. 이는 혈전만을 특이적으로 분해하는 기질특이성을 보였으며, 혈전분해효소로서의 이용가능성이 충분한 것으로 판단된다. A bacterium producing five fibrinolytic isozymes was isolated from black bean chung kuk. The bacterium was identified as Bacillus subtilis BB-1 by 16s rDNA sequence homology search. A gene out of five fibrinolytic genes in the Bacillus subtilis BB-1 was cloned by shot-gun method. A Cla I DNA fragment of B. subtilis BB-1 chromosome was cloned in to pBluescript II SK(-) and showed the fibrinolytic activity to bacterial cells. The Cla I DNA fragment was sequenced and the sequences did not show homology with gene for protease or fibrinolytic enzyme genes in other organisms. The Cla I DNA fragment was reduced to 2,142 bp by activity-guided PCR cloning method. The optimum pH and temperature of the enzyme were 5.0 and $35^{\circ}C$, respectively. Substrate specificity of the fibrinolytic enzyme was detected in skim milk, casein, gelatin and blood agar plates. The activity of the enzyme was not detected with these substrates. Taken together, this enzyme is a new fibrinolytic enzyme and may be used to prevent thrombosis and arteriosclerosis.

Antitumor Sterol Isolated from the Fruiting Body of Pleurotus eryngii

이영훈,박기훈,이병원,조용운,최영주,갈상완,Lee Young-Hoon,Park Ki-Hun,Lee Byong-Won,Cho Yong-Un,Choi Young-Ju,Gal Sang-Wan Korean Society of Life Science 2006 생명과학회지 Vol.16 No.2

새송이버섯으로부터 활성추적법으로 항암활성이 있는 물질인 에르고스테롤 프록사이드를 분리하였다. 이 스테롤의 구조는 분광법과 NMR법으로 확인하였으며 분자식은 $C_{28}H_{44}O_3$이었다. 폐암과 난소암에 $IC_{50}$값은 각각 $7{\mu}M$과 $14{\mu}M$이었다. DNA단편화 실험에서 이 화합물은 암세포의 chromosimal DNA 를 사닥다리모양으로 분해하였고, 세포 분열주기의 억제실험에서 G1단계를 억제함을 관찰하였다. Activity-guided fractionations led to the isolation of antitumor compound, ergosterol peroxide ($5{\alpha},\;8{\alpha}-epideoxy-24(R)-methylcholesta-6,\;22-dien-3{\beta}-ol$) from the fruiting body of Pleuratus eryngii that was cultivated artificially. This sterol structure was established by using spectroscopic methods ($^1H\;and\;^{13}C$ nuclear magnetic resonance and high resolution mass spectra). The purified compound showed a molecular formular of $C_{28}H_{44}O_3$ displaying characteristic features of epidioxy sterols. The 50% inhibitory concentrations ($IC_{50}$) of ergosterol peroxide against human lung cancer cell line (A549) and human ovarian cell line (SK-OV3) were $7{\mu}M\;and\;14{\mu}M$, respectively. In the DNA fragmentation assay, the compound showed the programmed cell death causing the chromosomal DNA fragmentation. It reveals that ergosterol peroxide arrests G1 phase of the cell division cycle.

초고속 자기부상철도 개발을 위한 시스템엔지니어링 계획서

이영훈,민성기,Lee, Young-Hoon,Min, Sung-Ki 한국시스템엔지니어링학회 2009 시스템엔지니어링학술지 Vol.5 No.1

The systems engineering (SE) process shall be applied to the project for successful development of ultra-high-speed (UHS) Maglev train system which is one of very large and complex systems. It is important to abolish technology differentials from the advanced developers such as Germany, Japan, etc. and to ensure discriminatory competitiveness of the application of systems engineering process for the development of the system based on appropriate concepts and requirements. General operation concept and stakeholder's requirements of UHS Maglev train system must be elicited with system concept for initiating the project. The management plan should be devised for all sorts of systems engineering activities of risk management, performance management, lifecycle cost management, etc. This paper would support to establish the systems engineering management plan (SEMP) for the program of UHS Maglev train system development with associated documents.

그래픽 모사기법을 이용한 하천 변천의 재현과 예측

이영훈,Lee, Young-Hoon 대한자원환경지질학회 1994 자원환경지질 Vol.27 No.6

A case study is presented where a fluvial system is modeled in three dimensions and compared to data gathered from a study of the Arkansas River. The data is unique in that it documents changes that affected a straight channel that was excavated within the river by the U.S. Army Corps of Engineers. Excavation plan maps and sequential aerial photographs show that the channel underwent massive deposition and channel migration as it returned to a more natural, meandering path. These records illustrate that stability of fluvial system can be disrupted either by catastrophic events such as floods or by subtle events such as the altering of a stream's equilibrium base level or sediment load. SEDSIM, Stanford's Sedimentary Basin Simulation Model, is modified and used to model the Arkansas River and the geologic processes that changed in response to changing hydraulic and geologic parameters resulting from the excavation of the channel. Geologic parameters such as fluid and sediment discharge, velocity, transport capacity, and sediment load are input into the model. These parameters regulate the frequency distribution and sizes of sediment grains that are eroded, transported and deposited. The experiments compare favorably with field data, recreating similar patterns of fluid flow and sedimentation. Therefore, simulations provide insight for understanding and spatial distribution of sediment bodies in fluvial deposits and the internal sedimentary structure of fluvial reservoirs. These techniques of graphic simulation can be contributed to support the development of the new design criteria compatible with natural stream processes, espacially drainage problem to minimize environmental disruption.

Fractal 차원과 Variogram을 이용한 암반 불연속면의 굴곡도 특성 서술

이영훈,Lee, Young-Hoon 대한자원환경지질학회 1994 자원환경지질 Vol.27 No.1

There has been considerable research dealing with the influence of surface roughness along surfaces of rock discontinuities in relation to the peak shear strength of rock masses. Concepts accepted recently for measuring such strength include estimation of a roughness coefficient such as developed by Barton's studies. The method for estimation the Joint Roughness Coefficient (JRC) value of a measured roughness profile is subjective. The aim of this research is to estimate the JRC value of the roughness of a surface profile in a rock mass system using an objective method. The study of roughness of surfaces has included measurement of fractal geometric characteristics. Once the irregularity of the surface has been described by the fractal dimension, the spatial variation of the surface irregularities can be described using variogram and drift analysis. An empirical relationships between the roughness profiles of selected JRC ranges and their fractal dimension with variogram and drift were derived. The application of analyses of fractal dimension, variogram and drift was novel for the analysis of roughness profiles. Also, an empirical equation was applied to experimental data.

조직 내 시스템엔지니어링 환경 구축방안

이영훈,Lee, Young Hoon 한국시스템엔지니어링학회 2009 시스템엔지니어링학술지 Vol.5 No.2

For the effective and efficient system development, it is provided of complex and diverse disciplines, analysis and design, points of views for decisions making, technical information management, assurance of products, and so on to be integrated and ensured. Systems engineering is conducted through a process and is implemented by tools based on relevant methodologies. Systems engineering environment should be provided to support the systems engineering process, method, and tools used on the project. In this paper, the essential environment in performing works in a organization applying systems engineering on the basis of system life cycle was reviewed. The present states on establishment of the systems engineering in Korea Railroad Research Institute were also examined with some reviews of other cases.

고열충격반응에 영향을 미치는 초파리 hsp70 유전자의 DNA 염기배열

이영훈,전은순,박충웅,Lee, Young-Hoon,Jeon, Eun-Soon,Park, Chung-Ung 생화학분자생물학회 1988 한국생화학회지 Vol.21 No.4

The DNA sequences required for heat shock regulated expression were examined in Drosophila melanogaster. Various hsp70-YP1 fusion constructs where D. melanogaster hsp70 gene and YPI gene were used as a heat shock promoter and as a target gene, respectively, were transfected into D. melanogaster Schneider line 2 cells and their heat inducibility was determined by Northern analysis. The sequences containing two HSEs between -89 and -38 of the hsp70 were not sufficient to induce YP1 RNA upon heat shock whereas the sequences between -89 and +34 including the TATA box and the transcription start region of the hsp70 besides the HSEs were able to drive YP1 transcription during heat shock. The small deletion of the sequences near the transcription start region of the YP1 gene was also observed to repress the heat induced transcription of YP1 RNA by the sequences between -89 and +34 of the hsp70. 초파리 Drosophila melalnogaster에서 고열충격 유발을 조절하는데 필요한 DNA 염기배열을 조사하였다. hsp 70 유전자를 고열충격 촉진자로, YP1 유전자를 표적유전자로 하여 hsp70-YP1 융합유전자를 제조하고 이들 융합유전자를 이용하여 초파리 Schneider line 2 세포를 형질변환시키고 Northern 분석을 함으로써 hsp70-YP1 융합유전자의 고열충격 유발성을 결정하였다. 두개의 HSE (heat shock element) 가 존재하는 hsp70 유전자의 -89에서 -38까지 염기배열만으로는 고열충격에 의해 YP1 RNA를 유발시킬 수 없었지만 두 개의 HSE 이외에 hsp70 유전자의 TATA 박스와 전사시작점을 포항하고 있는 -89에서 +34까지 염기배열은 고열충격에 의해 YP1 RNA를 유발시킬 수 있었다. 또한 YP1 유전자의 전사시 작정 근처의 염기배열이 조금 결실되었을 때 hsp70 유전자의 -89에서 +34까지 염기배열에 의한 YP1 RNA의 고열충격 유발이 억제되는 것이 관찰되었다.

Bacillus subtilis BB-1으로부터 나토키나아제 유전자 크로닝 및 대량발현

이영훈,이성호,박기훈,최영주,정영기,갈상완,Lee Young-Hoon,Lee Sung-Ho,Park Ki-Hoon,Choi Young-Ju,Jeong Yong-Kee,Gal Sang-Wan 한국생명과학회 2006 생명과학회지 Vol.16 No.2

흑두청국으로부터 분리된 혈전용해력이 우수한 Bacillus subtilis BB-1(KFCC 11344P)으로부터 혈전용해효소 유전자를 PCR법에 의해 크로닝하였고 이를 BCF-1으로 명명하였다. BCF-1의 DNA 염기서열결정 결과 1,145 bp 크기의 혈전용해 효소로, 일본의 natto로부터 분리된 nattokinase 유전자와 99%의 상동성을 보임을 확인하였다. 혈전용해효소 유전자의 발현을 위하여 Bacillus 발현계인 Bacillus-E. coli의 shuttle vector인 pEB vector에 크로닝 하고 host로서 B. subtilis 168에 형질전환시켜 대량 발현시켰다. 생산된 혈전용해효소의 최 적활성 pH와 온도는 7.0과 $35^{\circ}C$로 확인되었다, 기질에 대한 분해양상을 조사한 결과 fibrin에서만 특이적으로 강한 분해가 일어났으며, skim milk에서 아주 약한 분해능을 보였으나 blood agar, gelatin, casein에서는 전혀 분해능을 보이지 않았다. 특히 blood agar plate에서 분해능이 없는 것으로 보아 혈액 내에서의 적혈구 파괴현상과 같은 부작용에 대한 위험을 배제할 수 있을 것으로 사료된다. BCF-1에 의해 생산된 혈전용해효소는 fibrin 특이적으로 활성을 나타냄을 확인할 수 있으며, 이는 임상적이나 산업적으로 적용하였을 때 부작용에 대한 위험적인 문제는 배제될 수 있으리라 생각된다. A fibrinolytic enzyme gene was isolated from Bacillus subtilis BB-1 by PCR method. Primers for PCR cloning were designed according to pre-identified gene for fibrinolytic enzymes from B. subtilis. The primer sequences were 5'-CGG ATC CGT GAG AGG CAA AAA GGT G-3' and 5'-TGA ATT CTT AAT GTG CTG CTG CTT GTC C-3' as concensus sequences of the fibrinolytic genes of Bacillus species. The PCR product was 1,145 bp and the sequence homology was 99% with nattokinase gene isolated from Japanese natto. The cloned fibrinolytic gene was reconstructed in Bacillus-E. coli shuttle vector, pEB for bulk-production. The fibrinolytic enzyme was purified by FPLC from the cloned B. subtilis 168. The optimum pH and temperature of the enzyme were 7.0 and $35^{\circ}C$, respectively. The fibrinolytic enzyme did not show any activity toward to skim milk, gelatin, casein and blood agar plate. The enzyme specific polyclonal antibody was prepared in rabbit for further assays such as detection of the gene expression in plant cells. This means that the enzyme may be used for health-care such as thrombosis without any hamful effects in the blood vessel.

임프란트 식립시 초기 안정성이 골유착에 미치는 영향

이영훈,김여갑,최병준,Lee, Young-Hoon,Kim, Yeo-Gab,Choi, Byung-Jun 대한악안면성형재건외과학회 2008 Maxillofacial Plastic Reconstructive Surgery Vol.30 No.6

Purpose: The long-term experience of using osseointegrated implants for prosthetic rehabilitation of the edentulous patients shows that high success rates can be predictably achieved. Primary implant stability has been identified to be a prerequisite to achieve osseointegration. In this study, we set up the amount of removed bone so that it differed on implant installation site for each group. The influence of each initial stability on secondary stability and osseointegration was compared with time lapse using resonance frequency analysis and histomorphometric analysis. Materials and methods: A total 27 US $II^{(R)}$ (Osstem, Korea) implants were placed in the mandibular edentulous area of 3 beagle dogs. The implant site was prepared by the conventional technique with drills, and three experimental groups were divided into under-drilling group, normal-drilling group and over-drilling group. The Implant Stability Quotient (ISQ) was measured at intervals of immediately, 4, 8, 12 weeks after placement using $Osstell^{(R)}$ mentor RFA. After the animals were sacrificed, histomorphometric evaluation was executed for measuring BIC and BD. Results: 1) The under-drilled group showed most high ISQ value for whole experiment period. 2) Bone-to-implant-contact(BIC) showed the tendency to be increased gradually as the experiment period passed except the 8 weeks of the normal group. 3) The under-drilled group showed most high bone density(BD) level for whole experiment period, and it was expressed the aspect to be increased gradually according to an experiment period passage in the average of all group. 4) Resonance frequency analysis and histomorphometric analysis are presumed by generally proportional. Conclusions: As this research result, it seems that there are some correlation between resonance frequency analysis and histomorphometric analysis. As are accomplished osseointegration stably so that more superior at the region which the overpressure comes to add, it will be applicable method in clinical field.