• Molecules and Cells
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• 제26권2호
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• pp.146-151
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• 2008

The brown planthopper (BPH) is a major insect pest in rice, and damages these plants by sucking phloem-sap and transmitting viral diseases. Many BPH resistance genes have been identified in indica varieties and wild rice accessions, but none has yet been cloned. In the present study we report fine mapping of the region containing the Bph1 locus, which enabled us to perform marker-aided selection (MAS). We used 273 F8 recombinant inbred lines (RILs) derived from a cross between Cheongcheongbyeo, an indica type variety harboring Bph1 from Mudgo, and Hwayeongbyeo, a BPH susceptible japonica variety. By random amplification of polymorphic DNA (RAPD) analysis using 656 random 10-mer primers, three RAPD markers (OPH09, OPA10 and OPA15) linked to Bph1 were identified and converted to SCAR (sequence characterized amplified region) markers. These markers were found to be contained in two BAC clones derived from chromosome 12: OPH09 on OSJNBa0011B18, and both OPA10 and OPA15 on OSJNBa0040E10. By sequence analysis of ten additional BAC clones evenly distributed between OSJNBa0011B18 and OSJNBa0040E10, we developed 15 STS markers. Of these, pBPH4 and pBPH14 flanked Bph1 at distances of 0.2 cM and 0.8 cM, respectively. The STS markers pBPH9, pBPH19, pBPH20, and pBPH21 co-segregated with Bph1. These markers were shown to be very useful for marker-assisted selection (MAS) in breeding populations of 32 F6 RILs from a cross between Andabyeo and IR71190, and 32 F5 RILs from a cross between Andabyeo and Suwon452.

• BMB Reports
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• 제46권5호
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• pp.282-287
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• 2013

Cecropin A and papiliocin are novel 37-residue cecropin-like antimicrobial peptides isolated from insect. We have confirmed that papiliocin possess high bacterial cell selectivity and has an ${\alpha}$-helical structure from $Lys^3$ to $Lys^{21}$ and from $Ala^{25}$ to $Val^{35}$, linked by a hinge region. In this study, we demonstrated that both peptides showed high antimicrobial activities against multi-drug resistant Gram negative bacteria as well as fungi. Interactions between these cecropin-like peptides and phospholipid membrane were studied using CD, dye leakage experiments, and NMR experiments, showing that both peptides have strong permeabilizing activities against bacterial cell membranes and fungal membranes as well as $Trp^2$ and $Phe^5$ at the N-terminal helix play an important role in attracting cecropin-like peptides to the negatively charged bacterial cell membrane. Cecropin-like peptides can be potent peptide antibiotics against multi-drug resistant Gram negative bacteria and fungi.

• Journal of the Korean Wood Science and Technology
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• 제40권4호
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• pp.229-236
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• 2012

본 연구는 감마선 조사가 소나무(Pinus densiflrora), 느티나무(Zelkovas serrenata), 오동나무(Paulownia tomentosa)의 세포벽 열화, 셀룰로오스 결정화도, 휨 강도에 미치는 영향을 구명하기 위해 수행되었다. 각 목재 시편들을 100 kGy 선량의 감마선 조사 후 주사전자현미경(scanning electron microscope; SEM) 관찰 결과, 목재 세포벽 열화는 관찰되지 않았으며 휨 강도 역시 현저한 변화가 나타나지 않았다. 감마선 조사에 의한 셀룰로오스 결정화도 역시 유의한 변화가 관찰되지 않았다. 이러한 감마선에 대한 목재의 높은 안전성은 감마선 조사 기술을 재질손상 없이 목재를 가해 중인 균류와 충류 구제를 위해 적용할 수 있음을 의미한다.

• 한국동물학회지
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• 제11권4호
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• pp.103-117
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• 1968

1. 變態時期에 따른 배추흰나비(Pieris rapae)의 血球를 組織化學的方法 및 autoradiographic method 에 의거하여 硏究하였다. 2.幼蟲, 전용 및 용기에서 形態의變化와 細胞內容物을 土臺로 血球細胞를 prohemocyte, plasmatocyte, podocyte, granular hemocyte, spherule cell 및 oenocytoid 의 6型으로 分類했다. 3, 幼蟲基에서 용기까지의 血球를 組織化學的으로 硏究하고 血球細胞內의 蛋白質, 核酸, 多糖類 및 脂質을 檢出하였으며, 成長 및 變態에 따른 그들 物質의 消長을 밝혔다, 4. Granular hemocyte 와 spherule cell 의 大部分의 顆粒은 glycogen, neutral mucopolysaccharide, mucoprotein 으로 되었고 neutral fat와 phospholipid 의 顆粒도 나타났다, 5. 細胞分裂像(mitotic figure) 과 tritium-thymidine 攝取세포 (DNA 合成細胞)를 幼蟲 2齡부터 5齡까지의 各種 未分化血球에서 觀察하였고, 모든型의 血球가 prohemocyte에 起源함을 밝혔다. 6. Plasmatocyte 와 podocyte 의 細胞質突起가 전용기 와 용기에 매우 길게 伸張함을 보아 vermiform cell 은 plasmatocyte 의 더 分化한 型이라고 생각된다.

• Korean Chemical Engineering Research
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• 제44권1호
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• pp.40-45
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• 2006

원하는 유전자를 배큘로바이러스의 감염에 의하여 초파리 Drosophila melanogaster S2 세포에 도입하는 배큘로바이러스/S2 세포 발현 시스템은 강력하고 안전한 배큘로바이러스의 장점과 세포가 파괴되지 않는 S2 세포의 장점을 접목한 시스템이다. 본 연구에서는 외래 목적 단백질로 발현 모니터링이 손쉬운 녹색형광단백질(green fluorescent protein)을 발현시키는 재조합 배큘로바이러스를 이용하여 S2 세포에의 감염조건들에 대한 영향 연구를 수행하였다. 재조합 배큘로바이러스의 S2 세포에의 감염조건으로는 multiplicity of infection(MOI), 초기 세포 수, 배큘로바이러스 용액이 세포를 적시는 최소 부피, 배큘로바이러스를 첨가한 후 제거하기까지의 배양시간 그리고 배큘로바이러스 제거 후 S2 세포를 혈청이 존재하는 배지에서 키우는 배양시간을 선정하였다. MOI는 일반적으로 크게 하는 것이 높은 발현 수율을 위하여 좋은 결과를 보였으나 세포배양이 길어지는 경우 세포독성의 문제가 심각해 질 수 있고 실제적인 배양에서는 높은 MOI를 유지하는 것이 불가능하므로 사용하는 100 mm 배양접시에서 배큘로바이러스와 S2 세포가 접촉하는 동안의 변수들인 MOI를 적정의 값인 30으로, 배큘로바이러스 배양 시간을 1.5 시간으로 고정함으로써 배큘로바이러스가 숙주 세포인 S2에 대해 미치는 세포독성을 낮출 수 있었다. 또한, 배큘로바이러스 최소 부피는 배양접시에서 사용부피의 2.4％에서 가장 좋은 결과를 보였으며 배큘로바이러스 용액을 10배 농축시킴으로써 이 부피를 맞추기 위해 첨가시키는 새로운 배지의 비율을 높임으로써 결과적으로 배큘로바이러스에 의한 세포독성을 줄일 수 있었다. 이를 통하여 세포 파쇄 및 배양접시 표면에서 탈착되는 것을 막고 높은 생장 상태를 유지해서 감염 효율도 높일 수 있었다. 배큘로바이러스의 감염이 끝난 후의 관여하는 변수들인 감염 후 배양시간은 24시간에서 최대의 녹색형광단백질의 발현을 나타내었다.

• 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.

• Biomolecules & Therapeutics
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• 제7권4호
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• pp.307-314
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• 1999

The human muscarinic acetylcholine receptor (mAChR) subtypes Hml, Hm2 and Hm3 have been expressed in insect cells (Spodoptera frugiperda, Sf9) using the baculovirus expression system. Expression of relevant DNA, transcript and receptor proteins was identified by PCR, Northern blotting and [$^{3}H$]QNB binding, respectively. As assessed by [$^{3}H$]QNB binding sites, yields of muscarinic receptors in membrane preparations in this study were as about 5-20 times high as those in mammalian cells reported in previous studies. The [$^{3}H$]QNB competition binding studies with well-known subtype-selective mAChR antagonists showed that the receptors expressed in Sf9 cells retain the pharmacological characteristics expected for the ml , m2 and m3 muscarinic receptors. The ml-selective antagonist, pirenzepine, displayed a considerably higher affinity for Hml by 110-fold and 35-fold than for Hm2 and Hm3, respectively, The m2-selective methoctramine displayed a significantly higher affinity for Hm2 than for Hml and Hm3 (10- and 26-fold, respectively). p-F-HHSiD exhibited high affinity for Hm3 that is not significantly different from those for Hml, but 66-fold higher than its affinity for Hm2. The functional coupling of the recombinant receptors to second messenger systems was also examined. While both Hml and Hm3 stimulated phosphoinositide hydrolysis upon activation by carba-chol, Hm2 produced no response. On the other hand, activation of mAChRs induced the inhibition of forsko-lin-stimulated cyclic AMP formation in Hm2-expressing cells, whereas the significant dose-dependent increase in or poor response on cyclic AMP formation were produced in Hml or Hm3-expressing cells, respectively. These results indicate the differential coupling of recombinant Hml, Hm2 and Hm3 receptors expressed in SF9 cells to intracellular signalling system.

• 혜화의학회지
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• 제25권1호
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• pp.71-86
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• 2016

Objective : Hongyi (Formica yessensis) is the dried insect of fomicidae. In previous studies, it appeared possibilities on anti-thrombosis, preventing atherosclerosis, treating rheumatoid disease, and inhibiting hela cell. In this study, we investigated anti-inflammatory effects and mechanism of Hongyi. Methods : Hongyi A was extracted by water and made dried powder. Hongyi B was extracted by ethanol and made dried powder. We measured Nitric Oxide (NO) production on the mouse macrophages (RAW 264.7), mouse vascular endothelial cell (MOVAS) and human vascular endothelial cell (HUVEC) for anti-inflammatory effect. In addition, we conducted reverse transcription reaction (RT-PCR) for investigating the mechanism. Results : In RAW 264.7 macrophages stimulated by LPS, Hongyi A ($100{\mu}g/m{\ell}$) decreased NO production compared with LPS $2{\mu}g/ml$ control group with statistical significance (p<0.05). Hongyi A (50, $100{\mu}g/m{\ell}$) also decreased NO production compared with LPS $4{\mu}g/ml$ control group with statistical significance (p<0.01). Hongyi B (50, $100{\mu}g/m{\ell}$) decreased NO production compared with LPS $2{\mu}g/ml$ control group with statistical significance (p<0.01). Hongyi B (10, 50, $100{\mu}g/m{\ell}$) also decreased NO production compared with LPS $4{\mu}g/ml$ control group with statistical significance (p<0.01, p<0.001, p<0.001). In the MOVAS, Hongyi A and B increased NO production compared with control group. In the HUVEC, Hongyi B increased NO production compared with control group. The expression of NF-${\kappa}B$ in 12-hours MOVAS culture was decreased by Hongyi A and B (10, $50{\mu}g/ml$) compared with control group, but expression of $I{\kappa}B$ was increased. In the 24-hours MOVAS culture, expression of $I{\kappa}B$ was significantly increased. The expression of NF-${\kappa}B$ in 12-hours HUVEC culture was decreased by Hongyi A and B compared with control group, but expression of $I{\kappa}B$ was increased. Hongyi B also increased eNOS mRNA gene expression. Conclusions : Hongyi A and B showed anti-inflammatory effect in mouse macrophages with the activation of vascular endothelial cell through NO production in MOVAS and HUVEC repectively. Honyi B showed superior effect than Hongyi A, but additonal mechanism study should be conducted.

• Mycobiology
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• 제46권2호
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• pp.114-121
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• 2018

Mon1 is a guanine nucleotide exchange factor subunit that activates the Ypt7 Rab GTPase and is essential for vacuole trafficking and autophagy in eukaryotic organisms. Here, we identified and characterized the function of Mon1, an ortholog of Saccharomyces cerevisiae Mon1, in a human fungal pathogen, Cryptococcus neoformans. Mutation in mon1 resulted in hypersensitivity to thermal stress. The mon1 deletion mutant exhibited increased sensitivity to cell wall and endoplasmic reticulum stress. However, the mon1 deletion mutant showed more resistance to the antifungal agent fluconazole. In vivo studies demonstrated that compared to the wild-type strain, the mon1 deletion mutant attenuated virulence in the Galleria mellonella insect model. Moreover, the mon1 deletion mutant was avirulent in the murine inhalation model. These results demonstrate that Mon1 plays a crucial role in stress survival and pathogenicity in C. neoformans.

• 약학회지
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• 제42권3호
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• pp.248-256
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• 1998

We had already reported that we purified N-${\beta}$-alanyl-5-S-glutathionyl-3,4-dihydroxyphenylalanine (5-S-GAD), a novel antibacterial substance from the immunized adult Sarcoph aga peregrina (Flesh fly). We found that the antibacterial activity of synthetic 5-S-GAD is equal to that of authentic 5-S-GAD without a specificity of antibacterial activity against Gram positive and Gram negative. Significant synergism was detected between 5-S-GAD and streptomycin against streptomycin resistant strain E.coli K12 594. It has an antitumor activity against several tumor cell lines at a concentration of $100{\mu}M$. However, no cytotoxic activity against murine macrophage was detected at a concentration of $500{\mu}M$. Furthermore, haemolytic activity against sheep erythrocytes was not detected at the same concentration. We suggest that the S-conjugation of glutathion with dihydroxyphenylalanine might be important to increase antibacterial activity of dihydroxyphenylalanme.