• Title/Summary/Keyword: $CP_1$

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Inhibitory Effects of a Recombinant Viral Cystatin Protein on Insect Immune and Development (바이러스 유래 시스타틴 재조합 단백질의 곤충 면역 및 발육 억제효과)

  • Kim, Yeongtae;Eom, Seonghyun;Park, Jiyeong;Kim, Yonggyun
    • Korean journal of applied entomology
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    • v.53 no.4
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    • pp.331-338
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    • 2014
  • Cystatins (CSTs) are reversible and competitive inhibitors of C1A cysteine proteases, corresponding to papain-like cathepsins in plants and animals. A viral CST (CpBV-CST1) was identified from a polydnavirus, Cotesia plutellae bracovirus (CpBV). Our previous study indicated that a transient expression of CpBV-CST1 interfered with immune response and development of Plutella xylostella larvae. To directly demonstrate the protein function, this study produced a recombinant CpBV-CST1 protein (rCpBV-CST1) using bacterial expression system to determine its inhibitory activity against cysteine protease and to assess its physiological alteration in insect immune and development. The open reading frame of CpBV-CST1 encodes a polypeptide of 138 amino acids (${\approx}15kDa$). rCpBV-cystatin protein in BL21 STAR (DE3) competent cells containing a recombinant pGEX4T-3:CpBV-CST1 was over-expressed by 0.5 mM IPTG for 4 h. In biological activity assay, the purified rCpBV-CST1 showed a significant inhibition against papain activity. It inhibited a cellular immune response of hemocyte nodule formation in the beet armyworm, Spodoptera exigua. Moreover, its oral administration retarded larval development of the diamondback moth, Plutella xylostella in a dose-dependent manner. These results suggest that CpBV-CST1 may be applied to control insect pest populations.

$New η^3-Allyl-Alkenyl- and η^3-Allyl-Alkynyl-Ir-Cp^* Compounds from Reactions of [Cp^*Ir(η^3-CH_2CHCHPh)(NCMe)]^+ with Alkynes$

  • Jin, Jong Sik;Jong, Dae Seong;Kim, Mi Yeok;Lee, Hyeon Gwi
    • Bulletin of the Korean Chemical Society
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    • v.22 no.7
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    • pp.739-742
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    • 2001
  • Reactions of [Cp*Ir(η3-CH2CHCHPh)(NCMe)]OTf (1) with HC≡CR (R = H, CH2OH) in the presence of bases, B (B=NEt3, PPh3, AsPh3) produce stable Cp*Ir-η3-allyl-alkenyl compounds [Cp*Ir(η3-CH2CHCHPh)(-CH=CH-+B)]OTf (2) and [Cp*Ir(η3-CH2CHCHPh)(-C(CH2OH)=CH- +PPh3)]OTf (3), respectively in high yields. Cp*Ir-η3-allyl-alkynyl compounds Cp*Ir(η3-CH2CHCHPh(-C≡C-R') (4) and Cp*(η3-CH2CHCHPh)Ir-C≡C-p-C6H4-C≡C-Ir(η3-CH2CHCHPh)Cp* (5) have been prepared from reactions of 1 with HC≡CR'(R' = C6H5, p-C6H4CH3, C3H5, C6H9) and HC≡C-p-C6H4-C≡CH in the presence of NEt3.

A SERI technique reveals an immunosuppressive activity of a serine-rich protein encoded in Cotesia plutellae bracovirus

  • Barandoc, Karen P.;Park, Jay-Young;Kim, Yong-Gyun
    • BMB Reports
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    • v.43 no.4
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    • pp.279-283
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    • 2010
  • Polydnavirus genome is segmented and dispersed on host wasp chromosome. After replication, the segments form double- stranded circular DNAs and embedded in viral coat proteins. These viral particles are delivered into a parasitized host along with parasitoid eggs. A serine-rich protein (SRP) is predicted in a polydnavirus, Cotesia plutellae bracovirus (CpBV), genome in its segment no. 33 (CpBV-S33), creating CpBV-SRP1. This study explored its expression and physiological function in the diamondback moth, Plutella xylostella, larvae parasitized by C. plutellae. CpBV-SRP1 encodes 122 amino acids with 26 serines and several predicted phosphorylation sites. It is persistently expressed in all tested tissues of parasitized P. xylostella including hemocyte, fat body, and gut. Its physiological function was analyzed by injecting CpBV-S33 and inducing its expression in nonparasitized P. xylostella by a technique called SERI (segment expression and RNA interference). The expression of CpBV-SRP1 significantly impaired the spreading behavior and total cell count of hemocytes of treated larvae. Subsequent RNA interference of CpBV-SRP1 rescued the immunosuppressive response. This study reports the persistent expression of CpBV-SRP1 in a parasitized host and its parasitic role in suppressing the host immune response by altering hemocyte behavior and survival.

Synthesis of $Cp^*Ru(CO)Cl_2(Cp^*={\eta}^5-C_5Me_5)$ Complex and Reaction with Phosphines ($Cp^*Ru(CO)Cl_2(Cp^*={\eta}^5-C_5Me_5)$착물의 합성과 포스핀과의 반응)

  • Lee, Dong Hwan;Kim, Sng Il;Jun, Jin Hee;Oh, Yung Hee;Kam, Sang Kyu
    • Journal of the Korean Chemical Society
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    • v.41 no.12
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    • pp.639-644
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    • 1997
  • Novel carbonylruthenium (Ⅲ) complex Cp*Ru(CO)Cl2(2, Cp*=η5-C5Me5) was synthesized by the reaction of [Cp*RuCl2]2(1) with CO in toluene. The effective magnetic moment (Veff=1.81 B.M.) derived from the magnetic susceptibility measurement of the complex (2) was consistent with the presence of one "single" unpaired electron. Dibromocarbonylruthenium (Ⅲ) complex Cp*Ru(CO)Br2(3) was obtained by the reaction of complex (2) with KBr in toluene. Complex (2) was easily reduced by the reaction with phosphine in toluene to give the corresponding Ru (Ⅱ) complex Cp*Ru(CO)(PR3)Cl (4a∼4e, PR3=PMe3, PEt3, PMePh2, PPh3, PCy3).

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Effects of CpG Oligodeoxynucleotides on Immune Responses and Expression of Cytokine Genes in Cultured Olive Flounder Paralichthys olivaceus

  • Ahn, kyoung-Jin;Nam, Bo-Hye;Kim, Young-Ok;Kang, Jung-Ha;Kim, Bong-Seok;Jee, Young-Ju;Lee, Sang-Jun
    • Fisheries and Aquatic Sciences
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    • v.10 no.1
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    • pp.1-7
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    • 2007
  • The induction of cellular and humoral immunity and cytokine gene expression by synthetic CpG oligodexoynucleotides (CpG-ODNs) has not been investigated systematically in olive flounder Paralichthys olivaceus in vivo. We optimized the proper concentration of CpG-ODNs using an in vitro assay for the superoxide anion $(O_2^-)$. CpG-ODNs induced $O_2^-$ and nitric oxide (NO) production, lysozyme activity, and the proinflammatory cytokine gene expression of $IL-1{\beta}$ and $TNF-{\alpha}$ in olive flounder significantly in vivo, whereas non-CpG-ODNs did not produce these effects or produced them to a lesser extent. This implied that CpG-ODNs could stimulate cellular and humoral immunity and cytokine gene expression in olive flounder. This is the first evidence of NO production and the first study on the mRNA expression of the proinflammatory cytokine genes $IL-1{\beta}$ and $TNF-{\alpha}$ in olive flounder in response to CpG-ODNs. Comparison of the variation in NO production and lysozyme activity to that of other studies led us to postulate that a group-specific difference exists in the immune responses of olive flounder against CpG-ODNs. Furthermore, the detailed immunostimulatory spectrum of CpG-ODNs in olive flounder could be a useful index with which to analyze the effect of CpG-ODNs against the challenge test prior to field applications.

Effects of protease enzyme supplementation in weanling pigs' diet with different crude protein levels on growth performance and nutrient digestibility

  • Olivier, Munezero;In Ho, Kim
    • Journal of Animal Science and Technology
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    • v.64 no.5
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    • pp.854-862
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    • 2022
  • The purpose of this study is to determine the impact of different levels of crude protein (CP) diets supplemented with dietary protease on the growth performance and nutrient digestibility of the weanling pigs. In a 5-week study, 100 crossbred ([Landrace × Yorkshire] × Duroc) of weaner pigs that have an average initial body weight (BW) of 7.17±1.06 kg were assigned to one of four dietary treatments with 5 replications and 5 pigs (3 gilts and 2 castrated male pigs) per pen in a randomized complete block design. The dietary treatments were as follows: Phase 1: CON: basal diets (20.60% CP); low protein (LP): CON - 0.30% CP; PLP1: (CON - 0.30% CP) + 0.05% protease; PLP2: (CON - 0.50% CP) + 0.05% protease. Phase 2: CON: basal diets (18.88% CP); LP: CON - 0.30% CP; PLP1: (CON - 0.30% CP) + 0.05% protease; PLP2: (CON - 0.50% CP) + 0.05% protease. The addition of protease to low CP diets significantly increased the feed conversion ratio (FCR) (p = 0.039), BW (p = 0.046), average daily gain (ADG) (p = 0.049), and average daily feed intake (ADFI) tended to increase (p = 0.053) in the young pigs during phase 1. However, FCR tended to increase throughout the experiment but did not change during phase 2, whereas BW, ADG, and ADFI stayed unchanged throughout phase 2 and overall. There was no significant difference in dry matter, nitrogen (N), and gross energy of nutrient digestibility in all phases and overall in weaned pigs with low CP when protease was fed. In contrast, adding protease to the low CP diets increased the tendency of N digestibility (p = 0.059) during phase 1. It is concluded that dietary protease supplementation tended to increase N retention during the first phase of the weaning period, hence increasing piglet performance.

Immune Enhancement of Polysaccharide from Submerged Culture with Phellinus linteus in the Medium Supplemented with Ginseng Extract (수삼추출물 첨가 혼합배지에서 조제된 상황 균사체 심부배양물 다당획분의 면역활성 증진)

  • Kim, Hoon;Song, Ki-Yun;Jeong, Jae-Hyun;Jeong, Heon-Sang;Lee, Hyeon-Yong;Yu, Kwang-Won
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.1
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    • pp.20-28
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    • 2011
  • Crude polysaccharide (CP) was fractionated from the submerged culture (containing both mycelia and culture broth, SC) with Phellinus linteus (PL) in mushroom complete medium (MCM) supplemented with ginseng extract ($65^{\circ}$Bx, GE) to enhance the immune activity. PL-GE-15-CP from SC cultivated in MCM supplemented with GE-15% (v/v, a ratio of MCM volume to GE) showed significantly higher macrophage stimulation (1.45 fold of the saline control at $100{\mu}g$/mL) than PL-GE-5 and 10-CP with GE-5 and 10%, or PL-CP from SC without GE. The potent intestinal immune system modulating activity through Peyer's patch was also obtained by PL-GE-15-CP (1.46 fold). When PL-GE-15-CP further fractionated on DEAE-Sepharose CL-6B (Cl- form), PL-GE-15-CP-II was the significantly higher than others from PL-GE-15-CP or PL-CP on macrophage stimulation, interleukin (IL)-12 production and intestinal immune system modulation (1.54, 3.96 and 1.56 fold, respectively). PL-GE-15-CP-II also had higher anti-metastatic activity against colon 26-M3.1 carcinoma cell (57.3% inhibition of tumor control, $200{\mu}g$/mouse) rather than PL-CP-II. This active fraction (PL-GE-15-CP-II) mainly contained neutral sugar (82.45%) and uronic acid (12.99%), and component sugar analysis showed that PL-GE-15-CP-II consisted mainly of uronic acid, Ara, Man, Gal and Glc (molar ratio of 0.52:0.97:0.63:1.00:0.54). Furthermore, the activity of GE culture was higher compared with culture without GE, indicating that GE helped to enhance the immune activity of P. linteus; also, it is assumed that the polysaccharide plays an important role in immune enhancement.

Non-CpG Methylation of Pre-1 Sequence in Pig SCNT Blastocysts (돼지 체세포복제 배반포에서 Pre-1 영역의 Non-CpG 메틸화 양상)

  • Ko, Yeoung-Gyu;Im, Gi-Sun;Lee, Hwi-Cheul;Cho, Sang-Rae;Choi, Sun-Ho;Choe, Chang-Yong;Lee, Poong-Yeon;Cho, Chang-Yeon;Cho, Jae-Hyeon;Yoo, Young-Hee
    • Reproductive and Developmental Biology
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    • v.35 no.1
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    • pp.93-97
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    • 2011
  • Previously, we reported that the osmolarity conditions in the satellite region were affected CpG DNA methylation status while Pre-1 sequence was not affected CpG DNA methylation in pNT blastocyst stage. This study was conducted to investigate the DNA methylation status of repeat sequences in pig nuclear transfer (pNT) embryos produced under different osmolarity culture conditions. Control group of pNT embryos was cultured in PZM-3 for six days. Other two treatment groups of pNT embryos were cultured in modified PZM-3 with 138 mM NaCl or 0.05 M sucrose (mPZM-3, 320 mOsmol) for two days, and then cultured in PZM-3 (270 mOsmol) for four days. The DNA methylation status of the Pre-1 sequences in blastocysts was characterized using a bisulfite-sequencing method. Intriguingly, in the present study, we found the unique DNA methylation at several non-CpG sequences at the Pre-1 sequences in all groups. The non-CpG methylation was hypermethylated in all three groups, including in vivo group (86.90% of PZM-3; 83.87% of NaCl; 84.82% of sucrose; 90.94% of in vivo embryos). To determine whether certain non-CpG methylated sites were preferentially methylated, we also investigated the methylation degree of CpA, CpT and CpC. Excepting in vivo group, preference of methylation was CpT>CpC>CpA in all three groups investigated. These results indicate that DNA methylation of Pre-1 sequences was hypermethylated in CpG as well as non-CpG site, regardless modification of osmolarity in a culture media.

Critical Pathway for Operable Gastric Cancer (위암수술 환자에서의 Critical Pathway의 개발과 적용)

  • Song, Kyo-Young;Kim, Seung-Nam;Park, Cho-Hyun
    • Journal of Gastric Cancer
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    • v.5 no.2
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    • pp.95-100
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    • 2005
  • Purpose: Critical pathways (CP), also known as clinical pathways, are management plans that display goals for patients and have led to improved outcomes for many disease entities. This study was aimed at developing a critical pathway for the surgical treatment of gastric cancer patients and evaluating its usefulness. Materials and Methods: A CP was developed and implemented by a team of surgeons, nurses, nutritionists, and administrative officials. Among the 117 patients who received curative gastrectomies for gastric cancer at Kangnam St. Mary's Hospital, The Catholic University of Korea, between October 2003 and August 2004, 26 patients were treated according to the CP. We evaluated its usefulness by comparing the clinical characterisctics, postoperative progress, hospital stays, and costs between the CP and the non-CP groups. Patient satisfaction was also surveyed with questionnaires. Results: Of the initial 26 patients in the CP group, two were excluded from the final evaluation; one patient had a duodenal stump leakage, and the other had a gastric stasis postoperatively. In 8 patients, protocol violation occurred; six patients refused to be discharged on the $7^{th}$ postoperative day, one patient who had an gastric staisis postoperatively stayed for 2 additional days, and one patient who needed ICU care stayed for 4 additional days. The drop-out rate was $7.7\%$ (2/26), and the variance rate was $30.8\%$ (8/26). The mean hospital stay was 11.3 days ($10\~15$ days) for the CP group compared with 17.5 days ($9\∼68$ days) for the non-CP group, resulting in a difference of about 6 days (P<0.05). The mean hospital stays after surgery were 10.3 days ($7\∼68$ days) and 8.3 days ($7\∼12$ days) for the non-CP and the CP groups, respectively, but the difference was statistically not significant (P>0.05). The mean charge during the hospital stay was higher in the non-CP group ( $\\$ 6,292,200) than in the CP group ( $\\$ 4,863,685). The charge per hospital day was higher in the CP group ( $\\$ 430,414) than in the non-CP group ( $\\$ 359,554). Patient satisfaction was higher in the CP group than in the non-CP group. Conclusion: By developing and applying a critical pathway in the surgical treatment of stomach cancer patients, we could reduce the length of hospital stay as well as the cost. A multi-centered prospective study to establish a standard treatment pathway and to demonstrate its effectiveness is needed in the future.

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A Simple Purification Method and Chemical Properties of Potphyran from Porphyra yezoensis (Porphyran의 신속한 정제 방법 및 화학적 특성)

  • Park, Jin-Hee;Koo, Jae-Geun
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.41 no.6
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    • pp.409-413
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    • 2008
  • A simple method for the purification of porphyran from laver Porphyra yezoensis was developed to obtain information for the development of food materials with biological functionality. Crude porphyran (CP) was extracted from dried laver in boiling water for 3 h, and then fractionated using cetylpyridinium chloride into an acidic fraction (CP-F1) and a neutral fraction (CP-F2). CP-F1 was fractionated further by fractional ethanol precipitation. Fraction CP-F1-70, precipitated at an ethanol concentration of 61-70% was the major fraction containing 68.1% of the yield from the initial fraction CP-F1. The CP-F1-70 fraction displayed a single band on Sepharose CL-4B with a molecular mass of 550 kDa, indicating a homogeneous polysaccharide. The molar ratio of galactose, 3,6-anhydro-L-galactose, 6-0-methyl-D-galactose and ester sulfate of CP-F1-70 was 1:0.32:0.07:0.53. This method is very useful for rapid and large-scale preparation of purified porphyran because it is compatible with mass production.