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Analysis of molecular mechanism of cellular localization of various N-terminal mutants of Aplysia PDE4 in HEK293T cells (ApPDE4 long-form의 N-말단 돌연변이체들의 세포내타기팅과 타기팅 기전 분석)

  • Um, Su-Min;Jun, Yong-Woo;Kim, Kun-Hyung;Lee, Jin-A;Jang, Deok-Jin
    • Analytical Science and Technology
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    • v.29 no.1
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    • pp.10-18
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    • 2016
  • Phosphodiesterase (PDE) plays an important role in cAMP-mediated signaling within cells. We previously showed that the long-form of Aplysia PDE4 (ApPDE4) was localized in the plasma membrane and the presynaptic terminal in Aplysia sensory neurons, and the 16 N-terminal amino acid was sufficient for this targeting process. In this study, we characterized the cellular localization of various ApPDE4 mutants. We first identified the roles of each amino acid within the group of 16 N-terminal amino acids of long-form ApPDE4. As a result, we were able to identify various mutants that were localized to both the plasma membrane and the Golgi complex, Golgi only, or both the endoplasmic reticulum (ER) and the Golgi complex. To examine the role of palmitoylation on the cellular localization of ApPDE4 mutants, 2-bromo palmitate (2-BR) was used as a treatment. As a result, in the presence of 2-BR, the plasma membrane targeting of many mutants was impaired, indicating that palmitoylation was involved in the plasma membrane targeting of the mutants. We also found that PI4P play crucial roles in the Golgi targeting of (N16,C3S/VV/G)-mRFP, L(N16,C3S/LFS/R)-mRFP, and L(N16,EPL/R)-mRFP.

Comparative studies of various transfection processes for the optimal luminescence signal analysis (최적의 luminescence 신호 분석을 위한 유전자 전달 방법의 비교연구)

  • Park, Seohyun;Lee, Sunghou
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.17 no.11
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    • pp.640-647
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    • 2016
  • By minimizing fluorescence interference phenomena, aequorin-based luminescence technology can provide a relatively sensitive detection platform with integration of $G{\alpha}16$ protein in order to track internal calcium mobilization by G protein-coupled receptors (GPCR). In this type of cell-based functional assay format, it is essential to optimize the transfection process of a receptor and $G{\alpha}16$ protein. For this study, corticotropin releasing factor receptor subtype 2(CRF2) was set as a model system to generate three stable cells with CRF2 and $G{\alpha}16$ in addition to transiently transfected cells under three different conditions. Agonist (sauvagine) and antagonist (K41498) responses in those cells were analyzed to develop the optimum transfection process. As a result, the effective signal ratio in the dose response experiments of sauvagine and K41498 were at least 10-fold higher (z'=0.77) in CRF2-$G{\alpha}16$ stable cells. For the transient transfection cells, stable expression of $G{\alpha}16$ prior to the CRF2 represented a two-fold higher signal (z'=0.84) than the other cases of transient transfection. In conclusion, for the utilization of transient transfection processes to develop a cell-based GPCR functional assay system, it is suggested to introduce various target receptors after stable expression of $G{\alpha}16$ protein.

Folate Deficiency and FHIT Hypermethylation and HPV 16 Infection Promote Cervical Cancerization

  • Bai, Li-Xia;Wang, Jin-Tao;Ding, Ling;Jiang, Shi-Wen;Kang, Hui-Jie;Gao, Chen-Fei;Chen, Xiao;Chen, Chen;Zhou, Qin
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.21
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    • pp.9313-9317
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    • 2014
  • Fragile histidine triad (FHIT) is a suppressor gene related to cervical cancer through CpG island hypermethylation. Folate is a water-soluble B-vitamin and an important cofactor in one-carbon metabolism. It may play an essential role in cervical lesions through effects on DNA methylation. The purpose of this study was to observe effects of folate and FHIT methylation and HPV 16 on cervical cancer progression. In this study, DNA methylation of FHIT, serum folate level and HPV16 status were measured using methylation-specific polymerase chain reaction (MSP), radioimmunoassay (RIA) and polymerase chain reaction (PCR), respectively, in 310 women with a diagnosis of normal cervix (NC, n=109), cervical intraepithelial neoplasia (CIN, n=101) and squamous cell carcinoma of the cervix (SCC, n=101). There were significant differences in HPV16 status (${\chi}^2=36.64$, P<0.001), CpG island methylation of FHIT (${\chi}^2=71.31$, P<0.001) and serum folate level (F=4.57, P=0.011) across the cervical histologic groups. Interaction analysis showed that the ORs only with FHIT methylation (OR=11.47) or only with HPV 16 positive (OR=4.63) or with serum folate level lower than 3.19ng/ml (OR=1.68) in SCC group were all higher than the control status of HPV 16 negative and FHIT unmethylation and serum folate level more than 3.19ng/ml (OR=1). The ORs only with HPV 16 positive (OR=2.58) or with serum folate level lower than 3.19ng/ml (OR=1.28) in CIN group were all higher than the control status, but the OR only with FHIT methylation (OR=0.53) in CIN group was lower than the control status. HPV 16 positivity was associated with a 7.60-fold increased risk of SCC with folate deficiency and with a 1.84-fold increased risk of CIN. The patients with FHIT methylation and folate deficiency or with FHIT methylation and HPV 16 positive were SCC or CIN, and the patients with HPV 16 positive and FHIT methylation and folate deficiency were all SCC. In conclusion, HPV 16 infection, FHIT methylation and folate deficiency might promote cervical cancer progression. This suggests that FHIT may be an effective target for prevention and treatment of cervical cancer.

Optimum Dietary Protein and Lipid Levels on Growth in Parrot Fish (Oplegnathus fasciatus) (돌둠사료의 적정 단백질 및 지질 함량)

  • 강용진;이상민;황형규;배승철
    • Journal of Aquaculture
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    • v.11 no.1
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    • pp.1-10
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    • 1998
  • A feeding trial was conducted to investigate optimum dietary protein and lipid levels in diets for parrot fish (Oplegnathus fasciatus). Three groups of 25 fish averaging 7.0g were fed each of the experimental diets containing 30, 40, 50 or 60% crude protein (CP) with 8 or 16% crude lipid level ($4{\times}2$factorial desing) for 8 weeks. Weight gain and feed efficiency increased significantly (P<0.05) with dietary protein level up to 50% CP when diets contained 16% lipid level, and increased with dietary protein level up 60% CP when diets contained 8% lipid level (P<0.05). The broken-like model showed that an optimum dietary protein level was 46% when the diet contained 16% dietary lipid. Fish fed diets containing 16% lipid level showed significantly (P<0.05) higher weight gain, feed efficiency and protein efficiency ratio than did fish fed diets containing 8% lipid level when diets contained 40 or 50% dietary protein (P<0.05).

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Identification of Bacteria Causing Fermentation of Oriental Melon in Korea (참외 발효과를 유발하는 세균의 동정)

  • Choi, Jae-Eul;Cha, Sun-Kyung;Kim, Jin-Hee;Yuk, Jin-Ah;Hwang, Yong-Soo;Kwon, Soon-Wo
    • Research in Plant Disease
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    • v.9 no.4
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    • pp.189-195
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    • 2003
  • Bacteria causing fermentation in Oriental melon were identified as three independent groups on the basis of 16S rDNA sequence analysis. The 16S rDNA sequence of the strain CM2105 showed the highest identity (99.6%) with that of Microbacterium phyllosphaerae, and also indicated high sequence identity to that of M. holiorum (99.5%). The 16S rDNA sequences of the strain CM2101 and CM2121 matched at the high sequence similarity (98.9%, 98.8, respectively), to that of Pseudomonas pavonacea, and the DNA sequence of CM2126 showed high sequence identity to that of P. costantinii (99.5%), and P. grimontii (99.0%). The 16S rDNA sequence of the strain CM2113 showed the highest identity (99.7%) with that of Enterobacter cloacae. The 16S rDNA sequences, the physiological and biochemical analysis suggested that the strain CM2105 belonged to Microbacterium phyllosphaerae, CM2101, CM2121 and CM2126 to Pseudomonas spp., CM2113 to Enterobacter cloacae.

Changes in Development and Nutrient Composition of Pod after Flowering in Cowpea 〔Vigna unguiculata(L) Walp〕 (동부(Vigna unguiculatal(L) Walp)의 개화후 일수경과에 따른 협실비대 및 품질의 변화)

  • 김수동;차영훈;조진태;권규칠;손삼곤;박상일
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.31 no.1
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    • pp.68-73
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    • 1986
  • In order to identify the physiological maturity and to determine the proper time of harvesting as fresh seed for cooking with rice and germination ability according to the harvesting time in cowpea (Vigna unguiculata (L.) Walp), this experiment was conducted. The results obtained are as follows; 1. The length and width of pods became maximum size on 10 days after flowering(DAF) and thickness of them increased to 14 DAF and then decreased to 18 DAF. 2. The weight of pod wall reached the maximum on 14 DAF, the thickness of them increased to 8 DAF and decreased to 18 DAF and it was stabilized. 3. The length and thickness of the seeds became larger to 10 DAF, were not changed from 10 DAF to 16 DAF and then decreased after 16 DAF. 4. The seeds coloured 2 days earlier than the pod wall on 8 DAF and the grade increased by degrees to 16 DAF. 5. The moisture content of seeds and pod wall started to decrease on 8 DAF and the moisture content of pod wall was higher than that of seeds in the middle stage but the condition changed after 18 DAF. 6. When the weight of seeds reached the maximum on 16 DAF, the moisture content of them was 54.5%. 7. The content of total nitrogen, phosphate and oil decreased but the carbohydrate increased by degrees along the passing of DAF. The silicic acid did not exist and K$_2$O, CaO, MgO, protein etc. existed but did not show regular tendency. 8. The physiological maturity was 16 DAF and the proper time of harvesting as fresh seed for cooking with rice was 12 DAF to 16 DAF. 9. Germination was possible after 10 DAF but we could get the normal roots from the seeds harvested after 14 DAF.

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Change of Dry Matter Yield and Feed Values According to Different Growth Stages of Italian Ryegrass and Triticale Cultivated in the Central Northern Region (중북부 지역에서 생육단계에 따른 이탈리안 라이그라스와 트리티케일의 건물수량과 사료가치)

  • Lee, Hong-Ju;Byeon, Ji-Eun;Hwang, Sun-Goo;Ryoo, Jong-Won
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.40 no.1
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    • pp.50-56
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    • 2020
  • The field experiment was carried out to evaluate changes of dry matter productivity and feed value of Italian ryegrass and triticale at different growth stages in Wonju from 2018 to 2019. Italian ryegrass and triticale forages were harvested at an interval of seven days from May to June. The dry matter yield of Italian ryegrass with 7,870 kg/ha harvested on May 16 was significantly higher than that harvested on May 3 and May 9. The dry matter yield of triticale with 12,050 kg/ha harvested on June 7 was significantly higher than that harvested from May 3 to May 16. The crude protein yields harvested on May 16 were 890 kg/ha for Italian ryegrass and 1,103 kg/ha for triticale and were significantly higher than those harvested on May 3 and May 9. The total digestible nutrient yield(TDN) of Italian ryegrass with 5,223 kg/ha harvested on May 16 was significantly higher than that harvested on May 3 and May 9. The total digestible nutrient yield of triticale with 8,277 kg/ha harvested on June 7 was significantly higher than that harvested during May, but not different from that harvested after June 7. Considering the dry matter yield, crude protein yield and total digestible nutrient yield, it is suggested that the optimal harvesting times for Italian ryegrass and triticale seem to be May 16 and June 7, respectively.

Ontogenetic Color Variation of Abudefduf notatus (Pomacentridae: Perciformes) Revealed by 16S rRNA Sequences Analysis (미토콘드리아 16S rRNA 염기서열 분석에 의해 밝혀진 동갈자돔 치어의 성장에 따른 체색변이)

  • Song, Young Sun;Kim, Jin-Koo
    • Korean Journal of Ichthyology
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    • v.25 no.2
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    • pp.53-58
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    • 2013
  • Seven individuals (16.1~29.1 mm in length) which were estimated with fishes of genus Abudefduf were collected in Seogwipo, Jeju Island in the summer of 2011 and 2012. Among them, five individuals (20.8~29.1 mm SL) are similar to Abudefduf notatus, based on morphological characters such as yellow transverse band on the body, one small black spot on the base of the pectoral fin, and yellow caudal fin. On the other hand, two individuals (16.1 mm, 17.0 mm SL) differ from them in several light transverse bands and transparent rays of all fins except for pelvic fin, and anterior transparent head with only one (16.1 mm). According to the results of molecular analyses of 578 base pairs of mtDNA 16S rRNA sequences, these individuals correspond to A. notatus adult, with a high bootstrap value of 95% (genetic distance, d=0.002). Therefore, this study shows that the individuals more than 20.8 mm similar to adult body color but the 16.1 mm individual differs to that of adult. We confirm that this species changes to body color during their early life stages. This result regards as a survival strategy to protect themselves against their predator during their early life stages.

Studies on the Lipid and Fatty Acid Compositions of Ark-Shell, Anadara broughtonii (피조개 지질 및 지방산 조성에 관한 연구)

  • 문숙임
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.21 no.4
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    • pp.436-442
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    • 1992
  • The present study was designed to analyze the lipid and fatty acid compositions of ark shell, Anadara broughtonii. The crude lipid was extracted by Bligh and Dyer's method, and then fractionated by TLC and quantitatively analyzed by TLC scanner. Lipid extracted from ark shell was fractionated into neutral and polar lipid by column chromatography with silicic acid. The fatty acid composition of lipid fractions were determined by gas liquid chromatography. Total lipid content of ark shell was 0.83% base on wet weight. The content of unsaponifiable matter was 20.19%, and iodine value was 156.13. The main components of total lipids were triglyceride, diglyceride, hydrocarbon, and sterol ester. The fatty acid composition of total lipid chiefly consisted of $C_{17 : 0}$, $C_{16 : 0}$, $C_{18 : 1}$ and $CT_{16 : 1}$. The main fatty acids of neutral lipid were $C_{16 : 0}$, $C_{18 : 1}$, $C_{22 : 1}$, $C_{18 : 0}$ and $C_{16 : 1}$. The major fatty acids of polar lipid were $C_{16 : 0}$, $C_{18 : 2}$, $C_{20 : 5}$ and $C_{22 : 6}$. In total lipid fractionation, saturated acid contents were high in all (SA>MA> PA), in neutral lipid fractionation, menoenoic acid contents were high in all (MA > SA> PA), and in polar lipid fractionation, saturated acid con-tents were high in all (SA> PA> MA).

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Development of 1×16 Thermo-optic MZI Switch Using Multimode Interference Coupler (다중모드 간섭현상을 이용한 1×16 마하젠더 스위치 개발)

  • Kim, Sung-Won;Hong, Jong-Kyun;Lee, Sang-Sun
    • Korean Journal of Optics and Photonics
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    • v.17 no.5
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    • pp.469-474
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    • 2006
  • A $1{\times}16$ thermo-optic switch with small excess loss using multimode interference(MMI) couplers is designed, fabricated, and measured. This paper introduces the proposed $1{\times}16$ thermo-optic switch, and discusses the measurement results. The $1{\times}16$ thermo-optic switch is farmed as 4-stage which consists of 15 unit devices. The unit devices are the $2{\times}2$ thermo-optic switches with Mach-Zehnder interferometer(MZI) structure. The characteristics of the $1{\times}16$ thermo-optic switch depends strongly on each unit device. The unit deviceconsists of two 3-dB general interference MMI couplers and two single mode waveguide arms as a phase shifter. First of all, the 3-dB optical splitter and $2{\times}2$ MZI thermo-optic switch have been tested to confirm the characteristics of the unit devices of the $1{\times}16$ MZI thermo-optic switch. Using the measurement results of the unit devices, the $1{\times}16$ MZI thermo-optic switch can be produced with better characteristics. The resultant structure of the MMI coupler with the optical light source of wavelength of 1550nm for the $1{\times}16$ thermo-optic switch is that the width and the optimized length are $25{\mu}m\;and\;1580{\mu}m$, respectively. The smallest excess loss fur the unit device is -0.5dB and the average excess loss is -0.7dB.