저온온열치료에 의한 종양 내 저산소상태 개선효과를 18F-Fluoromisonidazole의 섭취 변화를 이용한 평가

이상욱(Sang-wook Lee),류진숙(Jin Sook Ryu),오승준(Seung Joon Oh),임기천(Ki Chun Im),천기정(Gi Jeong Chen),이소령(Ryung Lee),송도영(Do Young Song),임수정(Soo Jeong Im),문은숙(Eun Sook Moon),김종훈(Jong Hoon Kim),안승도(Seung Do Ahn),신 대한방사선종양학회 2004 Radiation Oncology Journal Vol.22 No.4

목 적: 종양 내에서 산소공급 부족현상으로 발생하는 저산소증 조직에서 저온온열치료(42oC)와 nicotinamide 에 의한 perfusion limited 저산소증의 개선 효과를 마우스 종양 모델을 이용하여 종양 내 [18F]FMISO 섭취변화 를 이용하여 증명할 수 있는지 알아보고자 하였다. 대상 및 방법: C3H 마우스에 [18F]FMISO를 정주하고 11개 장기에서 %ID/g을 구하여 biodistribution을 관찰하였다. 또한 같은 마우스에 동종 종양세포인 SCC7을 이식하여 종양모델을 만들고 저온온열치료(42oC)와 nicotinamide를 투여한 마우스와 대조군 마우스에서 [18F]FMISO의 섭취정도 차이를 %ID/g, autoradiography, PET scan을 시행하여 비교하고자 하였다. 결 과: 대조군에서 종양의 FMISO의 섭취는 5.1+/-2.28 %ID/g였고, 종양/근육, 종양/혈액의 섭취비는 2.2와 1.8이었다. 실험군에서는 각각 2.4+/-0.64 %ID/g, 1.4와 1.2를 나타내어 대조군보다 유의하게 낮았다(p<0.021). Autoradiography에서 대조군의 종양 내부에 FMISO가 섭취됨을 확인하였고, 저온온열치료와 nicotinamide를 투여한 실험군에서는 섭취가 감소된 것을 관찰하였다. 결 론: C3H 마우스와 동종 종양세포인 SCC-VII을 이용한 종양모델에서 [18F]FMISO가 종양내에 섭취가 되어 자산소증 종양모델로 적절함을 확인하였고, 저온온열치료(42oC)와 nicotinamide에 의한 perfusion limited 저산소증 개선효과를 [18F]FMISO의 종양 내 섭취가 감소하는 것을 통하여 확인할 수 있었다. Purpose: The aims of this study were to evaluate the change of [18F]fluoromisonidazole ([18F]FMISO) uptake in C3H mouse squamous cell carcinoma-VII (SCC-VII) treated with mild hyperthermia (42oC) and nicotinamide and to assess the biodistribution of the markers in normal tissues under similar conditions. Methods and Materials: [18F]FMISO was producedby our hospital. Female C3H mice with a C3H SCC-VII tumor grown on their extremities were used. Tumors were size matched. Non-anaesthetized, tumor-bearing mice underwent control or mild hyperthermia at 42oC for 60 min with nicotinamide (50 mg/kg i.p. injected) and were examined by gamma counter, autoradiography and animal PET scan 3 hours after tracer i.v. injected with breathing room air. The biodistribution of these agents were obtained at 3 h after [18F]FMISO injection. Blood, tumor, muscle, heart, lung, liver, kidney, brain, bone, spleen, and intestine were removed, counted for radioactivity and weighed. The tumor and liver were frozen and cut with a cryomicrotome into 10-μm sections. The spatial distribution of radioactivity from the tissue sections was determined with digital autoradiography. Results: The mild hyperthermia with nicotinamide treatment had only slight effects on the biodistribution of either marker in normal tissues. We observed that the whole tumor radioactivity uptake ratios were higher in the control mice than in the mild hyperthermia with nicotinamide treated mice for [18F]FMISO (1.56±1.03 vs. 0.67± 0.30; p=0.063). In addition, autoradiography and animal PET scan demonstrated that the area and intensity of [18F]FMISO uptake was significantly decreased. Conclusion: Mild hyperthermia and nicotinamide significantly improved tumor hypoxia using [18F]FMISO and this uptake reflected tumor hypoxic status.

Autoradiographic Verification of Transdermal Penetration of Oleic Acid-conjugated Peptide Nanosomes

이경은,정민교,엄재훈,정세희,하권수,박정해,이진성,한성식,최면,Lee, Kyung-Eun,Jung, Min-Kyo,Eum, Jai-Hoon,Jung, Se-Hui,Ha, Kwon-Soo,Park, Jeong-Hae,Lee, Jin-Sung,Han, Sung-Sik,Choe, Myeon Korean Society of Electron Microscopy 2010 Applied microscopy Vol.40 No.4

Short 펩타이드는 약용화장품(cosmeceuticals)으로 가능성이 있지만, 펩타이드의 친수성 성질 때문에 피부를 통한 전달이 어렵다. 따라서 이러한 short 펩타이드를 약용화장품으로 이용하기 위해서는 피부를 통과할 수 있는 성질로 바꾸어야만 한다. 이 연구에서는 프로콜라겐 type I의 신호펩타이드로 작용할 수 있는 KTTKS의 5개 펩타이드에 양친매적인 특성을 부여하기 위해 올레산을 결합시켰고, 그 후 나노좀으로 제조하였다. 제조된 나노좀은 동결투과전자현미경법을 이용하여 크기가 100에서 200 nm 사이의 리포좀의 형태임이 확인되었으며, 제타전위측정장치(ELS)를 통해 24주 동안 안정적인 크기를 유지하고 있음이 확인되었다. 이렇게 제조된 나노좀을 피부에 바른 후 자가방사기록법(autoradiography)을 이용하여 피부에 침투 여부를 확인하였다. 이 5개 펩타이드의 친수성 성질로 인하여 펩타이드만 피부에 도말하였을 때는 피부에 침투되지 않았으나, 펩타이드와 올레산을 결합하여 나노좀으로 제조된 경우에는 피부의 진피층까지 잘 도달한 것을 확인하였다. 이 연구를 통해 자가방사법이 피부를 통한 효과물질의 전달을 검증할 수 있는 좋은 방법임을 보여줌과 동시에 피부에 효과를 나타내는 친수성펩타이드의 피부를 통한 전달에 올레산 결합을 이용한 나노좀 형태가 효과적인 것을 보여주었다. Short peptides are potentially effective materials as cosmeceuticals, but their delivery across the skin can be problematic due to the ionic nature of peptides and the structure of the skin. For short peptide to be utilized as cosmeceuticals, its ability to penetrate the skin must be altered. In this study, we conjugated the widely used procollagen type I signal peptide, KTTKS, with oleic acid to improve the lipophilic properties of the peptide, and used the oleic acid-conjugated peptides to construct cosmeceutical nanosomes. Then we examined the penetration of cosmeceutical nanosomes prepared from isotope-labeled peptide into the skin after transdermal application using autoradiography. Because of its hydrophilic property of penta-peptide, the penta-peptide itself was not able to be penetrated through the stratum corneum of the skin. In contrast, nanosomes made from olecic acid conjugated penta-peptide were able to be penetrated through the stratum corneum effectively. Autoradiography showed the precise penetration points to dermal layer, demonstrating the appropriateness of this method for clarifying the mechanism of penetration of transdermal delivery systems.

Autoradiography를 이용한 자외선 조사 후 생쥐 표피세포의 DNA 합성에 관한 연구

신준우(Jun Woo Shin),김규한(Kyu Han Kim),윤재일(Jai Il Youn) 대한피부과학회 1990 대한피부과학회지 Vol.28 No.6

The status of DNA synthesis and the effect of UV radiation on the DNA synthesis were studied in mouse skin by microautoradiography. Mice exposed to 100mJ/cm of UVB were injected intradermally with tritiated thymidine, 5 minutes, 2, 6 and 24 hours after irradiation and biopsies were processed for light microscopic autoradiography, A total of 25 ICR female albino haired mice were used as subjects. We compared heavily labeled cells(>10 grains/nucleus) and sparsely labeled cells (3-10 grains/nucleus) in UVL irradiated skin with nonirradiated control skin. 1. Within 5 minutes after UVL exposure an apparent depression in the number of heavily labeled cells occurred. This reduction was statistically significant and remained so for at least six hours post,irradiation. By 24 hours after UVL exposure, the reduction was recovered to nearly control level. 2. By five minutes after UVL exposure, sparsely labeled cells were observed in basal cell layer and differentiated cell layer. The nurriber of sparsely labeled cells appeared highest at five minutes after UVL exposure and then tended to fall to nearly control level at 24 hours. This aberrant type of TdR H incorporation is thought to represent unscheduled DNA synthesis to repair pyrimidine dimers formed in UVL injured DNA molecules, (Kor J Dermatol 28(6):677 685, 1990)

實驗的 齒牙移動後 膠原酵素 投與가 齒根膜 纖維의 變化에 미치는 影響에 關한 自己放射法的 硏究

洪性埈,徐廷勳 대한치과교정학회 1990 대한치과교정학회지 Vol.20 No.2

The purpose of this study was to analyze the reorganization of periodontal ligament after collagenase treatment with autoradiography. The author compared the collagenase-treated experimental group and no-treated experimental group with control group. Fourty eight Sprague-Dawley rats were divided into nine groups, including normal control and immediate group. Closed coil springs were used between the upper incisors and the first molars with 100 grams. Collagenase and ³H-proline were adminstered and the samples were sacrificed and sectioned. After being dipped into the NTB-3 emulsion the samples were analyzed with light microscope under H/E stain. Data were analyzed by t-test and ANOVA. The results were as follows: 1) Generally collagenase-treated groups got more ³H-proline uptake than no-treated groups. 2) Compared with normal control group, collagenase-treated group had the same ³ H-proline uptake in amount at 21th day. 3) Among cemento-enamel junction, middle, apex areas, cemento-enamel junction area of collagenase-treated group arrived at normal control level earlier than no-treated group. 4) Cemento-enamel junction area had the most ³H-proline incorporation amount in no-treated group, but apex area had the most in collagenase group.

A STUDY ON THE BEHAVIOR OF BORON DISTRIBUTION INLOW CARBON STEEL BY PARTICLE TRACKING AUTORADIOGRAPHY

DONG JUN MUN,EUN JOO SHIN,구양모 한국원자력학회 2011 Nuclear Engineering and Technology Vol.43 No.1

The behavior of the non-equilibrium grain boundary segregation of boron in low carbon steel was studied through a particletracking autoradiography. The behavior of the non-equilibrium grain boundary segregation of boron during continuous coolingwas compared with the isothermal kinetics of the non-equilibrium grain boundary segregation of boron at the holdingtemperature using an effective time method. On the basis of the experiments, the cooling rate dependence of the non-equilibriumsegregation of boron was explained using the time dependence of the non-equilibrium segregation of boron in low carbonsteel. The experimental observations for the cooling rate dependence of the grain boundary segregation of boron are in goodagreement with the time dependence of the grain boundary segregation of boron. The mechanisms of the non-equilibriumsegregation of boron during cooling in low carbon steel are also discussed.

A STUDY ON THE BEHAVIOR OF BORON DISTRIBUTION IN LOW CARBON STEEL BY PARTICLE TRACKING AUTORADIOGRAPHY

Mun, Dong-Jun,Shin, Eun-Joo,Koo, Yang-Mo Korean Nuclear Society 2011 Nuclear Engineering and Technology Vol.43 No.1

The behavior of the non-equilibrium grain boundary segregation of boron in low carbon steel was studied through a particle tracking autoradiography. The behavior of the non-equilibrium grain boundary segregation of boron during continuous cooling was compared with the isothermal kinetics of the non-equilibrium grain boundary segregation of boron at the holding temperature using an effective time method. On the basis of the experiments, the cooling rate dependence of the non-equilibrium segregation of boron was explained using the time dependence of the non-equilibrium segregation of boron in low carbon steel. The experimental observations for the cooling rate dependence of the grain boundary segregation of boron are in good agreement with the time dependence of the grain boundary segregation of boron. The mechanisms of the non-equilibrium segregation of boron during cooling in low carbon steel are also discussed.

초파리 단안의 발생시기에 따른 각막형성세포의 형태

윤춘식,Yoon, Chun-Sik 한국현미경학회 1999 Applied microscopy Vol.29 No.2

초파리 단안의 발생단계에 따른 각막형성세포의 형태적인 변화를 관찰하고, 생산된 단백질의 이동방향을 조사하였다. 그 결과 우화 후 수일이 지나면서 각막형성세포는 그 기능이 급격하게 감소하는 것으로 보인다. 반면 우화 직후의 각막형성세포는 왕성한 분비활동을 하는 것으로 생각된다. 특히 자기방사섭의 결과에서는 각막형성세포에서 생산된 대부분의 단백질이 랜즈쪽으로 가는 것을 볼 때, 이 시기에서 이들 세포는 시세포에 기능하기보다는 렌즈형성에 주 기능을 할 것으로 사료된다. 번데기시기의 각막형성세포는 더욱 잘 발달되어 있었다. 렌즈가 형성되고 있는 시기의 각막형성세포에는 rER이 잘 발달되어 물질생산이 왕성하게 일어나고 있음을 보여주었다. 또한 분비과립과 미세소관이 많이 관찰되는데, 후자는 물질이 이동하는 통로를 제공하기 때문이다. 발생도중의 단안에서 새롭게 만들어진 렌즈의 가장 아래층의 저밀도의 불연속적 라멜라구조를 이루고 있고, 이 부분의 각막형성세포는 미세융모가 잘 발달되어 있어 물질을 분비하는 조직의 일반적인 특징을 보여준다. 결론적으로 각막형성세포는 렌즈가 왕성하게 형성되는 시기에는 잘 발달하고, 우화 후 완전히 형성된 추에는 그 기능이 급격하게 감소하면서 세포의 크기도 작아진 다는 것을 알 수 있었다. Morphological changes of Drosophila ocellar corneagenous cells were studied to the development with electron microscopy, and the movement of produced proteins was traced with autoradiography. Corneagenous cells of immediate postemergence showed very active secretion pattern. However, a few days after the emergence, the secretory activity of corneagenous cell was supposed to be dropped suddenly. In autoradiography, almost of proteins that produced by corneagenous cells moved toward lens. From this, it was supposed that the corneagenous cells do not function in photoreceptor cells rather in the formation of lens at the postemergence stage. Corneagenous cells of pupal stage were well developed. In the period of lens formation, rER of corneagenous cells were well developed and it suggested very active material metabolism. Granules and microtubules were also frequently observed and the later would be a pathway of the movement of materials. In conclusion, corneagenous cells were well developed at vigorous lens forming stage. After emergence, when the lens formation was completed, both the function and the size of corneagenous cells were reduced.

흰쥐 뇌의 신경흥분성 아미노산 수용체 개체발생에 대한 자가방사법적 연구

이혜성 대한해부학회 1991 Anatomy & Cell Biology Vol.24 No.3

Localization and Function of a Storage Protein in Testes of Summer Diapause Males in Tobacco Budworm, Heliothis virescens

Seo, Sook-Jae,Kim, Chang-Hwan 한국곤충학회 1992 Entomological Research Vol.22 No.1

담배나방의 사육조건을 낮시간 동안은 41$^{\circ}C$, 밤시간 동안 26$^{\circ}C$로 번갈아 조정함으로써 고온휴면(夏眼)을 유도하였다. 전기영동과 autoradiograph에 의해 정상온도에서 사육한 숫컷과 하면을 유도한 조건에서 사육한 숫컷의 저장단백질(82kd) 합성과 양적변화를 관찰한 결과 하면에 들어간 숫컷에서는 정자발육이 정지되고, 반면 저장단백질 합성은 대조구에 비해 증가하였다. 면역세포화학적 방법으로 합성된 저장단백질의 정세포내 위치를 추적한 결과 하면에 들어간 숫컷에서는 정세포의 핵에 축적되었다. 이와 같은 결과들로 보아 하면에 들어간 개체의 정소에서 아마도 저장단백질은 열쇼크단백질과 유사한 작용을 하여 정자발육을 정지시키는데 관여하는 것으로 추측된다. Tobacco budworm, Heliothis virescens, pupal diapause was induced by exposing fourth instar larvae through pupae to fluctuating temperature between 26$^{\circ}C$ and 41$^{\circ}C$. The synthesis and quantitative changes of a storage protein (Mr 82 Kd) in testes during development were determined using SDS-PAGE and autoradiography. The testes of diapause males, in which sperm development ceased, showed a high level of storage protein synthesis. By immunocytochemical method it was revealed that the synthesized storage protein was accumulated in the nucleus of spermatocyte of diapause male. These observations suggest that, in testes of summer diapause male, storage protein may be related to the arrest of sperm development by acting like heat shock protein or by assisting heat shock protein function.

탈수흰쥐에서 심방특수과립의 분비에 관한 전자현미경자기방사법적 연구

신해영(Hae-Young Shin),송창호(Chang Ho Song),김형태(Hyoung-Tae Kim),한의혁(Eui-Hyeog Han),임재원(Jae-Won Lim),이무삼(Moo Sam Lee) 대한해부학회 2004 Anatomy & Cell Biology Vol.37 No.3

이 연구는 탈수가 ANP의 합성과 분비 그리고 분비과정에 미칠 수 있는 영향을 전자현미경자기방사법을 이용하여 알아봄 으로써 심방의 내분비기능을 좀더 잘 이해하고자 시도되었다. 체중 50 g 정도의 Sprague-Dawley 계 수컷흰쥐를 대조군과 탈수 1일, 3일군으로 나누고, 각 실험군을 L-leucine3H 주입 후조직을 채취할 때까지 시간인 20분, 60분, 240분으로 다시 나누었다. 채취한 조직을 전자현미경 시료제작 과정을 거쳐 Epon 812에 포매한 다음 초박절편한 조직을 전자현미경자기방사법 과정을 거쳐 전자현미경으로 관찰하였다. 전자현미경관찰 결과 : 탈수동물의 심방특수과립 (ANP 함유과립)은 정상동물에서와 달리 크기가 다양하고 과립바탕질의 전자밀도가 낮았으며 윤곽이 뚜렷하지 않은 과립도 관찰되었다. 전자현미경자기방사법 결과 : 심방조직으로 들어간 L-leucine3H의 위치를 나타내는 은입자는 굽은 실 또는 실토리 모양으로 보였다. 이 은입자는 심방특수과립을 포함한 세포포함물과 세포소기관, 세포사이공간, 혈관에서 관찰되었다. 대조군에서는 L-leucine3H를 주입하고 20분 후에 골지장치의 방사능치가 높았고, 과립세포질세망과 심방특수과립의 방사 능치는 60분에 높았다. 그리고 세포막과 혈관의 방사능치는 240분 후에 높게 나타났다. 탈수 1일군과 3일군에서는 골지장치와 심방특수과립, 세포막, 세포사이공간의 방사능치는 20분 후에 높았고, 과립세포질세망과 혈관은 60분에 높게 나타났다. 심방특수과립의 동태 : 심방특수과립의 양은 대조군에 비해 탈수 1일군에서는 34.1%, 탈수 3일군에서는 27.4% 증가하였다. 또한 탈수 3일군에서 새로 합성된 심방특수과립의 양은 동위원소 주입 후 20분에 전체 심방특수과립의 85.02%이고, 60분에는 36.87%, 240분에는 20.45%로 급격하게 감소하였다. 이상의 결과로 탈수동물에서 심방근육세포 속에서는 ANP 합성이 매우 활발하게 일어나고 새로 합성된 ANP의 분비도 활 발하지만, 탈수 동안 심방내강으로 분비되는 ANP의 양이 줄어들고 세포사이에 축적되고 머무는 양과 시간이 증가한다는 것을 알 수 있었다. This study was performed to investigate for the effect of dehydration on the synthesis, secretion and secreted pathway of atrial specific granules contained ANP by electron microscopic autoradiography. Male Sprague-Dawley rats, body weigh of about 50 g (range 47 to 53 g), were divided into control, 1 day dehydration and 3 days dehydration groups. Each group was divided into four groups according to sacrificed time on 20 min, 60 min and 240 min after the injection of L-leucine3H. Tissues of the right atrium obtained from animals were processed for typical electron microscopic procedure, and then embedded in Epon 812. Ultrathin sections were followed for electron microscopic autoradiographic method. Atrial specific granules were various in size, and some granules had a lower electron densities and indistinct granular membrane in the dehydration groups compared with the control group. In the electron microscopic autoradiographs of atrial wall, silver grains indicated by means of the positions of labelled L-leucine3H over the cell inclusion included atrial specific granules, cell organelles, intercellular spaces and blood vesseles. In the control group, high specific radioactivity was observed in the Golgi apparatus at 20 min, and in the rough endoplasmic reticulums and atrial specific granules at 60 min after the injection of L-leucine3H. And high level of radioactivities were observed in the cell membranes and blood vesseles at 240 min after the injection of L-leucine3H. In the 1 day and 3 days dehydration groups, radioactivities of Golgi apparatus, atrial specific granules, cell membranes and intercellular spaces were high level at 20 min, and radioactivities of rough endoplasmic reticulums and blood vesseles were high level at 60 min after isotope injection. Stored atrial specific granules were increased to 34.1% in the 1 day dehydration group, 27.4% in the 3 days dehydration group compared with the control group. In the 3 days dehydration group, newly formed granules increased 85.02% at 20 min, but those decreased rapidly to 36.87% at 60 min, 20.45% at 240 min after the injection of Lleucine3H in atrial cardiocytes. This results suggest that total ANP increased rapidly in the atrial cardiocytes, and newly formed ANP secreted rapidly into the intercellular space in the condition of dehydration, and ANP from atrial cardiocytes remain in intercellular space for dehydration period.