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다양한 유리화 용액과 동결기구에서 유리 동결 생쥐 포배기 배아의 생존율 비교 (The Comparison of Survival Rates of Vitrified Mouse Blastocysts in Various Vitrification Solutions and Apparatuses)

  • 조동휴;이기숙;류철희;권중기;이정헌
    • 한국임상수의학회지
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    • 제29권1호
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    • pp.27-32
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    • 2012
  • The purpose of this study was to evaluate survival rates of vitrified mouse blastocysts in various vitrification solutions (cryoprotectants) and apparatuses. The mouse blastocysts were harvested from culture of mouse 2 cell embryo and were divided into three group (i) untreated (control); (ii) exposed to cryoprotectant agents; or (iii) cryopreserved by various vitrification apparatuses. Vitrification solutions are 40% ethylene glycol (EG) + 5.8 mg/mL ficoll + 0.5M sucrose (EFS solution), 3M glycerol + 3M EG (ES solution), 20% EG + 20% dimethyl sulfoxide (ED solution), 3M EG + 1.0 m sucrose (ES solution). Vitrification apparatuses consisted of 5 groups ; closed plastic straw (CPS), electron microscope (EM) grid, cryoloop, open pulled straw (OPS), and glass micropippete in plastic straw (GPS). The survival rates of control were 88.0%. The survival rates of exposed blastocysts in EFS, GE, ED, and ES solutions were 70.8%, 43.5% (P<0.01), 83.3% and 65.2%, respectively. The survival rates of vitrified blastocysts in CPS, EM grid, cryoloop, OPS and GPS were 56.5% (P< 0.01), 72.7%, 83.3%, 60.9% (P<0.05) and 54.2% (P<0.01), respectively. Among the vitrification solutions, the highest survival rate was seen in blastocysts vitrified in EG + DMSO (83.3%). The survival rate was not significantly different from that of the control (88%). Blastocysts cryopreserved with glycerol in all groups had an overall low survival rate of 43.5%. Survival rate of mouse blastocysts between vitrification apparatuses showed higher in cryoloop.

Novel splice isoforms of pig myoneurin and their diverse mRNA expression patterns

  • Guo, Xiaohong;Li, Meng;Gao, Pengfei;Cao, Guoqing;Cheng, Zhimin;Zhang, Wanfeng;Liu, Jianfeng;Liu, Xiaojun;Li, Bugao
    • Asian-Australasian Journal of Animal Sciences
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    • 제31권10호
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    • pp.1581-1590
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    • 2018
  • Objective: The aim of this study was to clone alternative splicing isoforms of pig myoneurin (MYNN), predict the structure and function of coding protein, and study temporal and spatial expression characteristics of each transcript. Methods: Alternative splice isoforms of MYNN were identified using RNA sequencing (RNA-seq) and cloning techniques. Quantitative real-time polymerase chain reaction (qPCR) was employed to detect expression patterns in 11 tissues of Large White (LW) and Mashen (MS) pigs, and to study developmental expression patterns in cerebellum (CE), stomach (ST), and longissimus dorsi (LD). Results: The results showed that MYNN had two alternatively spliced isoforms, MYNN-1 (GenBank accession number: KY470829) and MYNN-2 (GenBank accession number: KY670835). MYNN-1 coding sequence (CDS) is composed of 1,830 bp encoding 609 AA, whereas MYNN-2 CDS is composed of 1,746 bp encoding 581 AA. MYNN-2 was 84 bp less than MYNN-1 and lacked the sixth exon. MYNN-2 was found to have one $C_2H_2$ type zinc finger protein domain less than MYNN-1. Two variants were ubiquitously expressed in all pig tissues, and there were significant differences in expression of different tissues (p<0.05; p<0.01). The expression of MYNN-1 was significantly higher than that of MYNN-2 in almost tissues (p<0.05; p<0.01), which testified that MYNN-1 is the main variant. The expression of two isoforms decreased gradually with increase of age in ST and CE of MS pig, whereas increased gradually in LW pig. In LD, the expression of two isoforms increased first and then decreased with increase of age in MS pig, and decreased gradually in LW pig. Conclusion: Two transcripts of pig MYNN were successfully cloned and MYNN-1 was main variant. MYNN was highly expressed in ST, CE, and LD, and their expression was regular. We speculated that MYNN plays important roles in digestion/absorption and skeletal muscle growth, whereas the specific mechanisms require further elucidation.

Simultaneous Determination of Statins in Human Urine by Dilute-and-Shoot-Liquid Chromatography-Mass Spectrometry

  • Jang, Haejong;Mai, Xuan-Lan;Lee, Gunhee;Ahn, Jae Hyoung;Rhee, Jongsook;Truong, Quoc-Ky;Vinh, Dinh;Hong, Jongki;Kim, Kyeong Ho
    • Mass Spectrometry Letters
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    • 제9권4호
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    • pp.95-99
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    • 2018
  • An innovative, simple, and rapid assay method based on liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) was developed and validated for the simultaneous determination of eight statin drugs in human urine. A simple sample clean-up procedure using the "dilute and shoot" (DAS) approach enabled a fast and reliable analysis. The influence of the dilution factor was investigated to ensure detectability and reduce the matrix effect. Chromatographic separation was performed on a Phenomenex Kinetex C18 column ($50{\times}3.0mm$ i.d., $2.6{\mu}m$) using an elution gradient of mobile phase A composed of 0.1% acetic acid, and mobile phase B composed of acetonitrile, at a flow rate of 0.35 mL/min. Quantitation was performed on a triple quadrupole mass spectrometer operated in multiple reaction monitoring (MRM) mode using electrospray ionization in positive ion mode. The total chromatographic run time was 15 min. The method was validated for selectivity, sensitivity, recovery, linearity, accuracy, precision, and stability. The present method was successfully applied to the analysis of Rosuvastatin in urine samples after oral administration to healthy human subjects.

Effects of Abiotic Stresses on Cell Cycle Progression in Tobacco BY-2 Cells

  • Jang, Su Jin;Shin, Sung Hae;Yee, Sung Tae;Hwang, Baek;Im, Kyung Hoan;Park, Ky Young
    • Molecules and Cells
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    • 제20권1호
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    • pp.136-141
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    • 2005
  • Mild stresses such as high temperature ($30^{\circ}C$) or a low $H_2O_2$ concentration induced transient cell cycle arrest at G1/S or G2/M depending on the cell cycle stage at which the stress was applied. When stresses were introduced during G0 or G1, the G1/S checkpoint was mainly used; when stresses were introduced after S phase, G2/M was the primary checkpoint. The slowing of cell cycle progression was associated with transient delays in expression of A-, B-, and D-type cyclins. The delay in expression of NtcycA13, one of the A-type cyclins, was most pronounced. The levels of expression of Ntcyc29 (a cyclin B gene) and of CycD3-1 differed most depending on the applied stress, suggesting that different cellular adjustments to mild heat and a low concentration of $H_2O_2$ are reflected in the expression of these two cyclins.

Synthesis and Biological Evaluation of Novel IM3829 (4-(2-Cyclohexylethoxy)aniline) Derivatives as Potent Radiosensitizers

  • Ahn, Jiyeon;Nam, Ky-Youb;Lee, Sae-Lo-Oom;Ryu, Hwani;Choi, Hyun Kyung;Song, Jie-Young
    • Bulletin of the Korean Chemical Society
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    • 제35권12호
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    • pp.3623-3626
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    • 2014
  • Nuclear factor-erythroid 2-related factor 2 (Nrf2) regulates the expression of over 200 genes of antioxidant and phase II drug-metabolizing enzymes, and is highly expressed in non-small cell lung cancer (NSCLC). Nine derivatives of 4-(2-cyclohexylethoxy)aniline were designed. Our previous study demonstrated that IM3829 increases radiosensitivity of several lung cancer cells in vitro and in vivo. Here, biological effects of IM3829 derivatives (2a-2i) were evaluated. Compound 2g derivative effectively inhibits mRNA and protein expression of Nrf2 and HO-1. In addition, we observed over two fold enhancement in IR-induced cell death, from $2.90{\pm}0.22$ to $6.02{\pm}0.87$, in H1299 cancer cell-line. Among the nine derivatives, compound 2g derivative exhibited the highest enhancement of radiosensitizing effect via inhibition of Nrf2 activity.

분자역학을 사용한 단층 그래핀 시트의 모드 III 파괴인성 (Mode III Fracture Toughness of Single Layer Graphene Sheet Using Molecular Mechanics)

  • 웬민키;염영진
    • 대한기계학회논문집A
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    • 제38권2호
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    • pp.121-127
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    • 2014
  • 단층 그래핀 시트(Single layer graphene sheet, SLGS)의 찢어짐 모드(모드 III) 파괴 예측을 위한 원자 기반 미세결합요소모델이 개발되었다. 이 모델은 그래핀 시트의 최대 변형률 관계를 예측하기 위해 수정된 모스포텐셜을 사용한다. 면외 전단하중 조건에서 그래핀의 모드 III 파괴를 광범위한 분자역학(Molecular mechanics, MM) 시뮬레이션으로 조사하였다. 분자역학은 원자의 균열선단 근처 원자의 변위를 설명하기 위해 사용되었고, 선형탄성파괴역학은 이 영역 바깥의 영역을 설명하기 위해 사용되었다. 해석 결과 분자역학 방법이 SLGS의 전단 물성 계산뿐만 아니라 armchair 및 zigzag 방향 모드 III 파괴인성 연구에도 단순하면서도 신뢰할만하다는 것을 보여준다. SLGS 의 모드 III 파괴인성은 zigzag 방향에 대해 $0.86MPa{\sqrt{m}}$, armchair 방향에 대해 $0.93MPa{\sqrt{m}}$로 예측되었다.

Testosterone Relaxes Rabbit Seminal Vesicle by Calcium Channel Inhibition

  • Kim, Jong-Kok;Han, Woo-Ha;Lee, Moo-Yeol;Myung, Soon-Chul;Kim, Sae-Chul;Kim, Min-Ky
    • The Korean Journal of Physiology and Pharmacology
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    • 제12권2호
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    • pp.73-77
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    • 2008
  • Recent studies have documented that testosterone relaxes several smooth muscles by modulating $K^+$ channel activities. Smooth muscles of seminal vesicles playa fundamental role in ejaculation, which might involve testosterone. This study was aimed to assess the role of testosterone in seminal vesicular motility by studying its effects on contractile agents and on the ion channels of single vesicular myocytes in a rabbit model. The contractile responses of circular smooth muscle strips of rabbit seminal vesicles to norepinephrine ($10{\mu}M$), a high concentration of KCI (70 mM), and testosterone ($10{\mu}M$) were observed. Single vesicular myocytes of rabbit were isolated using proteolytic enzymes including collagenase and papain. Inside-out, attached, and whole-cell configurations were examined using the patch clamp technique. The applications of $10{\mu}M$ norepinephrine or 70 mM KCl induced tonic contractions, and $10{\mu}M$ testosterone (pharmacological concentration) evoked dose-dependent relaxations of these precontracted strips. Various $K^+$ channel blockers, such as tetraethylammonium (TEA; $10{\mu}M$), iberiotoxin ($0.1{\mu}M$), 4-aminopyridine (4-AP, $10{\mu}M$), or glibenclamide ($10{\mu}M$) rarely affected these relaxations. Single channel data (of inside-out and attached configurations) of BK channel activity were also hardly affected by testosterone ($10{\mu}M$). On the other hand, however, testosterone reduced L-type $Ca^{2+}$ currents significantly, and found to induce acute relaxation of seminal vesicular smooth muscle and this was mediated, at least in part, by $Ca^{2+}$ current inhibition in rabbit.

근위축성 측삭 경화증의 하 운동 신경원 과흥분성: 운동유발전위를 이용한 분석 (Lower Motor Neuron Hyperexcitability in Amyotrophic Lateral sclerosis: Analysis Using Motor Evoked Potentials)

  • 배종석;홍석찬;김민기;김병준
    • Annals of Clinical Neurophysiology
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    • 제5권1호
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    • pp.21-26
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    • 2003
  • Background & Objectives: Hyperexcitablity of motor system is a well-established characteristic pathophysiologic finding of amyotrophic lateral sclerosis (ALS). Whereas little is known about the source of excitability according to the progression of the disease. We evaluated the excitability and its source in advanced ALS patients using transcranial magnetic stimulation (TMS). Meterial & Methods: Motor evoked potentials (MEP) by TMS were recorded for abductor pollicis brevis muscles in 20 patients, 11 men and 9 women, with ALS. Mean age was $54.2{\pm}12.1years$, and mean disease duration was $13.9{\pm}13.4years$. Serial magnetic stimulations were applied to get the parameters; excitability threshold (ET), amplitude and latency of MEP. We also had a facilitated MEP (fMEP). Results: The parameters were analyzed according to the clinical settings. ET was higher in ALS(mean $63.5{\pm}18.1$) than normal control (mean $46.0{\pm}8.4$, p<0.01). Amplitudes of MEP were reduced in ALS ($2.6{\pm}3.6mV$; control $6.5{\pm}3.1mV$, p<0.01). Duration of the disease and ET showed significant inverse correlation (Spearson correlation coefficient = -0.57, p<0.01). Duration of the disease and fMEP/MEP ratio showed less but also significant inverse correlation (Spearson correlation coefficient, r = -0.52, p < 0.05). Conclusions: Lower ET in advanced ALS patients, in spite of decreased fMEP/MEP ratio, may indicate the hyperexcitability of lower motor neurons in these patients. This study suggests that lower motor neurons is hyperexcitable due to upper motor neuron dysfunction at advanced stage.

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형태적 수렴 특성을 이용한 하구 분류 (Classification of Estuaries based on Morphological Convergence)

  • 신현정;류호상;이관홍
    • 한국지형학회지
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    • 제19권3호
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    • pp.1-22
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    • 2012
  • 하구의 분류는 크게 지형경관적 특성에 근거한 정성적인 분류와 수괴의 물리적 특성에 근거한 정량적 분류로 구분될 수 있으나, 전자는 하구의 특성을 정량화하기 어려우며 후자는 자료부족으로 인해 폭넓은 적용이 어렵다. 하구의 형태적 수렴 특성은 자료의 획득이 상대적으로 용이하면서도 하구의 조석전파 특성과 연관시킬 수 있어 하구분류 기준으로 사용하기 적합하다. 본 연구는 국내 19개 하구에 대해 하구의 형태적 수렴특성을 이용하여 하구별 특징을 파악하고 분류를 시도한 것이다. 하구의 수심과 너비자료를 이용하여 수심과 너비의 감소율(${\nu}$)과 무차원화된 하구의 길이($y_0$)를 구하고, 1차원 수심적분 운동방정식과 연속방정식으로부터 유도한 전이함수 ${\xi}({\nu},ky)$${\nu}$$y_0$을 대입하여 하구별 조위/조석 진폭비와 조위/조류 위상차를 도출하였다. 이 결과를 토대로 19개 하구의 특징을 파악하고 3개 그룹을 구분하였다. 국내 19개 하구는 크게 조위 변동이 조류 변동보다 우세한 제1그룹, 하구와 조류 변동이 조위 변동보다 우세한 제2그룹, 그리고 1그룹과 2그룹의 전이 형태를 갖는 제3그룹 하구로 분류된다. 민감도 분석 결과 하구분류 알고리듬은 하구 길이의 변화에 큰 영향을 받는 것으로 나타났다. 하구 길이가 과대 혹은 과소 추정되었을 때 본래의 조위/조류 진폭비는 과소 혹은 과대 추정되며, 본래의 조위/조류 위상차는 과대 혹은 과소 추정된다. 그러므로 형태적 수렴 특성을 이용하여 하구분류를 하고자 할 때는 하구 길이의 적절한 산정이 무엇보다 중요하다. 하구분류 알고리듬은 수심의 변화를 무시하여도 동일한 분류 결과를 주는 것으로 확인되었다. 이는 하구길이의 적절한 산정이 전제될 경우 하구분류 알고리듬이 자료의 제약 없이 대부분의 국내하구에 적용할 수 있음을 의미한다.

Multi-Color Chip-LED용 어레이 렌즈 개발에 관한 연구 (Development of Array-Lens for Multi-Color Chip-LED)

  • 최병기;이동길;장경천
    • 한국공작기계학회논문집
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    • 제16권3호
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    • pp.50-55
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    • 2007
  • The purpose of this research is to enhance the luminance of the LED and to improve the implementation of color by mounting an array lens on the LED without special technology in process. The workmanship of key components considering the economical efficiency and the injection molding technology for high quality of the product are essential to achieve it. In this paper, the mold was computer-aided was designed and manufactured by CAM software (NX4) and high speed machining center. the applied final machining conditions were 3,000-5,000mm/min feed speed, 15,000-25,000rpm and ${\Phi}0.3mm$ ball end-mill. And the Flow analysis was performed using the mold flow software(MPI) in order to get uniformity of resin. Injection conditions acquired by the flow analysis and the injection experiment are as follows. The cylinder temperature is $220-260^{\circ}C$, the mold temperature is $70-80^{\circ}C$, the injection time is about 1.2sec, the injection pressure and velocity is each 7.8-14.7Mpa, and the injection velocity is 0.8-1.2m/sec.