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The Evaluation of Various Conditions in the Cryopreservation of Primordial Germ Cells on Korean Native Chicken (Ogye) (한국재래닭(오계)의 원시 생식 세포의 냉동 보존에 있어서 여러 조건의 평가)

  • Kim, Hyun;Cho, Young Moo;Han, Jae Yong;Choi, Sung Bok;Byun, Mi Jeong;Kim, Young Sin;Ko, Yeoung-Gyu;Seong, Hwan-Hoo;Kim, Sung Woo
    • Korean Journal of Poultry Science
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    • v.41 no.4
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    • pp.249-259
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    • 2014
  • Cryopreserving cells which are maintaining their viability are the very complex process. This study has been carried out in order to find the effects of cryopreservation steps and freezing media on the rates of viability of cryopreserved chicken primordial germ cells (PGCs). PGCs obtained from the germinal gonade of 5.5~6 day (stage 28) chick embryos of Korean Ogye (KO) and Commercial breeds (C), using the MACS method were suspended in a freezing medium containing a freezing and protecting agents (e.g. dimethyl sulfoxide (DMSO), ethylene glycol (EG) and propylene glycol (PG)). Gonads were harvested from stage 28 chick embryos and pooled in groups of 5, 10, 15, 20E embryos, contributing gonads to the cell suspension. The gonadal cells, including PGCs, were then frozen in 1 of the following cryoprotectant treatments : 2.5%, 5%, 10%, 15% and 0% cryoprotectant (DMSO, EG, PG) as a control. Effects of exposure to slow freezing and vitrification, with different concentrations of the cryoprotectant solution, were examined. After vitrification and slow freezing, survival rates of the frozen-thawed PGCs from the 10% EG plus FBS treatment were 85.63%, and 66.14% (p<0.05), respectively. The viability of PGCs after freeze-thawing was significantly higher for 10% EG plus FBS treatment than for 10% PG + FBS treatment (p<0.05) (85.63% vs 66.81%) by vitrification. This study established a method for preserving chicken PGCs that enables systematic storage and labeling of cryopreserved PGCs in liquid ($LN_2$) at a germplasm repository and ease of entry into a data base. In the future, the importance for this new technology is that poultry lines can be conserved while work is being conducted on improving the production of germline chimeras.

Spatial Distribution of Transparent Exopolymer Particles(TEP) and Their Relation to Carbon Species in the Euphotic Layer of the Northern East Sea (동해 북부해역 유광층에서 TEP 분포와 이산화탄소 인자와의 상호관련성)

  • Jeon, Hyun-Duck;Rho, Tae-Keun;Lee, Tong-Sup
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.17 no.2
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    • pp.33-44
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    • 2012
  • Transparent exopolymer particles (TEP) are formed by aggregation of polysaccharide products excreted by phytoplankton and have sticky character like gel. They play important role in the production of marine snow in water column. To study the distribution pattern of TEP concentration and its role in carbon cycle in the surface ocean, we measured pH, Total alkalinity (TA), and chlorophyll-a in addition to physical characteristics of seawater within the surface water column. TEP concentrations ranged from nearly undetectable values to $338{\mu}g\;Xeq\;l^{-1}$. They were considerably lower than previously reported values from costal sites, but showed similar values observed in other oceanic region during phytoplankton bloom periods. The spatial distribution of TEP concentrations were similar to those of chlorophyll-a, which indicate that the production of TEP were closely related to phytoplankton. Calculated total dissolved inorganic carbon ($TCO_2$) from the pH and TA was normalized to 35 psu of salinity ($NTCO_2$) and showed negative linear relationship with temperature. Biological drawdown of $NTCO_2$ ($NTCO_{2bio}$) was estimated from the difference between theoretical $NTCO_2$ values and observed $NTCO_2$. In the warm region located south of $40^{\circ}N$ along the $132.5^{\circ}N$ meridional lines, $NTCO_{2bio}$ showed negative value and TEP concentrations were high. This suggested that negative $NTCO_{2bio}$ may be attributed to the biological processes. At the stations located between 44 and $46^{\circ}N$, TEP concentrations showed high concentration at the chlorophyll-a maximum layer within the water column while they showed low concentration in the surface layer. Carbon content of TEP constituted about 40% of $NTCO_{2bio}$ at the chlorophylla maximum layer. In this study, we could not observe any positive and negative relationship between TEP concentration and $NTCO_2$ or pH. It is obvious that we should consider the importance of TEP in the biological carbon cycling processes within surface layer.

Isolation and functional analysis of three microsomal delta-12 fatty acid desaturase genes from Camelina sativa (L.) cv. CAME (카멜리나 (Camelina sativa L. cv. CAME)로부터 3 microsomal delta-12 fatty acid desaturase 유전자들의 분리 및 기능 분석)

  • Kim, Hyojin;Go, Young Sam;Kim, Augustine Yonghwi;Lee, Sanghyeob;Kim, Kyung-Nam;Lee, Geung-Joo;Kim, Gi-Jun;Suh, Mi Chung
    • Journal of Plant Biotechnology
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    • v.41 no.3
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    • pp.146-158
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    • 2014
  • Camelina sativa that belongs to Brassicaceae family is an emerging oilseed crop. Camelina seeds contain approximately 40% storage oils per seed dry weight, which are useful for human and animal diets and industrial applications. Microsomal delta-12 fatty acid desaturase2 (FAD2) catalyzes the conversion of oleic acid to linoleic acid. The polymorphisms of FAD2 genes are correlated with the levels of oleic acids in seed oils. Microsomal delta-12 fatty acid desaturase2 (FAD2) catalyzes the conversion of oleic acid to linoleic acid. The polymorphisms of FAD2 genes are correlated with the levels of oleic acids in seed oils. In this study, three CsFAD2 genes (CsFAD2-1, CsFAD2-2 and CsFAD2-3.1) were isolated from developing seeds of Camelina sativa (L.) cv. CAME. The nucleotide and deduced amino acid sequences of three CsFAD2 genes were compared with those from dicotyledon and monocotyledon plants including Camelina cultivars Sunesone and SRS933. Three histidine motifs (HECGH, HRRHH, and HVAHH) required for FAD activity and a hydrophobic valine or isoleucine residue, which is a SNP (single nucleotide polymorphism) marker related with enzyme activity are well conserved in three CsFAD2s. The expressions of CsFAD2-1 and CsFAD2-3.1 were ubiquitously detected in various Camelina organs, whereas the CsFAD2-2 transcripts were predominantly detected in flowers and developing seeds. The contents of oleic acids decreased, whereas the amounts of linoleic acid increased in dry seeds of transgenic fad2-2 lines expressing each CsFAD2 gene compared with fad2-2 mutant, indicating that three CsFAD2 genes are functionally active. The isolated CsFAD2 genes might be applicable in metabolic engineering of storage oils with high oleic acids in oilseed crops.

Qualitative and quantitative PCR detection of insect-resistant genetically modified rice Agb0101 developed in korea (해충저항성 유전자변형 벼 Agb0101에 대한 PCR 검정)

  • Shin, Kong-Sik;Lee, Jin-Hyoung;Lim, Myung-Ho;Woo, Hee-Jong;Qin, Yang;Suh, Seok-Cheol;Kweon, Soon-Jong;Cho, Hyun-Suk
    • Journal of Plant Biotechnology
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    • v.40 no.1
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    • pp.18-26
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    • 2013
  • Genetically modified (GM) rice Agb0101, which expresses the insecticidal toxin modified cry1Ac (mcry1Ac1) gene, was developed by the Rural Development Administration in Korea. To monitor the probable release of Agb0101 in the future, it is necessary to develop a reliable detection method. Here, we developed the PCR detection method for monitoring and tracing of GM rice. The primer pair (RBEgh-1/-2) from a starch branching enzyme (RBE4) gene was designed as an endogenous reference, giving rise to an expected PCR amplicon of 101 bp. For the qualitative PCR detection, construct- and event-specific primers were designed on the basis of integration sequence of T-DNA. Event-specific PCRs amplified specifically 5'- or 3'-junction region spanning the native genome DNA and the integrated gene construct, while none of amplified product was shown on crops, rice varieties, and other insect-resistant transgenic rice lines. The event-specific real-time PCR method was performed using TaqMan probe and plasmid pRBECrR containing both rice endogenous gene RBE4 sequence and 5'-junction sequence as the reference molecule. The absolute limit of quantification (LOQ) of real-time PCR was established with around 10 copies for one plasmid molecule pRBECrR. Thereafter, the different amounts of transgenic rice (1, 3, 5, and 10%, respectively) were quantified by using the established real-time PCR method, with a range below 19.55% of the accuracy expressed as bias, 0.06-0.40 of standard deviation (SD) and 3.80-7.01% of relative standard deviations (RSD), respectively. These results indicate that the qualitative and quantitative PCR methods could be used effectively to detect the event Agb0101 in monitoring and traceability.

CT Simulation Technique for Craniospinal Irradiation in Supine Position (전산화단층촬영모의치료장치를 이용한 배와위 두개척수 방사선치료 계획)

  • Lee, Suk;Kim, Yong-Bae;Kwon, Soo-Il;Chu, Sung-Sil;Suh, Chang-Ok
    • Radiation Oncology Journal
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    • v.20 no.2
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    • pp.165-171
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    • 2002
  • Purpose : In order to perform craniospinal irradiation (CSI) in the supine position on patients who are unable to lie in the prone position, a new simulation technique using a CT simulator was developed and its availability was evaluated. Materials and Method : A CT simulator and a 3-D conformal treatment planning system were used to develop CSI in the supine position. The head and neck were immobilized with a thermoplastic mask in the supine position and the entire body was immobilized with a Vac-Loc. A volumetrie image was then obtained using the CT simulator. In order to improve the reproducibility of the patients' setup, datum lines and points were marked on the head and the body. Virtual fluoroscopy was peformed with the removal of visual obstacles such as the treatment table or the immobilization devices. After the virtual simulation, the treatment isocenters of each field were marked on the body and the immobilization devices at the conventional simulation room. Each treatment field was confirmed by comparing the fluoroscopy images with the digitally reconstructed radiography (DRR)/digitally composite radiography (DCR) images from the virtual simulation. The port verification films from the first treatment were also compared with the DRR/DCR images for a geometrical verification. Results : CSI in the supine position was successfully peformed in 9 patients. It required less than 20 minutes to construct the immobilization device and to obtain the whole body volumetric images. This made it possible to not only reduce the patients' inconvenience, but also to eliminate the position change variables during the long conventional simulation process. In addition, by obtaining the CT volumetric image, critical organs, such as the eyeballs and spinal cord, were better defined, and the accuracy of the port designs and shielding was improved. The differences between the DRRs and the portal films were less than 3 mm in the vertebral contour. Conclusion : CSI in the supine position is feasible in patients who cannot lie on prone position, such as pediatric patienta under the age of 4 years, patients with a poor general condition, or patients with a tracheostomy.

Characteristics of Sea Water Intrusion Using Geostatistical Analysis of Geophysical Surveys at the Southeastern Coastal Area of Busan, Korea (지구물리 탐사자료의 지구통계학적 분석에 의한 부산 동남해안 지역의 해수침투 특성)

  • 심병완;정상용;김희준;성익환;김병우
    • Journal of Soil and Groundwater Environment
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    • v.7 no.3
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    • pp.3-17
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    • 2002
  • Data analysis of groundwater monitoring wells and geostatistical methods are used to identify the local characteristics of sea water intrusion and the range of sea water intrusion at the southeastern coastal area of Busan, Korea. Rainfall and groundwater level of two monitoring wells show a linear correlation because of the direct groundwater recharge by the precipitation. However, rainfall and electric conductivity have the inverse relationship because of the increase of groundwater. Electric conductivity rapidly increased at 24m depth and exceeded 20,000$\mu\textrm{s}$/cm near 26m depth in the monitoring wells. The variations of groundwater level and electric conductivity show that the interface between sea water and fresh water tends to move upward when groundwater level goes down. In the cross correlation analysis, groundwater level versus rainfall represents the largest cross correlation coefficient in 0 time lag but the cross correlation coefficient of electric conductivity versus rainfall is the largest when the time lag is 9 days. This suggests that the fluctuations of groundwater level respond to rainfall in a short time, but the interface between sea water and fresh water respond very slow to rainfall. Horizontal extents of sea water intrusion are estimated to 14 m from the east of Line 1, and 25 m from the southeast end of Line 2 in the inversion of dipole-dipole profiling data of two survey lines. The data of VES by the Schulumberger array in May and July show lognormal distributions. In the kriged apparent resistivity and earth resistivity distributions, the resistivities of July are increased comparing to those of May. This reflects that the concentration of sea water in aquifer is reduced due to the increased groundwater recharge from the rainfall in June and July. In analyzing the vertical and horizontal apparent resistivities and earth resistivity distributions, the geostatistical methods are very useful to identify the variations of earth resistivity distributions at the coastal area.

Relations between Polymorphism of NRAMP1 Gene and Susceptibility to Pulmonary Tuberculosis (NRAMP1 유전자 다형성과 폐결핵의 감수성과의 관계)

  • Lee, Ji Seok;Cho, Jin Hoon;Kim, Ki Uk;Park, Hye-Kyung;Kim, Yun Seong;Lee, Ho Seok;Kim, Yeong Dae;Jeon, Doo Soo;Park, Seung Kyu;Lee, Min Ki;Park, Soon Kew
    • Tuberculosis and Respiratory Diseases
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    • v.62 no.6
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    • pp.492-498
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    • 2007
  • Background: Several lines of evidence suggest that a host's genetic factors influence the outcome of exposure to Mycobacterium tuberculosis. The aim of this study was to determine whether polymorphism in NRAMP1 (natural resistance associated macrophage protein 1) gene is associated with the susceptibility or resistance to tuberculosis infection for patients with drug-sensitive pulmonary tuberculosis (DS-TB) and multi-drug resistant pulmonary tuberculosis (MDR-TB). Methods: Eight genetic polymorphisms of the NRAMP1 gene were investigated in patients suffering with DS-TB (n=100) or MDR-TB (n=102), and in healthy normal controls (NC, n=96). The genetic polymorphisms of NRAMP1 were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Results: The frequency of D543N A/G heterogygotes was significantly higher in the DS-TB subjects than the NCs (OR=2.10, 95% CI: 1.00 to 4.41, p=0.049). The frequency of 823C/T T/C heterozygotes was significantly higher in the DS-TB subjects (OR=2.79, 95% CI: 1.11 to 7.04, p=0.029) and the MDR-TB subject (OR=3.30, 95% CI 1.33 to 8.18, p=0.010) than in the NCs. However, the frequency of these genotypes was not different between the DS-TB and MDR-TB subjects. Conclusion: A significant association was found between NRAMP1 823 C/T polymorphism and pulmonary tuberculosis. This result suggests that NRAMP1 polymorphism may be involved in the development of pulmonary tuberculosis in Koreans.

Design of CMOS Multifunction ICs for X-band Phased Array Systems (CMOS 공정 기반의 X-대역 위상 배열 시스템용 다기능 집적 회로 설계)

  • Ku, Bon-Hyun;Hong, Song-Cheol
    • Journal of the Institute of Electronics Engineers of Korea TC
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    • v.46 no.12
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    • pp.6-13
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    • 2009
  • For X-band phased array systems, a power amplifier, a 6-bit phase shifter, a 6-bit digital attenuator, and a SPDT transmit/receive (T/R) switch are fabricated and measured. All circuits are demonstrated by using CMOS 0.18 um technology. The power amplifier has 2-stage differential and cascade structures. It provides 1-dB gain-compressed output power ($P_{1dB}$) of 20 dBm and power-added-efficiency (PAE) of 19 % at 8-11 GHz frequencies. The 6-bit phase shifter utilizes embedded switched filter structure which consists of nMOS transistors as a switch and meandered microstrip lines for desired inductances. It has $360^{\circ}$ phase-control range and $5.6^{\circ}$ phase resolution. At 8-11 GHz frequencies, it has RMS phase and amplitude errors are below $5^{\circ}$ and 0.8 dB, and insertion loss of $-15.7\;{\pm}\;1,1\;dB$. The 6-bit digital attenuator is comprised of embedded switched Pi-and T-type attenuators resistive networks and nMOS switches and employes compensation circuits for low insertion phase variation. It has max. attenuation of 31.5 dB and 0.5 dB amplitude resolution. Its RMS amplitude and phase errors are below 0.4 dB and $2^{\circ}$ at 8-11 GHz frequencies, and insertion loss is $-10.5\;{\pm}\;0.8\;dB$. The SPDT T/R switch has series and shunt transistor pairs on transmit and receive path, and only one inductance to reduce chip area. It shows insertion loss of -1.5 dB, return loss below -15 dB, and isolation about -30 dB. The fabricated chip areas are $1.28\;mm^2$, $1.9mm^2$, $0.34\;mm^2$, $0.02mm^2$, respectively.

Effect of Globe Growth and Chromogenic on Day and Night Temperature and the LED Light Treatment of Expert Grafted Cactus (Chamecereus silvestrii f. variegata) Cultivar 'Hee-Mang' (수출 접목선인장 산취 '희망'의 주.야 온도 및 LED광 처리가 모구 생육과 발색에 미치는 영향)

  • Nam, Sang-Yong;Park, Sun-Mi;Ahn, Dong-Hyun
    • Journal of Bio-Environment Control
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    • v.19 no.3
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    • pp.140-146
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    • 2010
  • This experiment was conducted to find the day and night temperature combination and the light-emitting diodes (LEDs) on the most effective globe coloration and growth of Chamecereus silvestrii f. variegata 'Hee-Mang'. The $L^*$value, $a^*$value and $b^*$value were all significant difference in the yellow globe color expression in the day and night temperature combination. Especially, the bright yellow color and visual value were the highest in the temperature combination, the day temperature of $20^{\circ}C$ and night temperature of $20^{\circ}C$. Globe color quality was lowered from the day temperature of $25^{\circ}C$ and night temperatures of $5^{\circ}C$ and $10^{\circ}C$ combination. In light-emitting diodes (LEDs) treatments, $a^*$value and $b^*$value are significant difference. A unique beautiful yellow coloration and globe quality were maintained the $a^*$value of +5.23, $b^*$value of +39.9 in a red LED. The optimum temperature range and light-emitting diodes (LEDs) on the globe color expression were the most effective the day temperature of 200e and night temperature of $20^{\circ}C$ and a red LED. In addition, the outer globe color quality of Chamecereus silvestrii f. variegata cultivation, rather than the light environment improvement is better in a proper temperature environment to keep. Especially, a unique globe color expression of yellow lines was most effective in a red LED.

Microarray Analysis of Long Non-coding RNA Expression Profile Associated with 5-Fluorouracil-Based Chemoradiation Resistance in Colorectal Cancer Cells

  • Xiong, Wei;Jiang, Yong-Xin;Ai, Yi-Qin;Liu, Shan;Wu, Xing-Rao;Cui, Jian-Guo;Qin, Ji-Yong;Liu, Yan;Xia, Yao-Xiong;Ju, Yun-He;He, Wen-Jie;Wang, Yong;Li, Yun-Fen;Hou, Yu;Wang, Li;Li, Wen-Hui
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.8
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    • pp.3395-3402
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    • 2015
  • Background: Preoperative 5-fluorouracil (5-FU)-based chemoradiotherapy is a standard treatment for locally advanced colorectal cancer (CRC). However, CRC cells often develop chemoradiation resistance (CRR). Recent studies have shown that long non-coding RNA (lncRNA) plays critical roles in a myriad of biological processes and human diseases, as well as chemotherapy resistance. Since the roles of lncRNAs in 5-FU-based CRR in human CRC cells remain unknown, they were investigated in this study. Materials and Methods: A 5-FU-based concurrent CRR cell model was established using human CRC cell line HCT116. Microarray expression profiling of lncRNAs and mRNAs was undertaken in parental HCT116 and 5-FU-based CRR cell lines. Results: In total, 2,662 differentially expressed lncRNAs and 2,398 mRNAs were identified in 5-FU-based CRR HCT116 cells when compared with those in parental HCT116. Moreover, 6 lncRNAs and 6 mRNAs found to be differentially expressed were validated by quantitative real time PCR (qRT-PCR). Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis for the differentially expressed mRNAs indicated involvement of many, such as Jak-STAT, PI3K-Akt and NF-kappa B signaling pathways. To better understand the molecular basis of 5-FU-based CRR in CRC cells, correlated expression networks were constructed based on 8 intergenic lncRNAs and their nearby coding genes. Conclusions: Changes in lncRNA expression are involved in 5-FU-based CRR in CRC cells. These findings may provide novel insight for the prognosis and prediction of response to therapy in CRC patients.