• Molecules and Cells
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• 제38권9호
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• pp.773-780
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• 2015

3D8 single chain variable fragment (scFv) is a recombinant monoclonal antibody with nuclease activity that was originally isolated from autoimmune-prone MRL mice. In a previous study, we analyzed the nuclease activity of 3D8 scFv and determined that a HeLa cell line expressing 3D8 scFv conferred resistance to herpes simplex virus type 1 (HSV-1) and pseudorabies virus (PRV). In this study, we demonstrate that 3D8 scFv could be delivered to target tissues and cells where it exerted a therapeutic effect against PRV. PRV was inoculated via intramuscular injection, and 3D8 scFv was injected intraperitoneally. The observed therapeutic effect of 3D8 scFv against PRV was also supported by results from quantitative reverse transcription polymerase chain reaction, southern hybridization, and immunohistochemical assays. Intraperitoneal injection of 5 and $10{\mu}g$ 3D8 scFv resulted in no detectable toxicity. The survival rate in C57BL/6 mice was 9% after intramuscular injection of 10 $LD_{50}$ PRV. In contrast, the 3D8 scFv-injected C57BL/6 mice showed survival rates of 57% ($5{\mu}g$) and 47% ($10{\mu}g$). The results indicate that 3D8 scFv could be utilized as an effective antiviral agent in several animal models.

• 대한수의학회지
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• 제35권2호
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• pp.245-254
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• 1995

The density of ATP-sensitive potassium($K_{APT}$) channels, that open as intracellular ATP concentration falls below a critical level, is very high in skeletal muscle surface membrane and those high density may imply that $K_{ATP}$ channels have very important physiological roles. To elucidate a role of $K_{ATP}$ in relation to fatigue, the modulating effects of potassium channel openers and blockers on the fatigue velocity(FV) of mouse extensor hallucis longus muscle(EHL) were investigated in vitro. Twitch contraction was induced by an electrical field stimulation (EFS: 24-48V, 20ms, 0.2-4Hz) and resulting contraction force was isometrically recorded. The twitch forces were gradually decreased to 25% of initial contraction force(ICF) in $37.52{\pm}1.55sec$($mean{\pm}s.e.m.$, n=135), indicating the fatigue phenomena. The mean velocity for development of the fatigue was measured during the period that twitch force decreased to half($FV_{0/0.5}$) and during the period from half to 25%($FV_{0.5/0.25}$) of ICF. The fatigue was induced once every one hour and the tissue response was stable for up to 4 hours. In control condition, ICF was $5.8{\pm}0.12g$ (n=144) and decreased to 50% of ICF with the mean fatigue velocity of $0.182{\pm}0.006g/sec$($FV_{0/0.5}$, n=135) and from 50% to 25% of ICF with $0.084{\pm}0.004g/sec$($FV_{0.5/0.25}$, n=135). Cromakalim($50{\mu}M$) significantly increased $FV_{0.5/0.25}$(n=4). Glibenclamide($IC_{50}>50{\mu}M$), $Ba^{2+}$($IC_{50}=10{\mu}M$), 4-aminopyridine($FV_{0/0.5}$, $IC_{50}=0.5mM$; $FV_{0.5/0.25}$, $IC_{50}=2mM$) decreased both $FV_{0/0.5}$ and $FV_{0.5/0.25}$ concentration-dependently up to 75%. $TEA^+$(30mM), E-4031($10{\mu}M$), tolbutamide(1mM) decreased $FV_{0.5/0.25}$, but apamin(300nM) and $TEA^+$(10mM) showed no significant effects. Our results suggest that activation of the $K_{ATP}$ channels may be major cause of $K^+$ outflux during development of the fatigue and the isolated EHL muscle could be an useful experimental preparation in studying the fatigue phenomena in skeletal muscle. In addition, the possibility of activation of delayed rectifier during the fatigue development remains to be studied further.

• 한국잠사곤충학회지
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• 제24권1호
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• pp.28-31
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• 1982

가잠의 바이러스성 연화병에 대한 저항성의 유전양식을 구명하기 위한 시험을 실시하여 다음과 같은 결과를 얻었다. 1. 유전력 검정 1) FV저항성의 주동인자는 불완전 우성에 의한 상가적효과를 보였다. 2) 미동인자의 효과는 특히 저항성이 강한 계통과 약한 계통과의 교배조에서 나타났다. 3) 유전력은 0.93으로서 거의 유전적 요인에 의한것이며 환경변이는 적다고 생각된다. 2. 원종간의 조합능력 검정 1) 일반조합능력효과는 고도의 유의차가 있었으며, 잠 108이 13.1로 가장 높았고 경추가 -17.7로 가장 낮았다. 2) 특수조합능력효과도 고도의 유의차가 있었으며, 금호$\times$무등에서 13.7로 가장 높았고 잠115$\times$경추에서 -14.8로 가장 낮았다.

• 환경생물
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• 제19권1호
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• pp.43-48
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• 2001

구상나무 침엽의 엽록소형광과 생육지의 환경요인의 일변화를 계절별로 조사하여 환경요인의 변화에 따른 구상나무의 환경 적응성을 규명하였다. 광계 II의 광화학적 효율, 즉 Fv/Fm은 겨울에는 0.19-0.36으로 매우 낮은데 반하여 여름에는 0.8-0.86으로 높았다. 낮시간의 Fv/Fm이 겨울에는 아침, 저녁보다 상대적으로 높아서 겨울철 한낮의 환경조건이 구상나무 잎의 광합성기구에 우호적으로 작용을 하는 것으로 판단된다. 여름에는 낮시간에 광억제가 다소 일어나지만 Fv/Fm이 전반적으로 높아 낮시간의 환경 조건이 구상나무에 광피해를 야기하지는 않는 것으로. 해석되었다. 비광화학적 엽록소형광 소멸 능력, 즉 NPQ는 겨울에는 0-0.01로 밤과 낮의 차이가 없이 매우 낮았고, 여름에는 낮시간에 0.76으로 높았다. 따라서 구상나무는 여름철 환경 스트레스로부터 광계 II를 보호하는 조절 기구가 잘 발달한 것으로 판단되었다. 겨울에는 Fv/Fm이 온도, 광량, 상대습도와 모두 정의 상관관계를 보였으나 NPQ는 환경요인 모두와 상관성이 없는 것으로 나타났다. 여름에는 Fv/Fm이 온도와 광량과는 음의 상관관계를 나타낸 반면에 상대습도와는 정의 상관관계를 보였다. 따라서 구상나무 생육지의 조건하에서 겨울에는 온도, 광량, 상대습도의 증가는 모두 광합성 효율을 증가시키는 반면에 여름에는 높은 온도와 광량이 광억제를 야기할 수 있는 것으로 사료된다.

• IMMUNE NETWORK
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• 제4권1호
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• pp.7-15
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• 2004

Background: The heat shock proteins (HSPs) play an important role in cellular protection mechanisms against physical or chemical stresses. In this study scFv antibodies specific for human HSP70.1 were isolated from a semi-synthetic human scFv library with the ultimate goal of developing anti-HSP70.1 intracellular antibody (intrabody) that may offer an attractive alternative to gene targeting to study the function of the protein in cells. Methods: A semi-synthetic human scFv display library ($5{\times}10^{8}$ size) was constructed using pCANTAB-5E vector and the selection of the library against bacterially expressed recombinant human HSP70.1 was attempted by panning. Results: Three positive clones specific for recombinant HSP70.1 were identified. All three clones used $V_{H}$ subgroup III. On the other hand, $V_{L}$ of two clones belonged to the kappa light chain subgroup I, but the other utilized $V_{k}$ subgroup IV Interestingly, these scFv molecules specifically reacted to the recombinant HSP70.1, yet failed to recognize native HSP70 induced in U937 human monocytic cells by heat treatment. Conclusion: Our results indicated that affinity selection of an scFv phage display library using recombinant antigens produced in E. coli might not guarantee the isolation of scFv antibody molecules specific for a native form of the antigen. Therefore, the source of target antigens needs to be chosen carefully in order to isolate biofunctional antibody molecules.

• Journal of Microbiology and Biotechnology
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• 제27권5호
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• pp.965-974
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• 2017

The single-chain variable fragment (scFv) against lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) is a promising molecule for its potential use in the diagnosis and immunotherapy of atherosclerosis. Producing this scFv in several milligram amounts could be the starting point for further engineering and application of the scFv. In this study, the abundant expression of the anti-LOX-1 scFv was attempted using Escherichia coli (E. coli) and Brevibacillus choshinensis (B. choshinensis). The scFv had limited soluble yield in E. coli, but it was efficiently secreted by B. choshinensis. The optimized fermentation was determined using the Plackett-Burman screening design and response surface methodology, under which the yield reached up to 1.5 g/l in a 5-L fermentor. Moreover, the properties of the scFvs obtained from the two expression systems were different. The antigen affinity, transition temperature, and particle diameter size were 1.01E-07 M, $55.2{\pm}0.3^{\circ}C$, and 9.388 nm for the scFv expressed by B. choshinensis, and 4.53E-07 M, $52.5{\pm}0.3^{\circ}C$, and 13.54 nm for the scFv expressed by E. coli. This study established an efficient scale-up production methodology for the anti-LOX-1 scFv, which will boost its use in LOX-1-based therapy.

• 한방재활의학과학회지
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• 제23권2호
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• pp.73-83
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• 2013

Objectives : This study was performed to investigate the effects of Foeniculum Vulgare(FV) extract on the anti-inflammatory of lipopolysaccharide(LPS) exposed rats. Methods : We divided LPS exposed Sprague-Dawley rats into 4 groups. They were control group, feed with 100 mg/kg, 200 mg/kg, 300 mg/kg FV groups. They were administered for 6 weeks. We measured the concentration of plasma interleukin-1${\beta}$(IL-1${\beta}$), plasma interleukin-6(IL-6), plasma tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), plasma interleukin-10(IL-10), the concentration of liver IL-1${\beta}$, IL-6, TNF-${\alpha}$ and IL-10. Results : 1. Plasma IL-1${\beta}$, plasma IL-6 and plasma TNF-${\alpha}$ concentration increased rapidly at 2hours after LPS injection and maintained high levels at 5hours after LPS injection. The concentration of these cytokines in the FV extract groups showed lower values than control group(P<0.05). 2. The concentration of Plasma IL-10 in FV extract groups showed higher values than control group at all times(P<0.05). 3. The concentration of liver IL-1${\beta}$ and IL-6 in FV extract groups showed lower values than control group(P<0.05). The concentration of liver TNF-${\alpha}$ in FV extract groups showed a tendency to decrease and that of liver IL-10 in FV extract groups showed a tendency to increase; however, these values showed no significantly different. Conclusions : In inflammatory response by LPS derivation, the FV gives positive effect.

• Journal of Plant Biology
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• 제38권4호
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• pp.399-407
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• 1995

보리(Hordeum vulgare L.) 유식물의 녹화시기에 따라 오존이 엽록체의 광합성 활성에 미치는 영향을 알아보기 위하여 황백화된 보리 유산물을 녹화하는 중에 오존(0.5 ppm과 1.0 ppm)을 4시간 동안 처리하여 엽록소 함량, 광계 I과 II 활성 및 엽록소 형광을 측정하였다. 녹화를 시작하므로 엽록체가 발달되는 초기에 오존이 미치는 영향은 24시간 녹화구에서 엽록소 함량을 많이 감소시켰으나 48시간 녹화한 경우에는 대조구와 유사하였다. 24시간 녹화구에서는 처리한 오존의 농도가 0.5 ppm에서 1.0 ppm으로 증가함에 따라 Fo, Fv, Fm 및 qE가 급격히 감소하였으나 Fv/Fo와 qPsms 감소하지 않았으며, 48시간 녹화구에서는 Fo, Fv, Fm 및 qE가 많이 회복되었다. 엽록체의 발달 증가에 오존이 미치는 영향은 오존의 농도가 0.5 ppm에서 1.0 ppm으로 증가함에 따라 광계 II 활성을 감소시켰으며 광계 I활성은 감소시키지 않았고, Fv와 Fv/Fm 비가 크게 감소하였으며, 형광 소멸 분석에서는 qP와 qR이 특히 많이 감소하였다.

• 한국양식학회지
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• 제10권1호
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• pp.33-37
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• 1997

미꾸라지를 대상으로 어류 유전자 이식을 위한 유용 model system으로 개발하기 위한 연구의 일환으로 잉어의$\beta$-actin 유전자의 promoter와 CAT 유전자가 융합되어 있는 외래 유전자 (pFV4CAT)를 미꾸라지에 이식, transgenic founder 미꾸라지를 생산하였다. PCR 분석결과, 이식된 외래 유전자는 부화 후 9개월된 성어에서 7.4-37.0%의 빈도로 존재하였으며 Southern blot 분석에 의해서 염색체 상의 삽입 가능성을 나타내었다. In situ immunohistochemical analysis 결과 이식된 외래 유전자는 미꾸라지 세포내에서 비교적 높은 빈도로 발현하였으며 그 발현 양상은 개체마다 다양하게 나타났다.

• Bulletin of the Korean Chemical Society
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• 제30권5호
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• pp.1101-1106
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• 2009

Multivalent and multi-specific antibodies can provide valuable tools for bio-medical research, diagnosis and therapy. In antigen-antibody interactions, the avidity of antibodies depends on the affinity and the number of binding sites.$^1$ As artificial multivalent antibody agents, single chain Fv-streptavidin fusion tetramer proteins $(scFv-SA)_4$ have been previously tested.$^{1,\;2}$ Although, the Fab domain is known to be more stable than scFv in animal models,$^{3,\;4}$ it has never been used to make a multivalent agent with a streptavidin fusion. In this study, we prepared tetra-valent $(Fab-cSA)_4$ by fusing Fab with core streptavidin (cSA). This molecule was made using inclusion body production, refolding and chromatography purification. Affinities of the Fab-cSA tetramer and a scFv-cSA tetramer to a cell surface antigen were compared by ELISA using biotin-HRP. The Fab-cSA tetramer showed higher binding avidity than the scFv-cSA tetramer. The higher binding avidity of the Fab-cSA tetramer demonstrates its potential as a therapeutic agent for target-specific antibody therapy.