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Production of a Monoclonal Antibody to Human $\alpha$-Fetopotein and Development of Monoclonal Antibody-Based Enzyme-Linked Immunosorbent Assays for Human $\alpha$-Fetoprotein (인간 $\alpha$-fetoprotein에 대한 모노클로날 항체의 제조 및 모노클로날 항체를 이용한 효소면역분석법의 개발)

  • Michung Yoon;Hyun-Hee Lee;Youngwon Lee
    • Biomedical Science Letters
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    • v.5 no.1
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    • pp.1-10
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    • 1999
  • This study was attempted to generate a monoclonal antibody against human $\alpha$-fetoprotein (AFP) and to produce an immunoassay, recognizing AFP in plasma and amniotic fluid. AFP was purified from human amniotic fluid and used to immunize mice. Spleens were taken from the mice and the cells were fused with mouse myeloma cells (Sp2/0-Ag-14) for the production of monoclonal antibodies by employing the hybridoma technology. As a result, a hybridoma cell line producing anti-AFP monoclonal antibody was cloned out and designated as MabF22. From isotyping analysis, it was found that monoclonal antibody MabF22 was IgG type with IgG1 heavy chain and k light chain. The binding specificity of MabF22 was analyzed by immunoblotting as well as by ELISA. MabF22 was highly specific, reacting with only AFP-containing samples. The binding affinity was determined by ELISA (free-capture mode) and Scatchard analysis. As a result, the value of Kd was 0.8$\times$10$^{-10}$M. The validity of the MabF22 for AFP assay was examined by two kinds of ELISAs, i.e., non-competitive and competitive ELISA. Both assays revealed that MabF22 reacted well with AFP in sample in a concentration-dependent manner. Standard curve and antibody titration curve were obtained by using purified AFP and MabF22. These results indicate that the monoclonal antibody produced in this study would be useful not only for research purposes but also for further development of immune-diagnostic kit for the measurement of AEP concentration.

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Data Qualification of Optical Emission Spectroscopy Spectra in Resist/Nitride/Oxide Etch: Coupon vs. Whole Wafer Etching

  • Kang, Dong-Hyun;Pak, Soo-Kyung;Park, George O.;Hong, Sang-Jeen
    • Proceedings of the Korean Vacuum Society Conference
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    • 2012.02a
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    • pp.433-433
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    • 2012
  • As the requirement in patterning geometry continuously shrinks down, the termination of etch process at the exact time became crucial for the success in nano patterning technology. By virtue of real-time optical emission spectroscopy (OES), etch end point detection (EPD) technique continuously develops; however, it also faced with difficulty in low open ratio etching, typically in self aligned contact (SAC) and one cylinder contact (OCS), because of very small amount of optical emission from by-product gas species in the bulk plasma glow discharge. In developing etching process, one may observe that coupon test is being performed. It consumes costs and time for preparing the patterned sample wafers every test in priority, so the coupon wafer test instead of the whole patterned wafer is beneficial for testing and developing etch process condition. We also can observe that etch open area is varied with the number of coupons on a dummy wafer. However, this can be a misleading in OES study. If the coupon wafer test are monitored using OES, we can conjecture the endpoint by experienced method, but considering by data, the materials for residual area by being etched open area are needed to consider. In this research, we compare and analysis the OES data for coupon wafer test results for monitoring about the conditions that the areas except the patterns on the coupon wafers for real-time process monitoring. In this research, we compared two cases, first one is etching the coupon wafers attached on the carrier wafer that is covered by the photoresist, and other case is etching the coupon wafers on the chuck. For comparing the emission intensity, we chose the four chemical species (SiF2, N2, CO, CN), and for comparing the etched profile, measured by scanning electron microscope (SEM). In addition, we adopted the Dynamic Time Warping (DTW) algorithm for analyzing the chose OES data patterns, and analysis the covariance and coefficient for statistical method. After the result, coupon wafers are over-etched for without carrier wafer groups, while with carrier wafer groups are under-etched. And the CN emission intensity has significant difference compare with OES raw data. Based on these results, it necessary to reasonable analysis of the OES data to adopt the pre-data processing and algorithms, and the result will influence the reliability for relation of coupon wafer test and whole wafer test.

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Effect of the supplementation of Coconopsis lanceolata extract on lipid metabolism amelioration in type 2 diabetes mouse model induced by high fat diet (울릉도 섬더덕 추출물의 급여가 제2형 당뇨 동물의 지질대사 개선에 미치는 영향)

  • Yun, Won-Kap;Bae, Hyun-Ji;Kim, You-Jeong;Kwon, O Jun;Im, Moo Hyeog;Cho, Hyun Duk;Kim, Taewan
    • Food Science and Preservation
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    • v.21 no.1
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    • pp.107-113
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    • 2014
  • The present study was designed to investigate the antihyperlipidemic effect of Coconopsis lanceolata extract in C57BL/6J mice. The mice were divided into four groups: normal diet group (ND), high fat diet group (HFD), positive control group with 0.05% metformin (PC), Coconopsis lanceolata extract group (UCL). After 5 weeks of feeding, average body weight of the UCL group mice was slightly decreased, while that of the HFD group significantly increased) Also, liver and adipose tissue weights in the UCL group significantly increased. The levels of trigliceride (TG) and total in the plasma of UCL-supplemented group were significantly lower than those of high fat diet group. On the other hand, HDL-cholesterol level was increased. Expression level of proteins related with adipogenesis such as SREBP-1c, ACC, and FAS in the liver of the UCL group mice was much lower comparing with the HFD group mice. In conclusion, the results showed that the Coconopsis lanceolata extract possesses significant antihyperlipidemic effects in C57BL/6J mice.

Effects of Calcium Citrate Isolated from Oyster Shell on Papain-induced Osteoarthritis in C57BL/6J Mice (굴 패각 구연산칼슘이 Papain으로 유도된 골관절염 C57BL/6J Mice에 미치는 영향)

  • Choi, Eun-Young;Kim, Hak-Ju;Han, Ji-Sook
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.10
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    • pp.1510-1518
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    • 2014
  • This study investigated the effects of calcium citrate on papain-induced osteoarthritis in C57BL/6J mice. Osteoarthritis was induced by injecting $6{\mu}L$ of papain into the knee joints of mice. Calcium citrate was made by crushing the centrifuged precipitate after reacting 0.5 M citric acid with 1 kg of oyster shell extract. The mice were divided into five groups (n=8). The normal group was untreated, whereas the papain group was induced to have osteoarthritis and treated with $200{\mu}L$ of water per day. The papain+DS group was treated with diclofenac sodium. The papain+calcium citrate groups were treated with calcium citrate at 150 and 300 mg/kg/bw for 28 days. Proteoglycan contents in articular cartilages were measured by safranin O/fast green staining and hematoxylin & eosin staining. Histopathological changes in cartilages were analyzed by the Rudolphi score approach. Contents of pro-inflammatory cytokines including TNF-${\alpha}$, IL-$1{\beta}$ and IL-6 in plasma, were measured by the ELISA method. Body weights among the treated groups were not significantly different compared with that of the normal group. Cartilage loss and joint instability in the calcium citrate group improved significantly (P<0.05) in a dose-dependent manner compared with the papain group. Further, proteoglycan content of the calcium citrate group was considerably (P<0.05) higher than that of the papain group. Osteoarthritis scores in the calcium citrate group were considerably (P<0.05) reduced compared with the papain group. In the group treated with calcium citrate, contents of TNF-${\alpha}$, IL-$1{\beta}$ and IL-6 in plasma were significantly (P<0.05) reduced in a dose-dependent manner in comparison with the normal group. Based on these results, we suggest that calcium citrate is effective for treatment of osteoarthritis.

Effects of Oyster Shell Extract on Papain-induced Osteoarthritis in C57BL/6J Mice (굴 패각 추출물이 Papain으로 유도된 골관절염 C57BL/6J Mice에 미치는 영향)

  • Lee, Se-Young;Kim, Hak-Ju;Han, Ji-Sook
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.8
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    • pp.1183-1189
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    • 2013
  • We investigated the effects of oyster shell extract (OSE) on papain-induced osteoarthritis in C57BL/6J mice. Osteoarthritis was induced in mice by a papain injection into the knee joint. The mice were divided into a total of five groups (n=8). The normal group was untreated, whereas the papain group (negative control) was induced with osteoarthritis and treated with water daily. The papain+DS group (positive control) was treated with diclofenac sodium. Papain+OSE groups were treated with OSE concentrations of 100 and 200 mg/kg/bw for 20 days. Proteoglycan content in articular cartilage was analyzed through safranine-O fast green staining and H&E staining. The histopathological changes in cartilage were measured by the Rudolphi score approach. The contents of tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), interleukin (IL)-$1{\beta}$, and IL-6 in plasma were analyzed by the ELISA method. After experiments, body weights of the treated groups were not significantly different compared with the normal group. Cartilage loss and joint instability significantly improved in a dose-dependent manner in the OSE-treated group compared with the papain group (P<0.05). Proteoglycan content was significantly higher in the OSE-treated group than the papain group (P<0.05). Osteoarthritis scores of the OSE-treated group were significantly decreased compared with the papain group (P<0.05). TNF-${\alpha}$, IL-$1{\beta}$, and IL-6 content in the plasma of the papain+OSE treated groups significantly decreased in a dose-dependent manner compared with the papain group (P<0.05). These results suggest that OSE treatment might have anti-arthritic effects on papain-induced osteoarthritis in C57BL/6J mice.

Pharmacokinetic Profiles of Isoniazid and Rifampicin in Korean Tuberculosis Patients (한국인 결핵환자에서 Isoniazid와 Rifampicin의 약동학)

  • Ahn, Seok-Jin;Park, Sang-Joon;Kang, Kyeong-Woo;Suh, Gee-Young;Chung, Man-Pyo;Kim, Ho-Joong;Kwon, O-Jung;Rhee, Chong-H.;Cha, Hee-Soo;Kim, Myoung-Min;Choi, Kyung-Eob
    • Tuberculosis and Respiratory Diseases
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    • v.47 no.4
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    • pp.442-450
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    • 1999
  • Background : Isoniazid(INH) and rifampicin(RFP) are the most effective anti-tuberculosis drugs which make the short-course chemotherapy possible. Although prescribed dosages of INH and RFP in Korea are different from those recommended by American Thoracic Society, there has been few study about pharmacokinetic profiles of INH and RFP in Korean patients who receive INH, RFP, ethambutol(EMB) and pyrazinamide(PZA) simultaneously. Methods : Among the patients with active tuberculosis from Dec. 1997 to July 1998, we selected 17 patients. After an overnight fast, patients were given INH 300mg, RFP 450mg, EMB 800mg and PZA 1500mg daily. Blood samples for the measurement of plasma INH(n=15) and RFP(n=17) level were drawn each at 0, 0.5, 1, 1.5, 2, 4, 6, 8 and 12hrs, and urine was also collected. INH and RFP level in the plasma and the urine were measured by high-performance liquid chromatography(HPLC). Pharmacokinetic parameters such as peak serum concentration(Cmax), time to reach to peak serum concentration(Tmax), half-life, elimination rate constant(Ke), total body clearance(CLtot), nonrenal clearance(CLnr), and renal clearance(CLr) were calculated. Results : 1) Pharmacokinetic parameters of INH were as follows: Cmax; $7.63{\pm}3.20{\mu}g/ml$, Tmax; $0.73{\pm}0.22hr$, half-life; $2.12{\pm}0.84hrs$, Ke; $0.83{\pm}0.15hrs^{-1}$, CLtot; $17.54{\pm}8.89L/hr$, CLnr; $14.74{\pm}8.35L/hr$, CLr; $2.79{\pm}1.31L/hr$. 2) Pharmacokinetic parameters of RFP were as follows: Cmax; $8.93{\pm}3.98{\mu}g/ml$, Tmax; $1.76{\pm}1.13hrs$, half-life; $2.27{\pm}0.54hrs$, Ke; $0.32{\pm}0.08hrs^{-1}$, CLtot; $14.63{\pm}6.60L/hr$, CLr; $1.04{\pm}0.55L/hr$, CLnr; $13.59{\pm}6.21L/hr$. 3) While the correlation between body weight and Cmax of INH was not statistically significant (r=-0.514, p value>0.05), Cmax of RFP was significantly affected by body weight of the patients(r=-0.662, p value<0.01). Conclusion : In Korean patients with tuberculosis, 300mg of INH will be sufficient to reach the ideal peak blood level even in the patients over 50kg of body weight However, 450mg of RFP will not be the adequate dose in the patients who weigh over 50~60kg.

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Effects of Dietary Germanium Biotite in Weaned, Growing and Finishing Pigs (이유자돈, 육성돈 및 비육돈에 있어 게르마늄흑운모의 급여 효과)

  • Kwon, O.S.;Kim, I.H.;Hong, J.W.;Lee, S.H.;Jung, Y.K.;Min, B.J.;Lee, W.B.;Shon, K.S.
    • Journal of Animal Science and Technology
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    • v.45 no.3
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    • pp.355-368
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    • 2003
  • In Exp. 1, this study was conducted to determine the effect of dietary germanium biotite on growth performance and nutrient digestibility in nursery pigs. A total of sixty crossbred pigs (initial body weight 15.09$\pm$0.18kg) were used in this experiment. This study was carried out for 28 days. The five treatments were control (CON; basal diet), GB0.1 (basal diet + germanium biotite 0.1%), GB0.3 (basal diet + germanium biotite 0.3%), GB0.6 (basal diet + germanium biotite 0.6%) and GB1.0 (basal diet + germanium biotite 1.0%). For overall period, ADG and Gain/feed were not significantly different among the treatments. In Exp. 2, a study was conducted to evaluate the effect of germanium biotite as a substitute for antibiotics in growing pigs. A total of fifty five crossbred pigs (initial body weight 32.47$\pm$0.9kg) were used in this experiment. The three treatments were negative control (NC: basal diet without antibiotic), positive control (PC: basal diet + 200ppm CTC) and GB0.3 (basal diet + germanium biotite 0.3%). Pigs fed PC (17%, 385 vs 451 g/d) and GB0.3 (14%, 385 vs 438 g/d) diets grew faster(P<0.05) than pigs fed NC diet. Pigs fed PC and GB0.3 diets resulted higher(P<0.05) ADFI than pigs fed CON diet. However, pigs fed GB0.3 diet had improved gain/feed compared to pigs fed NC diet(P<0.05). Apparent digestibility of DM and N by pigs fed PC and GB0.3 diets were greater(P<0.05) than those by pigs fed NC diet. In Exp. 3, a study was conducted to determine the effect of dietary germanium biotite on growth performance, plasma characteristics, backfat thickness and fecal ammonia gas concentration in finishing pigs. A total of seventy-two finishing pigs (initial body weight 78.56$\pm$1.32kg) were used in this experiment. The treatments included 1) Control (CON; basal diet) 2) GB1.0 (basal diet + germanium biotite 1.0%), 3) GB3.0 (basal diet + germanium biotite 3.0%). Pigs fed GB1.0 diet grew faster than pigs fed CON diet and GB0.3 diet (P<0.05). Also, pigs fed CON diet showed higher(p<0.05) ADFI than pigs fed GB3.0 diet. Pigs fed GB diets had improved gain/feed compared to pigs fed CON diet(P<0.05). Total?and VLDL concentrations in plasma of pigs fed GB diets treatments were significantly decreased compared to those in pig fed CON diet(P<0.05). However, HDL-cholesterol concentration in plasma of the pig was significantly increased compared to those in pigs fed CON diet (P<0.05). Pigs fed CON diet exerted higher(P<0.05) backfat thickness than pigs fed GB1.0 (5.4%, 27.19 vs 25.71mm) and GB3.0 (16.1%, 27.19 vs 22.81mm) diets. Feces from CON treatment were higher in fecal ammonia gas concentration than faces from pigs fed GB1.0 (64.1%, 17.00 vs 6.10mg/kg)and GB3.0 (61.8%, 17.00 vs 6.50mg/kg) treatments(P<0.05). In conclusion, the results suggest that the dietary addition of germanium biotite into diets for nursery pigs did not affect growth performance. The results also suggest the possibility of germanium biotite to replace antibiotic in diets for growing pigs. In finishing pigs, dietary supplementation of germanium biotite was an effective means for improving growth performance and for decreasing Total-and LDL+VLDL-plasma cholesterols, backfat and fecal ammonia gas concentration.

Biocompatibility and Surface Characteristics of (Si,Mn)-HA Coated Ti-Alloy by Plasma Electrolytic Oxidation (PEO법으로 (Si,Mn)-HA 코팅된 치과 임플란트용 Ti 합금의 생체적합성 및 표면특성)

  • Gang, Jeong-In;Son, Mi-Gyeong;Choe, Han-Cheol
    • Proceedings of the Korean Institute of Surface Engineering Conference
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    • 2017.05a
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    • pp.83-83
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    • 2017
  • 생체재료의 표면은 이식과 동시에 생체계면의 역할을 하게 되어, 일련의 생물학적 반응이 시작되고 진행되는 중요한 장소가 된다. 초기에 생체계면에서 일어나는 단백질 흡착이나 염증반응을 비롯한 생물학적 반응들은 궁극적으로 임플란트의 성패를 좌우할 만큼 중요하다. 골융합을 개선하기 위한 다른 방법으로 생체불활성의 타이타늄 (Ti)과 골조직의 능동적인 반응을 이루기 위해 생체활성 표면을 부여함으로서 계면에서의 골형성 반응을 증진시키는 방법이 이용된다. 생체불활성의 Ti과 Ti합금은 골조직과 직접적인 결합을 이루지 못하므로, 골조직과의 반응을 향상하기 위해 여러 종류의 생체활성 재료를 코팅하는 방법이 연구되어 왔고, 이 중 생체의 변화와 가장 유사한 하이드록시아파타이트 코팅이 가장 대중적인 방법으로 사용되었으며 이는 초기 골형성을 촉진하는 것으로 알려졌다. 치과용 임플란트의 표면형상과 화학조성이 골 융합에 영향을 미치는 가장 중요한 인자이므로 최근의 연구동향은 이들 두 가지 표면특성을 결합함으로서 결과적으로 최적의 골세포반응을 유도하고, 골융합 후 골조직과의 micromechanical interlocking에 의해 임플란트의 안정성에 중요한 역할을 하는 마이크론 단위의 표면조도와 표면 구조를 유지하면서, 부가적으로 골 조직 반응을 능동적으로 개선할 수 있는 생체활성 성분을 부여하여 골 융합에 상승효과를 이루기 위한 표면처리법에 관해 많은 연구가 요구되어지고 있다. 따라서 골을 구하는 원소인 망간과 실리콘으로 치환된 하이드록시아파타이트를 플라즈마 전해 산화법으로 코팅하여 세포와 잘 결합할 수 있는 표면을 제공함으로써 골 융합과 치유기간을 단축시킬 수 있을 것으로 사료된다. 실험방법은 시편은 치과 임플란트 제작 합금인 Ti-6Al-4V ELI disk (grade 5, Timet Co., USA; diameter, 10 mm, thickness, 3 mm)이며, calcium acetate monohydrate, calcium glycerophosphate, manganese(II) acetate tetrahydrate, sodium metasilicate을 설계조건에 따라 혼합 제조된 전해질 용액을 이용하여 플라즈마 전해 산화법으로 표면 코팅을 실시하였다. 각 시편의 플라즈마 전해시 전압은 280V로 인가하였고, 전류밀도는 70mA로 정전류를 공급하여 해당 인가전압 도달 후 3분 동안 정전압 방식을 유지하였다. 코팅된 피막 표면을 주사전자현미경과 X-선 회절분석을 통하여 미세구조 및 결정상을 관찰하였다. 또한 코팅된 표면의 생체활성 평가는 정량적으로 평가하기 위해 동전위시험과 AC 임피던스를 통하여 시행하였다. 분극거동을 확인하기 위해 potentiostat (Model PARSTAT 2273, EG&G, USA)을 이용하여 구강 내 환경과 유사한 $36.5{\pm}1^{\circ}C$의 0.9 wt.% NaCl에서 실시하였다. 전기화학적 부식 거동은 potentiodynamic 방법으로 조사하였고 인가전위는 -1500 mV에서 2000 mV까지 분당 1.67 mV/min 의 주사속도로 인가하여 시험을 수행하였다. 임피던스 측정은 potentiostat (Model PARSTAT 2273, EG&G, USA)을 이용하였으며, 측정에 사용한 주파수 영역은 10mHz ~ 100kHz 까지의 범위로 하여 조사하였고 ZSimWin(Princeton applied Research, USA) 소프트웨어를 사용하여 용액의 저항, 분극 저항 값을 산출하였다. 망간의 함량이 증가할수록 불규칙한 기공을 보였으며, 실리콘은 $TiO_2$ 산화막 형성을 저해하는 경향을 확인할 수 있었다. 단독으로 표면을 처리한 경우보다 두 가지 원소를 이용해 복합 표면처리를 시행한 경우가 내식성이 좋아 임플란트과의 골 유착에 긍정적인 영향을 미칠 것으로 사료된다.

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Labeling Efficiency of $^{99m}Tc$-Labeled RBC Due to Labeling Time Change (표지시간 변화에 의한 $^{99m}Tc$과 적혈구 표지효율)

  • Dong, Kyeong-Rae;Kim, Ho-Seong;Choi, Seong-Kwan
    • Journal of radiological science and technology
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    • v.30 no.3
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    • pp.259-263
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    • 2007
  • For the preparation of $^{99m}Tc$-labeled RBC, $10{\sim}20\;{\mu}g/kg$ of Stannous(II) chloride and $10{\sim}40\;min$ of preparation was used. For finding out the effect of contrast agent, the blood samples were collected in three days, seven days, and 1 months after the diagnostic procedure. In the normal volunteer, the concentration of reducing agent and preparation time did not effect on the radiochemical yield. But in the patients, 10 mg of Stannous(II) chloride and 60 min incubation times was shown high radiochemical yield. Contrast agent has a significant effect on the radiochemical yield. Although the blood samples which were collected after seven days of diagnostic procedure did not effect on the radiochemical yield of $^{99m}Tc$-labeled RBC, but the radiochemical yield of $^{99m}Tc$-labeled RBC which was prepared with a sample of high concentration of contrast agent in blood led to low radiochemical yield. For these samples, the modified method showed high radiochemical yield than previous in vivo preparation method. The recommended method is followed. Blood collecting was performed at 30 minutes after injection of reducing agent, and it is centrifuged for removal of plasma. After addition of $^{99m}TcO^-_4$, sample reservoir was rotated. After addition of normal saline, and it is centrifuged for separation of saline. Then $^{99m}Tc$-labeled RBC was obtained after removal of saline.

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A Study on the Change of Microstructures by Heat-treatment in Mo-Hf-C Alloys (Mo-Hf-C계 합금의 열처리에 따른 미세조직 변화에 관한 연구)

  • Yoon, Kook-Han;Kim, Hyeong-Ki;Lee, Chong-Mu;Park, Won-Koo;Choi, Ju
    • Korean Journal of Materials Research
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    • v.3 no.2
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    • pp.111-120
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    • 1993
  • Abstract In this study, the Mo-Hf-O ingots containing 0.31-1.14at % Hf and 0.08-1.00at % 0 were prepared by plasma arc melting. The change of microstructure depending on the condition of heat treatmen~ was analysed by optical microscophy, auger electron microscophy, and transmission electron microscophy. Molybdenum powder with the oxygen content of 830ppm was compacted, and then melted. The oxygen content of molybdenum ingots was detected to be 40 -130ppm. As the contents of Hf and 0 increased, the grain size of ingots decreased. When molybdenum igot containing l.14at % Hf and 1.00at % C was heat treated, p-molybdenum carbide in grains was transformed into ${\alpha}$-molybdenum carbide at 130$0^{\circ}C$. Between 140$0^{\circ}C$ and 150$0^{\circ}C$, the precipitation of hafnium carbide was due to the reaction of solute Hf and C, and the hafnium carbide was saturated at grain boundaries at 150$0^{\circ}C$. When the sample was heat treated from 150$0^{\circ}C$ to 170$0^{\circ}C$, Hafnium oxide more stable thermodynamically precipitated both at grain boundaries and in grains after hafnium carbide had been dissolved at grain boundaries.

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