렘 31 : 1-34에 나타난 새언약 연구

강남현 총신대학 1984 국내석사

Penicillium verruculosum D-xylanase 遺傳子의 클로닝

강남현 全南大學校 1995 국내석사

Xylan함유 배지로 배양한 P.verruculosum 균사체로부터 total RNA를 분리 하고 PolyATtract mRNA Isolation System II를 이용하여 mRNA를 추출하였다(5.7㎍/wet weight mycelium). 이 mRNA를 template로 해서 ZAP-cDNA^R Synthesis Kit를 사용하여 first 및 second strand 를 합성하였고, 합성된 cDNA양 말단에 EcoR I 및 Xho I site를 부가하였다. 이 cDNA를 Sephacryl spin column을 통해 size fraction하여 0.6∼2.3kb 단편을 회수했다. 이 단편을 Uni-ZAP XR vector에 ligation한 후 Gigapack II Gold Packaging Extract를 사용해서 packaging했다. 이 Phage를 host E.coli XL-I Blue MRF' 에 도입시켜 cDNA library를 만들었다. Phage titering결과 298.350pfu였다. Xylanase 항체를 이용한 immunoscreening 방법을 통하여 3개의 클론을 얻었다. 이 phage DNA 와 ExAssit/SOLA System helper phage 를 사용하여 클론된 유전자가 들어있는 pBluescript SK를 E.coli SOLA cell에 도입시켜 LB-ampicillin plates (50㎍/ml)에서 pBluescript SK를 함유하는 colony를 확인했다. 이들 세포를 IPTG 에 의해 유도발현 시킬경우 분자량 35kDa 단백질이 현저히 생산되었으며 이 단백질을 Western immunoblotting 한 결과 anti-xylanase IgG와만 결합하였다. 이 SOLA cell에서 plasmids를 추출한 다음 EcoR I 과 Xho I 으로 처리하여 얻은 1.37kbp의 삽입단편의 제한효소 지도작성을 위해 Acc I, Hinc II, sal I, Sac I, Kpn I, 으로 처리한 결과 Eco RI 부위로부터 490,320,20,20,50,270 및 200bp 의 단편으로 절단되는 제한효소절단부위가 인정되었다. 따라서 본 연구결과 P. verruculosum의 xylanase(35KD)를 암호하는 1.37kb의 cDNA가 클로닝 되었음이 입증되었다. Total RNA was extracted from the Penicillium verrucosum xylanase-induced cultures grown for 4days by using guanidium salts method and mRNA(5.7㎍ per 1g of wet mycelium) was purified with PolyATtract mRNA Isolation System II. cDNA was synthesized according to the direction of ZAP-cDNA Synthesis kit and then attached Eco R I adaptors and digested with Xho I. The fractions of 0.6∼2.3kb cDNA were collected by Sephacryl spin column centrifugation. After ligation of the cDNA fraction to the Uni-ZAP XR vector, the ligation mixture was packaged into Gigapack II Gold Packaging Extract and introduced into E. coli XL1-Blue MRF'. Xylanase cDNA clones were screened by immunoscreening with anti-xylanase IgG as described by the Promega Protocol. Three positive clones of the Uni-ZAP XR cDNA library were detected and single plaque was picked by the second and third screening. E. coli SOLR cell containing pBluescript SK(-) DNA was obtained from Uni-ZAP XR phage clone using in vivo excision with ExAssit/SOLR System helper phage. The significant amounts of protein coded by the xylanase cDNA was produced by the IPTG induction and analysed by Western immunoblotting with positive signal band when anti-xylanase IgG was applied. The phagemid DNAs were isolated and digested with EcoR I and Xho I. The insert DNA of three clones were about 0.7kb. 0.7kb and 1.37kb long. The 1.37kb insert DNA in pIS 2-1 contained six recognition sites of Acc I(0.49kb and 0.81kb), Hinc II (0.83kb), Sal I (0.85kb), Sac I (0.90kb),. Kpn I(1.17kb) from the EcoR I site. From the results. therefore, it was proved that 1.37kb size of cDNA coding for xylanase(MW. 35KD) of P.verrucosum was cloned in E.coli.

구강보호장치의 재료적인 특성이 하악골 충격 시 악골 및 두부에 미치는 영향에 관한 유한요소분석

강남현 경희대학교 2006 국내박사

예비인상용 하악 트레이 제작을 위한 한국인 무치악자 치조제궁의 형태에 관한 연구

강남현 경희대학교 대학원 1999 국내석사

It is very important to make a correct impression for restoring edentulous arch. Accurate impressions are depend on prelimimary tray to fit edaitulous arches exactly. Although many kinds of tray have been used for Korean edentulous patients over the year, most of them are based on weste arch shape and size. The propose of this study is to evaluate the size and shape of Korean mandibular edentulous arcades and to classify their she and size based on several linear and angular measurement. For this study, to 100 patients mandibular mstar models( 50 male's models and & female's models ) were duplicated with irreversible hydrocolloid impression meterial to fabricate the study models. And the study models were treasured with 3-demensional measuring device. After measuring the models mandibular edentulous arches were classified by arch shape and size according to each measurement and then obtained data were analysed with student t-test and ANOVA test for the statistical anaysis. The results were as follows: 1. In the avenge linear measurements at reference points. There are no significant sexual differanes in the linear measurements except in TAP, AW3 and AW4. 2. Rt and Lt interalvolar ridge width on 4/4 reference line was more useful thai other linear measurements to classify the edentulous arch size because its range between minimum value and broadest. 3. The edentulous arch size classified into six arch size group ; group 1(below 48mm), group 2(48~52mm), group 3(52-56mm), group 4(56~60mm), group 5(60-64mm) and group 6(above 64mm), and each size distribution was 3.0% 3.0% 23.0% 51.0% 18.0% and 2.0% respectively. 4. In only 56 models of total 100 models, arch shape could be clearly classified by visual method into three shape groups(V-shape: 15. O-shape: 13 and U-shape: 28). 5. Also, in total 100 modelss, arch shape was classified into three shape groups(V-shape: 29, O-shape: 44 and U-shape: 27) by data of visual classification. 6. The third classification of arch shape was classified into four shape group(V shape- Tapered type: 23, V shape-straght type 18, Ovoid shape-tapsed type: 28 and Ovoid shape -Straight type: 28) by anterior classified measurments(AA, WIR WOR, WLIR and WLOR) and posterior classifying measurments(LA, AA3A, AA4A, W2R, W3R WL3R and WLAR). 24

Anti-obesity Effect of Trans-anethole and Trans-cinnamic Acid by Induction of Fat Browning

강남현 대구대학교 대학원 2019 국내석사

최근 백색 지방세포의 갈색지방화 (browning)와 갈색 지방세포의 활성화를 이용하는 방법이 비만 예방을 위한 좋은 전략이 되고 있다. 특히 갈색 지방화 활성이 있는 천연 화합물들을 이용한 항비만 치료제 개발이 활기를 띠고 있다. 본 연구에서는 다양한 식물의 필수 지방에 존재하는 향미 물질인 trans-anethole (TA)과 계피껍질에 존재하는 trans-cinnamic acid (tCA)이 백색 지방세포를 갈색 지방화 세포로 유도한다는 사실을 입증하였다. TA와 tCA는 백색 지방세포에서 갈색 지방 마커 단백질 (PGC-1α, PRDM16, UCP1)의 함량과 갈색지방화된 지방세포 (베이지 지방)의 특이 유전자 (Cd137, Cited1, Tbx1, Tmen26)의 발현 수준을 증가시키고, HIB1B 갈색 지방 세포에서 갈색 지방 특이 적 유전자 (Cidea, Lhx8, Ppargc1, Prdm16, Ucp1 및 Zic1)의 발현을 증가시켰다. 또한 TA와 tCA는 3T3-L1 백색 지방 세포의 지방 산화를 증가시키고 지방 생성을 감소시켰으며, HIB1B 갈색 지방 세포를 활성화시켰다. 또한 지방대사에서는 TA와 tCA가 β3-AR 및 AMPK 신호 전달 경로의 활성화를 통해 3T3-L1 백색 지방 세포의 갈색 지방화를 유도한다는 사실을 알아냈다. 특히, TA는 β3-AR뿐만 아니라 AMPK 매개체인 SIRT1 경로의 조절을 통해서도 갈색 지방화를 유도하였다. 결론적으로, TA와 tCA는 백색지방세포의 갈색화 전환과 동시에 갈색지방을 활성화시키는 기능에 의하여 에너지 대사를 크게 증가시킴으로써 비만 치료제로서의 가능성을 보여주었다. Browning of white adipocytes and activation of brown adipocytes suggests attractive strategy for prevention of obesity. The use of natural compounds for browning is regarded as a safe and new strategy for anti-obesity. Here, we report that trans-anethole (TA), a flavoring substance present in the essential oils of various plants and trans-cinnamic acid (tCA), a class of cinnamon from the bark of Cinnamomum cassia, induced browning in white adipocytes. TA and tCA increased protein content of brown-fat specific markers (PGC-1α, PRDM16, and UCP1) and expression levels of beige-fat-specific genes (Cd137, Cited1, Tbx1, and Tmen26) in 3T3-L1 white adipocytes, as well as brown-fat-specific genes (Cidea, Lhx8, Ppargc1, Prdm16, Ucp1, and Zic1) in HIB1B brown adipocytes. In addition, TA and tCA increased fat oxidation, reduced adipogenesis and lipogenesis in 3T3-L1 adipocytes, and activated HIB1B adipocytes. Further, mechanistic study revealed that TA and tCA induced browning of 3T3-L1 adipocytes by activating the β3-AR and AMPK signaling pathways. Specially, TA induced browning via regulation of not only β3-AR but also the AMPK-mediated SIRT1 pathway. Moreover, in vivo study on TA revealed its efficiency as a potent anti-obesity agent by alleviating thermogenesis, suggesting its potential to treat obesity and its related metabolic disorders.

Gravitational effects on weld pool shape and microstructural evolution during gas tungsten arc and laser beam welding on 304 stainless steel, Ni, and Al-4 wt.% Cu alloy

강남현 Pennsylvania State University The Graduate School 2003 해외박사

본 연구는 Ni 200 합금 (99.5% Ni), 304 stainless steel, Al-4Cu 합금에서의 레이저 및 Gas Tungsten Arc Welding(GTAW)시 중력의 영향을 조사하였다. 중력벡터의 크기와 방향에 따른 용접부 형상, 미세조직 및 solute distribution의 거동을 연구하였다. 중력실험은 미국항공우주국(NASA) KC-135 비행기 실험을 통하여 무중력에서 1.8g까지의 중력크기 실험을 수행하였고, 중력방향에 따른 용접방향을 변화시키기 위한 ground 실험을 병행하면서 중력방향의 영향을 조사하였다. 중력벡터의 방위에 따른 실험은 용접 속도와 중력벡터의 방향을 체계적으로 변화시키면서 다음 3가지 형태의 용접 방식이 행하여졌다: welding upward in the direction opposing gravity (vertical-up 또는 ll-U weld), welding downward in the direction of gravity (vertical-down 또는 ll-D weld), and welding perpendicular to the direction of gravity (perpendicular 또는 ⊥ weld). 중력의 크기 및 방향에 따른 용접부 형상의 거동을 조사하여 “Stable" 용접부와 "Unstable" 용접부로 나눌 수 있었다. 일반적으로 높은 아크전류의 translational GTAW 공정에서 낮은 전류의 spot GTAW 공정에 비하여 중력방향에 따른 용접부 형상의 변화가 심하였다. 용접부 단면적은 용접부 형상의 안정성을 결정하는데 있어서 제1인자가 되지는 못했다. 304 STS GTAW의 "Stable" 용접부의 경우, ll-U 용접부는 용접풀 하단에서 less convexity 및 more free surface depression 현상을 보였다. 이로 인하여 ll-D 용접부에 비하여 10-20% 깊은 용입을 ll-U 용접부에서 볼 수 있었다. 용접풀 가운데에서의 용입 변화 이외에는 전체적인 용접부 형상에서 중력의 영향은 크지 않았다. 용접부의 크기가 커질수록 중력방향에 따른 용접부 형상의 차이가 많이 발생하였으며 이러한 상태를 "Unstable" 용접부로 규정하였다. 304 STS GTAW의 경우 ll-D 용접부를 ll-U 용접부와 비교하면 31% 용입 증가 및 28% 용융폭 감소를 보였다. 이러한 용접부 형상의 변화는 free surface deformation 및 convection flows 현상이 중력 방향에 따라 다르게 반응을 하였기 때문으로 생각한다. 중력방향에 따른 미세조직 거동을 조사하기 위하여 합금의 주요성분의 밀도차가 큰 Al-Cu 합금을 선택하여 GTAW를 수행하였다. 니켈 및 304 STS 경우와 같이 용접부 형상은 중력방향의 영향을 많이 받음을 알 수 있었다. 용접부 형상이 변한다는 것은 용접풀 내부의 convection flows의 변화를 의미하는 것으로 미세조직의 크기 및 성장방향을 결정하는 응고조직 앞에서의, 즉 solidification front에서의 응고속도(Vs) 및 열구배(GL)에 영향을 준 것으로 조사되었다. 응고방향, 미세조직 형상 및 primary dendrite arm spacing(λ1)은 ll-D 용접부의 경우 ll-U 용접부에 비하여 다른 현상을 보였다. ll-D 용접부 미세조직은 용접풀 표면에서 columnar 구조를 보였고, Vs의 굴골부를 용접풀의 가운데에서 발견하였다. 이는 용접풀 free surface의 변화에 따른 solidification front의 영향으로 설명이 가능하였다. 이러한 용접부 형상 및 미세조직에 대한 중력의 영향을 연구한 결과는 International Space Station(ISS)를 건설하거나 파이프 용접 등 거대한 구조물과 같은 열악한 용접 환경시 용접부의 크기와 질을 조절할 수 있는 중요한 데이터가 될 수 있을 것이다. 또한 수치해석(Numerical analysis)을 통한 용접부 해석은 실험자의 눈으로 확인하기 힘든 물리적/화학적 이론 및 현상을 검증할 수 있을 뿐 아니라 예측까지 가능하게 하는 중요한 역할이 기대된다. The objective of the present work was to investigate effects of gravitational (acceleration) level and orientation on Ni 200 alloy (99.5% Ni purity), 304 stainless steel, and Al-4 wt.% Cu alloy during gas tungsten arc welding (GTAW) and laser beam welding (LBW). Main characterization was focused on the weld pool shape, microstructure, and solute distribution as a function of gravitational level and orientation. This was accomplished in part through NASA's KC-135 aircraft experiment to produce a range of the gravitational level (microgravity to 1.8 g_(o)) and through a ground-based experiment. Welding for the ground-based experiment was conducted by varying the welding direction with respect to the gravity vector, i.e., welding upward opposing gravity (Ⅱ-U weld) on a vertical weld piece, welding downward with gravity (Ⅱ-D weld) on a vertical weld piece, and welding perpendicular to gravity (⊥ weld) on a horizontal weld piece. The welds were divided into two classes, i.e., 'stable' and 'unstable' welds, in view of the variation of weld pool shape as a function of gravitational level and orientation. In general, higher arc current and translational GTAW produced more significant effects of gravitational orientation on the weld pool shape than the case of lower arc current and spot welding. Cross-sectional area (CSA) was a secondary factor in determining the stability of weld pool shape. For the 'stable' weld of 304 stainless steel GTAW, the Ⅱ-U weld showed less convexity in the pool bottom and more depression of the free surface, therefore producing deeper penetration (10-20%) than the case of Ⅱ-D weld. Except for the penetration in the weld pool center, overall weld pool shape remained mostly constant with respect to gravity. As the weld pool size became larger, the weld pool shape was changed significantly as a function of gravitational orientation, i.e., the 'unstable' weld. The Ⅱ-D weld of 304 stainless steel showed 31% deeper penetration, 28% narrower width, and more hemispherical shape of the weld pool than the case of Ⅱ-U weld. Variations of the weld pool shape were due to the different degree of free surface deformation and convection flows with respect to the gravitational orientation. For GTAW on 304 stainless steel, gravitational level variation from low gravity (LG≈1.2 g_(o)) to high gravity (HG≈1.8 g_(o)) caused 10% increase in width and 10% decrease in depth while maintaining the overall weld pool volume. Furthermore, LBW on 304 stainless steels showed mostly constant shape of weld pool as a function of gravitational orientation. These results were because the weld pool size was not large enough to exhibit the gravity effects on the weld pool shape. For the LBW, there were other reasons of insignificant gravity effects; ⅰ) The keyhole formation dominated the weld pool shape; ⅱ) The laser beam diameter was not significantly changed as a function of the direction perpendicular to the weld piece, while the diameter of gas tungsten arc might be changed significantly due to its diverging phenomenon from the small electrode to the large weld piece. GTAW on Ni showed similar trends of weld pool shape compared with GTAW on 304 stainless steel, i.e., the weld pool became unstable by showing more penetration in the Ⅱ-D weld for slower arc translational velocity (V_(α)) and larger weld pool size. However, the Ni weld pool shape had greater stability of the weld pool shape with respect to the gravitational orientation than the case of 304 stainless steel, i.e., higher current boundary and no humping. These results were due to the combination of greater surface tension/thermal diffusivity and smaller viscosity for Ni compared with the properties of 304 stainless steel. Regardless of the gravitational level, the ferrite content and the distribution of the solutes (Cr and Ni) remained constant for GTAW on 304 stainless steel. However, for GTAW on Al-4 wt.% Cu alloys, the gravitational orientation changed the weld pool shape associated with convection flows. This variation on the convection flows influenced the shape of the trailing solid-liquid (s-1) interface. Correspondingly, the solidification morphology and primary dendrite spacing (λ_(1)) were modified because the solidification rate (V_(S)) and thermal gradient (G_(L)) were affected by the convection flow. For this reason, larger λ_(1) for the Ⅱ-U weld was observed near the solid-liquid boundary and surface than that of the perpendicular and Ⅱ-D weld. The Ⅱ-D weld exhibited different solidification morphology, e.g., more columnar structure near the weld pool surface and abnormal 'S' shape of the solidification rate (V_(S)) during its growth. In summary, gravity influenced the weld pool shape that was associated with convection flows and weld surface deformation for specific welding conditions. The variation of convection flows and weld pool shape played a role in modifying (V_(S)) and (G_(L)). Solidification orientation and morphology were affected because (V_(S)) and (G_(L)) were changed as a function of gravity. Studies of gravity on the welding process are expected to play a significant role in the space-station construction and circumferential pipe welding on the earth.