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  • 1
    Language: English
    In: eLife, 2013-12-03, Vol.2, p.e01096-e01096
    Description: Plant-herbivore interactions dominate the planet's terrestrial ecology. When it comes to host-plant specialization, insects are among the most versatile evolutionary innovators, able to disarm multiple chemical plant defenses. Sequestration is a widespread strategy to detoxify noxious metabolites, frequently for the insect's own benefit against predation. In this study, we describe the broad-spectrum ATP-binding cassette transporter CpMRP of the poplar leaf beetle, Chrysomela populi as the first candidate involved in the sequestration of phytochemicals in insects. CpMRP acts in the defensive glands of the larvae as a pacemaker for the irreversible shuttling of pre-selected metabolites from the hemolymph into defensive secretions. Silencing CpMRP in vivo creates a defenseless phenotype, indicating its role in the secretion process is crucial. In the defensive glands of related leaf beetle species, we identified sequences similar to CpMRP and assume therefore that exocrine gland-based defensive strategies, evolved by these insects to repel their enemies, rely on ABC transporters as a key element. DOI: http://dx.doi.org/10.7554/eLife.01096.001.
    Subject(s): ABC transporter ; ABC transporters ; Animals ; ATP-Binding Cassette Transporters - physiology ; Biochemistry ; Biosynthesis ; chemical defense ; Chrysomela lapponica ; Chrysomela populi ; Coleoptera - metabolism ; Ecology ; Exocrine glands ; fluorescence microscopy ; Glands ; Glucosides ; Glucosides - metabolism ; Hemolymph ; Host plants ; Insects ; Leaves ; Metabolites ; Morphology ; Other ; Phaedon cochleariae ; Phylogenetics ; Predation ; Proteins ; RNA interference ; Secretion ; Secretions ; Specialization
    ISSN: 2050-084X
    E-ISSN: 2050-084X
    Source: PubMed Central
    Source: DOAJ Directory of Open Access Journals - Not for CDI Discovery
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  • 2
    Language: English
    In: Frontiers in physiology, 2019-03-28, Vol.10, p.343-343
    Description: Herbivorous insects mainly rely on their sense of taste to decode the chemical composition of potential hosts in close range. Beetles for example contact and scan leaves with their tarsi, mouthparts and antennal tips, i.e., appendages equipped with gustatory sensilla, among other sensillum types. Gustatory neurons residing in such uniporous sensilla detect mainly non-volatile compounds that contribute to the behavioral distinction between edible and toxic plants. However, the identification of gustatory sensilla is challenging, because an appendage often possesses many sensilla of distinct morphological and physiological types. Using the specialized poplar leaf beetle (Chrysomela populi, Chrysomelidae), here we show that cuticular autofluorescence scanning combined with electron microscopy facilitates the identification of antennal gustatory sensilla and their differentiation into two subtypes. The gustatory function of sensilla chaetica was confirmed by single sensillum tiprecordings using sucrose, salicin and salt. Sucrose and salicin were found at higher concentrations in methanolic leaf extracts of poplar (Populus nigra) as host plant compared to willow (Salix viminalis) as control, and were found to stimulate feeding in feeding choice assays. These compounds may thus contribute to the observed preference for poplar over willow leaves. Moreover, these gustatory cues benefited the beetle's performance since weight gain was significantly higher when C. populi were reared on leaves of poplar compared to willow. Overall, our approach facilitates the identification of insect gustatory sensilla by taking advantage of their distinct fluorescent properties. This study also shows that a specialist beetle selects the plant species that provides optimal development, which is partly by sensing some of its characteristic non-volatile metabolites via antennal gustatory sensilla.
    Subject(s): antenna ; Beetles ; Chrysomela populi ; contact chemosensation ; cuticular autofluorescence ; Fluorescence ; gustation ; herbivory ; leaf beetle ; Life Sciences & Biomedicine ; Physiological aspects ; Physiology ; Science & Technology ; sensilla chaetica ; Taste
    ISSN: 1664-042X
    E-ISSN: 1664-042X
    Source: Web of Science - Science Citation Index Expanded - 2019〈img src="http://exlibris-pub.s3.amazonaws.com/fromwos-v2.jpg" /〉
    Source: PubMed Central
    Source: DOAJ Directory of Open Access Journals - Not for CDI Discovery
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  • 3
    Language: English
    In: PloS one, 2014, Vol.9 (6), p.e98637-e98637
    Description: Insects evolved ingenious adaptations to use extraordinary food sources. Particularly, the diet of herbivores enriched with noxious plant secondary metabolites requires detoxification mechanisms. Sequestration, which involves the uptake, transfer, and concentration of occasionally modified phytochemicals into specialized tissues or hemolymph, is one of the most successful detoxification strategies found in most insect orders. Due to the ability of ATP-binding cassette (ABC) carriers to transport a wide range of molecules including phytochemicals and xenobiotics, it is highly likely that they play a role in this sequestration process. To shed light on the role of ABC proteins in sequestration, we describe an inventory of putative ABC transporters in various tissues in the sequestering juvenile poplar leaf beetle, Chrysomela populi. In the transcriptome of C. populi, we predicted 65 ABC transporters. To link the proteins with a possible function, we performed comparative phylogenetic analyses with ABC transporters of other insects and of humans. While tissue-specific profiling of each ABC transporter subfamily suggests that ABCB, C and G influence the plant metabolite absorption in the gut, ABCC with 14 members is the preferred subfamily responsible for the excretion of these metabolites via Malpighian tubules. Moreover, salicin, which is sequestered from poplar plants, is translocated into the defensive glands for further deterrent production. In these glands and among all identified ABC transporters, an exceptionally high transcript level was observed only for Cpabc35 (Cpmrp). RNAi revealed the deficiency of other ABC pumps to compensate the function of CpABC35, demonstrating its key role during sequestration. We provide the first comprehensive phylogenetic study of the ABC family in a phytophagous beetle species. RNA-seq data from different larval tissues propose the importance of ABC pumps to achieve a homeostasis of plant-derived compounds and offer a basis for future analyses of their physiological function in sequestration processes.
    Subject(s): ABC transporter ; ABC transporters ; Adaptations ; Analysis ; Animals ; Arsenic ; ATP-Binding Cassette Transporters - genetics ; Beetles ; Biology and life sciences ; Cancer ; Cell adhesion & migration ; Chemical ecology ; Coleoptera - genetics ; Coleoptera - growth & development ; Detoxification ; Ecology and Environmental Sciences ; Excretion ; Food plants ; Food sources ; Gene expression ; Gene Expression Profiling ; Genomes ; Glands ; Hemolymph ; Herbivores ; Homeostasis ; Insects ; Larva - genetics ; Larvae ; Leaves ; Lipids ; Malpighian tubules ; Metabolism ; Metabolites ; Phylogeny ; Physiology ; Phytochemicals ; Plant metabolites ; Plants ; Poplar ; Profiling ; Proteins ; Pumps ; Real-Time Polymerase Chain Reaction ; Ribonucleic acid ; RNA ; RNA Interference ; RNA, Messenger - genetics ; RNA-mediated interference ; Salicin ; Secondary metabolites ; Sequestering ; Tissues ; Transcription ; Transporter ; Xenobiotics
    ISSN: 1932-6203
    E-ISSN: 1932-6203
    Source: Academic Search Ultimate
    Source: PubMed Central
    Source: DOAJ Directory of Open Access Journals - Not for CDI Discovery
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  • 4
    Language: English
    In: Scientific reports, 2016-04-12, Vol.6 (1), p.24082-24082
    Description: Extracellular Cu/Zn superoxide dismutases (SODs) are critical for balancing the level of reactive oxygen species in the extracellular matrix of eukaryotes. In the present study we have detected constitutive SOD activity in the haemolymph and defensive secretions of different leaf beetle species. Exemplarily, we have chosen the mustard leaf beetle, Phaedon cochleariae, as representative model organism to investigate the role of extracellular SODs in antimicrobial defence. Qualitative and quantitative proteome analyses resulted in the identification of two extracellular Cu/Zn SODs in the haemolymph and one in the defensive secretions of juvenile P. cochleariae. Furthermore, quantitative expression studies indicated fat body tissue and defensive glands as the main synthesis sites of these SODs. Silencing of the two SODs revealed one of them, PcSOD3.1, as the only relevant enzyme facilitating SOD activity in haemolymph and defensive secretions in vivo. Upon challenge with the entomopathogenic fungus, Metarhizium anisopliae, PcSOD3.1-deficient larvae exhibited a significantly higher mortality compared to other SOD-silenced groups. Hence, our results serve as a basis for further research on SOD regulated host-pathogen interactions. In defensive secretions PcSOD3.1-silencing affected neither deterrent production nor activity against fungal growth. Instead, we propose another antifungal mechanism based on MRJP/yellow proteins in the defensive exudates.
    Subject(s): Amino acids ; Animals ; Chemical ecology ; Coleoptera - growth & development ; Coleoptera - immunology ; Coleoptera - microbiology ; Entomopathogenic fungi ; Enzymes ; Eukaryotes ; Extracellular matrix ; Exudates ; Fat body ; Gene Silencing ; Glands ; Hemolymph ; Hemolymph - enzymology ; Hemolymph - immunology ; Host-pathogen interactions ; Insects ; Larva - immunology ; Larva - microbiology ; Metarhizium - immunology ; Metarhizium - pathogenicity ; Oxidation ; Oxidative stress ; Peptides ; Proteins ; Proteomes ; Reactive Oxygen Species - metabolism ; Secretions ; Superoxide Dismutase-1 - genetics ; Superoxide Dismutase-1 - metabolism ; Survival Analysis
    ISSN: 2045-2322
    E-ISSN: 2045-2322
    Source: Nature Open Access
    Source: Academic Search Ultimate
    Source: PubMed Central
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  • 5
    Language: English
    In: PloS one, 2013, Vol.8 (12), p.e84461-e84461
    Description: Phytophagous insects have emerged successfully on the planet also because of the development of diverse and often astonishing defensive strategies against their enemies. The larvae of the mustard leaf beetle Phaedon cochleariae, for example, secrete deterrents from specialized defensive glands on their back. The secretion process involves ATP-binding cassette transporters. Therefore, sugar as one of the major energy sources to fuel the ATP synthesis for the cellular metabolism and transport processes, has to be present in the defensive glands. However, the role of sugar transporters for the production of defensive secretions was not addressed until now. To identify sugar transporters in P. cochleariae, a transcript catalogue was created by Illumina sequencing of cDNA libraries. A total of 68,667 transcripts were identified and 68 proteins were annotated as either members of the solute carrier 2 (SLC2) family or trehalose transporters. Phylogenetic analyses revealed an extension of the mammalian GLUT6/8 class in insects as well as one group of transporters exhibiting distinctive conserved motifs only present in the insect order Coleoptera. RNA-seq data of samples derived from the defensive glands revealed six transcripts encoding sugar transporters with more than 3,000 counts. Two of them are exclusively expressed in the glandular tissue. Reduction in secretions production was accomplished by silencing two of four selected transporters. RNA-seq experiments of transporter-silenced larvae showed the down-regulation of the silenced transporter but concurrently the up-regulation of other SLC2 transporters suggesting an adaptive system to maintain sugar homeostasis in the defensive glands. We provide the first comprehensive phylogenetic study of the SLC2 family in a phytophagous beetle species. RNAi and RNA-seq experiments underline the importance of SLC2 transporters in defensive glands to achieve a chemical defense for successful competitive interaction in natural ecosystems.
    Subject(s): Adaptive systems ; Analysis ; Animals ; ATP ; ATP-binding protein ; Base Sequence ; Bayes Theorem ; Beetles ; Biology ; Chemical defense ; Chemistry ; Coleoptera - genetics ; Coleoptera - metabolism ; Deterrents ; DNA sequencing ; DNA, Complementary - genetics ; Energy sources ; Exocrine Glands - metabolism ; Fourier transforms ; Gas Chromatography-Mass Spectrometry ; Gene expression ; Gene Expression Profiling ; Gene Library ; Genomes ; Genomics ; Glands ; Homeostasis ; Insects ; Larva - metabolism ; Larvae ; Leaves ; Mammals ; Metabolism ; Metabolites ; Models, Genetic ; Molecular Sequence Data ; Monosaccharide Transport Proteins - genetics ; Monosaccharide Transport Proteins - metabolism ; Mustard ; Nucleotide sequencing ; Phylogenetics ; Phylogeny ; Physiological aspects ; Proteins ; Real-Time Polymerase Chain Reaction ; Ribonucleic acid ; RNA ; RNA Interference ; RNA-mediated interference ; Secretions ; Sequence Analysis, DNA ; Small intestine ; Sugar ; Transcription ; Transport processes ; Transporter ; Trehalose
    ISSN: 1932-6203
    E-ISSN: 1932-6203
    Source: Academic Search Ultimate
    Source: PubMed Central
    Source: DOAJ Directory of Open Access Journals - Not for CDI Discovery
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  • 6
    Language: English
    In: Ecology and evolution, 2018-08, Vol.8 (16), p.8055-8075
    Description: Due to its fundamental role in shaping host selection behavior, we have analyzed the chemosensory repertoire of Chrysomela lapponica. This specialized leaf beetle evolved distinct populations which shifted from the ancestral host plant, willow (Salix sp., Salicaceae), to birch (Betula rotundifolia, Betulaceae). We identified 114 chemosensory candidate genes in adult C. lapponica: 41 olfactory receptors (ORs), eight gustatory receptors, 17 ionotropic receptors, four sensory neuron membrane proteins, 32 odorant binding proteins (OBPs), and 12 chemosensory proteins (CSP) by RNA‐seq. Differential expression analyses in the antennae revealed significant upregulation of one minus‐C OBP (ClapOBP27) and one CSP (ClapCSP12) in the willow feeders. In contrast, one OR (ClapOR17), four minus‐C OBPs (ClapOBP02, 07, 13, 20), and one plus‐C OBP (ClapOBP32) were significantly upregulated in birch feeders. The differential expression pattern in the legs was more complex. To narrow down putative ligands acting as cues for host discrimination, the relative abundance and diversity of volatiles of the two host plant species were analyzed. In addition to salicylaldehyde (willow‐specific), both plant species differed mainly in their emission rate of terpenoids such as (E,E)‐α‐farnesene (high in willow) or 4,8‐dimethylnona‐1,3,7‐triene (high in birch). Qualitatively, the volatiles were similar between willow and birch leaves constituting an “olfactory bridge” for the beetles. Subsequent structural modeling of the three most differentially expressed OBPs and docking studies using 22 host volatiles indicated that ligands bind with varying affinity. We suggest that the evolution of particularly minus‐C OBPs and ORs in C. lapponica facilitated its host plant shift via chemosensation of the phytochemicals from birch as novel host plant. We test whether host plant shift of a specialist phytophagous beetle (Chrysomela lapponica) is accompanied by the modulation of its chemosensory repertoire. We present a comparative and tissue‐specific transcriptomic inventory of chemosensory genes (OR, OBP, CSP, GR, IR, and SNMP) from two distinct populations that differ in their preference for either willow or birch leaves. Combined with leaf volatile analyses and protein modeling, our results indicate that changes in the expression of mainly minus‐C OBPs and ORs are associated with host shift.
    Subject(s): chemosensory genes ; Chrysomela lapponica ; host plant shift ; leaf volatile analysis ; Original Research ; structural protein modeling ; transcriptomics
    ISSN: 2045-7758
    E-ISSN: 2045-7758
    Source: PubMed Central
    Source: DOAJ Directory of Open Access Journals - Not for CDI Discovery
    Source: ProQuest Central
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  • 7
    Language: English
    In: Journal of chemical ecology, 2016-03-31, Vol.42 (3), p.240-248
    Description: Juveniles of the leaf beetles in subtribe Chrysomelina have efficient defense strategies against predators. When disturbed, they transiently expose volatile deterrents in large droplets from nine pairs of defensive glands on their back. Here, we report on an additional line of defense consisting of the non-volatile isoxazolin-5-one glucoside and its 3-nitropropanoyl ester in the larval hemolymph. Because isoxazolin-5-one derivatives were not detectable in related leaf beetle taxa, they serve as a diagnostic marker for the Chrysomelina subtribe. Conjugation of isotopically labelled 3-nitropropionic acid to isoxazolin-5-one glucoside in vivo demonstrates its function as a carrier for the 3-nitropropanoyl esters. The previous identification of characteristic glucosides as precursors of the volatile deterrents underlines the general importance of glucosides for sequestration from food plants, and the subsequent transport in the hemolymph to the defense system. The combination of repellent volatiles with non-volatile toxic compounds in the hemolymph has the potential to create synergistic effects since the odorant stimulus may help predators learn to avoid some foods. The combination of the two defense lines has the advantage, that the hemolymph toxins provide reliable and durable protection, while the repellents may vary after a host plant change.
    Subject(s): 3-nitropropionate esters ; Agriculture ; Animals ; Aposematic warning ; Article ; Beetles ; Biochemistry ; Biochemistry, general ; Biological Microscopy ; Biomedical and Life Sciences ; Chemical defense ; Chrysomelidae ; Coleoptera - metabolism ; Coleoptera - physiology ; Ecology ; Entomology ; Esters ; Food processing plants ; general ; Hemolymph - metabolism ; Isoxazolin-5-one glucoside ; Life Sciences ; Nitro Compounds - metabolism ; Phylogeny ; Predatory Behavior ; Propionates - metabolism
    ISSN: 0098-0331
    E-ISSN: 1573-1561
    Source: Alma/SFX Local Collection
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  • 8
    Language: English
    In: Applied and Environmental Microbiology, 2004-02-01, Vol.70 (2), p.693-703
    Description: Classifications Services AEM Citing Articles Google Scholar PubMed Related Content Social Bookmarking CiteULike Delicious Digg Facebook Google+ Mendeley Reddit StumbleUpon Twitter current issue Spotlights in the Current Issue AEM About AEM Subscribers Authors Reviewers Advertisers Inquiries from the Press Permissions & Commercial Reprints ASM Journals Public Access Policy AEM RSS Feeds 1752 N Street N.W. • Washington DC 20036 202.737.3600 • 202.942.9355 fax • journals@asmusa.org Print ISSN: 0099-2240 Online ISSN: 1098-5336 Copyright © 2014 by the American Society for Microbiology.   For an alternate route to AEM .asm.org, visit: AEM       
    Subject(s): Anti-Bacterial Agents - pharmacology ; Antiporters - chemistry ; Antiporters - genetics ; Antiporters - metabolism ; Bacterial Proteins ; Berberine - pharmacology ; Biological and medical sciences ; Erwinia amylovora - drug effects ; Erwinia amylovora - genetics ; Erwinia amylovora - growth & development ; Erwinia amylovora - pathogenicity ; Ethidium - pharmacology ; Flowers - microbiology ; Fundamental and applied biological sciences. Psychology ; Gene Expression Regulation, Bacterial ; Malus - microbiology ; Microbial Sensitivity Tests ; Microbiology ; Molecular Sequence Data ; Norfloxacin - pharmacology ; Pantoea - growth & development ; Plant Diseases - microbiology ; Plant Microbiology ; Polymerase Chain Reaction ; Rosaceae - microbiology ; Virulence
    ISSN: 0099-2240
    E-ISSN: 1098-5336
    Source: HighWire Press (Free Journals)
    Source: Hellenic Academic Libraries Link
    Source: PubMed Central
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  • 9
    Language: English
    In: Natural product reports, 2017-05-10, Vol.34 (5), p.478-483
    Description: Contact chemosensation, or tasting, is a complex process governed by nonvolatile phytochemicals that tell host-seeking insects whether they should accept or reject a plant. During this process, insect gustatory receptors (GRs) contribute to deciphering a host plant's metabolic code. GRs recognise many different classes of nonvolatile compounds; some GRs are likely to be narrowly tuned and others, broadly tuned. Although primary and/or secondary plant metabolites influence the insect's feeding choice, their decoding by GRs is challenging, because metabolites in planta occur in complex mixtures that have additive or inhibitory effects; in diverse forms composed of structurally unrelated molecules; and at different concentrations depending on the plant species, its tissue and developmental stage. Future studies of the mechanism of insect herbivore GRs will benefit from functional characterisation taking into account the spatio-temporal dynamics and diversity of the plant's metabolome. Metabolic information, in turn, will help to elucidate the impact of single ligands and complex natural mixtures on the insect's feeding choice. Contact chemosensation, or tasting, enables insect herbivores to identify nonvolatile metabolites in complex mixtures present in plants. The interplay of primary and secondary plant metabolites with gustatory receptors is outlined.
    Subject(s): Animals ; Chemistry ; Herbivory ; Insecta - physiology ; Molecular Structure ; Phytochemicals - chemistry ; Phytochemicals - pharmacology
    ISSN: 0265-0568
    E-ISSN: 1460-4752
    Source: Alma/SFX Local Collection
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  • 10
    Language: English
    In: Proceedings of the National Academy of Sciences - PNAS, 2013-03-12, Vol.110 (11), p.4194-4199
    Description: Isoprenyl diphosphate synthases (IDSs) produce the ubiquitous branched-chain diphosphates of different lengths that are precursors of all major classes of terpenes. Typically, individual short-chain IDSs (scIDSs) make the C₁₀, C₁₅, and C₂₀ isoprenyl diphosphates separately. Here, we report that the product length synthesized by a single scIDS shifts depending on the divalent metal cofactor present. This previously undescribed mechanism of carbon chain-length determination was discovered for a scIDS from juvenile horseradish leaf beetles, Phaedon cochleariae. The recombinant enzyme P. cochleariae isoprenyl diphosphate synthase 1 (PcIDS1) yields 96% C₁₀-geranyl diphosphate (GDP) and only 4% C₁₅-farnesyl diphosphate (FDP) in the presence of Co²⁺ or Mn²⁺ as a cofactor, whereas it yields only 18% C₁₀ GDP but 82% C₁₅ FDP in the presence of Mg²⁺. In reaction with Co²⁺, PcIDS1 has a Km of 11.6 µM for dimethylallyl diphosphate as a cosubstrate and 24.3 µM for GDP. However, with Mg²⁺, PcIDS1 has a Km of 1.18 µM for GDP, suggesting that this substrate is favored by the enzyme under such conditions. RNAi targeting PcIDS1 revealed the participation of this enzyme in the de novo synthesis of defensive monoterpenoids in the beetle larvae. As an FDP synthase, PcIDS1 could be associated with the formation of sesquiterpenes, such as juvenile hormones. Detection of Co²⁺, Mn²⁺, or Mg²⁺ in the beetle larvae suggests flux control into C₁₀ vs. C₁₅ isoprenoids could be accomplished by these ions in vivo. The dependence of product chain length of scIDSs on metal cofactor identity introduces an additional regulation for these branch point enzymes of terpene metabolism.
    Subject(s): Alkyl and Aryl Transferases - chemistry ; Alkyl and Aryl Transferases - genetics ; Alkyl and Aryl Transferases - metabolism ; Amino Acid Sequence ; Animals ; Biological Sciences ; Biosynthesis ; Body tissues ; cobalt ; Cobalt - chemistry ; Cobalt - metabolism ; Coleoptera - enzymology ; Coleoptera - genetics ; Diphosphates ; Diphosphates - chemistry ; Diphosphates - metabolism ; Diterpenes - chemistry ; Diterpenes - metabolism ; Enzymes ; Fat body ; Insect biochemistry ; Insect larvae ; Insect Proteins - chemistry ; Insect Proteins - genetics ; Insect Proteins - metabolism ; Insects ; kinetic ; Kinetics ; Manganese - chemistry ; Manganese - metabolism ; Metal ions ; Molecular Sequence Data ; Physical Sciences ; Physiological aspects ; Polyisoprenyl Phosphates - biosynthesis ; Polyisoprenyl Phosphates - chemistry ; prenyltransferase ; Recombinant Proteins - chemistry ; Recombinant Proteins - genetics ; Recombinant Proteins - metabolism ; Research ; secretions ; Sesquiterpenes - chemistry ; silencing ; Terpenes ; Terpenoids
    ISSN: 0027-8424
    E-ISSN: 1091-6490
    Source: JSTOR Life Sciences
    Source: HighWire Press (Free Journals)
    Source: Hellenic Academic Libraries Link
    Source: PubMed Central
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