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Cloning and Expression Analysis of a Chitinase Gene Crchi1 from the Mycoparasitic Fungus Clonostachys rosea (syn. Gliocladium roseum)

  • Gan, Zhongwei;Yang, Jinkui;Tao, Nan;Yu, Zefen;Zhang, Ke-Qin
    • Journal of Microbiology
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    • v.45 no.5
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    • pp.422-430
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    • 2007
  • Clonostachys rosea (syn. Gliocladium roseum) is a well-known biocontrol agent and widely distributed around the world. In this study, an endochitinase gene Crchi1 was isolated from the mycoparasitic fungus C. rosea using the DNA walking strategy. The Crchi1 ORF is 1,746 bp long and interrupted by three introns. The cloned gene Crchi1 encodes 426 amino acid residues and shares a high degree of similarity with other chitinases from entomopathogenic and mycoparasitic fungi. Several putative binding sites for transcriptional regulation of Crchi1 in response to carbon (5'-SYGGRG-3') and nitrogen (5'-GATA-3') were identified in the upstream of Crchi1. Expression of Crchi1 gene in different carbon sources was analyzed using real-time PCR (RT-PCR). We found that the Crchi1 expression was suppressed by glucose but strongly stimulated by chitin or solubilized components of the cell wall from Rhizoctonia solani. Phylogenetic analysis of chitinases from entomopathogenic and mycoparasitic fungi suggests that these chitinases have probably evolved from a common ancestor.

Knowledge, Confidence, and Learning Needs Regarding Advance Directives among Hospital Nurses (상급종합병원 간호사의 사전연명의료의향서에 대한 지식, 자신감 및 교육요구도)

  • Jang, Nan-Soon;Park, Hae-Sook;Kim, Mi-Ra;Lee, Joo-Yeon;Cho, Yeo-Won;Kim, Kyoung-Mi;Son, Youn-Jung
    • Journal of Korean Critical Care Nursing
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    • v.11 no.1
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    • pp.35-45
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    • 2018
  • Purpose : Nurses' knowledge regarding advance directives may affect their administration of and confidence towards end of life care. This study aimed to describe the relationships of knowledge, confidence, and learning needs with advance directives among hospital nurses. Method : This cross-sectional study was performed at a tertiary university hospital in Seoul between September 25 and October 14, 2017. Convenience sampling was used to recruit nurses who provided bedside care and had at least 1 year of clinical experience. We used a validated self-report questionnaire. Results : The mean score of knowledge, confidence and learning needs were $5.00{\pm}1.73$, $29.81{\pm}7.52$, and $64.54{\pm}8.48$ respectively. Hospital nurses' knowledge, confidence and learning needs were significantly different according to age, job position, educational level and perceived advance directives. Knowledge regarding advance directives was significantly associated with confidence (r = .27, p < .001) and learning needs (r = .16, p = .005). Conclusion : Knowledge regarding advance directives was relatively low compared to the findings of previous studies. Therefore, nurses should be knowledgeable and encouraged to initiate advance directives. It is necessary to develop a standardized educational program regarding advance directives based on Korean cultures.

Implementation of FlexRay Communication Controller Protocol and its Application to a Robot System (FlexRay 프로토콜 설계 및 로봇 시스템 응용)

  • Kang, Hyun-Soo;Xu, Yi-Nan;Kim, Yong-Eun;Chung, Jin-Gyun
    • Journal of the Institute of Electronics Engineers of Korea TC
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    • v.45 no.6
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    • pp.1-7
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    • 2008
  • FlexRay is a new standard of network communication system which provides a high speed serial communication, time triggered bus and fault tolerant communication between electronic devices for future automotive applications. FlexRay communication controller (CC) is the core of the FlexRay protocol specification. In this paper, we first design the FlexRay CC protocol specification and function parts using SDL (Specification and Description Language). Then, the system is re-designed using Verilog HDL based on the SDL source. The FlexRay CC system was synthesized using Samsung $0.35\;{\mu}m$ technology. It is shown that the designed system can operate in the frequency range above 80 MHz. In addition, to show the validity of the designed FlexRay system the FlexRay system is combined with sound source localization system in Robot applications. The combined system is implemented using ALTERA Excalibur ARM EPXA4F672C3. It is shown that the implemented system operates successfully.

Implementation of FlexRay Protocol Specification and its Application to a Automobile Advance Alarm System (FlexRay 프로토콜 설계 및 자동차 경보 시스템 응용)

  • Xu, Yi-Nan;Yang, Sang-Hoon;Chung, Jin-Gyun
    • Journal of the Institute of Electronics Engineers of Korea TC
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    • v.45 no.8
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    • pp.98-105
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    • 2008
  • FlexRay is a high-speed communications protocol with high flexibility and reliability. It was devised by automotive manufacturers and semiconductor vendors and implemented as on vehicle LAN protocol using x-by-wire systems. FlexRay provides a high speed serial communication, time triggered bus and fault tolerant communication between electronic devices for automotive applications. In this paper, we first design the FlexRay communication controller, bus guardian protocol specification and function parts using SDL (Specification and Description Language). Then, the system is re-designed using Verilog HDL based on the SDL source. The FlexRay system was synthesized using Samsung $0.35{\mu}m$ technology. It is shown that the designed system can operate in the frequency range above 76 MHz. In addition, to show the validity of the designed FlexRay system, the FlexRay system is combined with automobile advance alarm system in vehicle applications. The FlexRay system is implemented using ALTERA Excalibur ARM EPXA4F672C3. It is shown that the implemented system operates successfully.

Expressional Patterns of Connexin Isoforms in the Rat Epididymal Fat during Postnatal Development

  • Lee, Ki-Ho;Kim, Nan Hee
    • Development and Reproduction
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    • v.22 no.1
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    • pp.29-38
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    • 2018
  • In the multicellular tissue, cell-cell interaction is important for a precise control of its function. The exchange of signaling molecules between adjacent cells via connexon allows the functional harmony of cells in the tissue. The present research was to determine the presence and expressional patterns of connexin (Cx) isoforms in the rat epididymal fat during postnatal development using quantitative real-time polymerase chain reaction (PCR) analysis. Of 13 Cx isoforms examined, expression of 11 Cx isoforms in the epididymal fat during postnatal development was detected. These Cx isoforms include Cx26, Cx31, Cx31.1, Cx32, Cx33, Cx36, Cx37, Cx40, Cx43, Cx45, and Cx50. Expressional levels of all Cx isoforms at 1 and 2 years of age were significantly higher than those at the early postnatal ages, such as 7 days, 14 days, and 24 days of ages. Except Cx33 and Cx43, the transcript levels of rest Cx isoforms at 1 year of age were significantly lower than that at 2 years of age. In addition, expressional patterns of Cx isoforms between 7 days and 5 months of ages generally varied according to the isoform. The existence of various Cx isoforms in the rat epididymal fat has been identified and expression of each Cx isoform in the epididymal fat during postnatal development has shown a particular pattern, distinguishable from the others. To our knowledges, this is the first report showing expressional patterns of Cx isoforms at transcript level in the epididymal fat at various postnatal ages.

Caffeine Treatment during Oocyte Aging Improves the Developmental Rate and Quality in Bovine Embryos Developing In Vitro

  • Choi, Hyun-Yong;Lee, Sung-Hyun;Xu, Yong-Nan;Lee, Seung-Eun;Kim, Nam-Hyung
    • Reproductive and Developmental Biology
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    • v.37 no.4
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    • pp.281-287
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    • 2013
  • In mammal, unfertilized oocytes remain in the oviduct or under in vitro culture, which is called "oocyte aging". This asynchrony negatively affects fertilization in pre- and post-implantation embryo development. Caffeine a phosphodiesterase inhibitor is known to rescue oocyte aging in several species. The objective of this study is to determine the cytoskeleton distribution in aged oocytes and the embryo developmental ability of aged oocytes in the present or absence of caffeine during maturation. Caffeine treatment increased the incidence of normal spindle assembly of aged oocytes (treatment, $67.57{\pm}4.11%$ aging, $44.61{\pm}6.4%$) and no significant differences compared to control group. Fluorescence values were compared using ROS (Reactive oxidation species) stain. Fluorescence values appear of control group intensity rate ($51.53.{\pm}3.80$), aging group ($68.10{\pm}5.54$) and treatment of caffeine ($45.04{\pm}2.98$). Aged oocytes that were derived from addition of caffeine to the IVM (in vitro maturation) medium had significantly increased 2-cell that developed to the blastocyst stage compared to the aging group. Blastocysts, derived from caffeine treatment group, significantly increased the total cell number compare aging ($90.44{\pm}10.18$ VS $67.88{\pm}7.72$). Apoptotic fragments of genomic DNA were measured in individual embryo using TUNEL assay. Blastocyst derived from caffeine treatment group decreased significantly the apoptotic index compared to blastocyst derived from aging group. In conclusion, we inferred that the caffeine treatment during oocyte aging can improve the developmental rate and quality in bovine embryos developing in vitro.

Effect of harvest dates on β-carotene content and forage quality of rye (Secale cereale L.) silage and hay

  • Zhao, Guo Qiang;Wei, Sheng Nan;Liu, Chang;Kim, Hak Jin;Kim, Jong Geun
    • Journal of Animal Science and Technology
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    • v.63 no.2
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    • pp.354-366
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    • 2021
  • Limited data about the effects of various factors on forage quality and β-carotene content of rye produced in Korea are available, so this study investigated the effects of two preservation methods. Samples were collected from rye harvested every 5 days between April 25 and May 31, and comparisons were done among rye silage wilted for different periods of time and hay of three growth stages of rye. For the silage, dry matter (DM), acid detergent fiber (ADF), and neutral detergent fiber (NDF) contents increased with advanced maturity of rye, whereas crude protein, in vitro dry matter digestibility (IVDMD), total digestible nutrients (TDN), relative feed value (RFV), and DM loss decreased (p < 0.0001). Wilting increased the DM content and pH value significantly (p < 0.0001). Silage harvested at the heading stage had the lowest pH value (4.45), propionic acid (0.83 g/kg DM), butyric acid (0 g/kg DM), and fungi and yeast populations (3.70 Log CFU/g of fresh matter [FM]); conversely, it had the highest lactic acid (9.7 g/kg DM), lactic acid bacteria (LAB) (6.87 Log CFU/g of FM), total microorganisms (TM) (7.33 Log CFU/g of FM), and Flieg's score (70) (p < 0.0001). Wilting elevated LAB and TM populations, but it had no consistent effect on other fermentation products. Both delayed harvest and prolonged wilting decreased β-carotene content. Rye silage harvested around May 9 (heading stage) with 24 h of wilting was preferred for highland, Pyeongchang. For rye hay, advanced maturity decreased DM loss, IVDMD, TDN, and RFV, but it increased DM, ADF, and NDF significantly (p < 0.05). β-carotene was decreased by delay of hay-making. Consequently, to attain lower DM loss and higher hay quality, the harvest date of May 9 (heading stage) is recommended.

Anti-Inflammatory Effect of Grateloupia imbricata Holmes Ethanol Extract on LPS-Induced RAW 264.7 Cells (꽃지누아리 에탄올 추출물의 LPS로 유도된 RAW 264.7 세포에 대한 항염증 효과)

  • Kim, Min-Ji;Bae, Nan-Yong;Kim, Koth-Bong-Woo-Ri;Park, Ji-Hye;Park, Sun-Hee;Choi, Jung-Su;Ahn, Dong-Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.2
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    • pp.181-187
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    • 2016
  • Algae is a potential resource with various biological activities. In this study, the anti-inflammatory effect of Grateloupia imbricata Holmes ethanol extract (GIHEE) from red algae was investigated in LPS-induced RAW 264.7 cells. As a result, reduced secretion of pro-inflammatory cytokines [tumor necrosis factors-${\alpha}$, interleukin (IL)-$1{\beta}$, and IL-6] and nitric oxide (NO) was observed in a dose-dependent manner. Expression of nuclear factor-kappaB (NF-${\kappa}B$) as well as inducible NO synthase and cyclooxygenase-2 proteins was reduced by GIHEE, suggesting that the anti-inflammatory activity of GIHEE is related to suppression of NF-${\kappa}B$ signaling pathways. In addition, GIHEE reduced phosphorylation of mitogen-activated protein kinases. These results suggest that GIHEE can be used as a potential anti-inflammatory therapeutic.

Anti-Inflammatory Effect of Chondrus nipponicus Yendo Ethanol Extract on Lipopolysaccharide-Induced Inflammatory Responses in RAW 264.7 Cells (LPS로 유도된 RAW 264.7 세포에 대한 가락진두발 에탄올 추출물의 항염증 효과)

  • Kim, Min-Ji;Bae, Nan-Yong;Kim, Koth-Bong-Woo-Ri;Park, Ji-Hye;Park, Sun-Hee;Jang, Mi-Ran;Ahn, Dong-Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.2
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    • pp.194-201
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    • 2016
  • The anti-inflammatory activity of ethanol extract from Chondrus nipponicus Yendo (CNYEE) was investigated by measuring production of a lipopolysaccharide-induced inflammatory response mediator. CNYEE had no cytotoxic effects on proliferation of macrophages compared to the control. CNYEE significantly inhibited (over 50%) NO production at $50{\mu}g/mL$, with inhibitory effects on expression levels of cytokines such as interleukin (IL)-6, tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), and IL-$1{\beta}$. In particular, IL-6 inhibitory activity of CNYEE was higher than 70% at $100{\mu}g/mL$. CNYEE also reduced protein expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and nuclear factor (NF)-${\kappa}B$ in a dose-dependent manner. CNYEE also significantly reduced phosphorylation of p38, extracellular signal-regulated kinase, and c-Jun N-terminal kinase. Therefore, these results suggest that CNYEE may have anti-inflammatory effects by modulating the NF-${\kappa}B$ and mitogen-activated protein kinases signaling pathways and may be used as an anti-inflammatory therapeutic material.

Immunohistochemical Study on the Suppression of Cell mediated immunity in Lymph node of mouse by Cyclosporin A -Based on the change of T lymphocytes, Il-2 receptors, and NK cells- (Cyclosporin A로 유도된 생쥐 림프절의 세포성 면역억제에 관한 면역조직화학적 연구 -T 림프구, IL-2 수용기 및 NK세포의 변화를 중심으로-)

  • Kim, Jin-Taek;Park, In-Sick;Ahn, Sang-Hyun;Choi, Nan-Hee;Kim, Dong-Hoan
    • The Journal of Dong Guk Oriental Medicine
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    • v.6 no.2
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    • pp.99-107
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    • 1998
  • Cyclosporin A(CsA) is a selective immunosuppressive agent that has been credited with improved survival of solid organ allografts. Lymph node of BALB/C mouse administered CsA immunohistochemically observed to understand immunosuppressive effects of CsA on T lymphocytes, IL-2 receptors, and natural killer NK cells in lymph node. CsA orally administered daily for 10days at the dose 45mg/kg/day/. The lymph node were obtained at day 3, 7, and 14 after CsA administration and embedded with paraffin, and then stained by following ABC method that used monoclonal antibody including L3T4(CD4), Ly2(CD8), IL-2R(CD25), and NK-1.1(CD56). There were little changes of reactive degree and number of helper T lymphocytes, cytotoxic T lymphocytes, IL-2 receptors, and NK cells at day 3 after CsA administration, but they began to decrease at day 7. These decrease were greatest at day 14. The helper T lymphocytes. cytotoxic T lymphocytes, IL-2 receptors, and NK cells distributed in paracortex and medullary sinus. These results indicated that the secretion of IL-2 began to decrease at day 7 after CsA administration and subsequently to suppress T lymphocytes and NK cell as components of cell-mediated immunity.

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