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In Vitro Fertilization and Development of Bovine Immature Oocytes by Ultra- Rapid Cooling (미성숙 소난자의 초급속 동결ㆍ융해 후 수정 및 체외 배발달)

  • Ryu, C. S.;K. S. Kil;Kim, N-H;D. H. Ko;Lee, H. T.;K. S. Chung
    • Korean Journal of Animal Reproduction
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    • v.23 no.3
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    • pp.221-227
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    • 1999
  • Successful cryopreservation of bovine immature oocytes can increase availably of oocytes for the in vitro fertilization or nuclear transfer. However, it was not reported successful development to the blastocyst stage following in vitro fertilization of cryopreserved bovine immature oocytes. The objective of this study was to determine the incidence of survival, meiotic maturation, fertilization and in vitro development of cryopreserved bovine immature by ultra rapid cooling methods. The oocytes were adversely affected by brief exposure to EFS40 solution in electron microscope grids and plunged directly into liquid nitrogen. After such ultra-rapid cooled immature oocytes were warmed, 78% of oocytes were matured to the metaphase II stage, 50% of oocytes were fertilized after insemination, and 5% of oocytes were developed to the blastocyst stage. Different sodium concentration of sodium ion in the freezing medium did not affect survival, maturation, fertilization and in vitro development of cryopreserved oocytes. These results suggested that immature bovine oocytes can be cryopreserved by ultra-rapid cooling methods.

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Heterologous Expression and Characterization of Glycogen Branching Enzyme from Synechocystis sp. PCC6803

  • Lee, Byung-Hoo;Yoo, Young-Hee;Ryu, Je-Hoon;Kim, Tae-Jip;Yoo, Sang-Ho
    • Journal of Microbiology and Biotechnology
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    • v.18 no.8
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    • pp.1386-1392
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    • 2008
  • A gene (sll0158) putatively encoding a glycogen branching enzyme (GBE, E.C. 2.4.1.18) was cloned from Synechocystis sp. PCC6803, and the recombinant protein expressed and characterized. The PCR-amplified putative GBE gene was ligated into a pET-21a plasmid vector harboring a T7 promoter, and the recombinant DNA transformed into a host cell, E. coli BL21(DE3). The IPTG-induced enzymes were then extracted and purified using Ni-NTA affinity chromatography. The putative GBE gene was found to be composed of 2,310 nucleotides and encoded 770 amino acids, corresponding to approx. 90.7 kDa, as confirmed by SDS-PAGE and MALDI-TOF-MS analyses. The optimal conditions for GBE activity were investigated by measuring the absorbance change in iodine affinity, and shown to be pH 8.0 and $30^{\circ}C$ in a 50 mM glycine-NaOH buffer. The action pattern of the GBE on amylose, an $\alpha$-(1,4)-linked linear glucan, was analyzed using high-performance anion-exchange chromatography (HPAEC) after isoamylolysis. As a result, the GBE displayed $\alpha$-glucosyl transferring activity by cleaving the $\alpha$-(1,4)-linkages and transferring the cleaved maltoglycosyl moiety to form new $\alpha$-(1,6)-branch linkages. A time-course study of the GBE reaction was carried out with biosynthetic amylose (BSAM; $M_p{\cong}$8,000), and the changes in the branch-chain length distribution were evaluated. When increasing the reaction time up to 48 h, the weight- and number-average DP ($DP_w$ and $DP_n$) decreased from 19.6 to 8.7 and from 17.6 to 7.8, respectively. The molecular size ($M_p$, peak $M_w{\cong}2.45-2.75{\times}10^5$) of the GBE-reacted product from BSAM reached the size of amylose (AM) in botanical starch, yet the product was highly soluble and stable in water, unlike AM molecules. Thus, GBE-generated products can provide new food and non-food applications, owing to their unique physical properties.

Difference of Neuronal Recovery by Incubation Condition after Transient Hypoxia (배양조건에 의한 일과성 저산소상태 후 신경세포회복의 차이)

  • Moon, Soo-Hyeon;Oh, Jae-Inn;Park, Youn-Kwan;Chung, Heung-Sub;Lee, Hoon-Kap;Lee, Ki-Chan
    • Journal of Korean Neurosurgical Society
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    • v.29 no.9
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    • pp.1161-1170
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    • 2000
  • Objective : The transverse hippocampal slice is one of the most commonly studied in vitro models of mammalian brain physiology. However, despite its broad usage, there has been no standardization of slice preparation techniques or recording condition. It is well known that variations in recording conditions can result in profound different effects to neuronal responses. Evoked field potentials, recorded extracellularly, were used to investigate the effects of variations in hippocampal slice preparation protocol on hypoxia responses of CA1 neurones. Material & Methods : Before hypoxic injury, hippocampal slices were incubated for 4 hours. During incubation period, the slices were placed in a incubation chamber($21^{\circ}C$) for recovery from preparation injury and then transferred to recording chamber($34^{\circ}C$) for more recovery and baseline electric recording with current stimulation(0.1Hz). Various time periods in incubation chamber and recording chamber were applied to each experimental group(group 1=60min : 180min, group 2=90min : 150min, group 3=180min : 60min, time in incubation chamber : time in recording chamber) before 10 min hypoxia produced by replacing 95% $O_2$+5% $CO_2$ mixed gas to 95% $N_2$+5% $CO_2$ gas. Calcium, Magnesium ions and several drugs effecting on glutamate receptor also were studied. Recoveries from hypoxic injury of hippocampal slices were estimated by percent recovery of population spike(PS). Statistic analysis of study were performed using paired t-test. Results : The percent recovery of PS after 10min hypoxia was considerably enhanced by increasing the period of current stimulation during incubation period before hypoxic injury. Temperature effect on the result of this experiment was also studied(group 4) but the result from this showed no statistic significance. Low magnesium ion concentration of artificial CSF(Mg-free aCSF) during incubation period enhanced the recovery of PS but low calcium (calcium-free) and high magnesium ion concentration(2mM) reduced it after hypoxic injury. L-glutamate($100{\mu}M$) and AP-5($50{\mu}M$) had no effect on the recovery of PS but CNQX($10{\mu}M$) in artificial CSF during incubation period markedly enhanced the recovery of PS. Co-treatment of AP-5($50{\mu}M$), CNQX($10{\mu}M$) and high magnesium concentration(2mM) enhanced recovery of PS in immediate following period of hypoxic injury but the effect of cotreatment after then decayed rapidly and lost statistic significance. Conclusions : Judging from above results, the condition of baseline recording is important in observing the recovery of population spike after hypoxia, and the time and the condition should be controled more strictly to obtain reliable results.

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Study on Characteristics of the Visual Perception of Historic Cultural Landscape - A Case of Gyeongbok Palace - (역사문화경관의 시지각적 특성에 관한 연구 - 경복궁 내부의 조망점을 중심으로 -)

  • Suh, Joo-Hwan;Kim, Su-Yeon
    • Journal of the Korean Institute of Traditional Landscape Architecture
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    • v.31 no.1
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    • pp.108-117
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    • 2013
  • Modern city doesn't have its own identity that differentiates itself from others despite the fact that each city has a historical value. This is due to the demolition of historical and regional values. As such problems are connected to urban problems, they become even more serious. Thus, the researchers of this study set Gyeongbokgung(Palace) as an object to look into landscape factors and found out the relations between psychological variables and preference. Then, the researchers analyzed the quantitative relations between the physical attributes of historic cultural landscape and human response. As such, the researchers aimed to set and propose a confirmed standard in terms of identity and regional & historical values of historic cultural landscape. Since this study was performed with an object of historic cultural landscape that was reproduced based on original landscape in the past, a future study should be on the difference in preference based on the proportion of modern landscape. Based on such studies, a new plan should be made for forming the urban historic cultural landscape.

Relative Effectiveness of Replacement Diets for Mass Production of the Brackish Water Flea Diaphanosoma celebensis (기수산 물벼룩(Diaphanosoma celebensis)의 대량생산을 위한 대체 먹이원의 효과)

  • Park, Jin-Chul;Kwon, O-Nam;Park, Heum-Gi
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.45 no.2
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    • pp.146-153
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    • 2012
  • This study examined the effect of formulated diets (composed of condensed freshwater $Chlorella$ or $Schizochytrium$ sp., $Crypthecodinium$ sp., and EPA oil sources derived from baker's yeast) on the population growth and fatty acid composition of the brackish water flea ($Diaphanosoma$ $celebensis$) to determine diet effectiveness for mass production. Results indicated that the highest maximum density, fecundity, and population specific growth rate (SGR) of the flea were observed using a diet of $Tetraselmis$ $suecica$ (control). However, results using condensed freshwater $Chlorella$ and the mixed treatment of $Schizochytrium$ sp.+$Crypthecodinium$ sp.+EPA oil did not significantly differ from the control treatment ($P$>0.05). Thus, $T.$ $suecica$ could be replaced with a combination diet of $Schizochytrium$ sp., $Crypthecodinium$ sp., and EPA oil in condensed freshwater $Chlorella$ without growth interruption of $D.$ $celebensis$. As the percentage of combination diet (mix of $Schizochytrium$ sp., $Crypthecodinium$ sp., and EPA oil) in condensed freshwater $Chlorella$ increased, the level of ARA, EPA, DHA, and n-3 HUFA in $D.$ $celebensis$ also increased. However, the population density of $D.$ $celebensis$ sharply decreased with the use of treated supplements C5M5 (CHL [50%] and $Crypthecodinium$ sp.+ $Schizochytrium$ sp. + EPA oil [MIX 50%]). The optimum proportions of CHL and MIX for $D.$ $celebensis$ within the mixture were 70% and 30%, respectively. Based on the results, the combination of $Schizochytrium$ sp., $Crypthecodinium$ sp., and EPA oil in condensed freshwater $Chlorella$ appears to be a stable and effective food source for mass production of $D.$ $celebensis$.

Catalog use study : with reference to universities in Daegu (대학도서관의 목록이용연구)

  • 최달현
    • Journal of Korean Library and Information Science Society
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    • v.9
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    • pp.241-266
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    • 1982
  • This paper gives a summary and overview of a survey conducted at the catalogues of five universities in Daegu during November 1982. The major objective of this study was to secure information about user needs in order to improve the catalogue in Korean university libraries. Data was collected by a combined method of questionnaire and interview. A total of 379 respondents were taken on a randomly selected sample of catalogue users. Results of the survey can be summarized as follows: 1. Eighty-eight percent of the students answered that they had used the library more than twice a month. Nevertheless the number of students consulting the catalogue with the same frequency was only 220, or a n.0, pproximately 58 percent. Those who used the library most tended to use the catalogue more than those who rarely came to the library. 2. Those who had been shown how to use their own catalogue were only 32.5percent of which the students found the instruction sufficient for them to be able to use the catalogue were only 37.5 percent. In particular, they stated that instructions by printed materials and/or library orientation were so poor that they didn't give much help in using the catalogue. This problem makes many libraries to review their own method of instruction in order to encourage patrons to use the catalogue more effectively. 3. Most of the students consulted the catalogue in order to locate library materials. Known-item searches and subject searches were 84 and 16 percent respectively. While 70.5 percent of the students used the author-title catalogue without any difficulties, only 35.5 percent of those stated that using the classed catalogue was easy. 4. It was surprising that the number of students using title access in the search was far greater than that of students using author access. In contrast with this, other studies conducted by many earlier overseas investigators revealed that the great majority of patrons tended to use the latter first. Therefore, we should put more emphasis on the title entries in the catalogue itself as well as cataloging rules. 5. Most useful bibliographic elements in the entry were author, title, call number, date and publisher whereas edition, series statement and the location of publisher were rarely used compared with the other elements. Content note was the most desirable element in the entry to be involved, for many catalogers were used not to describe it on the note area. 6. The chief reason given for not using the catalogue was "I can manage without it" with "It's difficult to understand contents of the card entry." The other one was "It's useless to search materials by the catalogue because I've failed so many times to obtain them out of the stock." In response to this, circulation and acquisition system should be improved not to make such complaints any more.

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IN VITRO DEVELOPMENT OF BOVINE ONE-CELL EMBRYOS FERTILIZED IN VITRO IN SERUM- AND FEEDER CELL-FREE CULTURE SYSTEMS

  • Ohboshi, S.;Hanada, K.;Zhao, J.;Hattori, M.;Fujihara, N.;Umetsu, R.;Yoshida, T.;Tomogane, H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.9 no.5
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    • pp.583-590
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    • 1996
  • The purpose of this study was to evaluate some factors in the bovine embryonic development from one-cell to blastocyst using modified synthetic oviduct fluid medium (mSOFM), after maturation and in vitro fertilization of the oocytes. The embryonic development to the blastocyst stage was assessed at 7-10 days after in vitro fertilization, and the total cells in the blastocysts were counted by staining nuclei with fluorochrome. Some commercial calf sera (CS) and a superovulated cow serum had different effects on the embryonic development to the blastocyst stage (8.6-21.4%), dependent upon their product lots, although the development might not be affected at least by serum progesterone levels. ${\beta}$-Mercaptoethanol (${\beta}$-ME) supplemented into mSOFM was effective to the embryonic development (27.8%), as well as the co-culture system with cumulus cells (19.5%). In a serum- and feeder cell-free culture using mSOFM containing several growth factors and ${\beta}$-ME instead of CS plus co-cultured cumulus cells, bovine serum albumin (BSA, fraction V), but not polyvinyl alcohol (PVA), was highly effective in embryonic development to the blastocyst stage, almost comparable to CS in the serum-contained culture (CS, BSA and PVA; 27.8, 19.5 and 5.7%, respectively). However, fatty acid free BSA rather reduced the number of developed blastocysts, compared with fraction V BSA (7.3 vs 29.4%). In the serum- and feeder cell-free culture, supplement of glucose to the medium (final 2.0 mM) stimulated the cell proliferation of developing embryos 120 hr after in vitro fertilization. These results indicated that a serum-free medium supplemented with ${\beta}$-ME could successfully support the development of bovine one-cell embryos to the blastocyst stage. Moreover, supplement of glucose and fatty acids to the medium might support preferably the development and cell proliferation of embryos.

Dosimetric Validation of the Acuros XB Advanced Dose Calculation Algorithm for Volumetric Modulated Arc Therapy Plans

  • Park, So-Yeon;Park, Jong Min;Choi, Chang Heon;Chun, Minsoo;Kim, Jung-in
    • Progress in Medical Physics
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    • v.27 no.4
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    • pp.180-188
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    • 2016
  • Acuros XB advanced dose calculation algorithm (AXB, Varian Medical Systems, Palo Alto, CA) has been released recently and provided the advantages of speed and accuracy for dose calculation. For clinical use, it is important to investigate the dosimetric performance of AXB compared to the calculation algorithm of the previous version, Anisotropic Analytical Algorithm (AAA, Varian Medical Systems, Palo Alto, CA). Ten volumetric modulated arc therapy (VMAT) plans for each of the following cases were included: head and neck (H&N), prostate, spine, and lung. The spine and lung cases were treated with stereotactic body radiation therapy (SBRT) technique. For all cases, the dose distributions were calculated using AAA and two dose reporting modes in AXB (dose-to-water, $AXB_w$, and dose-to-medium, $AXB_m$) with same plan parameters. For dosimetric evaluation, the dose-volumetric parameters were calculated for each planning target volume (PTV) and interested normal organs. The differences between AAA and AXB were statistically calculated with paired t-test. As a general trend, $AXB_w$ and $AXB_m$ showed dose underestimation as compared with AAA, which did not exceed within -3.5% and -4.5%, respectively. The maximum dose of PTV calculated by $AXB_w$ and $AXB_m$ was tended to be overestimated with the relative dose difference ranged from 1.6% to 4.6% for all cases. The absolute mean values of the relative dose differences were $1.1{\pm}1.2%$ and $2.0{\pm}1.2%$ when comparing between AAA and $AXB_w$, and AAA and $AXB_m$, respectively. For almost dose-volumetric parameters of PTV, the relative dose differences are statistically significant while there are no statistical significance for normal tissues. Both $AXB_w$ and $AXB_m$ was tended to underestimate dose for PTV and normal tissues compared to AAA. For analyzing two dose reporting modes in AXB, the dose distribution calculated by $AXB_w$ was similar to those of AAA when comparing the dose distributions between AAA and $AXB_m$.

Deoxynivalenol- and zearalenone-contaminated feeds alter gene expression profiles in the livers of piglets

  • Reddy, Kondreddy Eswar;Jeong, Jin young;Lee, Yookyung;Lee, Hyun-Jeong;Kim, Min Seok;Kim, Dong-Wook;Jung, Hyun Jung;Choe, Changyong;Oh, Young Kyoon;Lee, Sung Dae
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.4
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    • pp.595-606
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    • 2018
  • Objective: The Fusarium mycotoxins of deoxynivalenol (DON) and zerolenone (ZEN) cause health hazards for both humans and farm animals. Therefore, the main intention of this study was to reveal DON and ZEN effects on the mRNA expression of pro-inflammatory cytokines and other immune related genes in the liver of piglets. Methods: In the present study, 15 six-week-old piglets were randomly assigned to the following three different dietary treatments for 4 weeks: control diet, diet containing 8 mg DON/kg feed, and diet containing 0.8 mg ZEN/kg feed. After 4 weeks, liver samples were collected and sequenced using RNA-Seq to investigate the effects of the mycotoxins on genes and gene networks associated with the immune systems of the piglets. Results: Our analysis identified a total of 249 differentially expressed genes (DEGs), which included 99 upregulated and 150 downregulated genes in both the DON and ZEN dietary treatment groups. After biological pathway analysis, the DEGs were determined to be significantly enriched in gene ontology terms associated with many biological pathways, including immune response and cellular and metabolic processes. Consistent with inflammatory stimulation due to the mycotoxin-contaminated diet, the following Kyoto encyclopedia of genes and genomes pathways, which were related to disease and immune responses, were found to be enriched in the DEGs: allograft rejection pathway, cell adhesion molecules, graft-versus-host disease, autoimmune thyroid disease (AITD), type I diabetes mellitus, human T-cell leukemia lymphoma virus infection, and viral carcinogenesis. Genome-wide expression analysis revealed that DON and ZEN treatments downregulated the expression of the majority of the DEGs that were associated with inflammatory cytokines (interleukin 10 receptor, beta, chemokine [C-X-C motif] ligand 9), proliferation (insulin-like growth factor 1, major facilitator superfamily domain containing 2A, insulin-like growth factor binding protein 2, lipase G, and salt inducible kinase 1), and other immune response networks (paired immunoglobulin-like type 2 receptor beta, Src-like-adaptor-1 [SLA1], SLA3, SLA5, SLA7, claudin 4, nicotinamide N-methyltransferase, thyrotropin-releasing hormone degrading enzyme, ubiquitin D, histone $H_2B$ type 1, and serum amyloid A). Conclusion: In summary, our results demonstrated that high concentrations DON and ZEN disrupt immune-related processes in the liver.

Changes in Mechanical Properties and Magnetic Parameters of Neutron Irradiated Mn-Mo-Ni Low Alloy Steels (중성자에 조사된 Mn-Mo-Ni 저합금강의 기계적 및 자기적 성질 변화)

  • Jang, Gi-Ok;Ji, Se-Hwan;Park, Seung-Sik;Kim, Byeong-Cheol;Kim, Jong-O
    • Korean Journal of Materials Research
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    • v.8 no.11
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    • pp.1020-1025
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    • 1998
  • Irradiation-induced changes in mechanical properties and magnetic parameters were measured and compared to explore possible correlations for Mn-Mo-Ni low alloy steel surveillance specimens which were irradiated to a neutron fluence of $2.3\times10^{19}n/cm^2$(E>1.0MeV) in a typical pressurized water reactor environment at about $288^{\circ}C$. For mechanical property parameters, microvickers hardness, tensile and Charpy impact test were performed and Barkhausen noise amplitude, coercivity, remanence, maximum induction were measured for magnetic parameters. respectively. Results of mechanical property measurements showed an increase in yield and tensile strength, microvickers hardness. 41J indexed $RT_{NDT}$ and a decrease in upper shelf energy irrespective of base and weld metals. However, in the case of tensile properties, the changes in weld metal were negligible compared to the base metal. In the case of magnetic measurements, it is found that magnetic remanence, BN amplitude. BN energy have dropped significantly but coercivity(H,) has increased rapidly after irradiation. In this study. the measurements conducted on surveillance specimens of Mn-Mo-Ni low alloy steel showed that there were strong correlations between mechanical properties and magnetic properties.

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