• Title/Summary/Keyword: Insect immune

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Kinetics of Binding of LPS to Recombinant CD14, TLR4, and MD-2 Proteins

  • Shin, Han Jae;Lee, Hayyoung;Park, Jong Dae;Hyun, Hak Chul;Sohn, Hyung Ok;Lee, Dong Wook;Kim, Young Sang
    • Molecules and Cells
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    • v.24 no.1
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    • pp.119-124
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    • 2007
  • TLR4 together with CD14 and MD-2 forms a pattern recognition receptor that plays an initiating role in the innate immune response to Gram-negative bacteria. Here, we employed the surface plasmon resonance technique to investigate the kinetics of binding of LPS to recombinant CD14, MD-2 and TLR4 proteins produced in insect cells. The dissociation constants ($K_D$) of LPS for immobilized CD14 and MD-2 were $8.7{\mu}m$, and $2.3{\mu}m$, respectively. The association rate constant ($K_{on}$) of LPS for MD-2 was $5.61{\times}10^3M^{-1}S^{-1}$, and the dissociation rate constant ($K_{off}$) was $1.28{\times}10^2S^{-1}$, revealing slow association and fast dissociation with an affinity constant $K_D$ of $2.33{\times}10^6M$ at $25^{\circ}C$. These affinities are consistent with the current view that CD14 conveys LPS to the TLR4/MD-2 complex.

Development of a High Efficient "Dual Bt-Plus" Insecticide Using a Primary Form of an Entomopathogenic Bacterium, Xenorhabdus nematophila

  • Eom, Seonghyeon;Park, Youngjin;Kim, Hyeonghwan;Kim, Yonggyun
    • Journal of Microbiology and Biotechnology
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    • v.24 no.4
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    • pp.507-521
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    • 2014
  • A phase variation has been reported in an entomopathogenic bacterium, Xenorhabdus nematophila. Compared with a wild-type primary form, a secondary form usually loses several physiological and biochemical characters. This study showed that the phase variation of X. nematophila caused a significant alteration in its immunosuppressive activity and subsequent entomopathogenicity. A secondary form of X. nematophila was detected in laboratory colonies and exhibited significant differences in dye absorption and entomopathogenicity. In addition, the secondary form was different in its production of eicosanoid-biosynthesis inhibitors (EBIs) compared with the primary form of X. nematophila. Production of oxindole and p-hydroxypropionic acid was significantly reduced in the culture broth of the secondary form of X. nematophila. The reduced EBI production resulted in significant suppression in the inhibitory effects on cellular nodule formation and phenoloxidase activity. Culture broth of the primary form of X. nematophila enhanced the pathogenicity of Bacillus thuringiensis ( Bt) significantly more than the culture broth of the secondary form. Furthermore, this study developed a highly efficient "Dual Bt-Plus: to control both lepidopteran insect pests Plutella xylostella and Spodoptera exigua, by mixing two effective Bt strains along with the addition of potent bacterial metabolites or 100-fold concentrated X. nematophila culture broth.

Classification of NOVCs and AVOCS for Healing Substance Measurement System Based on Gas Sensors Array in Forest Environment (가스센서 어레이를 이용한 산림환경 내 치유물질 측정시스템을 통한 자연적 휘발성 유기화합물(NVOCs)과 인위적 휘발성 유기화합물(AVOCs) 분류)

  • Joon-Boo Yu;Hyung-Gi Byun
    • Journal of Sensor Science and Technology
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    • v.32 no.2
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    • pp.95-99
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    • 2023
  • Forest healing is an activity that enhances immunity and human health using various elements of nature, such as fragrance and scenery. Particularly, phytoncide composed of terpene, a natural volatile substance emitted by forest plants, activates the immune function and is an important raw material in health-related products, such as antibacterial and insect repellents. Moreover, phytoncide is used as a measure to evaluate the impact of the forest atmosphere on the human body. This study aims to implement a highly sensitive gas sensor system that can measure phytoncide in real-time, which is an essential element for realizing a forest healing environment. A gas generation apparatus was implemented by using an adsorption tube in consideration of filed applicability in a laboratory atmosphere to enable the measurement of α-pinene and limonene, which are among the main components of phytoncide. Throughout the experimental trials, the sensitivity of gas sensor arrays to α-pinene and limonene was confirmed. In addition, the classification results demonstrated the AVOCs and NVOCs can be well discriminated using PCA. The primary results confirmed the possibility of developing a high-sensitivity gas sensor system for phytoncide sensing in real time.

Comparative transcriptome analysis of Cordyceps militaris grown on germinated soybean media

  • Yoo, Chang-Hyuk;Choi, Jaehyuk
    • Journal of Mushroom
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    • v.20 no.1
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    • pp.7-12
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    • 2022
  • The ascomycete fungus Cordyceps militaris infects lepidopteran insect pupae, forming characteristic fruiting bodies called "Dong Chung Ha Cho" in Korean. They have been used as medicines owing to their anti-allergic, anti-inflammatory, and immune-enhancing effects. This fungus can be grown on the geminated soybeans Rhynchosia nulubilis, which also contains several novel isoflavones. We performed a comparative transcriptome analysis to determine core gene sets or pathways contributing to biologically active products such as isoflavone. Initially, we sequenced 2-week-old fungal cultures on different soybean agar media, where different amounts of water agar were implemented to show different surface topology. We selected 830 upregulated and 188 downregulated genes by comparing linear models of the samples (two-fold change threshold). Gene ontology analysis identified that the "IMP biosynthesis" term was significantly found in the upregulated gene sets. The pathway is involved in the synthesis of cordycepin, the reference chemical for C. militaris. This finding in the transcriptome data is consistent with the previous observation: increased cordycepin concentrations in the C. militaris cultured on germinated soybean.

Pyriproxyfen Inhibits Hemocytic Phagocytosis of the Beet Armyworm, Spodoptera exigua (파밤나방(Spodoptera exigua)의 혈구세포 식균반응에 대한 피리프록시펜의 억제효과 Nalini Madanagopal)

  • Madanagopal, Nalini;Lee, Yong-Joon;Kim, Yong-Gyun
    • The Korean Journal of Pesticide Science
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    • v.11 no.3
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    • pp.164-170
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    • 2007
  • The concept of innate immunity in insects which refers to the first line of host defense constitutes the humoral and cellular components which are involved in recognition and actively participate in the elimination of the intruding foreign micro- or macro-organisms. Several recent studies suggest that juvenile hormone (JH) modulates the cellular immune reactions in response to pathogen. In this study, pyriproxyfen (a JH agonist as an insect growth regulator) was tested in its any inhibitory effect on the immune reactions of the beet armyworm, Spodoptera exigua. To this end, five different hemocyte morphotypes of final instar S. exigua were identified by phase contrast microscopy. Plasmatocytes and granular cells, which constitute about 90% of the total hemocyte count, were prominently distinguished based on their basophilic/acidophilic nature using Giemsa stain. The role of pyriproxyfen on the functional ability of hemocytes was analyzed using FITC-labeled Providencia vermicola for the phagocytic potential of the hemocytes. Both granular cells and plasmatocytes exhibited phagocytosis behavior. Pyriproxyfen significantly inhibited the phagocytosis of both cell types, proposing its novel action as an immunosuppressant.

Comparative Analysis of Host Insect Immunodepression Induced by Two Entomopathogenic Bacteria, Xenorhabdus nematophilus and Staphylococcus gallinarum, with Differential Pathogenicities (병원력 차이를 보이는 두 곤충병원세균(Xenorhabdus nematophilus와 Staphylococcus gallinarum)의 면역저하 능력 비교 분석)

  • 박영진;김길호;김용균
    • Korean journal of applied entomology
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    • v.42 no.4
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    • pp.353-360
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    • 2003
  • Immunodepression can be required for entomopathogenic bacteria to induce their potent pathogenicities to the target insects. Here, we raise a hypothesis that the capacity of a pathogenic bacterium to induce the target insect immunodepression has positive relationship with the degree of pathogenicity. X. nematophilus had 1,200 times as potent as another entomopathogenic bacterium, Staphylococcus gallinarum against the fifth instar larvae of silkworm, Bombyx mori, when they were Injected into the hemocoel. Although both bacteria had significant cytotokic effect on the hemocytes of B. mori, X. nematophilus gave faster and greater cytotoxicity than did S. gallinarum. In cellular immune reactions, B. mori could form 20 hemocyte nodules against the bacterial injection with 5${\times}$10$\^$5/ cells. The number of the hemocyte nodules was significantly depressed when live X. nematophilus was inject-ed, but not in S. gallinarum. Activation of prophenoloxidase (proPO) was depressed in the bacterial injection. The depression of PO activation was significantly greater in X. nematophilus infection than in S. gallinarum injection. Lysozyme activity was induced by the injection of S. gallinarum at 4 h after the treatment, but not induced in X. nematophilus at all the time. These results showed that X. nemato-philus induced greater immunodepression against B. mori and resulted in higher pathogenicity than did S. gallinarum. Therefore, this study suggests that the immunodepression induced by entomopathogenic bacteria has positive relationship with their pathogenicity.

Sound Stress Alters Physiological Processes in Digestion and Immunity and Enhances Insecticide Susceptibility of Spodoptera exigua (스트레스 음파에 따른 파밤나방(Spodoptera exigua ) 소화 및 면역 생리작용 저하와 살충제 감수성 제고 효과)

  • Park, Jung-A;Seok, Jung-Kyun;Prasad, Surakasi Venkara;Kim, Yong-Gun
    • Korean journal of applied entomology
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    • v.50 no.1
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    • pp.39-46
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    • 2011
  • This study analyzed effects of different sound treatments in frequencies and intensities on digestion and immune physiological processes of the beet armyworm, Spodoptera exigua larvae. Without effect on egg hatch, sound treatments with 100-5,000 Hz at 95 dB suppressed feeding behavior and inhibited a digestive enzyme activity. In addition, two dimensional electrophoresis of midgut luminal proteins indicated a marked difference of the sound-treated larvae. In response to 5,000 Hz at 95 dB, larvae showed a significant decrease in hemocyte nodule formation against fungal challenge along with significant suppression in phospholipase $A_2$ activity in hemocyte and plasma. With increase of sound frequencies, the treated larvae showed an enhanced susceptibility to insecticides. Such sound frequency effect was significantly modulated with different sound intensities. These results suggest that sound treatment may give adverse stress to physiological processes of S. exigua larvae and may be applied to a nonchemical insect pest control.

Study on Development of Novel Biopesticides Using Entomopathogenic Bacterial Culture Broth of Xenorhabdus and Photorhabdus (Xenorhabdus 및 Photorhabdus 세균 배양액을 이용한 생물농약 개발에 관한 연구)

  • Seo, Sam-Yeol;Kim, Yong-Gyun
    • Korean journal of applied entomology
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    • v.49 no.3
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    • pp.241-249
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    • 2010
  • Two groups of entomopathogenic bacteria, Xenorhabdus and Photorhabdus, are known to suppress insect immune responses by inhibiting eicosanoid biosynthesis. This study used these bacterial culture broths to develop novel biochemical insecticides against the diamondback moth, Plutella xylostella. Though the bacterial culture broths alone showed little insecticidal activity, they significantly enhanced pathogenicity of Bacillus thuringiensis against the fourth instar larvae of P. xylostella. Sterilization of the bacterial culture broth by autoclaving or $0.2\;{\mu}m$ membrane filtering did not influence the synergistic effect on the pathogenicity of B. thuringiensis. Three metablites identified in the culture broth of X. nematophila also showed similar synergistic effects. In field test, both entomopathogenic bacterial culture broth also enhanced the control efficacy of B. thuringiensis against P. xylostella.

Quality Evaluation of Wind Vectors from UHF Wind Profiler using Radiosonde Measurements (라디오존데 관측자료를 이용한 UHF 윈드프로파일러 바람관측자료의 품질평가)

  • Kim, Kwang-Ho;Kim, Min-Seong;Seo, Seong-Woon;Kim, Park-Sa;Kang, Dong-Hwan;Kwon, Byung Hyuk
    • Journal of Environmental Science International
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    • v.24 no.1
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    • pp.133-150
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    • 2015
  • Wind profiler provides vertical profiles of three-dimensional wind vectors with high spatiotemporal resolution. The wind vectors is useful to analyze severe weather phenomena and to validate the various products from numerical weather prediction model. However, the wind measurements are not immune to ground clutter, bird, insect, and aircraft. Therefore, quality of wind vectors from wind profiler must be quantitatively evaluated prior to its application. In this study, wind vectors from UHF wind profiler at Ganwon Regional Meteorological Administration was quantitatively evaluated using 27 radiosonde measurements that were launched every two or three hours according to rainfall intensity during Intensive Observation Period (IOP) from June to July 2013. In comparison between two measurements, wind vectors from wind profiler was relatively underestimated. In addition, the accuracy and quality of wind vectors from wind profiler decrease with increasing beam height. The accuracy and quality of the wind vectors for rainy periods during IOP were higher than for the clear-air measurements. The moderate rainfall intensity lead to multi-peaks in Doppler spectrum. It results in overestimation of vertical air motion, whereas wind vectors from wind profilers shows good agreement with those from radiosonde measurements for light rainfall intensity.

Activated Phenoloxidase Interacts with A Novel Glycine-rich Protein on the Yeast Two-hybrid System

  • Lee, Sun-Woo;Lee, Hyun-Seong;Kim, Eun-Jun;Yoo, Mi-Ae;Lee, Bok-Luel
    • BMB Reports
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    • v.34 no.1
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    • pp.15-20
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    • 2001
  • One of the innate immune reactions in invertebrates is the pro-phenoloxidase (pro-PO) activation system that is involved in the generation of superoxide, melanin synthesis, and the subsequent sequestration of foreign matter entering the hemocoel of the invertebrates. However, the molecular mechanism of this biological reaction is still obscure. To expand our understanding of the biological roles of the pro-PO activation system in invertebrates, we performed a yeast two-hybrid screening by using three regions of pro-PO as bait and a yeast two-hybrid cDNA library from Tenebrio molitor larvae as prey We isolated a novel partial cDNA clone that encodes a glycine-rich protein that interacted with the active phenoloxidase (termed phenoloxidase interacting protein, POIP). POIP consists of two domains: One is an N-terminal unique domain and the other is a C-terminal glycine-rich domain. The C-terminal glycine-rich domain showed sequential homology with those of insect antifungal proteins. Also, the yeast two-hybrid screen in a reverse orientation (using POIP as bait) yielded PO, suggesting that the PO-POIP interaction is specific. By using a 315 bP PCR fragment of the N-terminal unique region of POIP, we cloned the full-length cDNA of POIP from the Tenebruo cDNA library constructed by using E. coli injected larvae. The interaction analysis between PO, and a truncated fragment lacking the N-terminal unique region of POIP, indicated that the N-terminal unique region is necessary for interaction between PO and POIP. The expression level of the POIP mRNA is increased by bacterial injection into T. molitor larvae. This suggests that POIP might be engaged in the humoral defense reaction.

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