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Sanae Sekihara, Toshio Shibata, Mai Hyakkendani, Shun-Ichiro Kawabata
We recently reported that transglutaminase (TG) suppresses immune deficiency pathway-controlled antimicrobial peptides (IMD-AMPs), thereby conferring immune tolerance to gut microbes, and that RNAi of the TG gene in flies decreases the lifespan compared with non-TG-RNAi flies. Here, analysis of the bacterial composition of the Drosophila gut by next-generation sequencing revealed that gut microbiota comprising one dominant genus of Acetobacter in non-TG-RNAi flies was shifted to that comprising two dominant genera of Acetobacter and Providencia in TG-RNAi flies...
October 19, 2016: Journal of Biological Chemistry
Peter Kupferschmied, Tiancong Chai, Pascale Flury, Jochen Blom, Theo H M Smits, Monika Maurhofer, Christoph Keel
Some plant-beneficial pseudomonads can invade and kill pest insects in addition to their ability to protect plants from phytopathogens. We explored the genetic basis of O-polysaccharide (O-PS, O-antigen) biosynthesis in the representative insecticidal strains Pseudomonas protegens CHA0 and Pseudomonas chlororaphis PCL1391 and investigated its role in insect pathogenicity. Both strains produce two distinct forms of O-PS, but differ in the organization of their O-PS biosynthesis clusters. Biosynthesis of the dominant O-PS in both strains depends on a gene cluster similar to the O-specific antigen (OSA) cluster of Pseudomonas aeruginosa...
October 11, 2016: Environmental Microbiology
T V Shevchuk, N S Zakharchenko, S V Tarlachkov, O V Furs, O V Dyachenko, Y I Buryanov
Transgenic kalanchoe plants (Kalanchoe pinnata L.) expressing the antimicrobial peptide cecropin P1 gene (cecP1) under the control of the 35S cauliflower mosaic virus 35S RNA promoter and the selective neomycin phosphotransferase II (nptII) gene under the control of the nopaline synthase gene promoter were studied. The 35S promoter methylation and the cecropin P1 biosynthesis levels were compared in plants growing on media with and without kanamycin. The low level of active 35S promoter methylation further decreases upon cultivation on kanamycin-containing medium, while cecropin P1 synthesis increases...
September 2016: Biochemistry. Biokhimii︠a︡
Andrey Yu Yakovlev, Alexander P Nesin, Nina P Simonenko, Natalia A Gordya, Dmitriy V Tulin, Anastasia A Kruglikova, Sergey I Chernysh
Antimicrobial peptides accumulated in the hemolymph in response to infection are a key element of insect innate immunity. The involvement of the fat body and hemocytes in the antimicrobial peptide synthesis is widely acknowledged, although release of the peptides present in the hemolymph from the immune cells was not directly verified so far. Here, we studied the presence of antimicrobial peptides in the culture medium of fat body cells and hemocytes isolated from the blue blowfly Calliphora vicina using complex of liquid chromatography, mass spectrometry, and antimicrobial activity assays...
September 1, 2016: In Vitro Cellular & Developmental Biology. Animal
Chunmiao Bo, Chaozhan Wang, Yinmao Wei
Immobilized metal affinity chromatography (IMAC) has been widely used for the specific separation of biopolymers. However, leakage of metal ions from IMAC adsorbents is of concern in IMAC. In this study, we designed a novel tridenate bis(5-methyltetrazolium)amine (BMTA) to reduce the leakage of metal ions by improving the affinity to immobilized metal ions. The ligand was bonded onto silica via three-step reaction to prepare a high-performance IMAC stationary phase. The chromatographic behaviors of ribonuclease A, cytochrome c, and lysozyme on the Cu(II)-, Ni(II)-, and Zn(II)-chelated stationary phase were investigated with respect to pH effect and elution with an imidazole gradient...
November 2016: Analytical and Bioanalytical Chemistry
Zhen-Ping Li, Da-Wei Chen, Yan-Qiu Pan, Li Deng
Two isoforms of piscidin from Malabar grouper (Epinephelus malabaricus), EmPis-1 and EmPis-2, were cloned and studied. EmPis-1 and EmPis-2 showed the different in the 3'UTR features of mRNA and gene expression patterns. AUUUA-motif-containing ARE was found in mRNA of EmPis-1, but not in that of EmPis-2. EmPis-1 and EmPis-2 expressed not only in the potential sites of pathogen entry, but also in grouper's immune-related tissues such as head kidney (HD), peripheral blood leukocytes (PBL) and spleen. The expression level of EmPis-1 was higher than that of EmPis-2 in most fish tissues...
October 2016: Fish & Shellfish Immunology
Feng-Yu Lin, Yan Gao, Hao Wang, Qiu-Xia Zhang, Chang-Lin Zeng, Hai-Peng Liu
It is well-known that anti-lipopolysacchride factors (ALFs) are involved in the recognition and elimination of invading pathogens. In this study, the full-length ALF cDNA sequence of the red claw crayfish Cherax quadricarinatus (termed CqALF) was cloned from a suppression subtractive hybridization library constructed using red claw crayfish hematopoietic tissue cell (Hpt cell) cultures following challenge with white spot syndrome virus (WSSV). The full-length cDNA sequence of CqALF was 863 bp, and the open reading frame encoded 123 amino acids with a signal peptide in the N-terminus and a conserved LPS-binding domain...
October 2016: Fish & Shellfish Immunology
André F Ferreira, Akhilesh Rai, Lino Ferreira, Pedro N Simões
The immobilization of gold nanoparticles (AuNPs) with antimicrobial peptides (AMPs) is a new and promising way to enhance both the activity and targeting capabilities of AMPs. However, a full understanding of the adsorption process underlying these materials is still lacking. Cecropin-melittin is a peptide with a broad antimicrobial activity while displaying low hemolytic properties, whose conjugation with AuNPs has not been studied before. In this context, we report the investigation of the adsorption process of the cecropin-melittin peptide, with (CM-SH) and without (CM) cysteine at its C-terminus, onto a gold surface based on all-atom MD simulations...
July 28, 2016: European Biophysics Journal: EBJ
Jian Xu, Fei Zhong, Yonghong Zhang, Jianlou Zhang, Shanshan Huo, Hongyu Lin, Liyue Wang, Dan Cui, Xiujin Li
Probiotics and antimicrobial peptides (AMPs) in animal gastrointestinal tract play the crucial roles in microbiota maintenance, immune regulation and pathogen defending. Generating probiotics strain engineered for expressing AMPs should be beneficial for animal growth and production. Here, we generated B. subtilis strain engineered for expressing porcine β-defensin-2 (pBD-2) and cecropin P1 (CP1) fusion antimicrobial peptide using their codon-optimized genes mediated by pMK4 prokaryotic expression vector. Results showed that the B...
June 30, 2016: Asian-Australasian Journal of Animal Sciences
Jiaxin Zhang, Ali Movahedi, Zhiheng Wei, Ming Sang, Xiaolong Wu, Mengyang Wang, Hui Wei, Huixin Pan, Tongming Yin, Qiang Zhuge
The antimicrobial peptide ABP-dHC-cecropin A is a small cationic peptide with potent activity against a wide range of bacterial species. Evidence of antifungal activity has also been suggested; however, evaluation of this peptide has been limited due to the low expression of cecropin proteins in Escherichia coli. To improve the expression level of ABP-dHC-cecropin A in E. coli, tandem repeats of the ABP-dHC-cecropin A gene were constructed and expressed as fusion proteins (SUMO-nABP-dHC-cecropin, n = 1, 2, 3, 4) via pSUMO-nABP-dHC-cecropin A vectors (n = 1, 2, 3, 4)...
September 15, 2016: Analytical Biochemistry
Jiawen Xu, Christophe Morisseau, Jun Yang, Kin Sing Stephen Lee, Shizuo G Kamita, Bruce D Hammock
Epoxide hydrolases (EHs) are enzymes that play roles in metabolizing xenobiotic epoxides from the environment, and in regulating lipid signaling molecules, such as juvenile hormones in insects and epoxy fatty acids in mammals. In this study we fed mosquitoes with an epoxide hydrolase inhibitor AUDA during artificial blood feeding, and we found the inhibitor increased the concentration of epoxy fatty acids in the midgut of female mosquitoes. We also observed ingestion of AUDA triggered early expression of defensin A, cecropin A and cecropin B2 at 6 h after blood feeding...
September 2016: Insect Biochemistry and Molecular Biology
Xu-Biao Wei, Ru-Juan Wu, Da-Yong Si, Xiu-Dong Liao, Lu-Lu Zhang, Ri-Jun Zhang
Hybridizing different antimicrobial peptides (AMPs) is a particularly successful approach to obtain novel AMPs with increased antimicrobial activity but minimized cytotoxicity. The hybrid peptide cecropin A (1-8)-LL37 (17-30) (C-L) combining the hydrophobic N-terminal fragment of cecropin A (C) with the core antimicrobial fragment of LL37 (L) was designed and synthesized. C-L showed higher antibacterial activity against all indicator strains than C and L, and no hemolytic activity to sheep erythrocytes was observed...
2016: International Journal of Molecular Sciences
JiEun Yun, Dong Gun Lee
Cecropin A, isolated from the giant silk moth Hyalophora cecropia, is a 37-mer peptide that exerts potent antimicrobial effects. We investigated cecropin A-induced apoptosis associated with ion balance and redox state of Candida albicans. The antifungal effect of cecropin A, associated with ion movement was verified by significant increase of cell viability following pretreatment of ion channel blockers. Cecropin A induced undesired ion movement such as calcium accumulation and potassium leakage. Furthermore, the reduction of phosphatidylserine (PS) externalization was detected following pretreatment of ion channel blockers...
August 2016: IUBMB Life
Xiaoqiao Zhou, Xiaowen Li, Xiliang Wang, Xiue Jin, Deshi Shi, Jun Wang, Dingren Bi
Cecropins are peptide antibiotics used as drugs and feed additives. Cecropin B can inhibit the expression of CYP3A29, but the underlying mechanisms remain unclear. The present study was designed to determine the mechanisms responsible for the effects of cecropin B on CYP3A29 expression, focusing on the Toll-like receptors (TLRs) and NF-κB pathways. Our results indicated that the CYP3A29 expression was inhibited by cecropin B, which was regulated by pregnane X receptor (PXR) in a time- and dose-dependent manner...
2016: Scientific Reports
Akanksha Kaushal, Kajal Gupta, Ruhee Shah, Monique L van Hoek
Francisella tularensis is the cause of the zoonotic disease tularemia. In Sweden and Scandinavia, epidemiological studies have implicated mosquitoes as a vector. Prior research has demonstrated the presence of Francisella DNA in infected mosquitoes but has not shown definitive transmission of tularemia from a mosquito to a mammalian host. We hypothesized that antimicrobial peptides, an important component of the innate immune system of higher organisms, may play a role in mosquito host-defense to Francisella...
October 2016: Developmental and Comparative Immunology
H Tanaka, A Sagisaka, N Suzuki, M Yamakawa
E26 transformation-specific (Ets) family transcription factors are known to play roles in various biological phenomena, including immunity, in vertebrates. However, the mechanisms by which Ets proteins contribute to immunity in invertebrates remain poorly understood. In this study, we identified a cDNA encoding BmEts2, which is a putative orthologue of Drosophila Yan and human translocation-ets-leukemia/Ets-variant gene 6, from the silkworm Bombyx mori. Expression of the BmEts2 gene was significantly increased in the fat bodies of silkworm larvae in response to injection with Escherichia coli and Staphylococcus aureus...
October 2016: Insect Molecular Biology
Ingrid Faye, Bo G Lindberg
Four decades ago, immunological research was dominated by the field of lymphoid biology. It was commonly accepted that multicellular eukaryotes defend themselves through phagocytosis. The lack of lymphoid cells in insects and other simpler animals, however, led to the common notion that they might simply lack the capacity defend themselves with humoral factors. This view was challenged by microbiologist Hans G. Boman and co-workers in a series of publications that led to the advent of antimicrobial peptides as a universal arm of the immune system...
May 26, 2016: Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences
Carson Gill, Simon Bahrndorff, Carl Lowenberger
The house fly, Musca domestica, has been implicated as a vector of Campylobacter spp., a major cause of human disease. Little is known whether house flies serve as biological amplifying hosts or mechanical vectors for Campylobacter jejuni. We investigated the period after C. jejuni had been ingested by house flies in which viable C. jejuni colonies could be isolated from whole bodies, the vomitus and the excreta of adult M. domestica and evaluated the activation of innate immune responses of house flies to ingested C...
May 2, 2016: Insect Science
Gongqing Wu, Li Xu, Yunhong Yi
Larvae of Galleria mellonella are useful models for studying the innate immunity of invertebrates or for evaluating the virulence of microbial pathogens. In this work, we demonstrated that prior exposure of G. mellonella larvae to high doses (1×10(4), 1×10(5) or 1×10(6) cells/larva) of heat-killed Photorhabdus luminescens TT01 increases the resistance of larvae to a lethal dose (50 cells/larva) of viable P. luminescens TT01 infection administered 48h later. We also found that the changes in immune protection level were highly correlated to the changes in levels of cellular and humoral immune parameters when priming the larvae with different doses of heat-killed P...
June 2016: Immunology Letters
Mohammad Reza Bolouri Moghaddam, Miray Tonk, Christine Schreiber, Denise Salzig, Peter Czermak, Andreas Vilcinskas, Mohammad Rahnamaeian
Antimicrobial peptides (AMPs) are ubiquitous components of the insect innate immune system. The model insect Galleria mellonella has at least 18 AMPs, some of which are still uncharacterized in terms of antimicrobial activity. To determine why G. mellonella secretes a repertoire of distinct AMPs following an immune challenge, we selected three different AMPs: cecropin A (CecA), gallerimycin and cobatoxin. We found that cobatoxin was active against Micrococcus luteus at a minimum inhibitory concentration (MIC) of 120 μm, but at 60 μm when co-presented with 4 μm CecA...
September 1, 2016: Biological Chemistry
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