Escherichia coli에서 효소활성을 지닌 Human HtrA3 단백질 제조와 HtrA Serine Protease 1, 2와의 효소활성 비교

김지환,김구영,남민경,김상수,임향숙,Kim, Ji-Hwan,Kim, Goo-Young,Nam, Min-Kyung,Kim, Sang-Soo,Rhim, Hyang-Shuk 한국미생물학회 2009 미생물학회지 Vol.45 No.4

본 연구에서는 Human HtrA3 (HtrA3)의 효소활성을 분자수준에서 연구하기 위해 HtrA간의 상동성과 기존에 알려진 maturation site들을 비교 분석하여 예상 mature HtrA3인 M1-HtrA3와 M2-HtrA3를 발현하는 construct를 제작하였다. pGEX system을 통해 Top10 균주에서 발현, 정제한 M1-HtrA3 단백질은 $10{\mu}g$/L를 정제할 수 있었으며 발현량 대비 1%를 회수할 수 있었다. M2-HtrA3는 M1보다 5배 가량 많은 양을 정제할 수 있었으며 발현량 대비 회수율은 3배 정도 더 높았다. $\beta$-Casein을 이용한 in vitro cleavage test를 통해 M1, M2 form 모두 protease 활성을 갖는 것을 확인하였다. 또한, $\beta$-casein cleavage를 통해 HtrA serein protease들 간의 상대적인 활성을 비교한 결과, HtrA3와 HtrA2는 HtrA1보다 약 2배 더 높은 proteolytic cleavage 활성을 보였다. 본 연구에서 정립한 protease 활성을 지닌 HtrA3의 제작과 정제 조건은 HtrA3의 substrate를 탐색을 용이하게 할 수 있을 것이며, HtrA3 연관된 질환의 발병기전과 세포 신호전달을 이해하는 연구에 활용될 수 있을 것이다. To elucidate HtrA3's functional roles in the HtrA3 mediated cellular processes, it is necessary to investigate its biochemical characteristics. In the present study, we constructed the plasmids encoding putative mature HtrA3 proteins (M1-HtrA3 and M2-HtrA3) based on the putative maturation sites of highly homologous HtrA1 and mouse HtrA3. We used the pGEX bacterial expression system to develop a simple and rapid purification for the recombinant HtrA3 protein. Although yields of the mature HtrA3 proteins were slightly low as 10~50 ${\mu}g$/L, the amounts and purity of M1- and M2-HtrA3 were enough to investigate their proteolytic activities. The putative mature HtrA3 proteins have proteolytic activity which could cleave $\beta$-casein as an exogenous substrate. We compared the proteolytic activity between the HtrA family, HtrA1, HtrA2, and HtrA3. The cleavage activity of HtrA3 and HtrA2 were 2 folds higher than that of HtrA1, respectively. Our study provides a method for generating useful reagents to identify natural substrates of HtrA3 in the further studies.

플렉서블 디스플레이용 저온공정을 갖는 대향 타겟식 스퍼터링 장치를 이용한 ZrO₂ 가스 차단막의 특성

김지환,조도현,손선영,김화민,김종재,Kim, Ji-Hwan,Cho, Do-Hyun,Sohn, Sun-Young,Kim, Hwa-Min,Kim, Jong-Jae 한국전기전자재료학회 2009 전기전자재료학회논문지 Vol.22 No.5

$ZrO_2$ film was deposited by facing target sputtering (FTS) system on polyethylene naphthalate (PEN) substrate as a gas barrier layer for flexible organic light emitting devices (FOLEDs), In order to control the heat of the FTS system caused by the ion bombardment in the cathode compared with the conventional sputtering system, the process characteristics of the FTS apparatus are investigated under various sputtering conditions such as the distance between two targets ($d_{TT}$), the distance between the target and the substrate ($d_{TS}$), and the deposition time. The $ZrO_2$ film by the FTS system can reduce the damage on the films because the ion bombardment with high-energy particles like gamma-electrons, Moreover, the $ZrO_2$ film with optimized condition ($d_{TT}$=140 mm) as a function of the distance from center to edge showed a very uniform thickness below 5 % for a deposition time of 3 hours, which can improve the interface property between the anode and the plastics substrate for flexible displays, It is concluded that the $ZrO_2$ film prepared by the FTS system can be applied as a gas barrier layer or an interlayer between the anode and the plastic substrate with good properties of an uniform thickness and a low deposition-temperature.

Analysis of stress distribution of tooth restored with metal-ceramic crown covering abfraction lesion according to its finish line location under occlusal load

김지환,윤철욱,김태현,김한성,우대곤,이근우,심준성,Kim, Jee-Hwan,Yoon, Chol-Wook,Kim, Taehyeon,Kim, Han-Sung,Woo, Dae-Gon,Lee, Keun-Woo,Shim, June-Sung The Korean Academy of Prosthodonitics 2014 대한치과보철학회지 Vol.52 No.4

목적: 다양한 원인으로 발생된 Abfraction 병소가 있는 치아를 금속도재관으로 수복 할 경우, 변연의 위치에 따라 예후가 다양해질 수 있다. 수복물의 장기적 성공을 검증하기 위해서는 응력분포의 분석이 필요하지만 금속도재관이 abfraction 병소의 적응증인 경우 금속도재관이 응력 분포에 미치는 영향을 밝히는 연구는 아직 없다. 본 연구의 목적은 abfraction 병소가 있는 치아를 수복하는 금속도재관이 바람직한 응력 분포를 나타낼 수 있는 조건을 알아 보고자 함이다. 재료 및 방법: 치아, 주위 조직과 금속 도재관의 외형을 반영한 2차원 유한 요소모델을 제작하고 144 N의 편심 교합력 하에서의 응력분포를 유한요소 분석법으로 분석하였다. 금속 도재관의 변연 위치를 쐐기모양의 결손부의 하연에(Group 0), 그리고 결손부의 하연보다 1 mm 하방(Group 1)과 2mm 하방(Group2)에 위치시켰다. 결과: Group 0에서 von Mises stress는 금속 도재관의 변연과 결손부의 첨부에 집중 되었고 협측 변연의 응력은 설측 부위로 분포되었다. Group 1과 Group 2에서의 응력분포양상은 비슷하게 나타났다. 응력은 결손부의 첨부에 집중되지만 협측과 설측의 응력띠는 서로 분리된 양상으로 나타났다. 결론: 금속도재관의 변연을 Abfraction 병소의 하연에 설정할 경우, 금속도재관은 Abfraction 병소의 첨부에 응력을 집중시킨다. Purpose: When the full veneer crown was treated in the tooth with abfraction lesion due to various causes, the prognosis of it may be compromised according to the location of the finish line, but there is few study about the location of its buccal finish line. The purpose of this study was to investigate the effect of location of the finish line of the full veneer crown on stress distribution of the tooth with abfraction lesion. Materials and methods: The two dimensional finite element model was developed to express tooth, surrounding tissue and full veneer crown. The stress distribution under eccentric 144 N occlusal load was analyzed using finite element analysis. The location of finish line was set just at the lower border of the lesion (Group 0), 1 mm (Group 1) and 2 mm (Group 2) below the lower border of the lesion. Results: In the Group 0, von Mises stress was concentrated at the finish line and the apex of the lesion. Also, the stress at the bucal finish line propagated to the lingual side. In the Group 1 and Group 2, stress distribution was similar each other. Stress was concentrated at the apex of lesion, but the stress at the buccal finish line did not propagate to the lingual side. That implied decrease of the possibility of horizontal crown fracture. Conclusion: Full veneer crown alleviated the stress concentrated at the apex of the abfraction lesion, when the finish line of full veneer crown was set below the lower border of abfraction lesion.

한국산 녹조식물 해캄속(SPIROGYRA)의 수리표현학

김지환,김영환,이인규,Kim, Ji-Hwan,Kim, Yeong-Hwan,Lee, In-Gyu 한국조류학회(藻類) 2003 ALGAE Vol.18 No.4

The morphotaxonomic characters of the genus Spirogyra were investigated to clarify their taxonomic category and the variation range among species on the basis of comparative morphological and numerical analyses by unialgal cultures and field samples collected from various freshwater habitats in Korea. 25 characters selected on morphological feature of the species were examined on 568 individuals for morphological comparisons and numerical analyses. Width, length and their length/width ratio of vegetative cell, shape of septum, chloroplast number, maximum width, width, length and their length/width ratio of female gametangium, length of male gametangium, size and shape of zygospore, and cell wall ornamentation of the spore showed a comparatively high vector in principal component analysis. In cluster analysis, 15 taxa analysed were divided into 8 major groups by the average taxonomic distance 1.0 level. Considering the morphology and numerical analysis, Spirogyra crassoidea could not be recognized as an independent species, therefore it was treated as a variety of S. ellipsospora. S. koreana (nom. invalidum), recognized as a new taxon, is under the investigation for its clear taxonomic category.

Ag 코팅 Cu 플레이크의 제조에서 전처리 및 Ag 코팅 공정 변화의 효과

김지환,이종현,Kim, Ji Hwan,Lee, Jong-Hyun 한국재료학회 2014 한국재료학회지 Vol.24 No.11

To elucidate the effects of a pretreatment process on the uniformity of Ag electroless plating on Cu flakes, pretreatment time was mainly considered with a mixed solution of 0.15 M ammonium hydroxide and 0.0375 M ammonium sulphate. Optical inspection of Ag-coated Cu flakes determined that the optimal pretreatment time is 120 s. Repetition of the sequence in which Ag plating was done immediately after the pretreatment of 120 s clearly enhanced the plating uniformity. Scanning electron microscopy revealed that holes were formed irregularly on some Cu flakes during the period from the asdropping of an Ag precursor solution to 5 min. The hole formation was judged to be due to continuous removal of Cu on the local surfaces by the repetitive formation and elimination of $Cu_2O$ or $Cu(OH)_2$ layers. However, the increase of the amount of Ag coating suppressed the hole creation and increasingly enhanced the antioxidant property.

특정 호흡수에서 행한 통제호흡이 심박변이도에 미치는 영향

김지환,김병수,박성식,이용재,금나래,배효상,Kim, Ji-Hwan,Kim, Byoung-Soo,Park, Seong-Sik,Lee, Yong-Jae,Keum, Na-Rae,Bae, Hyo-Sang 사상체질의학회 2016 사상체질의학회지 Vol.28 No.2

Objectives We observed what effects of Paced Breathing(PB) in specific respiration rate have on heart rate variability (HRV) according to Sasang Constitution.Methods HRV of 72 healthy participants in sitting position was recorded while they carried out usual breathing, 0.2Hz, 0.1Hz, and 0.05Hz PB each 5 minutes in consecutive order. Five minute of relaxation was permitted between each breathing. Finally, HRV indices were statictically analyzed of 32 participants (SOEUM: 11, SOYANG: 10, TAEEUM: 11) after data out of accord with respiration rate or outliers were excluded.Results and Conclusions According to respiration rates, there was no statistical significance of HRV among Sasang Constitution. Regardless of Sasang Constituion, 0.2Hz PB increased mean heart rate and decreased natural logarithmic low frequency(lnLF) oscillation of HRV without the change of natural logarithmic high frequency(lnHF), while 0.1Hz PB increased lnLF and standard deviation of N-N interval(SDNN), and slightly decreased lnHF without the change of mean heart rate. 0.05 Hz PB also showed the same effect as 0.1Hz PB, but was impracticable.

단일염기다형성을 이용한 치과 질환 유전체 연구

김지환,이재훈,Kim, Jee-Hwan,Lee, Jae-Hoon 대한치과보철학회 2011 대한치과보철학회지 Vol.49 No.4

Genetic association study has been progressed in medicine along with advance in genetic technology. It focused on the individual differences in genotype due to errors occurring during DNA duplication, which can cause vulnerability to specific diseases. Polymorphism defines the varieties in phenotype due to those genetic variations. Polymorphism due to change in one DNA base sequence is called as a Single Nucleotide Polymorphism. In the near future, the evaluation of relative risk to specific disease according to SNP will be essential part of fundamental of medicine for the diagnosis, treatment and prevention. Dental caries and periodontal diseases has been first subject to genetic association study in dentistry and broaden out to other areas like bone formation and resorption. This article presents the current state of genetic association study and its application to dentistry. DNA 복제과정에서의 오류(error)에 의한 유전적 변이(genetic variation)를 통해 개개인이 가지고 있는 유전변이형을 조사하고 특정 질환에 대한 감수성의 차이를 밝히는 유전체 연관성 연구가 의학계 전반에 활발히 진행되고 있다. 개인 간의 DNA 상에 존재하는 염기서열의 차이를 보이는 유전적 변이를 통해 개인간의 형질이 다르게 표현 되는 것을 다형성이라고 하는데 다형성의 원인 중 염기 서열 한 쌍의 변이에 의하여 다른 형질로 표현되는 것을 단일염기다형성(Single Nucleotide Polymorphism; SNP)이라고 정의한다. 유전자 분석 기술의 놀랄만한 발전과 컴퓨터를 이용한 분석 프로그램의 개발에 의해 SNP과 질병의 연관성에 관한 연구는 의학계 모든 분야에서 가속화 되고 있다. 최근 급격하게 빨리 진척되는 연구들에 힘입어 특정 질병에 대한 특정 유전자가 가지는 위험도를 분석하고 환자를 유전적 위험도에 따라 분류하여 진단, 예방 및 치료하는 환자 맞춤식의 진료가 모든 의약학 분야에 적용 될 것으로 생각 된다. 치과 영역에서는 충치와 치주 질환과의 관련성에 대해서 초기 단계의 연구가 진행되고 있는 수준이다. 본 종설에서는 유전체 질병 연구의 현황과 치과 영역에서 시작된 연구를 소개 하고자 한다.

알렌드로네이트의 투여기간이 발치와의 치유에 미치는 영향: 예비실험

김지환,홍종환,최현민,박영범,문홍석,Kim, Jee-Hwan,Hong, Jong-Hwan,Choi, Hyun-Min,Park, Young-Bum,Moon, Hong-Seok 대한치과보철학회 2013 대한치과보철학회지 Vol.51 No.3

연구 목적: 본 연구에서는 알렌드로네이트 투여기간의 차이가 백서의 상악 제 1대구치 발치와의 초기 창상치유에 미치는 영향을 알아보고자 하였다. 연구 재료 및 방법: 4주령 Sprague Dawley Rat (Body weight 130-140 g) 15마리를 이용하여 알렌드로네이트(Merial Inc., Parramatta, Australia) 투여군(실험군)과 투여하지 않은 대조군으로 나누고 실험군은 다시 투여기간에 따라 1주 투여군부터 2주 투여군, 4주 투여군, 6주 투여군까지 총 4개의 실험군으로 나눴다. 실험군에 정해진 시기(발치 5 주전, 발치 3주전, 발치 1주전, 발치 시)부터 발치 1주 후까지 알렌드로네이트를 1 mg/kg의 농도로 주 3회 피하주사 한 후 6주 투여군과 대조군에서 한 개체씩을 이용하여 미세배열(microarray)분석을 실시하였고, 다른 시편들을 이용하여 조직학적 분석을 시행하였다. 발치와 내에서의 신생골의 형성 비율과 발치와 주위 골에서 활성을 잃은 뼈에 대해 분석하였고, 파골세포 표지자인 TRAP 염색을 하여 분석하였다. 통계적인 분석으로 크루스컬-왈리스 검정(Kruskal Wallis test)을 실시하였다(${\alpha}=.05$). 결과:발치 후 1주 시점에서의 발치와 내에서의 신생골의 생성 비율은 대조군보다 ($16.77%{\pm}1.36%$) 4주 투여군($14.99%{\pm}6.26%$)에서 더 낮게 나오는 경향은 있었지만 유의 하지는 않았다. 투여기간이 증가할수록 활성을 잃은 골수강(empty lacuna)의 수는 증가하였고, TRAP positive cell 수는 감소하는 경향이 있으나 유의하지는 않았다. 결론: 본 예비실험을 통해서 알렌드로네이트의 투여기간이 증가할수록 신생골의 생성 비율이 감소되고 활성을 잃은 뼈들이 많아지며 파골세포의 수는 줄어드는 경향이 있음을 알 수 있었으나 통계적으로 유의차는 없었다. Purpose: The purpose of this study was to investigate the effect of different administration duration of alendronate on initial wound healing and new bone formation of extraction socket in rats. Materials and methods: Fifteen male Sprague-Dawley rats (body weight 130-140 g, 4 weeks old, male) were divided into control group (no alendronate administration) and experimental group (alendronate administration). Experimental group was subdivided into 1 week administrated group, 2 week administrated group, 4 week administrated group and 6 week administrated group according to duration of administration. For the experimental groups, during the designated time period (at the time of extraction, 1 week before extraction, 3 week before extraction and 5 week before extraction) till 1 week after extraction, rats were subcutaneously injected with Alendronate at the dose of 1.0 mg/Kg three times a week. Each specimen from 6 week experimental group and control group were used for microarray analysis, and other specimens were used for histological analysis. The rate of new bone formation within the extraction site and bone loss activity was analyzed using TRAP staining. Statistical analysis was performed using Kruskal Wallis test. (${\alpha}=.05$) Results: After one week from the time of extraction, the rate of new bone formation within extraction site for the control group ($16.77%{\pm}1.36%$) compared to the 4 week experimental group ($14.99%{\pm}6.26%$) was lower. However, no statistically significant difference was found. Increase in the number of inactive lacuna (empty lacuna) and decrease in the number of TRAP positive cell were identified with increased duration of administration. There was no significant difference. Conclusion: The results of this study showed as the duration of Alendronate administration increased the rate of new bone formation decreased with loss of bone activity and reduced number of osteoclast.

조각대황탕(皂角大黃湯) 적용 병증의 형성과 처방구성의 변천에 관한 고찰

김지환,Kim, Ji-Hwan 사상체질의학회 2017 사상체질의학회지 Vol.29 No.3

Objectives The purpose of this study is to understand the formation of Jogakdaehwang-tang(JD-T)'s patholoy and the compositional change of it in Sasang Constitutional Medicine(SCM). Methods The pathologic condition in which JD-T was used and the change of its composition was studied in SCM Results and Conclusions 1. Jogak and Daehwang in "Dongmuyoogo Haedong" were presented with Ungdam because of the common purpose flowing the energy of lung. Therapeutic method for exogenous Ungibyeong in "Dongyisusebowon sangchobongwon" was subdivided into Hangwul's treated by Ungdam, and the heat pattern featuring contagious cold and constipation, treated by Jogakhwanggeumdaehwang-tang(JHD-T). 2. JD-T in "Dongyisusebowon Gabogubon" was used for Yangdok and Onbyeong which are exogenous disease causing by heat. The composition of JD-T has been completed as it included Mahwang, Gilgyeong, and Seungma derived from prescriptions relevant with Yangdok. 3. JD-T in "Dongyisusebowon Sinchukbon(Sinchukbon)" was included in pathology of Ganyeol, and it substituted Galgeunhaegi-tang(GH-T) for Onbyeong, not for Yangdok, so Galgeun and Seungma became the main drug in JD-T. Galgeunseung-gitang(GS-T) was created for treating heat pattern owing to Daehwang, its main drug, and moreover Jogak's effect is too strong. Therefore, Mahwang, Hwanggeum and Gilgyeong was excluded from the composition of JD-T. 4. The origins of JD-T could be Yiseonggugo-hwan, Daesiho-tang, Jowiseungchung-tang, Heukno-hwan, and GH-T. The change of JD-T went for them throughout JHD-T to JD-T. JD-T in "Sinchukbon" consists of Galgeun and Daehwang which directly gets rid of severe heat from small intestine, and Jogak and Seungma which help to eliminate mild heat existing from small intestine throughout nose to flesh.

서브 마이크론급 구형 동분말의 볼 밀링을 통한 플레이크 동분말의 제조

김지환,이종현,Kim, Ji Hwan,Lee, Jong-Hyun 한국마이크로전자및패키징학회 2014 마이크로전자 및 패키징학회지 Vol.21 No.4

직경 수 마이크론급의 Ag 코팅 Cu 플레이크를 제조하기 위한 선행공정으로 습식 화학적 합성법으로 제조된 서브마이크론급의 Cu 입자를 볼 밀링 공정을 통해 프레이크화 하였다. 입자들의 산화 및 응집을 막기 위해 볼 밀링 유체로는 에틸렌글리콜을 사용하였고, 에틸아세테이트 표면개질제도 첨가하였다. 용기의 회전수에 따른 실험 결과를 통해 회전수에 따른 회전 모드의 변화가 밀링 후 Cu 입자들의 평균적인 형상과 형상 균일도를 크게 변화시킴을 확인할 수 있었다. 또한 첨가한 지르코니아 볼의 직경 역시 Cu 입자들의 플레이크화 균일도를 결정하는 대표 공정변수임을 확인할 수 있었다. 그 결과 다소간의 응집체를 포함한 서브마이크론급의 Cu 입자를 사용했음에도 불구하고, 회전수, 표면개질제 첨가량, 그리고 지르코니아 볼의 직경 등의 대표 공정변수들을 최적화한 볼 밀링 공정을 통해 분산성이 우수한 수 마이크론급의 Cu 플레이크를 성공적으로 제조할 수 있었다. As a preceding process for preparing several micrometer sized Ag-coated Cu flakes, ball milling of submicrometer-sized Cu particles synthesized through a wet chemical method was performed in order to convert the particles into flakes. To suppress oxidation and aggregation of the particles during ball milling, ethylene glycol and ethyl acetate were used as a medium and a surface modifying agent, respectively. Results obtained with different rotation speeds of a jar indicated that the rotation speed changes a rotating mode, and strikingly alters the final shapes and shape uniformity of Cu particles after milling. The diameter of zirconia ball was also confirmed. Although there was aggregates in the initial submicrometer-sized Cu particles, therefore, well-dispersed Cu flakes with a size of several micrometers were successfully prepared by ball milling through optimization of rotation speed, amount of ethyl acetate, and diameter of zirconia ball.