• Title/Summary/Keyword: ES cells

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Effect of Dimethylformamide on Post-Thaw Motility, Acrosome Integrity, and DNA Structure of Frozen Boar Sperm

  • Hwang, You-Jin;Yang, Jae-Hun;Kim, Sang-Ok;Kim, Bo-Kyung;Choi, Seon-Kyu;Park, Choon-Keun;Kim, Dae-Young
    • Journal of Embryo Transfer
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    • v.24 no.4
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    • pp.275-279
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    • 2009
  • The beneficial effect of glycerol as a cryoprotectant, especially for sperm cryopreservation, has been shown in many studies. However, glycerol is toxic to living cells, and boar sperm in particular show greater sensitivity to glycerol than sperm from other domestic animals. Amides have been studied as alternative cryoprotectants for freezing stallion sperm. Sperm frozen in methylformamide or dimethylformamide as cryoprotectants show similar motility when thawed compared with sperm frozen in glycerol. We evaluated the cryoprotective effects of dimethylformamide on boar sperm freezing. To test the effect of amides, the concentration of boar semen was adjusted to $10^9sperm/mL$, and seminal plasma was removed using Hulsen solution. After centrifugation, the pellet was diluted in modified-Modena B extender. Lactose-egg yolk (LEY) extender was used as the cooling extender. The freezing extender was madeed aaddition of the optimal amount of glycerol and amides to LEY-Glycerol-Orvus ES Paste extender, and this extender was used for the second dilution. Diluted sperm were frozen in liquid nitrogen using the 0.5 mL straw method. Sperm frozen in extender with glycerol as a cderol were compared with those frozen in extender including the different amides. Sperm were tested for motility, viability, the sperm chromatin structure assay, and normal apical ridge after thawing. The percent of motile sperm diluted in glycerol was as high as that in the stallion study (61%). Dimethylformamide showed positive effects on sperm quality and was better than glycerol. Methylformamide provided similar sperm quality as glycerol. Therefore, dimethylformamide is useful for reducing cryoinjury in boar sperm and is expected to be useful as an alternative cryoprotectant.

Disruption of the Tff1 gene in mice using CRISPR/Cas9 promotes body weight reduction and gastric tumorigenesis

  • Kim, Hyejeong;Jeong, Haengdueng;Cho, Yejin;Lee, Jaehoon;Nam, Ki Taek;Lee, Han-Woong
    • Laboraroty Animal Research
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    • v.34 no.4
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    • pp.257-263
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    • 2018
  • Trefoil factor 1 (TFF1, also known as pS2) is strongly expressed in the gastrointestinal mucosa and plays a critical role in the differentiation of gastric glands. Since approximately 50% of all human gastric cancers are associated with decreased TFF1 expression, it is considered a tumor suppressor gene. Tff1 deficiency in mice results in histological changes in the antral and pyloric gastric mucosa, with severe hyperplasia and dysplasia of epithelial cells, resulting in the development of antropyloric adenoma. Here, we generated Tff1-knockout (KO) mice, without a neomycin resistant ($Neo^R$) cassette, using the clustered regularly interspaced short palindromic repeats/CRISPR-associated nuclease 9 (CRSIPR/Cas9) system. Though our Tff1-KO mice showed phenotypes very similar to the previous embryonic stem (ES)-cell-based KO mice, they differed from the previous reports in that a reduction in body weight was observed in males. These results demonstrate that these newly established Tff1-KO mice are useful tools for investigating genetic and environmental factors influencing gastric cancer, without the effects of artificial gene insertion. Furthermore, these findings suggest a novel hypothesis that Tff1 expression influences gender differences.

High Efficiency Solar Cell(I)-Fabrication and Characteristics of $N^+PP^+$ Cells (고효율 태양전지(I)-$N^+PP^+$ 전지의 제조 및 특성)

  • 강진영;안병태
    • Journal of the Korean Institute of Telematics and Electronics
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    • v.18 no.3
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    • pp.42-51
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    • 1981
  • Boron was predeposited into p (100) Si wafer at 94$0^{\circ}C$ for 60minutes to make the back surface field. High tempreature diffusion process at 1145$^{\circ}C$ for 3 hours was immediately followed without removing boron glass to obtain high surface concentration Back boron was annealed at 110$0^{\circ}C$ for 40minutes after boron glass was removed. N+ layer was formed by predepositing with POCI3 source at 90$0^{\circ}C$ for 7~15 minutes and annealed at 80$0^{\circ}C$ for 60min1es under dry Of ambient. The triple metal layers were made by evaporating Ti, Pd, Ag in that order onto front and back of diffused wafer to form the front grid and back electrode respectively. Silver was electroplated on front and back to increase the metal thickness form 1~2$\mu$m to 3~4$\mu$m and the metal electrodes are alloyed in N2 /H2 ambient at 55$0^{\circ}C$ and followed by silicon nitride antireflection film deposition process. Under artificial illumination of 100mW/$\textrm{cm}^2$ fabricated N+PP+ cells showed typically the open circuit voltage of 0.59V and short circuit current of 103 mA with fill factor of 0.80 from the whole cell area of 3.36$\textrm{cm}^2$. These numbers can be used to get the actual total area(active area) conversion efficiency of 14.4%(16.2%) which has been improved from the provious N+P cell with 11% total area efficiency by adding P+ back.

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Effect of Gas Atmosphere on In Vitro Development of Bovine Embryos Derived from In Vitro Fertilization (배양기내 GAS 분압의 조성이 소 체외수정란의 체외발육에 미치는 영향에 대한 연구)

  • 이원유;신태영;이병천;황우석
    • Journal of Embryo Transfer
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    • v.10 no.2
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    • pp.121-129
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    • 1995
  • To examine the critical effect of oxygen concentration on embryonic development, in vitro fertilized embryos were cultured in media(TCM199 vs. SOF) supplemented sera(1O% FCS vs. 10% HS) with and without bovine oviduct epithelial cells under two gas atmosphere (5% $CO_2$ in air vs. 5% $CO_2$, 5% $O_2$, 90% $N_2$). Oocytes, obtained from abattoir ovaries, were matured in EGF containing TCM199 medium co-cultured with BOEC for 24 hours, followed by exposure to frozen-thawed, heparin4reated spermatozoa in TALP for 30 hours. And then early embryos(1~2 cell) were cultured in both TCM199 and SOF supplemented with 10% FCS or 10% RS under 5% $CO_2$ in air or 5% COi, 5% $O_2$, 90% $N_2$. Development to morulae and blastocysts was recorded on days 7, after the start of in vitro fertilization. The developmental rates of in vitro fertilized embryos to morulae and blastocysts cultured in SOF with BOEC under 5% $CO_2$, 5% $O_2$, 90% $N_2$(24.4%) were significantly(p<0.05) higher than cultured in SOF with BOEC under 5% $CO_2$ in air(14.1%) at seven days after in vitro fertilization. When early bovine embryos were cultured in TCM 199 and SOF under two different gas atmosphere, there were no significant differences in the developmental rates to morulae and blastocysts between supplements of 10% FCS and 10% HS. The rates of development to morulae and blastocysts were significantly(p<0.01) higher in TCM 199 with BOEC(24.7%) than TCM199 without BOEC(10.9%) under 5% $CO_2$ in air, otherwise SOF without BOEC(36.4%) were significantly (p<0.05) higher than in SOF with BOEC (24.4%) under 5% $CO_2$, 5% $O_2$, 90% $N_2$. In summary, these experiments have proved that the culture system in SOF supplemented 10% ES is effective on in vitro development of early bovine embryos under 5% $CO_2$, 5% $O_2$, 90% $N_2$. In addition, it is effective to development of bovine embryos that TCM 199 should be co-cultured with BOEC and SOF should be cultured without somatic cells under two different gas atmosphere.

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Subsets of Inflammatory Cytokine Gene Polymorphisms are Associated with Risk of Carcinogenic Liver Fluke Opisthorchis viverrini-Associated Advanced Periductal Fibrosis and Cholangiocarcinoma

  • Surapaitoon, Arpa;Suttiprapa, Sutas;Mairiang, Eimorn;Khuntikeo, Narong;Pairojkul, Chawalit;Bethony, Jeffrey;Brindley, Paul J.;Sripa, Banchob
    • Parasites, Hosts and Diseases
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    • v.55 no.3
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    • pp.295-304
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    • 2017
  • Opisthorchis viverrini infection induces chronic inflammation, and a minor proportion of infected individuals develop advanced periductal fibrosis (APF) and cholangiocarcinoma (CCA). Inflammatory cytokines and/or their gene polymorphisms may link to these biliary pathologies. We therefore investigated associations among cytokine gene polymorphisms and cytokine production in 510 Thai cases infected with O. viverrini who presented with APF+ or APF-, as established by abdominal ultrasonography as well as in patients diagnosed with CCA. Levels of pro-inflammatory and anti-inflammatory cytokines were determined in culture supernatants after stimulation of peripheral blood mononuclear cells (PBMCs) with O. viverrini excretory-secretory (ES) products. Pro-inflammatory cytokines, IL-$1{\beta}$, IL-6, IFN-${\gamma}$, LT-${\alpha}$, and TNF-${\alpha}$ were significantly increased in CCA patients compared with non-CCA (APF- and APF+) cases. Polymorphisms in genes encoding IL-$1{\beta}$-511C/T, IL-6-174G/C, IFN-${\gamma}$+874T/A, LT-${\alpha}$+252A/G, and TNF-${\alpha}$-308G/A were then investigated by using PCR-RFLP or allele specific-PCR (AS-PCR) analyses. In the CCA cases, LT-${\alpha}$+252A/G and TNF-${\alpha}$-308G/A heterozygous and homozygous variants showed significantly higher levels of these cytokines than the wild type. By contrast, levels of cytokines in wild type of IFN-${\gamma}$+874T/A were significantly higher than the variants in CCA cases. IFN-${\gamma}$+874T/A polymorphisms were associated with advanced periductal fibrosis, whereas IL-6-174G/C polymorphisms were associated with CCA. To our knowledge, these findings provide the first demonstration that O. viverrini infected individuals carrying several specific cytokine gene polymorphisms are susceptible to develop fibrosis and CCA.

Antioxidant activities of solvent extracts from different Glehnia Radix parts and their inhibitory effect against nitric oxide production in Raw 264.7 cell (해방풍 부위별 용매추출물의 항산화 활성 및 nitric oxide 생성 억제)

  • Gu, Yul-Ri;Kim, Sun-Won;Son, Yong-Won;Hong, Joo-Heon
    • Food Science and Preservation
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    • v.24 no.1
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    • pp.116-124
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    • 2017
  • In this study, we compared the antioxidant and immuno-modulatory effects of water (HR, HL, HS), 70% ethanol (ER, EL, ES), and 70% methanol (MR, ML, MS) extracts of different parts (root, leaf, and seed) from Glehnia Radix. Yields were 17.40-30.17% for water extract, 11.47-28.67% for 70% ethanol extract, and 10.73-30.57% for 70% methanol extract, respectively. The total polyphenol and flavonoid contents of EL were 10.79 g/100 g and 2.01 g/100 g, respectively. The DPPH and ABTS radical scavenging activities of EL at $1,000{\mu}g/mL$ were 84.70% and 57.64%, respectively. The superoxide radical scavenging and ferric-reducing antioxidant power of EL at $1,000{\mu}g/mL$ were 84.05% and $975.28{\mu}M$, respectively. Moreover, 70% ethanol and 70% methanol extracts of root from Glehnia Radix significantly inhibited production of NO in LPS-stimulated macrophage RAW 264.7 cells without cytotoxicity. These results suggest that 70% ethanol and 70% methanol extracts of Glehnia Radix leaf may be a useful functional food material in the food industry.

Sodium Sulfur Battery for Energy Storage System (대용량 에너지 저장시스템을 위한 나트륨 유황전지)

  • Kim, Dul-Sun;Kang, Sungwhan;Kim, Jun-Young;Ahn, Jou-Hyeon;Lee, Chang-Hui;Jung, Keeyoung;Park, Yoon-Cheol;Kim, Goun;Cho, Namung
    • Journal of the Korean Electrochemical Society
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    • v.16 no.3
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    • pp.111-122
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    • 2013
  • Sodium sulfur (NAS) battery is a high energy storage system (ESS). These days, as the use of renewable green energy like wind energy, solar energy and ocean energy is rapidly increasing, the demand of ESS is increasing and NAS battery is considered to be one of the most promising ESS. Since NAS battery has a high energy density(3 times of lead acid battery), long cycle life and no self-charge and discharge, it is a good candidate for ESS. A NAS battery consists of sulfur as the positive electrode, sodium as the negative electrode and ${\beta}$"-alumina as the electrolyte and a separator simultaneously. Since sulfur is an insulator, carbon felt should be used as conductor with sulfur and so the composition and property of the cathode could largely influence the cell performance and life cycle. Therefore, in this paper, the composition of NAS battery, the property of carbon felt and sodium polysulfides ($Na_2S_x$, intermediates of discharge), and the effects of these factors on cycle performance of cells are described in detail.

Antimelanogenic Effect and Whitening of Anthocyanin Rich Fraction from Seeds of Liriope platyphylla (맥문동 종실 안토시아닌 분획물의 멜라닌 생성 억제 및 미백 효과)

  • Choung, Myoung Gun;Hwang, Young Sun;Kim, Gi Ppeum;Ahn, Kyung Geun;Shim, Hoon Seob;Hong, Seung Beom;Choi, Jae Hoo;Yu, Chang Yeon;Chung, Ill Min;Kim, Seung Hyun;Lim, Jung Dae
    • Korean Journal of Medicinal Crop Science
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    • v.21 no.5
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    • pp.361-371
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    • 2013
  • This study was performed to determine the antimelanogenic effect and tyrosinase inhibitory activities of anthocyanin rich fraction (AN-SLP) from Liriope platyphylla Wang et Tang seeds. Anthocyanins isolated from L. platyphylla seeds revealed the presence of four major anthocyanin components, which were tentatively identified as delphinidin-3-Oglucoside, delphinidin-3-O-rutinoside, petunidin-3-O-rutinoside, and malvidin-3-O-rutinoside using semipreparative HPLC, $^1H$-NMR, $^{13}C$ NMR, FAB-MS and LC/ES-MS. The inhibitory effect of AN-SLP on tyrosinase activity was studied using in vitro (against mushroom tyrosinase) and ex vivo (against B16 melanoma cell tyrosinase) models. Cellular tyrosinase activity was decreased by AN-SLP treatment in B 16 melanoma cells through dose dependent manner, but AN-SLP did not inhibit mushroom tyrosinase and L-DOPA oxidation directly. AN-SLP showed melanin inhibition by 53.2% at 50 ${\mu}g/m{\ell}$ which was 0.7 times more efficient than the antimelanogenic effect of commercial arbutin and kojic acid (36.5%) also did not show cell toxicity. Additionally, AN-SLP inhibited the activity of ${\alpha}$-glucosidase and the glycosylation of tyrosinase in melanoma cell. The resulting unsaturated glycosylation of tyrosinase makes it unstable and disturb correct transportation. From theses results, we conclude that AN-SLP could be used as anti-melanogenic agent for skin whitening.

The Effect of Brown Tinted or UV-A blocking Ophthalmic Lens Against the Photooxidation of A2E, a Lipofuscin in Retina (망막 내 노인성 형광색소의 광산화에 미치는 Brown 착색렌즈와 UV-A 차단 안경렌즈의 영향)

  • Park, Sang-Il
    • Journal of Korean Ophthalmic Optics Society
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    • v.17 no.1
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    • pp.91-97
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    • 2012
  • Purpose: This purpose of study is to investigate the effect of UV-A-blocking or brown-tinted ophthalmic lens against A2E photooxidation which known as one of the etiologies of AMD(Age-related macular degenaration). Methods: The photooxidation of A2E, synthetic product of two molecules of all-trans-retinal and ethanolamine, was induced by the exposure to blue light (420~470 nm, $94mW/cm^2$) for 3 minutes. The inhibitory effect of UVblocking or brown-tinted ophthalmic lens against A2E photooxidation was evaluated by UV absorbance and HPLC analysis of remained A2E after the exposure to blue light. Results: UV-blocking ophthalmic lens could not inhibit A2E photooxidation induced by blue light irradiation. There was no difference in A2E photooxidation in the presence of brown-tinted ophthalmic lens to block 15% of visible ray, however, those lenses blocking 55% or 86% of visible ray showed the inhibitory effect of A2E photooxidation as 9.98% and 16.55%, respectively. By HPLC analysis, the amount of residual A2E which was not blocked by any lens was $199.29{\pm}26.53{\mu}M$, however, the inhibitory effect against A2E photooxidation was shown in the presence of brown-tinted lens. The remained A2Es were $264.58{\pm}31.91{\mu}M$ and $402.93{\pm}28.68{\mu}M$ in brown-tinted lenses of 55% and 86% blocking visible ray, respectively. However, there was no inhibitory effect against A2E photooxidation in the case of UV-blocking lens by HPLC analysis. Conclusions: In this study, brown-tinted ophthalmic lens was confirmed to have the inhibitory effect against the photooxidation of A2E, a causing substance of AMD onset.

Effect of lonizing Radiation on the Host Resistance Against Listeria Monocytogenes Infection and the Cytokine Production in Mice (방사선조사후 마우스에서의 Cytokine 생산능 및 Listeria monecytogenes에 대한 저항성의 변화)

  • Oh, Yoon-Kyeong;Chang, Mee-Young;Kang, In-Chol;Oh, Jong-Suk;Lee, Hyun-Chul
    • Radiation Oncology Journal
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    • v.15 no.3
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    • pp.175-186
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    • 1997
  • Purpose : To evaluate the qualitative immunologic changes by ionizing radiation. we studied the altered capacities of the macrophages and lymphocytes to produce cytokines in conjunction with resistance to Listeria monocytegenes (LM) infection in mice Materials and Methods : BALB/c mice and Listeria monocytogenes were used. The mice were infected intraperitoneally with $10^5LM$ at 1 day after irradiation (300cGy) and sacrificed at 1, 3, 5 days after infection, and then the numbers of viable LM per spleen in the irradiated and control group were counted. Tumor necrosis factor-alpha ($TNF-\alpha$), interferon-gamma ($IFN-\gamma$). interleukin-2 (IL-2), and nitric oxide (NO) were assessed after irradiation. Results : Under gamma-ray irradiation with a dose range of 100-850cGy, the number of total splenocytes decreased markedly in a dose-dependent manner, while peritoneal macrophages did so slightly Cultured peritoneal macrophages produced more $TNF-\alpha$ in the presence of lipopolysaccharide (LPS) during the 24 hours after in vitro irradiation, but their capacity of $TNF-\alpha$ Production showed a decreased tendency at 5 days after in vivo total body irradiation. With 100cGy and 300cGy irradiation, cultured peritoneal macrophages produced more NO in the presence of LPS during the 24 hours after in vitro irradiation than without irradiation. Activated splenocytes from irradiated mice (300cGy) exhibited a decreased capacity to Produce IL-2 and $IFN-\gamma$ with Concavalin-A stimulation at 3 days after irradiation. When BALB/c mice were irradiated to the total body with a dose of 300cGy, they showed enhanced resistance during early innate phase, but a significant inhibition of resistance to LM was found in the late innate and acquired T-cell dependent phases. Conclusion : These results su99es1 that increased early innate and decreased late innate and acquired immunity to LM infection by ionizing radiation (300cGy) may be related to the biphasic altered capacity of the macrophages to produce $TNF-\alpha$ and the decreased capacities of the lymphocytes to produce IL-2 and $IFN-\gamma$ in addition to a marked decrease in the total number of cells.

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