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Effect of Matrigel for Bone Graft using Hydroxyapatite/Poly $\varepsilon$-caprolactone Scaffold in a Rat Calvarial Defect Model (랫드의 두개골결손부 모델에서 HA/PCL 지지체를 사용한 골이식 시 Matrigel의 효과)

  • Kim, Se-Eun; Shim, Kyung-Mi;Kim, Seung-Eon;Choi, Seok-Hwa;Bae, Chun-Sik;Han, Ho-Jae;Kang, Seong-Soo
    • Journal of Veterinary Clinics
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    • v.27 no.4
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    • pp.325-329
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    • 2010
  • The osteogenic potential of hydroxyapatite/poly $\varepsilon$-caprolactone composite (HA/PCL) scaffolds with matrigel was evaluated in a rat calvarial defect model. Calvarial defect formation was surgically created in Sprague Dawley rats (n = 18). HA/PCL scaffold was grafted with matrigel (M-HA/PCL group, n = 6) or without matrigel (HA/PCL group, n = 6). A critical defect group (CD group, n = 6) did not received a graft. Four weeks after surgery, bone formation was evaluated with radiography, micro computed tomography (micro CT) scanning, and histologically. No bone tissue formation was radiographically evident in the CD group. Bone tissue was radiographically evident in the HA/PCL and M-HA/PCL groups, however, there was more bone-similar opacity in the M-HA/PCL group. Micro CT analysis revealed that the bone volume of the M-HA/PCL group was higher than the HA/PCL group, however, no significant difference was found between the HA/PCL and M-HA/PCL groups. Bone mineral density in the M-HA/ PCL group was significantly higher than in the HA/PCL group (p < 0.05). Histologically, new bone was formed only from existing bone in the CD group, showing concavity without bone formation in the defect. In the HA/PCL group, new bone formation was only derived from existing bone, while in the M-HA/PCL group the largest bone formation was observed, with new bone tissue forming at the periphery of existing bone and around the HA/PCL scaffold with matrigel. The results indicate that the combination of HA/PCL scaffold with matrigel may be an effective means of enhancing bone formation in critical-sized bone defects.

Quantitative Measurement of Membrane $Na^+-K^+$ ATPase Activity using Thallium-201 : Comparison with Rubidium-86 (Thallium-201을 이용한 세포막 $Na^+-K^+$ ATPase 활성도 측정: Rubidium-86 측정법과의 비교)

  • Lee, Jae-Tae;Lee, In-Kyu;Sohn, Sang-Kyun;Lee, Kyu-Bo
    • The Korean Journal of Nuclear Medicine
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    • v.32 no.2
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    • pp.121-128
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    • 1998
  • Purpose: $Na^+-K^+$ ATPase activity has been estimated by the degree of inhibition of cation transport by cardiac glycosides (ouabain) using Rb-86 as a substrate. The biological characteristisc of T1-201 is known to be similar to those of potassium as a transport substrate in the presence of glucose, insulin or phobol myristate acetate (PMA). The purpose of this study was to measure ouabain sensitive $Na^+-K^+$ ATPase activity using T1-201 and compare with that using Rb-86. Materials and Methods: Smooth muscle cells isolated from rat aorta or human placental umbilical artery were cultured, and used to measure cellular $Na^+-K^+$ ATPase activity. $Na^+-K^+$ ATPase activity was measured as a percentage decrease in cellular uptake of T1-201 or Rb-86 by ouabain under the presence of glucose, insulin or PMA in media. Results: $Na^+-K^+$ ATPase activity measured with T1-201, as a transport substrate, was not different from those measured with Rb-86 in rat or human smooth muscle cell preparation. Incubation with high concentration glucose resulted in about 30% decrease in enzyme activity. In contrast, insulin or PMA resulted in 10-70% or 28% increases from baseline activity, respectively. Conclusion: These results suggests that 71-201 could replace Rb-86 in measurement of ouabain sensitive $Na^+-K^+$ ATPase activity in vitro. High level of glucose concentration decreased cellular $Na^+-K^+$ ATPase activity, but insulin or PMA increased it.

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Normal Limits of Left Ventricular Volumes and Ejection Fraction Measured by Gated Myocardial Perfusion SPECT: Comparison of Tc-99m MIBI and Tl-201 (심근 게이트 SPECT로 측정한 좌심실 용적과 구혈률의 정상 값 확립: Tl-201과 Tc-99m MIBI 게이트 SPECT의 비교)

  • Hyun, In-Young;Seo, Jeong-Kee;Kwan, Jun;Park, Keum-Soo;Choe, Won-Sick;Lee, Woo-Hyung
    • The Korean Journal of Nuclear Medicine
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    • v.37 no.3
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    • pp.147-152
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    • 2003
  • Purpose: We evaluated radioisotope and sex-specific differences of normal limits for left ventricle volumes (LWs) and ejection fraction (EF) using myocardial perfusion gated SPECT (g-SPECT). Materials and Methods: Rest Tl-201/post-stress Tc-99m MIBI g-SPECT measurements with acquisitions of 8-frame were evaluated for 70 patients (mean age $55{\pm}14.56%$ female) who either had < 10% pretest likelihood of CAD (n = 12) or had normal coronary angiography (EF > 50%) (n = 58). LVEF, LWs were automatically determined by quantitative gated SPECT using QGS program. Results: Similar results were obtained for mean LVEF between Tc-99m MIBI ($62%{\pm}7%$ and Tl-201 ($63%{\pm}8%$) g-SPECT measurements. In Contrast, Tl-201 g-SPECT had significantly lower LWs values ($EDV;\;74{\pm}23mL,\;ESV;\;28{\pm}14mL$) than Tc-99m MIBI g-SPECT ($EDV;\;82{\pm}25mL,\;ESV;\;32{\pm}15mL$) (p<0.05). Women had significantly lower EDV ($Tc-99m\;MIBI;\;71{\pm}18mL,\;Tl-201;\;65{\pm}17mL$), and ESV values ($Tc-99m\;MIBI;\;27{\pm}10mL,\;Tl-201;\;23{\pm}8mL$) compared with EDV ($Tc-99m\;MIBI;\;96{\pm}27mL,\;Tl-201;\;85{\pm}24mL$), and ESV Values ($Tc-99m\;MIBI;\;40{\pm}17mL,\;Tl-201;\;36{\pm}16mL$) of men (p<0.05). Women had significantly higher LV EF Values ($65%{\pm}7%$) than men ($60%{\pm}8%$) by Tl-201 gated SPECT (p<0.05). Conclusion: These data suggest significant differeuces in normal limits for LWs and EF, according to genders and radiopharmaceutical. Therefore, the evaluation of cardiac function in patients should consider radioisotope and sex-matched normal values.

Production and Evaluation of Immunoreactivity of Poly Lysine-Tagged Single Chain Fragment Variable (ScFv) Lym-1 Antibody for Direct Conjugation to Fluorescence Dye (형광 물질 직접 표지를 위한 Poly Lysine 도입 Lym-1 단일사슬 항체의 제조 및 면역반응성 평가)

  • Jung, Jae-Ho;Choi, Tae-Hyun;Woo, Kwang-Sun;Chung, Wee-Sup;Kang, Joo-Hyun;Jeong, Su-Young;Choi, Chang-Woon;Lim, Sang-Moo;Cheon, Gi-Jeong
    • Nuclear Medicine and Molecular Imaging
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    • v.43 no.5
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    • pp.487-494
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    • 2009
  • Purpose: Small size of recombinant scFv antibody has many advantages such as rapid blood clearances and improved targeting antibodies to tumor region. On the other hand owing to small size, number of amino group is insufficient in conjugation with chelator and fluorescence labeling. This study is to introduce poly lysine tag to the C-terminal end of scFv lym-1 sequence for fluorescence chelator conjugation. Materials and Methods: Poly lysine scFv lym-1 gene, cloned into pET-22b (+) vector, was expressed in E. coli BL21 (DE3) strain. Antibody purification was performed with Ni-NTA column and then size exclusion column chromatography. Expression and purification levels of poly lysine tagged scFv lym-1 antibody were confirmed by western blot analysis. I-124, I-125, I-131 and Tc-99m were used for radiolabeling of purified poly lysine scFv lym-1. Flow cytometry analysis of FIT( conjugated poly lysine scFv lym-1 was performed for confirmation of immunoreactivity of human Burkitt's lymphoma cells. Results: Poly lysine scFv lym-1 antibody was purified through two steps and identified as molecular weight of 48 KDa. Radiolabeling yields of I-124, I-125, I-131 and Tc-99m into poly lysine scFv lym-1 were >99%, >99%, >95% and >99%, respectively. Flow cytometry analysis of poly lysine scFv and scFv lym-1 was showed similar immunoreactivity to human Burkitt's lymphoma cells. Conclusion: Poly lysine tag was useful for the sufficient number of amino groups to scFv lym-1 antibody for chelator conjugation with minimizing loss of immunoreactivity.

The Effect on The Result, in Case of the In-vitro Test Performance after an Imaging Test (핵의학 영상검사 후 시행된 핵의학 검체검사에서의 영향)

  • Moon, Ki Choon;Kwon, Won Hyun;Kim, Jung In;Lee, In Won
    • The Korean Journal of Nuclear Medicine Technology
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    • v.18 no.1
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    • pp.149-152
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    • 2014
  • Purpose: At our hospital blood is collected from a patient before an imaging test, with the concern of any effect possible when a nuclear medicine imaging test and an in-vitro test are carried out at the same time. However, occasionally, the blood collection is performed after an imaging test, with the reasons that the patient is not properly guided or the patient doesn't follow the guide correctly. In that case, we prefer to gather blood again after a few days. The purpose of this study is not only to see whether there is any effect of an imaging test on the result of the in-vitro test performed with the blood collected after the imaging test, but also to study how many days waiting after each test is appropriate to take a blood sample, if the effect exists. Materials and Methods: From September to October 2013, blood were collected from 13 patients in our hospital regardless of age and sex each time before and after the injection of the radioactive isotope from the tests : PET-CT, Gated Myocardial SPECT, and DTPA GFR Scan. Considering a half-life, AFP, CA19-9, CEA, TSH, and T3 were carried out right after the blood collection. In case of an iodine therapy, blood were taken each time before and after taking radioactive iodine, and, after AFP, CA19-9, and CEA, the difference between them in consistency and in cpm were compared. Results: With 10 patients after the imaging tests and 3 patients after the iodine therapy, their serum cpm was over 10,000. Over time, the cpm decreased in accordance with the half-life ($^{18}F$ 110minutes, $^{99}mTc$ 6hours, $^{201}Tl$ 72hours, $^{131}I$ 7days). Between the two cases, one before and the other after the injection of the radioactive isotope, the cpm and the results of AFP, CA19-9, CEA, TSH, and T3 from three patients each test, PET-CT, Gated Myocardial SPECT, and DTPA GFR Scan, were very similar. In addition, in case of an iodine therapy, there was also not a meaningful difference in the cpm and the results of AFP, CA19-9, and CEA, from three patients in an iodine therapy, between the two cases, one before and the other after taking the radioactive iodine. Conclusion: In case a blood collection was performed after the imaging test which required a radioactive isotope injection, the cpm increased, differently according to the kind of the radioactive isotope. However, the results of the in-vitro tests like AFP, CA19-9, CEA, TSH, T3, etc were nearly not affected. As the result, it's considered that there will not be any significant effect also from other tests, as the result from the performed seven tests.

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Abosrbed Dose Measurements and Phantom Image Ecaluation at Minimum CT Dose for Pediatric SPECT/CT Scan (소아 SPECT/CT 검사를 위한 최저조건에서의 피폭선량측정 및 팬텀의 영상평가)

  • Park, Chan Rok;Choi, Jin Wook;Cho, Seong Wook;Kim, Jin Eui
    • The Korean Journal of Nuclear Medicine Technology
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    • v.18 no.1
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    • pp.82-88
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    • 2014
  • Purpose: The purpose of study was to evaluate radiation dose for pediatric patients by changing tube voltage (kVp) and tube current (mA) at minimum conditions. By evaluating radiation dose, we want to provide dose reduction for pediatric patients and maintain good quality of SPECT/CT images. Materials and Methods: Discovery NM/CT 670 Scanne was used as SPECT/CT. Tube voltages are 80 and 100 kvP. Tube currents are 10, 15, 20, 25 mA. Using PMMA (Polymethyl methacrylate) Phantom, radiation dose which were calculated at center and peripheral dose and SNRD (Signal to Noise Ratio Dose) were evaluated. Using the CT performance phantom, spatial resolution was evaluated as the MTF (Modulation Transfer Function) graph. Jaszczak phantom was used for SPECT image evaluation by CNR (Contrast to Noise to Ratio). Results: Radiation dose using the PMMA phantom was higher peripheral dose than center dose about 7%. SNRD were 7.8, 8.2, 8.3, 8.8, 8.8, 9.9, 9.8, 9.6 for 80 kVp 10, 15, 20, 25 mA, 100 kVp 10, 15, 20, 25 mA. We can distinguish 35, 45, 70, 71, 52, 58, 90, 110 linepair for 80 kVp 10, 15, 20, 25 mA, 100 kVp 10, 15, 20, 25 mA at resolution with MTF. CNR of SPECT images using CT attenuation map were 57.8, 57.7, 57.1, 56.7, 56.6, 56.7, 56.7, 56.7% for 80 kVp 10, 15, 20, 25 mA, 100 kVp 10, 15, 20, 25 mA. Conclusion: In this study, radiation dose for pediatric patients showed decreased low dose condition. And SNRD value was similar in all condition. Resolution showed higher value at 100kVp than 80kVp. for CNR, there was no significant difference. we should take additional study to prove better quality and dose reduction.

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Real-Time RT-PCR for Validation of Reovirus Type 3 Safety During the Manufacture of Mammalian Cell Culture-Derived Biopharmaceuticals (세포배양 유래 생물의약품 생산 공정에서 Reovirus Type 3 안전성 검증을 위한 Real-Time RT-PCR)

  • Lee, Dong-Hyuck;Jeong, Hyo-Sun;Kim, Tae-Eun;Oh, Seon-Hwan;Lee, Jung-Suk;Kim, In-Seop
    • Korean Journal of Microbiology
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    • v.44 no.3
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    • pp.228-236
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    • 2008
  • Validation of viral safety is essential in ensuring the safety of mammalian cell culture-derived biopharmaceuticals, because numerous adventitious viruses have been contaminated during the manufacture of the products. Mammalian cells are highly susceptible to Reovirus type 3 (Reo-3), and there are several reports of Reo-3 contamination during the manufacture of biopharmaceuticals. In order to establish the validation system for the Reo-3 safety, a real-time RT-PCR method was developed for quantitative detection of Reo-3 in cell lines, raw materials, manufacturing processes, and final products as well as Reo-3 clearance validation. Specific primers for amplification of Reo-3 RNA was selected, and Reo-3 RNA was quantified by use of SYBR Green I. The sensitivity of the assay was calculated to be $3.2{\times}10^0\;TCID_{50}/ml$. The real-time RT-PCR method was proven to be reproducible and very specific to Reo-3. The established real-time RT-PCR assay was successfully applied to the validation of Chinese hamster ovary (CHO) cell artificially infected with Reo-3. Reo-3 RNA could be quantified in CHO cell as well as culture supernatant. When the real-time RT-PCR assay was applied to the validation of virus removal during a virus filtration process, the result was similar to that of virus infectivity assay. Therefore, it was concluded that this rapid, specific, sensitive, and robust assay could replace infectivity assay for detection and clearance validation of Reo-3.

Regional Analysis of Forest Eire Occurrence Factors in Kangwon Province (강원도 지역 산불발생인자의 지역별 유형화)

  • 이시영;한상열;안상현;오정수;조명희;김명수
    • Korean Journal of Agricultural and Forest Meteorology
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    • v.3 no.3
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    • pp.135-142
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    • 2001
  • This study attempts to categorizes the factors of forest fire occurrences based on regional meteorologic data and general forest no characteristics of 18 cities and guns in Kangwon province. lo accomplish this goal, some statistical analyses such as analysis of variance, correspondence analysis and multidimensional scaling were adopted. To reveal the forest fires pattern of study region, a categorization process was conducted by employing the quantification approach which modified and quantified the metric-data of fire occurrence dates. Also, The fire occurrence similarity was compared by using multidimensional scaling for each study region. The major results are summarized as follows: It was found that the meteorological factors emerged as different to each region are average and maximum temperature, minimum dew point temperature and average and maximum wind speed. In the result of correspondence analysis representing relationships between fire causes and study regions, Kangrung is caused by arsonist, Chulwon, Hwachen and Yanggu caused by military factor, Sokcho and Chunchen caused by the debris burning, and Samchuk caused by general man-caused fires, respectively. Finally, the forest fire occurrence pattern of this study regions were divided into five areas such as, group I including Samchuk, Kangryung, Chunchen, Wonju, Hongchen and Hhoingsung, group II including Donghae, Taebaek, Yangyang and Pyongchang, group III including Jungsun, Chulwon and Whachen, group Ⅵ including Gosung, Injae and Yanggu, and group V including Shokcho and Youngwol.

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Comparison in Seed and Sprout Quality under Different Cropping Patterns in Mungbean (재배방식에 따른 녹두 종실과 나물의 품질변화)

  • Kim, Dong-Kwan;Son, Dong-Mo;Choi, Jin-Gyung;Shin, Hae-Ryong;Chon, Sang-Uk;Lee, Kyung-Dong;Jung, Ki-Yeol;Rim, Yo-Sup
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.56 no.3
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    • pp.212-218
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    • 2011
  • This study was performed to determine the relative quality of mungbeans harvested in bulk after applying a labor-saving cultivation (LSC) method, compared to mungbeans harvested three different times under the conventional cultivation condition. There was no significant difference in starch, crude protein, and vitexin or isovitexin content of seed according to the cropping system or harvest time. The mungbeans grown under the LSC method had the highest crude fat content, followed by mungbeans from the third-, the second- and the first-harvest mungbeans under the conventional cultivation. No significant difference was found in the composite ratio of saturated fatty acids to unsaturated fatty acids according to cropping system or harvest time. The second-harvest mungbeans grown under the conventional cultivation condition had 17 different types of fatty acids, while the third-harvest mungbeans grown under the conventional cultivation and those grown under the LSC condition had the fewest types of fatty acids with 12. Of the major saturated fatty acids, palmitic acid and arachidonic acid had the highest composite ratio in the first conventional cultivation followed by the second, the third and the LSC. However, stearic acid showed the opposite tendency. Of the major unsaturated fatty acids, linoleic acid had the highest composite ratio in the first conventional cultivation, followed by the second and third conventional cultivation and the LSC. Amylogram characteristics of the mungbeans were significantly different according to cropping system and harvest times. The mungbeans harvested after the first conventional cultivation had significantly higher pasting temperature, peak viscosity, holding strength viscosity, final viscosity and breakdown, while mungbeans harvested after the third conventional cultivation had significantly higher setback viscosity. In contrast, the mungbeans harvested under the LSC methods had a significantly lower amylogram value. When harvest rate, color values and amino acid content of sprout were measured, mungbeans grown under the LSC conditions had a low harvest rate of sprout, but had Hunter's color values and amino acid content of sprout similar to those of mungbeans grown under the conventional cultivation condition.

Geochemical Characteristics of Black Slate and Coaly Slate from the Uranium Deposit in Deokpyeong Area (덕평리 지역 우라늄광상의 흑색점판암과 탄질점판암의 지구화학적 특성)

  • Shin, Dong-Bok;Kim, Su-Jeong
    • Economic and Environmental Geology
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    • v.44 no.5
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    • pp.373-386
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    • 2011
  • Geochemical study was performed on black slates and interbedded U-bearing coaly slates in Deokpyeongri area, the representative uranium mineralized district of the Ogcheon Metamorphic Belt, to discuss the genetic environments of the uranium deposit. REE concentration (254 ppm) of the black slates is higher than that (169 ppm) of the coaly slates and NASC-normalized REE patterns of the coaly slates show remarkable positive Eu anomaly. l11e redox-sensitive trace elements such as V, Cr, Co, Ni, Mo and U in the coaly slates are highly enriched compared to the black slates, especially for V of 24 times, Mo of 62 times, and U of 60 times. In additions, Pd and Pt are also enriched in the coaly slates. Positive Eu anomaly and the noticeable enrichment of the elements listed above compared to those of NASC indicate that those elements were not derived from common seawater but deposited under high temperature and reducing environment of submarine hydrothermal activities. Wide compositional ranges of major elements ($SiO_2/Al_2O_3$: 3.98~11.88, $Al_2O_3/Na_2O$: 25.6~139.06, $K_2O/Na_2O$: 6.80~46.85) also suggest that the source rocks of the sediments are mixtures of sedimentary rocks and igneous rocks. Higher sulfur contents in the coaly slates, 2.6 wt.%, than those in the black slates, 0.6 wt.% also indicates that the former was influenced by hydrothermal activities containing much sulfur. These geochemical characteristics are similar to the genetic environments of South China type PGE deposits (Mo-Ni-Zn-PGE) which is geotectonically correlated with the Ogcheon Metamorphic Belt and is known as sedimentary-exhalative deposits. In conclusions, the uranium and other metallic elements mineralization seems to have occurred in the sedimentary basin that was affected by submarine hydrothermal activities and rich in organic materials under oxygen-poor environments as well.