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Effect of Okadaic Acids (OA) on Nuclear Maturation and Mitochondrial Activity of Hanwoo COCs during in vitro Maturation (소 난구복합체의 체외성숙시 Okadaic Acid (OA)가 핵성숙 및 Mitochondria 활성에 미치는 영향)

  • Choi S. H.;Han M. H.;Cho S. R.;Kim H. J.;Choe C. Y.;Son D. S.;Kim Y. K.;Lee M. H.;Jeoung Y. G.;Chung Y. H.
    • Journal of Embryo Transfer
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    • v.20 no.3
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    • pp.303-309
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    • 2005
  • This study was conducted to examine the effects of OA on metaphase of meiosis II and the mitochondrial activity of cytoplasm in bovine cumulus oocytes complexes(COCs) during in vitro maturation. Hanwoo COCs were collected from the slaughterhouse cow ovaries and matured in TCM199 supplemented with $0.1\%$ PVA, 0.2 uM, 2 uM, 20 uM OA for the maturation rate of OA concentration. For the maturation effects between OA and cycloheximide(CX), COCs were matured in TCM199 with 25 ug/mL CX, 25 ug/mL CX (6 hrs culture) plus 2 uM OA or 2 uM OA only at a atmosphere $5\%\;CO_2,\;95\%$ air $39^{\circ}C$ for 6, 12, 24 hrs. To evaluate the nuclear types of matured COCs, cumulus cells were removedby $0.5\%$ hyaluronidase sol. and oocytes were fixed in 1:3 acetic acid ethyl alcohol for 30 sec. and then stained with $0.1\%$ basic Fuchsin sol. For the detection of fluoriscent intensity (FI) of matures oocytes, cumulus cells were removed same as performed above and were stained with 20 nM mite tracker for 20 min. at $39^{\circ}C$. Mitochondrial activity of FI in matured oocytes was imaged by laser conforcal microscopy (Fluoview, Olympus, Japan) and were measured scanned face on 5 um from median to endpoint of oocytes. Statical analysis of nuclear types observed the three replicates was carried out with ANOVA and Fisher's protected least significant difference test using the STATVIEW program. FI of matures oocytes was compared the multiples of the least intensity among the measured oocytes. Maturing in TCM199 supplemented with $0.1\%$ PVA, 0.2 uM, 2 uM, 20 uM OA, metaphase B were showed 72.0, 50.0, 70.0, $68.8\%$, respectively and there were different significant(p<0.05). In the case of treatment with OA and CX, metaphase were $73.8\%,\;8.2\%,\;45.5\%,\;73.7\%$ in $0.1\%$ PVA-TCM199, 25 ug/mL CX, 25 ug/mL CX plus OA or 2uM OA only, respeclively. FI was revealed the increasing tendency during the process of maturation. Whereas FI in CX was decreased about 3 times compared to the other treatments of 6 hrs maturation. We conclude that OA regulates bovine COCs maturation and induces the mitochondrial activity during the process of maturation.

GafChromic Film Dosimetry for Stereotactic Radiosurgery with a Linear Accelerator (선형가속기를 이용한 정위방사선 치료 시 GafChromic Film을 이용한 선량측정)

  • Han Seung Hee;Cho Byung Chul;Park Suk Won;Oh Do Hoon;Park Hee Chul;Bae Hoon Sik
    • Radiation Oncology Journal
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    • v.21 no.2
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    • pp.167-173
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    • 2003
  • Purpose: The purpose of this study was to evaluate whether a GafChromic film applied to stereotactic radiosurgery with a linear accelerator could provide information on the value for acceptance testing and quality control on the absolute dose and relative dose measurements and/or calculation of treatment planning system. Materials and methods: A spherical acrylic phantom, simulating a patient's head, was constructed from three points. The absolute and relative dose distributions could be measured by inserting a GafChromic film into the phantom. We tested the use of a calibrated GafChromic film (MD-55-2, Nuclear Associate, USA) for measuring the optical density. These measurements were achieved by irradiating the films with a dose of 0-112 Gy employing 6 MV photon. To verify the accuracy of the prescribed dose delivery to a target isocenter using a five arc beams (irradiated in 3 Gy per one beam) setup, calculated by the Linapel planning system the absolute dose and relative dose distribution using a GafChromic film were measured. All the irradiated films were digitized with a Lumiscan 75 laser digitizer and processed with the RIT113 film dosimetry system. Results: We verified the linearity of the Optical Density of a MD-55-2 GafChromic film, and measured the depth dose profile of the beam. The absolute dose delivered to the target was close to the prescribed dose of Linapel within an accuracy for the GafChromic film dosimetry (of $\pm$3$\%$), with a measurement uncertainty of $\pm$1 mm for the 50$\~$90$\%$ isodose lines. Conclusion: Our results have shown that the absolute dose and relative dose distribution curves obtained from a GafChromic film can provide information on the value for acceptance. To conclude the GafChromic flim is a convenient and useful dosimetry tool for linac based radiosurgery.

Development of Decision Tree Software and Protein Profiling using Surface Enhanced laser Desorption/lonization - Time of Flight - Mass Spectrometry (SELDI-TOF-MS) in Papillary Thyroid Cancer (의사결정트리 프로그램 개발 및 갑상선유두암에서 질량분석법을 이용한 단백질 패턴 분석)

  • Yoon, Joon-Kee;Lee, Jun;An, Young-Sil;Park, Bok-Nam;Yoon, Seok-Nam
    • Nuclear Medicine and Molecular Imaging
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    • v.41 no.4
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    • pp.299-308
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    • 2007
  • Purpose: The aim of this study was to develop a bioinformatics software and to test it in serum samples of papillary thyroid cancer using mass spectrometry (SELDI-TOF-MS). Materials and Methods: Development of 'Protein analysis' software performing decision tree analysis was done by customizing C4.5. Sixty-one serum samples from 27 papillary thyroid cancer, 17 autoimmune thyroiditis, 17 controls were applied to 2 types of protein chips, CM10 (weak cation exchange) and IMAC3 (metal binding - Cu). Mass spectrometry was performed to reveal the protein expression profiles. Decision trees were generated using 'Protein analysis' software, and automatically detected biomarker candidates. Validation analysis was performed for CM10 chip by random sampling. Results: Decision tree software, which can perform training and validation from profiling data, was developed. For CM10 and IMAC3 chips, 23 of 113 and 8 of 41 protein peaks were significantly different among 3 groups (p<0.05), respectively. Decision tree correctly classified 3 groups with an error rate of 3.3% for CM10 and 2.0% for IMAC3, and 4 and 7 biomarker candidates were detected respectively. In 2 group comparisons, all cancer samples were correctly discriminated from non-cancer samples (error rate = 0%) for CM10 by single node and for IMAC3 by multiple nodes. Validation results from 5 test sets revealed SELDI-TOF-MS and decision tree correctly differentiated cancers from non-cancers (54/55, 98%), while predictability was moderate in 3 group classification (36/55, 65%). Conclusion: Our in-house software was able to successfully build decision trees and detect biomarker candidates, therefore it could be useful for biomarker discovery and clinical follow up of papillary thyroid cancer.

The Role of Endoscopy for Tumorous Conditions of the Upper Gastrointestinal Tract in Children (내시경으로 진단된 소아 상부 위장관의 종양성 질환에 대한 고찰)

  • Kim, Hye Young;Park, Jae Hong
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.8 no.1
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    • pp.31-40
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    • 2005
  • Purpose: This study aimed to provide, as a basic material, the experiences of endoscopy in diagnosis and treatment of tumorous conditions in the upper gastrointestinal tract in children. Methods: The objects were 26 patients diagnosed as having tumorous conditions in the upper gastrointestinal tract among 1,283 patients who underwent upper gastrointestinal endoscopic examination at the Department of Pediatrics, Pusan National University Hospital, from January 1994 to July 2004 retrospectively. The characteristics of patients, the chief complaints for endoscopic examination, the sorts of tumors diagnosed, the endoscopic findings of tumors, and the treatment of tumors were analysed. Results: 1) Eleven male and fifteen female were included, whose mean age was $6.93{\pm}4.02years$. 2) The chief complaints for endoscopic examination were abdominal pain (80.7%), vomiting or nausea (30.8%), and gastrointestinal beeding (30.7%) in order. 3) Six cases of ectopic pancreas, five cases of sentinel polyp, three cases of papilloma and vallecular cyst, two cases of Brunner's gland hyperplasia and gastric submucosal tumor, one case of gastrointestinal stromal tumor, duodenal intramural hematoma, T cell lymphoma, lipoma, and Peutz-Jeghers syndrome were diagnosed by endoscopy with or without biopsy. 4) The location of tumors was in the pharynx (19.2%), esophagus (7.7%), gastro-esophageal junction (23.0%), stomach (30.7%) and duodeneum (26.9%). 5) The size of tumors was less than 10 mm in 53.8%, 10~20 mm in 26.9%, more than 20 mm 19.2%. 6) Treatments for tumors included resection by laser, surgical resection, endoscopic polypectomy with a forcep or snare, and observation 7) There was no significant complication. Conclusion: Various and not a few tumors were found in the upper gastrointestinal tract. The endoscopy was accurate, effective, and safe means for diagnosis and treatment of those lesions in children.

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Agricultural Geography of Rice Culture in California (미국 캘리포니아주(州)의 벼농사에 관한 농업지리학적 연구)

  • Lee, Jeon;Huh, Moo-Yul
    • Journal of the Korean association of regional geographers
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    • v.2 no.1
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    • pp.51-67
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    • 1996
  • There are three main rice-growing regions in the United States: the prairie region along the Mississippi River Valley in eastern Arkansas; the Gulf Coast prairie region in southwestern Louisiana and southeastern Texas; and the Central Valley of California. The Central Valley of California is producing about 23% of the US rice(Fig. 1). In California. most of the crop has been produced in the Colusa, Sutter, Butte, Glenn Counties of the Sacramento Valley since 1912, when rice was commercially grown for the first time in the state(Fig. 2). Roughly speaking, the average annual area sown to rice in California is about 300,000 acres to 400,000 acres during the last forty years(Fig. 3). California rice is grown under a Mediterranean climate characterized by warm, dry, clear days, and a long growing season favorable to high photosynthetic rates and high rice yields. The average rice yield per acre is probably higher in California than in any other rice-growing regions of the world(Fig. 4). A dependable supply of irrigation water must be available for a successful rice culture. Most of the irrigation water for California rice comes from the winter rain and snow-fed reservoir of the Sierra Nevada mountain ranges. Less than 10 percent of rice irrigation water is pumped from wells in areas where surface water is not sufficient. It is also essential to have good surface drainage if maximum yields are to be produced. Rice production in California is highly mechanized, requiring only about four hours of labor per acre. Mechanization of rice culture in California includes laser-leveler technology, large tractors, self-propelled combines for harvesting, and aircraft for seeding, pest control, and some fertilization. The principal varieties grown in California are medium-grain japonica types with origins from the cooler rice climates of the northern latitudes (Table 1). Long-grain varieties grown in the American South are not well adapted to California's cooler environment. Nearly all the rice grown recently in California are improved into semidwarf varieties. Choice of variety depends on environment, planting date, quality desired, marketing, and harvesting scheduling. The Rice Experiment Station at Biggs is owned, financed, and administered by the rice industry. The station was established in 1912, as a direct result of the foresight and effort of Charles Edward Chambliss of the United States Department of Agriculture. Now, The station's major effort is the development of improved rice varieties for California.

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Mineralogical, Micro-textural, and Geochemical Characteristics for the Carbonate Rocks of the Lower Makgol Formation in Seokgaejae Section (석개재 지역 하부 막골층 탄산염암의 광물조성, 미세구조 및 지화학적 특성)

  • Park, Chaewon;Kim, Ha;Song, Yungoo
    • Economic and Environmental Geology
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    • v.51 no.4
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    • pp.323-343
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    • 2018
  • This study defines the mineralogical, micro-textural and geochemical characteristics for the carbonate rocks and discusses the fluids that have affected the depositional environment of the Lower Makgol Formation in Seokgaejae section. Based on analysis of X-ray Diffraction (XRD), Scanning Electron Microscope-Energy Dispersive X-ray Spectrometry (SEM-EDS), Electron Probe Micro Analyzer-Wavelength Dispersive X-ray Spectrometry (EPMA-WDS) and Laser Ablation-Inductively Coupled Plasma-Mass Spectrometry (LA-ICP-MS), carbonate miorofacies in the basal and the lower members of the Makgol Formation are distinguished and classified into four types. Type 1 dolomite (xenotopic interlocking texture) and Type 2 dolomite (idiotopic interlocking texture) have relatively high Mg/Ca ratio, flat REE pattern, low Fe and Mn. Extensively interlocking textures in these dolomites indicate constant supply of Mg ion from hypersaline brine. Type 3 and Type 4 dolomite (scattered and loosely-aggregated texture) have relatively moderate Mg/Ca ratio, MREE enriched pattern, low to high Fe and Mn. These partial dolomitization indicate limited supply of Mg ion under the influx of meteoric water with seawater. Also, the evidence of Fe-bearing minerals, recrystallization and relatively high Fe and Mn in Type 4 indicates the influence of secondary diagenetic fluids under suboxic conditions. Integrating geochemical data with mineralogical and micro-textural evidence, the discrepancy between the basal and the lower members of the Makgol Formation indicates different sedimentary environment. It suggest that hypersaline brine have an influence on the basal member, while mixing meteoric water with seawater have an effect on the lower member of the Makgol Formation.

[ $Ca^{2+}\;and\;K^+$ ] Concentrations Change during Early Embryonic Development in Mouse (생쥐 초기 배 발달 동안 변화되는 칼슘과 포타슘 이온)

  • Kang D.W.;Hur C.G.;Choi C.R.;Park J.Y.;Hong S.G.;Han J.H.
    • Journal of Embryo Transfer
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    • v.21 no.1
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    • pp.35-43
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    • 2006
  • Ions play important roles in various cellular processes including fertilization and differentiation. However, it is little known whether how ions are regulated during early embryonic development in mammalian animals. In this study, we examined changes in $Ca^{2+}\;and\;K^+$ concentrations in embryos and oviduct during mouse early embryonic development using patch clamp technique and confocal laser scanning microscopy. The intracellular calcium concentration in each stage embryos did not markedly change. At 56h afier hCG injection when 8-cell embryos could be Isolated from oviduct, $K^+$ concentration in oviduct increased by 26% compared with that at 14h after injection of hCG During early embryonic development, membrane potential was depolarized (from -38 mV to -16 mV), and $Ca^{2+}$ currents decreased, indicating that some $K^+$ channel might control membrane potential in oocytes. To record the changes in membrane potential induced by influx of $Ca^{2+}$ in mouse oocytes, we applied 5 mM $Ca^{2+}$ to the bath solution. The membrane potential transiently hyperpolarized and then recovered. In order to classify $K^+$ channels that cause hyperpolarization, we first applied TEA and apamin, general $K^+$ channel blockers, to the bath solution. Interestingly, the hyperpolarization of membrane potential still appeared in oocytes pretreated with TEA and apamin. This result suggest that the $K^+$ channel that induces hyperpolarization could belong to another $K^+$ channel such as two-pore domain $K^+(K_{2P})$channel that a.e insensitive to TEA and apamin. From these results, we suggest that the changes in $Ca^{2+}\;and\;K^+$ concentrations play a critical role in cell proliferation, differentiation and reproduction as well as early embryonic development, and $K_{2P}$ channels could be involved in regulation of membrane potential in ovulated oocytes.

The Production of Sex Determined Cattle by Embryonic Sexing Using Fluorescence In Situ Hybridization Technique (FISH 기법을 이용한 소 수정란의 성감별과 산자 생산)

  • Sohn, S.H.;Park, H.
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2007.05a
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    • pp.39-50
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    • 2007
  • Sexing from bovine embryos fertilized in vitro implicates a possibility of the sex controlled cattle production. This study was carried out to produce the sex determined cattle through the embryonic sexing by fluorescence in situ hybridization (FISH) technique. FISH was achieved in in vitro fertilized bovine embryos using a bovine Y-specific DNA probe constructed from the btDYZ-1 sequence. Using this probe, a male-specific signal was detected on 100% of Y-chromosome bearing metaphase specimens. The analyzable rate of embryonic sexing by FISH technique was about 93% (365/393) regardless of embryonic stages. As tested single blastomere by FISH and then karyotype with their biopsied embryos, the accuracy of sex determination with FISH was 97.6%. We tried the embryo transfer with sex determined embryos on 15 cattle. Among them, the 5 cattle delivered calf with expected sex last year.

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Study on the Analytical Method and Monitoring of the Oxidized Polyethylene Wax in Foods (식품 중 oxidized polyethylene wax 분석법 연구 및 함유량 실태 조사)

  • Choi, Seung-Hyun;Kim, Jae-Min;Choi, Sun-il;Jung, Tae-Dong;Cho, Bong-Yeon;Lee, Jin-Ha;Lee, Gunyoung;Lim, Ho Soo;Yun, Sang Soon;Lee, Ok-Hwan
    • Journal of Food Hygiene and Safety
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    • v.32 no.4
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    • pp.284-289
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    • 2017
  • Oxidized polyethylene wax (OPEW) is, one of the food additives, used as a coating agent in citrus fruits and nuts. OPEW is authorized to quantum satis in EU, USA, and is acceptable less than 250 mg/kg in Australia and New Zealand. But OPEW is unauthorized as a food additive in Korea. This study was to establish the analytical method of OPEW and demonstrate the effective application of various food samples. We first conducted to compare the various analytical method including acid value (AV), high temperature gel permeation chromatography (HT-GPC), matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF/MS), gas chromatography flame ionization detector (GC-FID) and fourier transform infrared spectroscopy (FT-IR). This result indicated that FT-IR spectrum of OPEW treated food sample displayed absorption bands for carbonyl group (C=O, $1714cm^{-1}$), ester group (C-O, $1463cm^{-1}$), aliphatic group (C-H, $2916cm^{-1}$). Furthermore, IR spectrum of OPEW treated food sample showed similar tendency with IR spectrum of OPEW standard. Therefore, it is confirmed that analytical method using FT-IR can be detected on analysis of OPEW in food. As a result of monitoring of 111 samples using established analytical method, OPEW was not detected in the food samples.

Studies on the In Vitro Development of Cloned Embryos by Somatic Cell Nuclear Transfer in Korean Native Goats (재래산양의 체세포 핵이식에 의한 복제수정란의 체외발달에 관한 연구)

  • Park H. S.;Kim T. S.;Jung S. Y.;Lee Y. H.;Jung J. Y.
    • Journal of Embryo Transfer
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    • v.20 no.2
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    • pp.105-112
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    • 2005
  • The present study was conducted to examine some factors affecting in vitro development of oocytes from somatic cell nuclear transfer (SCNT) in Korean native goats. Recipient oocytes were surgically collected after superovulation by using CIDR and FSH, PMSG, hCG and estrous synchronization in Korean Native goats. For nuclear transfer, the fibroblasts from caprine ear cells and fetal fibroblasts were surgically harvested and were cultured in vitro until cell confluency in serum-starvation condition (TCM-199 + $0.5\%$ FBS) for 3 to 5 days. The zona pellucidae of matured oocytes were partially drilled by laser irradiation. A single somatic cell was individually transferred into each enucleated oocyte. The reconstructed oocytes were then electrically fused and activated. Activated NT embryos were cultured in mSOF medium supplemented with $0.8\%\;BSA\;6\~7\;day\;at\;39^{\circ}C,\;5\%\;CO_2,\;5\%\;O_2,\;90\%\;N_2$ in air. There were no significant difference in the number of embryos cleaved and 4-cell development between the fibroblast nuclei from mature ear cells and fetal cells, but the rate of 8-cell development was higher (P<0.05) in ear cells $(40.5\%)$ than in fetal cells $(55.5\%)$. However, the embryo development to morula or blastocyst was not significantly different between both the groups$(6.7\%\;vs\;16.0\%)$, respectively. The number of embryo cleaved $(79.0\%)$ were higher (P<0.05) in the oocytes activated with ionomycin+6-DMAP than in the oocytes activated electrically $(9.5\%)$. The development of fused embryos to morula or blastocyst was found $15.6\%$ in ionomycin+6-DMAP, but no morula or blastocysts were developed in electrical stimulation. The development rate of SCNT embryos to morula or blastocyst was love. (P<0.05) in SCNT embryos $(19.0\%\;vs\;0.0\%)$ than that in parthenotes $(66.1\%\;vs\;59.1\%)$. In the parthenotes, the cleavage rate and development to morula or blastocyst were significantly higher (P<0.05) as $86.8\%\;and\;50.0\%$ in ovulated oocytes than in follicular oocytes $(69.0\%\;vs\;23.6\%)$, respectively. These results suggest that some factors Including superovulation treatment, oocyte source, maturation of follicular oocytes, activation method and culture condition may affect in vitro developmental capability of embryos produced by somatic cell nuclear transfer in Korean Native goats, and the fusion rate be greatly low compared with other species.