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p66Shc in sheep preimplantation embryos: Expression and regulation of oxidative stress through the manganese superoxide dismutase-reactive oxygen species metabolic pathway

  • Tong Zhang;Jiaxin Zhang;Ruilan Li
    • Animal Bioscience
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    • v.36 no.7
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    • pp.1022-1033
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    • 2023
  • Objective: p66Shc, a 66 kDa protein isoform encoded by the proto-oncogene SHC, is an essential intracellular redox homeostasis regulatory enzyme that is involved in the regulation of cellular oxidative stress, apoptosis induction and the occurrence of multiple age-related diseases. This study investigated the expression profile and functional characteristics of p66Shc during preimplantation embryo development in sheep. Methods: The expression pattern of p66Shc during preimplantation embryo development in sheep at the mRNA and protein levels were studied by quantitative real-time polymerase chain reaction (RT-qPCR) and immunofluorescence staining. The effect of p66Shc knockdown on the developmental potential were evaluated by cleavage rate, morula rate and blastocyst rate. The effect of p66Shc deficiency on reactive oxygen species (ROS) production, DNA oxidative damage and the expression of antioxidant enzymes (e.g., catalase and manganese superoxide dismutase [MnSOD]) were also investigated by immunofluorescence staining. Results: Our results showed that p66Shc mRNA and protein were expressed in all stages of sheep early embryos and that p66Shc mRNA was significantly downregulated in the 4-to 8-cell stage (p<0.05) and significantly upregulated in the morula and blastocyst stages after embryonic genome activation (EGA) (p<0.05). Immunofluorescence staining showed that the p66Shc protein was mainly located in the peripheral region of the blastomere cytoplasm at different stages of preimplantation embryonic development. Notably, serine (Ser36)-phosphorylated p66Shc localized only in the cytoplasm during the 2- to 8-cell stage prior to EGA, while phosphorylated (Ser36) p66Shc localized not only in the cytoplasm but also predominantly in the nucleus after EGA. RNAi-mediated silencing of p66Shc via microinjection of p66Shc siRNA into sheep zygotes resulted in significant decreases in p66Shc mRNA and protein levels (p<0.05). Knockdown of p66Shc resulted in significant declines in the levels of intracellular ROS (p<0.05) and the DNA damage marker 8-hydroxy2'-deoxyguanosine (p<0.05), markedly increased MnSOD levels (p<0.05) and resulted in a tendency to develop to the morula stage. Conclusion: These results indicate that p66Shc is involved in the metabolic regulation of ROS production and DNA oxidative damage during sheep early embryonic development.

Amino Acid Concentrations in the Blastocoelic Fluid of In Vitro-Produced Bovine Blastocysts (체외생산된 소 배반포강 내의 아미노산 농도)

  • Jung, Y.G.;Wei, H.;Yoon, J.T.;Fukui, Y.
    • Journal of Embryo Transfer
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    • v.13 no.3
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    • pp.251-259
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    • 1998
  • Concentrations of free amino acids in the BF of IVP bovine BL and HBL were examined in this study. The embryos derived from IVF oocytes were cultured in a SOFM containing BSA, EAA and NEAA. BF was aspirated from BL (180 h of age after insemination) and HBL (216 h of age after insemination), and introduced into drops of SOFM (30$\mu$l/drop) containing PVA through micromanipulation. The medium containing BF was then subjected to measurement of 20 amino acids by an automatic amino acid analyzer. The concentrations of isoleucine, leucine and methionine were higher (p〈0.05) in the BF from HBL than from BL, and no difference was found in aspartate or glutamate concentrations between BL and HBL, while threonine, alanine (p〈0.01) and the rest of the amino acids (p〈0.001) were significantly higher in the BF from HBL than from BL. Cystine was not found in either BL or HBL. A high concentration of glutamine was found in the BF from both BL and HBL, although it was not added to the culture medium. These results indicate that bovine BF contains several EAA (methionine in BL and isoleucine, leucine and methionine in HBL) and NEAA (alanine, glutamate, glycine, proline, serine and aspartate in BL, and glutamate and aspartate in HBL), and there is significant differences in the amino acid concentration in the BF between BL and HBL derived by WP.

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하계 전기, 전자연합학술회의 및 산학협동 심포지엄 초록

  • 대한전기학회
    • 전기의세계
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    • v.27 no.5
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    • pp.33-54
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    • 1978
  • (차례) 1.산학협동심포지업 (1)우리나라에서의 연구개발과 산학협동 (2)산학협동과 산업계의 역할 (3)산학협동의 현황과 진로 2.학술회의A (1)전력게통의 계층구조와 협조원리에 관한 연구 (2)2중층괴상회전자 유도전동기의 이론해석 (3)초고주파가열장치에 사용하는 철공진변압기의 해석적 설계 (4)한국전기기시험연구소 대전력단락 시험설비설계 (5)직류전동기제어를 위한 Thyristor Chopper정류회로에 관한 연구 (6)선로의 개폐정보를 포함하는 전력계통의 상태추정 (7)단일신경세포에 대한 ITEM 신호 특성 3.학술회의B (1)MMM-1 Computer System의 설계 및 제작 (2)Adaptive Delta Modulation System의 성능비교 연구 (3)6GHZ FMD마이크로파 무선전송장치의 개발 (4)적선도에 의한 회로망함수의 결정 (5)동맥혈압의 해석과 그의 전기적 유사모델 (6)피부감각의 정보전달 특성에 관하여 (7)선형직접회로의 공정설계 및 그 특성 조성 (8)DH L.D의 전기적포화현상에 관한 이론적 해석 (9)Potocoupler를 이용한 Isolator 4.학술회의C (1)Al-Al$_{2}$O$_{3}$ -Al박막구조의 전기적 특성 (2)이종금속에 샌드위치된 고분자물질의 단락전조 (3)유전체가 일부체워진 직 6면체의 캐비티의 다중모오드 해석 (4)반도체 가스 검지소자의 제조 및 그의 전기적 특성 (5)실리콘 산화공정에 대한 실험적 고찰 (6)진공증착법에 의한 InSb 박막제도에서 열처리효과 (7)(Ba$_{1}$-xBix) Tio$_{3}$ PTC thermistor의 첨가량의 최적건안 (8)금속박막증착시 두께조절 5.특별강연회 (1)일본에 있어서의 절력계통공학연구 (2)Linear Motor의 최근개발동향량도 높았다. valine과 leucine 및 aspartic acid, glycine과 glutamic acid, leucine과 aspartic acid 간에는 고도의 정상관, glycine과 serine, valine과 phenylalanine, threonine과 proline, phenylalanine과 arginine, methionine과 glutamic acid, histidine과 lysine 간에는 유의 정상관, 그리고 isoleucine과 lysine 간에는 유의한 부상관이 있었다. 4. lysine 함량은 단백질 함량과 정산곤, isoleucine 함량은 단빅질 함량과 부상관을 보였으며, alanine, valine, leucine 함량은 지방함량과 각각 유의한 정산관을 보였다. 5. 대두 단백질은 7.5% acrylamide gel 전기영동에 의해 품종에 따라 12~16개의 구성분으로 분리되었으며, 이들중 주구성분들은 상대이동도가 0.06(a), 0.14(b). 0.24(d) 이었고, 구성분 b의 함량이 품종간에 가장 변이가 컸으며, 구성분 b는 그밖의 주요 구성분들의 함량과 부의 상관이 있었고, 구성분 a는 단백질 함량과 정상관이 있었다. 6. 종실단백질 구성분들의 조합 특성 면에서 공시 86품종은 11개 유형군으로 분류되었으며, 우리나라와 일본품종은 미국품종에 비해 단백질구성분 조성이 훨씬 다양하였다. 7. 이동도가 매우 빠른 단백질 구성분 o(Rm 0.77) p(Rm 0.81)를 모두 갖고 있는 품종은 3품종, 모두 갖고 있지 않은 품종은 1품종이었고, 나머지 82품종은 o나 p중 한 구성분을 갖고 있었으며 그 분포율은 30 : 65 이었는데 미국계 품종은 우리나라 품종에 비해 구성분 o를 간고 있는 비율이 현저히 적었다. 8. 대두 종실은 개화후 22일까지 완만히, 그 이후 20~30일간 급속히

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Effects of Temperature and Salinity on the Hemolymp Characteristic and Composition of Free Amino Acid of the Abalone, Haliotis discuss hannai (수온과 염분이 참전복, Haliotis discuss hannai의 혈림프액 성상과 유리아미노산 조성에 미치는 영향)

  • Yang, Sung Jin;Lee, Jeong Young;Hwang, Hyung Kyu;Min, Byung Hwa;Shin, Yun Kyung;Myeong, Jeong In
    • Journal of Marine Life Science
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    • v.1 no.1
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    • pp.50-55
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    • 2016
  • Glucose was continuously increased as time was elapsed under the temperature at 6℃ and 4℃. NH3 was significantly increased at 4℃. AST was increased as temperature and salinity were decreased which indicated that it was affected by temperature, salinity and interaction of temperature×salinity. ALT was significantly increased as temperature was decreased. Free amino acid contents, excluding aspartic acid, threonine, serine and glycine were increased as temperature was decreased. This decreasing tendency was also observed when salinity was decreased. Taurine, aspartic acid, glutamic acid and arginine were significantly increased as salinity was decreased, while glycine, alanine, methionine, tyrosine, and histidine were decreased as salinity was decreased. The result of this study suggested that transport condition to minimize the stress is considered in the range of 30~34 psu, 8~10℃.

Study of sweat content analysis and latent fingerprint developing (땀의 성분 분석과 잠재지문 현출에 관한 연구)

  • Choi, Mi-Jung;Sun, Yale-Shik;Kim, Chang-Seong;Choi, Man-Sik;Sung, Nack-Do;Park, Sung-Woo
    • Analytical Science and Technology
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    • v.20 no.2
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    • pp.147-154
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    • 2007
  • Sweat contents were investigated for using those data to forensic purpose. The experiments of identifying sweat contents were as follow: 1) measurement of amino acids (aspartic acid, serine, glycine etc) by HPLC, 2) anions ($Cl^-$, $F^-$, and $SO{_4}^{2-}$) by IC and 3) trace elements (Cu, Zn, Li, B, etc.) by ICP-MS. Amino acid contents in sweat are varied with individual samples and glycine, threonine, alanine, valine and histidine are detected as the prime one. The detected anions are $Cl^-$(2167~4073 ppm) and $F^-$(454~582 ppm) mostly. Trace elements of Rb, Zn and Cu are detected and those concentrations are relatively very high. The compositions of sweat can be influenced by various factors (diet, anthropometric, characteristics, physical fitness, age, gender and the state of the health).

Clinical Assessment of Lip Balm Containing Exfoliation Complex to Improve Wrinkles, Elasticity, and Hydration of Lips (입술의 주름, 탄력, 보습 개선을 위한 각질 박리 복합물이 함유된 립밤의 임상 효능 평가)

  • Jiye Park;Jae Young Shin;Jinyong Lee;Myoung Jin An;Nae Gyu Kang
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.49 no.4
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    • pp.355-364
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    • 2023
  • Lips have a unique desquamation process and moisturizing properties that are structurally different from ordinary skin. In particular, the turnover cycle of the stratum corneum is fast and the outermost stratum corneum is thin, so the amount of keratin is relatively high, and there are no skin appendages, so it is very vulnerable to maintaining moisture. In this study, we set three targets for lip care : stratum corneum, moisture, and barrier, and aimed to identify the potential of three target-specific ingredients for lips improvement. We confirmed the exfoliating, moisturizing, and barrier improvement efficacy of three target-specific ingredients which also have mild exfoliating effect. Specifically, we verified that Bacillus clausii extract improved skin exfoliation, gluconolactone improved skin moisture retention, and serine reinforced skin barrier function. To test the in vivo efficacy of the complex composed of three target-specific ingredients on the human lips, the lip balm manufactured with non-irritating range of concentration was applied, and we confirmed that it was effective in improving lip exfoliation·moisturizing·elasticity·wrinkles. Consequently, it was confirmed that the beauty and health indicators of the lips could be improved through the exfoliation-moisturization-barrier care of the lips and these ingredients were applied to LG H&H LIPCERIN products.

Culture Conditions of E. coli Harboring Human O-Linked N-Acetyl-${\beta}$-Glucosaminidase Gene and Enzymatic Properties (사람의 O-linked-N-acetyl-${\beta}$-D-glucosaminidase 유전자를 함유한 대장균의 배양조건과 효소학적 특성)

  • 강대욱;조용권;서현효
    • Korean Journal of Microbiology
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    • v.40 no.2
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    • pp.147-153
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    • 2004
  • Protein modification by N-acetyl-${\beta}$-D-glucosamine (O-G1cNAc) on the hydroxyl groups of Ser or Thr ubiq-uitously occurs in eukaryotic cells and is involved in many cellular phenomena. The level of O-G1cNAc-mod-ified protein is regulated by OGT and O-GlcNAcase enzymes. We have tried to produce recombinant O-GlcNAcase in E. coli as an effort to establish in vitro screening system for modulators of O-GlcNAcase. The culture conditions for improvement of O-GlcNAcase productivity, were as follows: induction temperature, $30^{\circ}C$; the concentration of L-arabinose, 0.02% and induction time, 5 hr. Under these culture conditions, E. coli cells containing O-GlcNAcase gene had no enzyme activity until up to 3 hr culture. However, O-GlcNAcase activity dramatically increased from 3 to 5 hr culture. It almost maintained the same level after 5 hr culture. Western blot analysis verified the amount of expressed O-GlcNAcase increased with culture time, being con-sistent with activity data. The optimal reaction condition determined in this study was as follows: protein quan-tity, $5{\mu}g$; reaction time, 30 min; reaction temperature, $45^{\circ}C$; substrate concentration, 2 mM; reaction pH, 6.5. Methanol had little effect on O-GlcNAcase activity and 90% of activity were retained at 10%. Only 15% resid-ual activity were detected at 5% of chloroform.

Inhibitory Effect of Chloroform Extract of Marine Algae Hizikia Fusifomis on Angiogenesis (Hizikia fusiformis 클로로포름 추출물의 in vitro 및 in vivo 혈관신생 억제 연구)

  • Myeong-Eun Jegal;Yu-Seon Han;Shi-Young Park;Ji-Hyeok Lee;Eui-Yeun Yi;Yung-Jin Kim
    • Journal of Life Science
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    • v.34 no.6
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    • pp.399-407
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    • 2024
  • Angiogenesis is the process by which new blood vessels form from existing blood vessels. This phenomenon occurs during growth, healing, and menstrual cycle changes. Angiogenesis is a complex and multifaceted process that is important for the continued growth of primary tumors, metastasis promotion, the support of metastatic tumors, and cancer progression. Impaired angiogenesis can lead to cancer, autoimmune diseases, rheumatoid arthritis, cardiovascular disease, and delayed wound healing. Currently, there are only a handful of effective antiangiogenic drugs. Recent studies have shown that natural marine products exhibit antiangiogenic effects. In a previous study, we reported that the hexane extract of H. fusiformis (HFH) could inhibit the development of new blood vessels both in vitro and in vivo. The aim of this study was to describe the inhibitory effect of chloroform extracts of H. fusiformis on angiogenesis. To investigate how chloroform extract prevents blood vessel growth, we examined its effects on HUVEC, including cell migration, invasion, and tube formation. In a mouse Matrigel plug assay, H. fusiformis chloroform extract (HFC) also inhibited angiogenesis in vivo. Certain proteins associated with blood vessel growth were reduced after HFC treatment. These proteins include vascular endothelial growth factor (VEGF), mitogen-activated protein kinase (MAPK)/extracellular signal transduction kinase, and serine/threonine kinase 1 (AKT). These studies have shown that the chloroform extract of H. fusiformis can inhibit blood vessel growth both in vitro and in vivo.

Impairment of Mitochondrial ATP Synthesis Induces RIPK3-dependent Necroptosis in Lung Epithelial Cells During Lung Injury by Lung Inflammation

  • Su Hwan Lee;Ju Hye Shin;Min Woo Park;Junhyung Kim;Kyung Soo Chung;Sungwon Na;Ji-Hwan Ryu;Jin Hwa Lee;Moo Suk Park;Young Sam Kim;Jong-Seok Moon
    • IMMUNE NETWORK
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    • v.22 no.2
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    • pp.18.1-18.15
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    • 2022
  • Dysfunction of mitochondrial metabolism is implicated in cellular injury and cell death. While mitochondrial dysfunction is associated with lung injury by lung inflammation, the mechanism by which the impairment of mitochondrial ATP synthesis regulates necroptosis during acute lung injury (ALI) by lung inflammation is unclear. Here, we showed that the impairment of mitochondrial ATP synthesis induces receptor interacting serine/threonine kinase 3 (RIPK3)-dependent necroptosis during lung injury by lung inflammation. We found that the impairment of mitochondrial ATP synthesis by oligomycin, an inhibitor of ATP synthase, resulted in increased lung injury and RIPK3 levels in lung tissues during lung inflammation by LPS in mice. The elevated RIPK3 and RIPK3 phosphorylation levels by oligomycin resulted in high mixed lineage kinase domain-like (MLKL) phosphorylation, the terminal molecule in necroptotic cell death pathway, in lung epithelial cells during lung inflammation. Moreover, the levels of protein in bronchoalveolar lavage fluid (BALF) were increased by the activation of necroptosis via oligomycin during lung inflammation. Furthermore, the levels of ATP5A, a catalytic subunit of the mitochondrial ATP synthase complex for ATP synthesis, were reduced in lung epithelial cells of lung tissues from patients with acute respiratory distress syndrome (ARDS), the most severe form of ALI. The levels of RIPK3, RIPK3 phosphorylation and MLKL phosphorylation were elevated in lung epithelial cells in patients with ARDS. Our results suggest that the impairment of mitochondrial ATP synthesis induces RIPK3-dependent necroptosis in lung epithelial cells during lung injury by lung inflammation.

The Biochemical Studies on Stored Soy-sauce (저장(貯藏)간장의 생화학적(生化學的) 연구(硏究))

  • Chang, Chi-Hyun
    • Applied Biological Chemistry
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    • v.9
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    • pp.9-27
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    • 1968
  • Studies were carried out in order to elucidate chemical components and microflora in three types of soy-sauce, 12-year aged soy-sauce prepared by improved method. 7-year aged and 20-year aged soy-sauce prepared by ordinary method. They results are summarized as follows: 1. The followings are found to be the important factors affecting the quality of soy sauce. a. Organic acids, reducing sugars and free amino acids were increased in the course of storage. b. In the aged soy-sauces under study non-volatile organic acid increased while volatile organic acid decreased and the total acidity was dependent only upon tie latter. c. It was found that suit concentration decreased during the storage. 2. The results of investigation of microflora in the stored soy-sauce are shown as follows. Soy-sauce Improved Ordinary Microbe 12-Y. 20-Y. 7-Y. Aerobic bacteria colony/1ml. 6 123 2 Halophilic lactic acid bacteria colony/1ml. 4 6 10 Osmophilic yeast colony/1ml. $828{\times}10^4$ 248 - b. In the stored soy-sauces, aerobic bacteria are incapable of growing due to drop in pH value and the influence of salt concentration. c. Halophilic lactic acid bacteria are incapable of growing due to drop in pH value, even the salt concentrations decreased during the storage. d. Osmophilic yeast are still growing in low pH value and in the decreasing salt concentration during the strage. 3. The results of amino acid analysis by paper partition chromatographic and calorimetric methods are shown as follows. a. Fourteen kinds of amino acid and thirteen amino acids were detected in the soy-sauce of 12-year aged improved soy-sauce and 7-year aged and 20-year aged ordinary one, respectively. b. The contents of aspartic acid, glutamic acid, serine, valine, leucine, lysine, histidine and methionine increased in the 20-year aged ordinary soy-sauce compared to the 7-year aged one. On the other hand those of alanine, tyrosine, phenylalanine and cystine decreased. 4. The results of sugar analysis by paper chromatography are as follows. a. In the 12-year aged improved soy-sauce, galactose, glucose, arabinose, xylose, rhamnose, maltose and an unknown were detected, and their amounts were in the above order except maltose and an unknown. b. Both in the 7 and 12-year aged ordinary soy-sauces, galactose, arabinose, xylose, glucose and rhamnose were detected and the amounts of the sugars were in the above order. c. In the non-aged ordinary soy-sauce, glucose was not detected but detected from 7-year and 20-year aged ordinary soy-sauce. 5. The results of organic acid analysis by paper chromatography were as follows. a. As volatile acids, acetic, propionic and butyric acids were detected in the 7-year aged ordinary soy-sauce. On the other hand in both the 20-year aged ordinary soy-sauce and the 12-year aged improved ones, only acetic acid was abundant while propionic and butyric acids were round in trace. It was found that propionic and butyric acids, as the unpleasant flavor components, decreased during the storage. b. In the ordinary soy-sauce, citric acid were produced during the storage and lack, malic and tartaric acids increased in the course of aging while succinic, glycolic, fumaric and malonic acids were shown to decrease. Glutaric and oxalic acids disappeared. Citric acid was produced also in the improved soy-sauce, but lactic, tartaric, succinic, malic, and glycolic acids decreased, while both malonic and glutaric acids disappeared. From the above results the citric acid production was considered to be a favorable factor for the taste. c. In the aged soy-sauces, pyruvic, α-ketoglutaric and probably acetoacetic and oxaloacetic acids (both in trace) were present and their amounts were in the above order. All of the α-keto acid abruptly decreased during the storage.

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