• 제목/요약/키워드: binding treatment

검색결과 1,247건 처리시간 0.025초

가압가열 처리에 의한 시판 돈육 햄과 베이컨의 알레르겐성 저감화 효과 (Reduction of Allergenicity of Domestic Pork Ham and Bacon by Autoclave Treatment)

  • 김서진;김꽃봉우리;송유진;이소영;윤소영;이소정;이청조;김규언;안동현
    • 한국축산식품학회지
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    • 제30권1호
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    • pp.133-140
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    • 2010
  • 대표적인 돈육 가공품인 햄과 베이컨에 가압가열 처리를 한 후 알레르겐성 변화를 살펴봄으로써 가압가열 처리법을 응용한 hypoallergenic pork의 개발 가능성을 알아보았다. 구입한 돈육 햄과 베이컨 중 ci-ELISA를 실시하여 가장 결합력이 높은 2개의 제품에 대해 가압가열 처리를 하였다. 먼저, 돈육 햄의 알레르겐성에 대한 가압가열 처리 영향을 살펴본 결과, 돈육 햄은 30분 가압가열 처리구의 경우 p-IgG와의 결합력이 조금 감소하였으며 환자혈청을 사용하였을 때는 10분과 30분 처리구 모두 무처리구보다 결합력이 다소 낮게 나타났다. 베이컨은 무처리구에서 p-IgG와의 결합력이 각각 60% 및 91%로 높았지만, 가압가열처리에 의해 약 16% 및 11% 이하로 크게 감소하였다. 환자혈청을 사용한 경우에도 베이컨 sample b와 c의 무처리구는 95% 및 126%의 높은 결합력을 나타내었지만, 가압가열 처리에 의하여 각각 22% 및 34% 이하로 크게 감소하였다. Immunoblotting 결과에서도 베이컨 두 제품 모두가압가열 처리구의 PSA band가 p-IgG와 반응하지 않았다. 특히, sample c는 무처리구의 PSA band가 2명의 환자혈청과 약하게 반응하였지만, 가압가열 5분 처리구에서는 PSA band가 나타나지 않아 베이컨의 알레르겐성이 가압가열 처리에 의해 효과적으로 감소되었음을 확인하였다. 이상의 결과를 종합해 볼 때 돈육 가공품의 종류와 처리조건에 따라 알레르겐성 감소 정도는 다르게 나타났다. 가압가열처리에 의해 돈육 햄의 알레르겐성은 큰 변화를 나타내지 않았지만, 조금 감소하는 경향을 보였으며 베이컨은 알레르겐성이 크게 감소하였음을 알 수 있었다. 따라서 본 연구는 돼지고기 민감성 환자들을 위한 돈육 가공품 개발에 가압가열 처리법이 적용 가능하다는 것을 시사하였다.

I-123 IPT SPECT를 이용한 주의력결핍 과잉행동장애 아동에서의 methylphenidate 투여 전후의 기저 신경절 도파민 운반체 밀도 변화 측정 (Dopamine Transporter Density of the Basal Ganglia in Children with Attention Deficit Hyperactivity Disorder Assessed with I-123 IPT PECT)

  • 류원기;김태훈;유영훈;윤미진;천근아;지대윤;김종호;최태현;이종두
    • 대한핵의학회지
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    • 제37권4호
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    • pp.235-244
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    • 2003
  • 목적: 주의력결핍 과잉행동장애 (Attention Deficit Hyperactivity disorder : 이하 ADHD)는 도파민계의 이상 기능으로 생기는 대표적 소아정신과 질환이다. Methylphenidate는 dopamine transporter(DAT)를 차단함으로써 ADHD 증상을 호전시키는 약물로 널리 알려져 있다. 따라서 본 연구에서는 ADHD 아동들을 대상으로 I-123 IPT SPECT를 이용하여 methylphenidate 투여 전후의 DAT density 양상을 비교해 보고자 한다. 대상 및 방법: 연구대상은 9명의 ADHD 아동과 7명의 정상 대조군이었다. ADHD 아동군과 정상 대조군에게 모두 약물 비노출 상태에서 $[^{123}I]IPT$를 정맥 주사후 2시간이 경과한 상태에서 SPECT를 촬영하였다. 이후 ADHD 아동군을 대상으로 methylphenidate 0.7mg/kg/d을 투여한 후 약 8주가 경과한 상태에서 $[^{123}I]IPT$ SPECT를 재촬영 하였다. Methylphenidate 투여 전과 후의 좌, 우측 기저 신경절 DAT 특이결합/비특이결합 비율을 구한 뒤 비교하였고, 약물 투여 후의 ADHD 증상 척도로 측정한 증상 호전도와 좌, 우측 기저 신경절 DAT 특이결합/비특이결합 비율 변화사이의 상관관계를 알아보았다. 결과: ADHD 아동군에서 약물 비노출 상태와 정상 대조군 사이의 좌, 우측 기저 신경절 DAT특이결합/비특이결합 비율을 비교한 결과 DAT 결합 비율이 정상 대조군에 비해서 유의하게 증가되었다. (Right : z=2.057, p=0.041 ; Left z=2.096, p=0.032). 또한 ADHD 아동들에게 methylphenidate 투여 전과 후 상태에서의 기저 신경절 DAT 특이결합/비특이결합 비율을 비교한 결과, methylphenidate 투여 후 상태가 methylphenidate 투여 전 상태에 비해 좌, 우측 DAT 결합비율이 유의하게 낮아진 것을 관찰할 수 있었다. (우측 : t=3.239, p=0.018 ; 좌측 : t=3.133, p=0.020). ADHD 증상의 호전도와 좌, 우측 기저 신경절 DAT 특이결합/비특이결합 비율 변하사이에는 유의한 상관관계를 보이지 않았다. 결론: 이러한 결과는 주의력결핍 과잉행동장애의 치료제인 methylphenidate가 작용하는 기전을 이용하여 주의력결핍 과잉행동장애의 병태생리와 연관된 도파민계 기능이상 가능성을 지지한다고 생각된다.

Effect of Quercetin in the UV-Irradiated Human Keratinocyte HaCaT Cells and A Model of Its Binding To p38 MAPK

  • Jnawali, Hum Nath;Lee, Eunjung;Shin, Areum;Park, Young Guen;Kim, Yangmee
    • Bulletin of the Korean Chemical Society
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    • 제35권9호
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    • pp.2787-2790
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    • 2014
  • Quercetin is a major dietary flavonoid found in onions, apples, tea, and red wine, and potentially has beneficial effects on disease prevention. We carried out this study to investigate the effect of quercetin on UVB-induced matrix metalloproteinase-1 (MMP-1) expression in human keratinocyte HaCaT cells and to further understand the mechanisms of its action. The anti-inflammatory activity of quercetin was investigated and quercetin significantly suppressed the NO production in LPS-stimulated RAW264.7 mouse macrophages. Post treatment of quercetin decreased UV irradiation-induced phosphorylation of JNK, p38 MAPK, and ERK by 91%, 21%, and 17%, respectively. MMP-1 is mainly responsible for the degradation of dermal collagen during the aging process of human skin and quercetin suppressed the UVB-induced MMP-1 by 94%. Binding studies revealed that quercetin binds to p38 with high binding affinity ($1.85{\times}10^6M^{-1}$). The binding model showed that the 4'-hydroxy groups of the B-ring of quercetin participated in hydrogen bonding interactions with the side chains of Lys53, Glu71, and Asp168 and the 5-hydroxy group of the A-ring formed a hydrogen bond with the backbone amide of Met109. The major finding of this study shows that quercetin inhibits phosphorylation of JNK, p38 MAPK, and ERK pathway leading to the prevention of MMP-1 expression in human keratinocyte HaCaT cells. Therefore, our findings suggested the potentials of quercetin as a skin anti-photoaging agent.

냉동절편을 이용한 돼지 자궁내 에스트로겐 수용체의 측정 (A Method for the Determination of Estrogen Receptor Level in Frozen Sections of Porcine Uterus)

  • 윤용달;박철홍;이영근
    • Clinical and Experimental Reproductive Medicine
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    • 제16권2호
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    • pp.131-138
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    • 1989
  • The present study was designed to develop a new method for the determination of estrogen receptor in porcine uterus using frozen sections. Cryostat sections were incubated with $^3H$-estradiol($^3H$-$E_2$) in the presence or absence of diethylstilbestrol(DES) and the radioactivity of 3H-E2 bound to estrogen receptor(ER) was detected. The level of specific estrogen receptor was determined by Scatchard analysis. The highest ratio of specific binding against total binding was achieved in 3 sec. tions(5mm x 5mm) which was corresponded to lOO${\mu}$/ml protein concentration. Optimal binding was obtained during incubation with $^3H$-$E_2$ for 30 minutes at 23$^{\circ}C$ after treatment of sections with acetone for 20 seconds. Three time-washing of sections was proved to be appropriate for the removal of unbound 3H-E2. 200-fold molar excess of DES was substituted for the binding of $^3H$-$E_2$ to ER sufficiently(binding efficiency of 54.8%). ER was saturated with 4nM of $^3H$-$E_2$ and its dissociation constant was 0.1nM. ER assay using frozen sections(Histological radioreceptor assay, HRRA) was significantly correlated with radioreceptor assay for estradiol(RRA, 0.976 , p

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Phosphate Binding Protein이 세포질에 발현된 재조합 박테리아를 이용한 인 제거 (Phosphate Removal Using Recombinant Bacteria with Cytoplasmic Phosphate Binding Protein)

  • 최석순;하정협;차형준
    • 공업화학
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    • 제24권5호
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    • pp.558-561
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    • 2013
  • 본 연구에서는 인 결합 단백질(phosphate-binding protein)이 세포질(cytoplasm)에서 발현되는 재조합 대장균(Escherichia coli)을 제조하였으며, 이를 이용하여 수중에 함유된 인의 제거에 대하여 고찰하였다. 이 균주를 6 h 동안 배양하여 1.0, 1.5, 2.0 mg/L의 인을 처리하였을 때, 각각 90, 49, 41%의 제거효율을 도출하였으며, 우수한 인 제거 능력(specific phosphate removal)을 나타내었다. 또한, 세포의 Optical density를 2.5, 5.0으로 조절하여 운전하였을 때, 인의 제거효율이 향상됨을 알 수 있었으며, 2.0 mg/L 인을 80%까지 제거시킬 수 있었다. 이 연구를 통하여 얻어진 새로운 생물기술은 수계의 부영양화 문제를 해결하는데 효과적으로 사용될 수 있을 것이다.

Protein Kinase C Receptor Binding Assay for the Detection of Chemopreventive Agents from Natural Products

  • An, Geon-Ho;Suh, Young-Bae;Son, Kun-Ho;Chang, Il-Moo;Mar, Woong-Chon
    • Natural Product Sciences
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    • 제3권1호
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    • pp.29-37
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    • 1997
  • Protein Kinase C (PKC) is generally believed to play a central role in signal transduction, cellular growth control, gene expression, and tumor promotion. And it has been suggested that inhibitors of PKC might play important roles for the prevention and treatment of cancer. In order to investigate the possible inhibitors of PKC from natural products, PKC receptor binding assay was performed using bovine brain particulate as a source of PKC and the amount of $[^3H]Phorbol$ 12,13-dibutyrate (PDBu) bound to PKC was measured in the presence of test materials. Total methanol extracts from 100 kinds of natural products were partitioned into 3 fractions (n-hexane, ethyl acetate and aqueous layer) and their binding ability to the regulatory domain of PKC was evaluated. The ethyl acetate fractions of Morus alba $(roots,\;IC_{50}:\;156.6\;{\mu}g/ml)$, Rehmannia glutinosa $(roots,\;IC_{50}:\;134.3\;{\mu}g/ml)$, Lysimachia foenum-graecum $(roots,\;IC_{50}:\;167.8\;{\mu}g/ml)$, Polygonum cuspidata $(roots,\;IC_{50}:\;157.3\;{\mu}g/ml)$, Cnidium officinale $(aerial\;parts,\;IC_{50}:\;145.2\;{\mu}g/ml)$, and the hexane $(IC_{50}:\;179.3\;{\mu}g/ml)$ and the EtOAc fraction of Symplocarpus nipponicus $(roots,\;IC_{50}:\;155.9\;{\mu}g/ml)$ showed inhibitory activity of $[^3H]PDBu$ binding to PKC.

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Sedative-Hypnotic and Receptor Binding Studies of Fermented Marine Organisms

  • Joung, Hye-Young;Kang, Young Mi;Lee, Bae-Jin;Chung, Sun Yong;Kim, Kyung-Soo;Shim, Insop
    • Biomolecules & Therapeutics
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    • 제23권5호
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    • pp.479-485
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    • 2015
  • This study was performed to investigate the sedative-hypnotic activity of ${\gamma}$-aminobutyric acid (GABA)-enriched fermented marine organisms (FMO), including sea tangle (FST) and oyster (FO) by Lactobacillus brevis BJ20 (L. brevis BJ20). FST and FO were tested for their binding activity of the $GABA_A$-benzodiazepine and 5-$HT_{2C}$ receptors, which are well-known molecular targets for sleep aids. We also measured the sleep latency and sleep duration during pentobarbital-induced sleep in mice after oral administration of FST and FO. In $GABA_A$ and 5-$HT_{2C}$ receptor binding assays, FST displayed an effective concentration-dependent binding affinity to $GABA_A$ receptor, similar to the binding affinity to 5-$HT_{2C}$ receptor. FO exhibited higher affinity to 5-$HT_{2C}$ receptor, compared with the $GABA_A$ receptor. The oral administration of FST and FO produced a dose-dependent decrease in sleep latency and increase in sleep duration in pentobarbital-induced hypnosis. The data demonstrate that FST and FO possess sedativehypnotic activity possibly by modulating $GABA_A$ and 5-$HT_{2C}$ receptors. We propose that FST and FO might be effective agents for treatment of insomnia.

사람 NC-37 세포에서 Benzo(a)pyrene과 Doxorubicin에 의한 Nuclei내전과 c-myc 유전자의 발현에 대한 연구 (Studies on the Nuclei Adduction and Expression of c-myc Gene by Benzo(a)pyrene and Doxorubicin in Human NC-37 Cells)

  • 김호찬;정인철;조무연
    • 생명과학회지
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    • 제8권4호
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    • pp.400-409
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    • 1998
  • Formation of adduct was studied in benzo(a)pyrene(BP)- and doxorubicin(Dx)-treated human NC-37 cells and isolated nuclei. Major adducts formed were determined by fluorescence absorption spectrophotometery and DNA-lin-ked protein assay. When isolated nuclei were exposed to carcinogens BP and DMBA, and anticancer drugs m-AMSA, ellipticine and Dx, varying degrees of adduct formation occured between DNA-protein complex and these drugs. When the mixture was centrifuged 1.7 M sucrose solution, binding BP and DMBA appeared to be similar between the sediment and the supernatant. When the sediment was centrifuged again with 0.35% polymin-P, the amount of BP bound was 2-fold greater in the protein(1077$\pm$55cpm) than in DNA fraction (470$\pm$20cpm), whereas that of DMBA was 1.6-fold greater in the DNA than in protein fraction. In the case of m-AMSA, ellipticine and Dx, the amount of binding was slightly greater in supernatant than in sediment in centrifugation with 1.7 M sucrose, and more than 3 times greater in the DNA- than in protein- fraction in centrifugation with 0.35% polymin P. DNA fractions which associated with a subset of nonhistone chromosomal protein were isolated from NC-37 cells exposed to $^{3}$H-BP and $^{14}$C-Dx. They were separated into two distince components DNA-S and DNA-P by centrifugation with 2M Nacl chromatin extraction. The results indicated that the amount of $^{3}$H-BP bound was 6.0-fold greater in DNA-P as compared with DNA-S, while that of $^{14}$C-Dx binding appreaed to be 6.2-fold greater in DNA-S than in DNA-P fraction. When $^{3}$H-BP binding wasdetermined in the presence of cold Dx, the amount of binding was reduced only in the DNA-P fraction, indicating that the interaction between DNA and protein is decreased. Gene expression by these drugs, BP treated cells were increased to compare with nomal cells but reduced by treatment with BP-Dx. These results suggest that the protein moiety which tightly bound to DNA-P fraction may play an important role in the regulation of gene expression.

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유전자변형 콩과 자연 콩의 알레르기 유발원 비교 (Comparison of Allergens in Genetically Modified Soybean with Conventional Soybean)

  • 박재현;정승태;김재희;김지영;노건웅
    • 약학회지
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    • 제45권3호
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    • pp.293-301
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    • 2001
  • Genetically modified organism (GMO) using recombinant DNA technique has been exponentially increased, however there are still arguments for the safety of GM foods. The objective of this research was to compare the allergens of GM soybean(Roundup Ready$^{TM}$) with conventional soybeans. Each soybean extracts were prepared as crude extracts, heated extracts, and as heated and simulated gastric quid (SGF)-digested samples to characterize the stability of allergens to physicochemical treatment. Positive sera from 20 soybean-sensitive patients and control sera from 5 normal subjects were used to identify the endogenous allergens in soybeans. Specific-IgE binding activities to each soybean preparations were evaluated by ELISA and immunoblot technique. In ELISA result, IgE binding activities of positive sera to soy crude extracts generally showed two fold higher mean value than those of control sera, how-ever there was no significant difference between GM soybean and natural soybean varieties. Extracted proteins form each of the soybean preparations were separated with SDS-PAGE. The band pattern of GM soybean was very similar to those of natural soybean varieties. Immunoblots for the different soybeans revealed no differences in IgE-binding protein patterns, moreover, disclosed five prominent IgE-binding bands (75, 70, 50, 44 and 34 kDa) in crude extracts, four (75, 70, 44 and 34 kDa) in heated preparations, one (50 kDa) in heated and SGF-digested preparations. These IgE binding bands were consistent with previously reported results on the soybean. These results indicate that GM soybean (Roundup Ready$^{TM}$) is no different from natural soybean in terms of its allergen.gen.

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CND41, a DNA-binding protein in chloroplast nucleoid, and its function

  • Sato, Fumihiko;Murakami, Shinya;Chatani, Hiroshi;Nakano, Takeshi
    • 한국식물학회:학술대회논문집
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    • 한국식물학회 1999년도 제13회 식물생명공학심포지움 New Approaches to Understand Gene Function in Plants and Application to Plant Biotechnology
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    • pp.51-56
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    • 1999
  • Plastids, which are organelles unique to plant cells, bear their own genome that is organized into DNA-protein complexes (nucleoids). Regulation of gene expression in the plastid has been extensively investigated because this organelle plays an important role in photosynthesis. Few attempts, however, have been made to characterize the regulation of plastid gene expression at the chromosomal structure, using plastid nucleoids. In this report, we summarize the recent progress in the characterization of DNA-binding proteins in plastids, with special emphasis on CND41, a DNA binding protein, which we recently identified in the choloroplast nucleoids from photomixotrophically cultured tobacco cells. CND41 is a protein of 502 amino acids which consisted of a transit peptide of 120 amino acids and a mature protein of 382 amino acids. The N-terminal of the 'mature' protein has lysine-rich region which is essential for DNA-binding. CNA41 also showed significant identities to some aspartyl proteases. Protease activity of purified CND41 has been recently confirmed and characterized. On the other hand, characterization of accumulation of CND41 both in wild type and transgenic tobacco with reduced amount of CND41 suggests that CND41 is a negative regulator in chloroplast gene expression. Further investigation indicated that gene expression of CND41 is cell-specifically and developmentally regulated as well as sugar-induced expression. The reduction of CND41 expression in transgenic tobacco also brought the stunted plant growth due to the reduced cell length in stem. GA3 treatment on apical meristem reversed the dwarf phenotype in the transformants. Effects of CND41 expression on GA biosynthesis will be discussed.

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