• Animal cells and systems
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• 제3권2호
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• pp.173-180
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• 1999

A blue biliprotein, insecticyanin (INS), has been purified from the last instar larval hemolymph of Agrius convolvuli by ultracentrifugation, Sephadex G-100 gel permeation chromatography, and preparative electrophoresis. The molecular mass of INS was estimated to be 26 kDa and the N-terminal sequence of INS revealed high similarity to that of Manduca sexta. Results of Western blotting and autoradiography indicated that INS is synthesized by the epidermis and released into the hemolymph. In contrast to the INS reported in other insects, Agrius convolvuli INS contained a small amount of lipid, predominately consisting of triacylglycerol. Subcellular localization of INS was determined using protein-A gold particles linked to secondary antibodies (anti-rabbit Ig). INS was heavily accumulated in the cytoplasmic inclusion body (CIB). CIBs showed a variety of shapes from rod to globule and generally surrounded the nucleus. They were mostly located near the basement membrane and especially abundant in the intersegmental membrane.

• BMB Reports
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• 제45권2호
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• pp.85-90
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• 2012

Our previous study demonstrated that CopA3, a disulfide dimer of the coprisin peptide analogue (LLCIALRKK), has antibacterial activity. In this study, we assessed whether CopA3 caused cellular toxicity in various mammalian cell lines. CopA3 selectively caused a marked decrease in cell viability in Jurkat T, U937, and AML-2 cells (human leukemia cells), but was not cytotoxic to Caki or Hela cells. Fragmentation of DNA, a marker of apoptosis, was also confirmed in the leukemia cell lines, but not in the other cells. CopA3-induced apoptosis in leukemia cells was mediated by apoptosis inducing factor (AIF), indicating induction of a caspase-independent signaling pathway.

• Asian-Australasian Journal of Animal Sciences
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• 제6권4호
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• pp.515-519
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• 1993

Young goats were fed low quality roughage ad libitum and supplements of insect-damaged Lablab purpureus (var. Highworth) seed fed at approximately 3, 6 or 12 g/kg liveweight (LW), or sweet lupin seed (Lupinus angustifolius var. Uniharvest) fed at 12 g/kg LW. Roughage intake was not changed by 3 or 6 g/kg LW levels of Lablab or by 12 g/kg LW lupin supplement, but was reduced (p<0.05) by 35% by 12 g/kg LW Lablab supplement. Organic matter (OM) digestibility was increased by all supplements, and digestible OM intake was increased by the 6 g/kg LW Lablab and 12 g/kg LW lupin supplements. LW gain and feed conversion ratio were not changed by 3 or 6 g/kg LW Lablab or the 12 g/kg LW lupin, but were reduced (p<0.05) by 12 g/kg LW Lablab supplement. It was concluded that young goats could efficiently utilize supplements of Lablab purpureus seed fed at levels of up to 6 g/kg LW. However, when 12 g/kg of the Lablab seed was fed, poor performance suggested that the goats were adversely affected by anti-nutritional factors which were not neutralized by rumen fermentation.

• Molecules and Cells
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• 제43권6호
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• pp.530-538
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• 2020

The Gustatory system enables animals to detect toxic bitter chemicals, which is critical for insects to survive food induced toxicity. Cucurbitacin is widely present in plants such as cucumber and gourds that acts as an anti-herbivore chemical and an insecticide. Cucurbitacin has a harmful effect on insect larvae as well. Although various beneficial effects of cucurbitacin such as alleviating hyperglycemia have also been documented, it is not clear what kinds of molecular sensors are required to detect cucurbitacin in nature. Cucurbitacin B, a major bitter component of bitter melon, was applied to induce action potentials from sensilla of a mouth part of the fly, labellum. Here we identify that only Gr33a is required for activating bitter-sensing gustatory receptor neurons by cucurbitacin B among available 26 Grs, 23 Irs, 11 Trp mutants, and 26 Gr-RNAi lines. We further investigated the difference between control and Gr33a mutant by analyzing binary food choice assay. We also measured toxic effect of Cucurbitacin B over 0.01 mM range. Our findings uncover the molecular sensor of cucurbitacin B in Drosophila melanogaster. We propose that the discarded shell of Cucurbitaceae can be developed to make a new insecticide.

• Animal cells and systems
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• 제16권6호
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• pp.455-461
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• 2012

Amongst the various antimicrobial peptides, lysozyme plays a central role in initiating and maintaining the antibacterial defense response of insect. Here we propose the biosynthesis and refolding of recombinant lysozyme in Escherichia coli expressed in inclusion body form. The Agrius lysozyme gene was amplified using gene specific primers and then ligated into the pGEX-4T-1 vector, which contained the glutathione S-transferase (GST) gene as a fusion partner. A recombinant lysozyme was expressed in E. coli Rosetta cells using a pGEX-4T-1 expression vector, and the fusion protein was induced by ioporpyl-${\beta}$-D-thiogalactopyranoside (IPTG). The recombinant protein produced as an inclusion body was resolubilized in solubilization buffer, and the resultant solution was dialyzed in refolding buffer. After thrombin cleavage, the recombinant lysozyme was purified by ion exchange chromatography and reverse phase chromatography. The recombinant lysozyme was subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis and immunoreactivity against the anti-Agrius lysozyme was observed by western blot analysis of this protein. The recombinant lysozyme displayed antibacterial activity against Bacillus megaterium and Micrococcus luteus, which was confirmed by the inhibition zone assay.

• 생물환경조절학회지
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• 제23권4호
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• pp.293-302
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• 2014

본 연구에서는 배추 재배에 적합한 규격과 생육 환경을 조성해 줄 수 있는 비가림하우스를 개발하고자 하였다. 전국 53개 배추 비가림재배 농가를 대상으로 비가림하우스 구조실태 및 구조개선 희망사항을 조사하여 비가림하우스 폭과 높이를 설정하였다. 비가림하우스 규격은 농기계 작업의 용이성, 농가의 의견 등을 고려하여 폭 6m, 처마높이 1.6m, 지붕높이 3.2m로 결정하였다. 서까래 규격별 구조안전성과 설치비를 분석한 후 설치비가 가장 적게 드는 Ø$25.4{\times}1.5t$ 파이프를 서까래로 하고 그 간격이 90cm인 모델을 기본 규격으로 결정하였다. 이 규격은 풍속 $27m{\cdot}s^{-1}$, 적설 17cm에 안전하기 때문에 이보다 기상하중이 큰 지역에는 적용하기가 곤란한데, 이를 해소하기 위해 피복재를 용마루까지 열어 골조 피해를 예방할 수 있는 구조로 설계하였다. 비가림하우스 양 측면에 있는 수동개폐기를 돌려 하우스밴드를 느슨하게 풀어주고 제어반에서 열림버튼을 누르면 개폐모터가 가이드 파이프를 따라 올라가면서 피복재가 용마루까지 개방된다. 피복재를 완전 개방할 경우 해충으로 인한 피해가 우려되므로 농가에서는 이를 막기 위해 방충망을 설치할 수 있다. 배추를 재배하는 기간에 태풍이 지나갈 수 있기 때문에 방충망이 구조안전에 미치는 영향을 분석하였다. $40m{\cdot}s^{-1}$의 바람이 방충망으로 덮여있는 비가림하우스 측면에 수직으로 작용하는 조건에 대해 유동-구조 연성해석 기법을 이용하여 구조안전성을 분석하였다. 유동해석 결과, 피복재 부분은 바람의 영향을 그대로 받기 때문에 피복재 표면에 압력이 크게 작용하였다. 방충망 부분에도 풍하중이 작용하였으나 피복재 부분보다는 압력이 작게 작용하고 분포가 균일하였다. 유동해석에서 도출된 압력 데이터를 적용하여 구조해석한 결과, 최대응력은 파이프의 끝단 즉, 지면부분에서 나타났으며, 그 값은 54.6Mpa이었다. 구조안전 판단 기준인 파이프의 허용응력 215MPa 이내여서 구조적으로 안전한 것으로 판단되었다.

• Parasites, Hosts and Diseases
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• 제56권5호
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• pp.437-446
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• 2018

To investigate the prevalence of Toxoplasma gondii in pork on the market in Korea, an in-house enzyme-linked immunosorbent assay for tissue fluid (CAU-tf-ELISA) was developed using a soluble extract of T. gondii RH strain tachyzoites. As the standard positive controls, the piglets were experimentally infected with T. gondii: Group A (1,000 cysts-containing bradyzoites), Group B (500 cysts-containing bradyzoites) and Group C ($1.0{\times}10^3$ or $1.0{\times}10^4$ tachyzoites). The CAU-tf-ELISA demonstrated infection intensity-dependent positivity toward tissue fluids with average cut-off value 0.15: 100% for Group A, 93.8% for Group B and 40.6% for Group C. When tissue-specific cut-off values 0.066-0.199 were applied, CAU-tf-ELISA showed 96.7% sensitivity, 100% specificity, 100% positive and 90.0% negative predictive values. When compared with the same tissue fluids, performance of CAU-tf-ELISA was better than that of a commercial ELISA kit. Of the 583 Korea domestic pork samples tested, anti-T. gondii antibodies were detected from 9.1% of whole samples and 37.9% from skirt meat highest among pork parts. In the 386 imported frozen pork samples, 1.8% (skirt meat and shoulder blade) were positive for anti-T. gondii antibodies. In Korea, prevalence of anti-T. gondii antibodies in the pork on retail markets appeared high, suggesting that regulations on pig farming and facilities are necessary to supply safe pork on the tables.

• 생명과학회지
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• 제25권5호
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• pp.607-614
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• 2015

암은 전 세계 사망률과 질병률의 가장 큰 원인이다. Cordycepin (3'-deoxyadenosine)은 동양의 전통의학에서 널리 사용되고 있는 동충하초의 주요 기능성 구성요소이자 아데노신 유사체로 알려져 있다. 지난 10년간 cordycepin은 in vitro 및 in vivo 모델에서 면역활성 기능뿐 만 아니라 항염증, 항산화 및 항암 등 다양한 약리학적 특성을 가진다고 보고되어왔다. 최근 들어 많은 연구들은 cordycepin을 화학예방요법 작용제 측면에서 흥미로운 특성을 보고하였고, 실험적인 증거들에 의해 세포사멸 촉진, 세포주기 정지 유도, 세포 내 신호 전달 경로 조절, 암세포의 침윤 및 전이 억제를 통해 암의 증식을 지연시킨다고 보고되어 왔다. Cordycpin은 많은 암 세포에서 retinoblastoma protein (RB)의 인산화를 막고 cyclin-dependent kinases (Cdks) inhibitors를 활성화시켜 G2/M기의 진행을 막는 효력이 있음이 밝혀졌다. 또한, 세포 사멸을 유도하기 위해 세포 내/외부에 존재하는 경로를 활성화시켜 활성 산소종을 생성하고 하위에 존재하는 kinase cascade 반응을 개시한다. 아울러 cordycepin은 또 다른 세포 사멸인 autophagy와 같은 대체 경로를 활성화 시킬 수도 있으며, nuclear factor-kappa B 및 activated protein-1 신호 경로를 포함한 다양한 기전을 통하여 암세포 분리, 이주, 침윤 및 전이 또한 억제 할 수 있다. 본 총설에서는 cordycepin의 항암 작용 기전을 요약하고, 다양한 암 발생의 치료제로서 가능성을 논의하고자 한다.

• 한국식품과학회지
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• 제51권6호
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• pp.537-542
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• 2019

국내에서 식용으로 유통 및 판매가 가능한 7종의 곤충 중 누에(Bombyx mori), 갈색거저리(Tenebrio molitor) 유충, 흰점박이꽃무지(Protaetia brevitarsis) 유충, 쌍별귀뚜라미(Gryllus bimaculatus)의 4종을 대상으로 단백질을 분리하고, 유청 단백질과 비교하여 항산화 및 영양 특성을 분석하였다. 원물시료에서 분리한 단백질의 수율은 65.0-75.0%로 나타났다. 각 단백질의 아미노산을 분석한 결과, 모든 곤충 시료에서 필수 아미노산인 Phe, Trp은 유청 단백질보다 높은 함량을 보였다. 근육생성에 도움이 되는 세 종류의 BCAA아미노산 중 Val은 모든 곤충단백질에서 유청단백과 유사한 함량을 보였으며, BCAA함량은 곤충분리단백질 중 쌍별 귀뚜라미와 갈색거저리 유충에서 그 함량이 각각 16.8과 16.4%로 가장 높아 유청단백(22.2-23%)의 73-76% 수준인 것으로 나타났다. 곤충 단백질의 총 폴리페놀함량은 유청단백보다 유의적으로 많았으며 곤충단백중에는 누에의 함량이 유의적으로 낮게 측정되었다. 항산화활성 또한 곤충단백이 유청단백보다 유의적으로 높았으며, 특히 갈색거저리 유충에서의 활성이 가장 높게 측정되었다. 관능검사결과에서는 시판 단백질파우더와 동일한 조성으로 제조했을 경우, 유청단백의 25%의 대체가 가능할 것으로 보이나 맛과 향이 대조군에 비해 많이 떨어지므로 보완할 필요가 있다. 곤충단백질은 유청단백질에 비해 BCAA함량은 다소 낮았지만 폴리페놀 함량이 높고 항산화활성이 있으므로 아미노산 조성을 참고하여 특정목적 단백질 제품으로 사용될 가능성이 높음이 확인되었다.

• BMB Reports
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• 제40권5호
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• pp.783-790
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• 2007

Receptor binding plays an important role in determining host specificity of the Bacillus thuringiensis Cry $\delta$-endotoxins. Mutations in domains II and III have suggested the participation of certain residues in receptor recognition and insect specificity. In the present study, we expressed the cloned domain II-III fragment of Cry4Ba and examined its binding characteristics to mosquito-larval midgut proteins. The 43-kDa Cry4Ba-domain II-III protein over-expressed in Escherichia coli as inclusion bodies was only soluble when carbonate buffer, pH 10.0 was supplemented with 4M urea. After renaturation via stepwise dialysis and subsequent purification, the refolded domain II-III protein, which specifically reacts with anti Cry4Ba-domain III monoclonal antibody, predominantly exists as a $\beta$-sheet structure determined by circular dichroism spectroscopy. In vitro binding analysis to both histological midgut tissue sections and brush border membrane proteins prepared from susceptible Aedes aegypti mosquito-larvae revealed that the isolated Cry4Ba-domain II-III protein showed binding functionality comparable to the 65-kDa full-length active toxin. Altogether, the data present the 43-kDa Cry4Ba fragment comprising domains II and III that was produced in isolation was able to retain its receptor-binding characteristics to the target larval midgut proteins.