• Title/Summary/Keyword: insect cells

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Susceptibility of Anthonomus grandis (Cotton Boll Weevil) and Spodoptera frugiperda (Fall Armyworm) to a Cry1Ia-type Toxin from a Brazilian Bacillus thuringiensis Strain

  • Grossi-De-Sa, Maria Fatima;De Magalhaes, Mariana Quezado;Silva, Marilia Santos;Silva, Shirley Margareth.Buffon;Dias, Simoni Campos;Nakasu, Erich Yukio Tempel;Brunetta, Patricia Sanglard Felipe;Oliveira, Gustavo Ramos;De Oliveira Neto, Osmundo Brilhante;De Oliveira, Raquel Sampaio;Soares, Luis Henrique Barros;Ayub, Marco Antonio Zachia;Siqueira, Herbert Alvaro Abreu;Figueira, Edson L.Z.
    • BMB Reports
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    • v.40 no.5
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    • pp.773-782
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    • 2007
  • Different isolates of the soil bacterium Bacillus thuringiensis produce multiple crystal (Cry) proteins toxic to a variety of insects, nematodes and protozoans. These insecticidal Cry toxins are known to be active against specific insect orders, being harmless to mammals, birds, amphibians, and reptiles. Due to these characteristics, genes encoding several Cry toxins have been engineered in order to be expressed by a variety of crop plants to control insectpests. The cotton boll weevil, Anthonomus grandis, and the fall armyworm, Spodoptera frugiperda, are the major economically devastating pests of cotton crop in Brazil, causing severe losses, mainly due to their endophytic habit, which results in damages to the cotton boll and floral bud structures. A cry1Ia-type gene, designated cry1Ia12, was isolated and cloned from the Bt S811 strain. Nucleotide sequencing of the cry1Ia12 gene revealed an open reading frame of 2160 bp, encoding a protein of 719 amino acid residues in length, with a predicted molecular mass of 81 kDa. The amino acid sequence of Cry1Ia12 is 99% identical to the known Cry1Ia proteins and differs from them only in one or two amino acid residues positioned along the three domains involved in the insecticidal activity of the toxin. The recombinant Cry1Ia12 protein, corresponding to the cry1Ia12 gene expressed in Escherichia coli cells, showed moderate toxicity towards first instar larvae of both cotton boll weevil and fall armyworm. The highest concentration of the recombinant Cry1Ia12 tested to achieve the maximum toxicities against cotton boll weevil larvae and fall armyworm larvae were 230 ${\mu}g/mL$ and 5 ${\mu}g/mL$, respectively. The herein demonstrated insecticidal activity of the recombinant Cry1Ia12 toxin against cotton boll weevil and fall armyworm larvae opens promising perspectives for the genetic engineering of cotton crop resistant to both these devastating pests in Brazil.

The anti-imflammatory effect and the mechanism of Formica yessensis extraction (홍의 추출물의 항염작용 및 그 기전 연구)

  • Kim, Jong-Min;Kim, Seung-Hyung;Yang, Won-Kyung;Jung, Taek-Geun;Kim, Se-Ran;Hwang, Sung-Joon;Yoo, Hwa-Seung
    • Journal of Haehwa Medicine
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    • v.25 no.1
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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.

Anti-cancer Properties and Relevant Mechanisms of Cordycepin, an Active Ingredient of the Insect Fungus Cordyceps spp., (동충하초 유래 cordycepin의 항암 활성 기전 최근 연구 동향)

  • Jeong, Jin-Woo;Choi, Yung Hyun
    • Journal of Life Science
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    • v.25 no.5
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    • pp.607-614
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    • 2015
  • Cancers are the largest cause of mortality and morbidity all over the world. Cordycepin, an adenosine analog, is a major functional component of the Cordyceps species, which has been widely used in traditional Oriental medicine. Over the last decade, this compound has been reported to possess many pharmacological properties, such as an ability to enhance immune function, as well as anti-inflammatory, antioxidant and anti-cancer effects. Recently, numerous studies have reported interesting properties of cordycepin as a chemopreventive agent as well. There is an accumulating body of experimental evidences suggesting that cordycepin impedes cancer progression by promoting apoptosis, inducing cell cycle arrest, modulating intracellular signaling pathways, and inhibiting invasion and metastasis of cancer cells. In many cancer cell lines, cordycepin inhibits growth and cell cycle progression by inducing arrest of the G2/M phase, resulting from the inhibition of retinoblastoma protein phosphorylation and induction of cyclin-dependent kinase inhibitors. To induce apoptosis, cordycepin activates the extrinsic and intrinsic pathways, which promotes reactive oxygen species generation and the downstream activation of kinase cascades. Cordycepin also can activate alternative pathways to cell death such autophagy. In addition, cordycepin can inhibit the pro-metastatic processes of cancer cell detachment, migration, and invasion through a variety of mechanisms, including the nuclear factor-kappa B and activated protein-1 signaling pathways. In this review, we summarized the variety of action mechanisms by which cordycepin may mediate chemopreventive effects on cancer and discussed the potential of this natural product as a promising therapeutic inhibitor of cancer development.

Nucleotide sequence analysis and expression of NSP4 gene of human rotaviruses isolated in Korea (국내에서 분리된 사람 로타바이러스의 NSP4 유전자 염기서열 분석 및 발현)

  • Jung, Dong-hyuk;Song, Yun-kyung;Kim, Kyung-mi;Park, Hyo-sun;Back, Myoung-soon;Kang, Shien-young
    • Korean Journal of Veterinary Research
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    • v.42 no.1
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    • pp.89-100
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    • 2002
  • The nonstructural glycoprotein NSP4, encoded by the 10th gene of rotavirus, has been known to play important roles in viral assembly and pathogenesis. The NSP4 genes of human rotavirus Korean isolates, designated as CBNU/HR-1, CBNU/HR-2, CBNU/HR-3, and CBNU/HR-4, were cloned, sequenced and characterized. Also, the NSP4 gene of the CBNU/HR-1 was expressed in a baculovirus-insect cell system. The sequence data indicated that the NSP4 genes of human rotavirus Korean isolates were 750 or 751 bases in length and encoded one open reading frame of 175 amino acids. Two glycosylation sites were recognized in the NSP4 gene of human rotavirus isolates tested. The NSP4 of CBNU/HR-1, CBNU/HR-3, and CBNU/HR-4 exhibited a high degree of amino acid sequence homology with that of NSP4 genotype B viruses, but a low degree of amino acid sequence homology with that of NSP4 genotype A viruses. However, the NSP4 of CBNU/HR-2 exhibited a high degree of amino acid sequence homology with that of NSP4 genotype A viruses, but a low degree of amino acid sequence homology with that of NSP4 genotype B viruses. The Sf9 cells infected with recombinant baculovirus, inserted with NSP4 gene of CBNU/HR-1, produced specific cytopathic effects and the expressed NSP4 was detected by immunofluorescence staining using NSP4-specific monoclonal antibody(MAb). The expressed NSP4 migrated at 16-26 kDa on SDS-PAGE and reacted with NSP4-specific MAb by Western blotting.

Anti-neuroinflammatory Effects of a Locusta migratoria Ethanol Extract in LPS-stimulated BV-2 Microglia (LPS로 자극된 미세아교세포에서 풀무치 에탄올 추출물의 신경염증 억제 효능)

  • Lee, Hwa Jeong;Seo, Minchul;Lee, Joon Ha;Kim, In-Woo;Kim, Sun Young;Hwang, Jae-Sam;Kim, Mi-Ae
    • Journal of Life Science
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    • v.28 no.11
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    • pp.1332-1338
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    • 2018
  • Activated microglia, induced by various pathogens, protect neurons and maintain homeostasis of the central nervous system (CNS). However, severe activation causes neurodegenerative disorders such as Alzheimer's disease and Parkinson's disease because of the secretion of various neurotoxic molecules, such as nitric oxide (NO), prostaglandin (PG), and pro-inflammatory cytokines. Because chronic microglial activation endangers neuronal survival, negative regulators of microglial activation have been identified as potential therapeutic candidates for treatment of many neurological diseases. One potential source of these regulators is Locusta migratoria, a grasshopper of the Acrididae, usually 4-6 cm in size, belonging to the family of large insects in Acrididae. This grasshopper is an edible insect resource that can be consumed by humans as protein source or used for animal feed. The aim of the present study was to examine the inhibitory effects of a L. migratoria ethanol extract (LME) on the production of inflammatory mediators in LPS-stimulated BV-2 microglia cells. The extract significantly inhibited the NO, iNOS, COX-2, and pro-inflammatory cytokine ($TNF-{\alpha}$, IL-6 and $IL-1{\beta}$) levels in BV-2 microglia cell. Because the inhibition of microglial activation may be an effective solution for treating brain disorders like Alzheimer's and Parkinson's diseases, these results suggest that LME may be a potential therapeutic agent for the treatment of brain disorders induced by neuroinflammation.

Research Trends on the Therapeutic Potential of Cordycepin, an Active Ingredient of the Insect Fungus Cordyceps spp., for the Prevention of Sarcopenia (동충하초(Cordyceps spp.)의 유효 생리활성 성분인 cordycepin의 근감소증 예방에 대한 연구 동향)

  • Kim, Sung Ok;Choi, Yung Hyun
    • Journal of Life Science
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    • v.32 no.6
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    • pp.482-490
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    • 2022
  • Sarcopenia, a geriatric and multifactorial syndrome characterized by progressive systemic skeletal muscle disorder, may be associated with many comorbidities. Sarcopenia caused by a decrease in muscle mass and muscle strength is accompanied by the aggravation of various pathological conditions, and as life expectancy increases, its prevalence will continue to increase in the future. During the aging process, chronic oxidative stress and increased inflammatory responses act as major contributors to skeletal muscle loss. In addition, disruption of autophagy and apoptosis signals associated with dysfunction of mitochondria, which are essential for energy metabolism, accelerates the loss of muscle proteins. The pharmacological effect of cordycepin, a major physiologically active substance in the genus Cordyceps, which has been widely used for the prevention and treatment of various diseases for a long time, is directly related to its antioxidant and anti-inflammatory actions. In this review, we present the correlation between apoptosis, autophagy, protein catabolism, and satellite cell activity important for muscle regeneration using cordycepin for the prevention and treatment of sarcopenia. Although there have been few studies so far on the use of cordycepin for sarcopenia, previous studies suggest that cordycepin may contribute to inhibiting the age-related weakening of mitochondrial function and blocking the breakdown of muscle proteins. In addition, the protective effect of cordycepin on muscle cell damage is considered to be closely related to its antioxidant and anti-inflammatory activities. Therefore, it is considered that more continuous basic research is needed, focusing on the molecular biological mechanism of cordycepin, which is involved in the anti-aging of muscle cells.

Anti-skinaging effects of Gryllus bimaculatus on ERM-CZ100-exposed human diploid fibroblasts (미세먼지 유발 피부노화에 대한 쌍별귀뚜라미의 예방 효과)

  • Kyong Kim;Chae-Heon Lee;Eun-Young Park;Yoon Sin Oh
    • Journal of Nutrition and Health
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    • v.56 no.6
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    • pp.615-628
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    • 2023
  • Purpose: Increasing levels of domestic fine dust (DFD) have emerged as a serious problem that threatens public health by causing chronic respiratory diseases and skin aging. The present study was performed to investigate the inhibitory effects of Gryllus bimaculatus (the two-spotted cricket), which has recently attracted attention as an edible insect in South Korea, on DFD-induced aging and inflammation. Methods: To verify that DFD causes skin aging and investigate the anti-aging effect of an aqueous ethanolic-Gryllus bimaculatus extract (AE-GBE), human diploid fibroblasts (HDF) were treated with 100 ㎍/mL of European reference material (ERM)-CZ100 dust for 24 hrs in the presence or absence of 100 ㎍/ml AE-GBE. Aging and cellular toxicities were assessed by measuring reactive oxygen species (ROS) levels, DNA fragmentation, and β-galactosidase activity. The protein levels of cyclooxygenase (COX) 2, matrix metalloproteinase (MMP)-1, and collagen were measured by western blot, and the mRNA expressions of inflammation-related genes were assayed by quantitative reverse transcriptase polymerase chain reaction. Results: Treatment with ERM-CZ100 induced an aged phenotype in HDF cells, as evidenced by increased ROS levels, DNA fragmentation, and senescence-associated β-galactosidase activity, but cotreatment with AE-GBE significantly reduced these inductions. The mRNA expressions of pro-inflammatory cytokines, such as interleukin (IL)-1β, IL-6, and tumor necrosis factor-α, induced by ERM-CZ100 were also reduced by AE-GBE cotreatment, which also reduced COX2 expression. Moreover, ERM-CZ100-induced MMP-1 expression and reduced collagen type I expression were recovered by AE-GBE treatment. Conclusion: These results suggest that AE-GBE is a potential treatment for domestic fine dust-induced skin inflammation and inflammaging.

Studies on Witches' Broom of Ligustrum ovalifolium Hasskarl Caused by Mycoplasma-like Organism (MLO) (Mycoplasma성(性) 왕쥐똥나무 빗자루병(病)에 관(關)한 연구(硏究))

  • Chai, Jyung-Ki;Kim, Young-Ho
    • Journal of Korean Society of Forest Science
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    • v.78 no.2
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    • pp.103-118
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    • 1989
  • The occurrence of witches' broom in Ligustrum ovalifolium was first noticed in Korea by author in 1984. The present study was carried out with particular emphasis on the symptomatology, etiology, transmission of the disease and antibiotic treatments. The infected tissue was observed by the fluorescence and electron microscopy and its biochemical characteristics were compared with healthy one by electrophoresis. The results are summarized as follows : 1. symptoms of the infected trees were characterized by the dwarfing of the organs, yellowing and brooming of the foliage. 2. The observation by the trans electron microscopy on the witches' broom of L. ovalifolium revealed the occurrence of numerous mycoplasma-like organisms(MLOs) in the phloem tissue cells of the midribs of infected leaves. 3. The MLOs were surrounded by a single unit membrane, and they appeared to be multiplied by binary fission. 4. The presence of crystals unidentified in the phloem parenchyma cells was noticed by electron rnicroscopy, 5. The disease was able to be transmitted by budding, crown, and greenwood graftings to L. ovalifolium, L. obtusifolium, L, japonicum and also transmitted, even when the stocks and scions were not completely grafted. 6. Insect transmission on L. ovalifolium and L, obtzrsifolium was carried by Hishimonus sellatus. 7. The infected roots dipped in the 1,000 ppm of teracyclin solution was only temporarily effective in controlling the disease. 8. Infected plant with MLOs showed specific fluorescent reactions in phloems with DAPI stain. 9. The protein and peroxidase separated by electrophoresis showed strikingly distinctive difference between the healthy and diseased leaves.

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Expression of Bombyx mori Transferrin Gene in Response to Oxidative Stress or Microbes (미생물 및 산화적 스트레스에 의한 누에 트랜스페린 발현)

  • Yun, Eun-Young;Kwon, O-Yu;Hwang, Jae-Sam;Ahn, Mi-Young;Goo, Tae-Won
    • Journal of Life Science
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    • v.21 no.11
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    • pp.1607-1611
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    • 2011
  • To analyze the role of Bombyx mori transferrin (BmTf) in response to microbes or oxidative stress, we investigated the level of BmTf transcripts in B. mori treated with various microbes and oxidative stress inducers. BmTf mRNA was mainly expressed in the epidermis and fat in the bodies of B. mori injected with Escherichia coli, and up regulated in response to microbes such as bacteria, fungi, or viruses, but was hardly altered in response to oxidative stress inducers such as $H_2O_2$, Cu, or $FeCl_3$. We also confirmed that BmTf mRNA expression was increased in Bm5 cells treated with ERK, PLC, PKA, PI3K, MAPK, or JNK inhibitors, respectively. To identify the major inducer of BmTf expression, we analyzed the amount of serum iron in the hemolymph of B. mori after injection or feeding with E. coli or $FeCl_3$. The results showed that the amount of serum iron was not changed by injection and feeding with E. coli, although BmTf mRNA was increased by injection with E. coli. On the contrary, injection and feeding with $FeCl_3$ significantly increased the amount of serum iron, although they did not alter the BmTf mRNA level. On the basis of these results, we assume that up-regulation of BmTf in B. mori is closely related to the defense of microorganism, and BmTf may be expressed at the basal constitutive level when it plays a role in iron metabolism by maintaining iron homeostasis and in the insect defense mechanism against oxidative stress.

A New Mid-late Maturing Rice Variety, 'Honong' with High-Quality and Multiple Disease Resistance (중만생 고품질 복합내병성 벼 신품종 '호농')

  • Kim, Ki-Young;Shin, Mun-Sik;Ko, Jae-Kwon;Nam, Jeong-Kwon;Ha, Ki-Yong;Baek, Man-Gee;Ko, Jong-Cheol;Kim, Bo-Kyeong;Park, Hyun-Su;Shin, Woon-Chul;Kang, Hyun-Jung;Choung, Jin-Il;Kim, Woo-Jae;Mo, Young-Jun;Kim, Jung-Gon
    • Korean Journal of Breeding Science
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    • v.42 no.3
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    • pp.252-256
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    • 2010
  • 'Honong' is a new japonica rice variety developed and registered by the rice breeding team of Department of Rice and Winter Cereal Crop, NICS, RDA in 2009. This variety was derived from mutagen MNU (N-methyl-N-nitrosourea) treatment on fertilized egg cells of Unbong31. This variety has about 126 days growth duration from transplanting to harvesting in west-southern coast, Honam and Youngnam plain of Korea. It has 79 cm in culm length and is tolerant to lodging. In reaction to biotic and abiotic stresses, it show resistance to blast, bacterial blight pathogen races from $K_1$ to $K_3$ and stripe virus, but susceptible to other major diseases and insect pests. The milled rice of 'Honong' exhibits translucent, relatively clear non-glutinous endosperm and medium short grain. It has higher amylose content (20.3%) and lower protein content (6.3%), and good palatability of cooked rice compared with Nampyeong. The milled rice yield performance of this variety is about 5.44MT/ha in local adaptability test for three years. 'Honong' would be adaptable to west-southern coast, Honam and Youngnam plain of Korea.