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1.
Proc Natl Acad Sci U S A ; 119(42): e2211254119, 2022 10 18.
Artigo em Inglês | MEDLINE | ID: mdl-36227916

RESUMO

Iridoid monoterpenes, widely distributed in plants and insects, have many ecological functions. While the biosynthesis of iridoids has been extensively studied in plants, little is known about how insects synthesize these natural products. Here, we elucidated the biosynthesis of the iridoids cis-trans-nepetalactol and cis-trans-nepetalactone in the pea aphid Acyrthosiphon pisum (Harris), where they act as sex pheromones. The exclusive production of iridoids in hind legs of sexual female aphids allowed us to identify iridoid genes by searching for genes specifically expressed in this tissue. Biochemical characterization of candidate enzymes revealed that the iridoid pathway in aphids proceeds through the same sequence of intermediates as described for plants. The six identified aphid enzymes are unrelated to their counterparts in plants, conclusively demonstrating an independent evolution of the entire iridoid pathway in plants and insects. In contrast to the plant pathway, at least three of the aphid iridoid enzymes are likely membrane bound. We demonstrated that a lipid environment facilitates the cyclization of a reactive enol intermediate to the iridoid cyclopentanoid-pyran scaffold in vitro, suggesting that membranes are an essential component of the aphid iridoid pathway. Altogether, our discovery of this complex insect metabolic pathway establishes the genetic and biochemical basis for the formation of iridoid sex pheromones in aphids, and this discovery also serves as a foundation for understanding the convergent evolution of complex metabolic pathways between kingdoms.


Assuntos
Afídeos , Produtos Biológicos , Atrativos Sexuais , Animais , Afídeos/genética , Afídeos/metabolismo , Produtos Biológicos/metabolismo , Iridoides/química , Iridoides/metabolismo , Lipídeos , Monoterpenos/metabolismo , Feromônios/metabolismo , Plantas/metabolismo , Atrativos Sexuais/genética , Atrativos Sexuais/metabolismo
2.
Public Health Nurs ; 2024 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-38946470

RESUMO

OBJECTIVE: Social residents become increasingly concerned about Alzheimer's dementia (AD) as a global public health crisis. China's AD population is the largest and growing fastest. However, no study has examined Chinese social residents' knowledge and attitudes concerning Alzheimer's illness. This study examined Chinese social residents' AD knowledge and attitudes using the Alzheimer's Disease Knowledge Scale (ADKS) and dementia attitudes scale (DAS). DESIGN: Cross-sectional survey. SAMPLE: 338 social residents over 18 years old from various Chinese regions were recruited using convenient sampling. MEASUREMENTS: The ADKS (Chinese) and the Dementia Attitude Scale (Chinese) were used to assess their knowledge and attitude regarding AD. RESULTS: A total of 328 respondents (97.04%) completed the survey. ADKS = 19.44 ± 3.33; DAS = 86.98 ± 12.7. Age and education levels can have a substantial impact on ADKS scores, and education levels can have a substantial impact on DAS scores. CONCLUSIONS: Low levels of awareness and acceptance of AD exist among Chinese residents. The results indicate that China must immediately implement comprehensive AD education for its social residents.

3.
BMC Genomics ; 24(1): 781, 2023 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-38102559

RESUMO

BACKGROUND: Odorant-binding proteins (OBPs) are essential in insect's daily behaviors mediated by olfactory perception. Megachile saussurei Radoszkowski (Hymenoptera, Megachilidae) is a principal insect pollinating alfalfa (Medicago sativa) in Northwestern China. The olfactory function have been less conducted, which provides a lot of possibilities for our research. RESULTS: Our results showed that 20 OBPs were identified in total. Multiple sequence alignment analysis indicated MsauOBPs were highly conserved with a 6-cysteine motif pattern and all belonged to the classic subfamily, coding 113-196 amino acids and sharing 41.32%-99.12% amino acid identity with known OBPs of other bees. Phylogenetic analysis indicated there were certain homologies existed among MsauOBPs and most sequences were clustered with that of Osmia cornuta (Hymenoptera, Megachilidae). Expression analysis showed the identified OBPs were mostly enriched in antennae instead of other four body parts, especially the MsauOBP2, MsauOBP3, MsauOBP4, MsauOBP8, MsauOBP11 and MsauOBP17, in which the MsauOBP2, MsauOBP4 and MsauOBP8 presented obvious tissue-biased expression pattern. Molecular docking results indicated MsauOBP4 might be the most significant protein in recognizing alfalfa flower volatile 3-Octanone, while MsauOBP13 might be the most crucial protein identifying (Z)-3-hexenyl acetate. It was also found the lysine was a momentous hydrophilic amino acid in docking simulations. CONCLUSION: In this study, we identified and analyzed 20 OBPs of M. saussurei. The certain homology existed among these OBPs, while some degree of divergence could also be noticed, indicating the complex functions that different MsauOBPs performed. Besides, the M. saussurei and Osmia cornuta were very likely to share similar physiological functions as most of their OBPs were clustered together. MsauOBP4 might be the key protein in recognizing 3-Octanone, while MsauOBP13 might be the key protein in binding (Z)-3-hexenyl acetate. These two proteins might contribute to the alfalfa-locating during the pollination process. The relevant results may help determine the highly specific and effective attractants for M. saussurei in alfalfa pollination and reveal the molecular mechanism of odor-evoked pollinating behavior between these two species.


Assuntos
Himenópteros , Receptores Odorantes , Abelhas , Animais , Himenópteros/metabolismo , Odorantes , Sequência de Aminoácidos , Filogenia , Simulação de Acoplamento Molecular , Perfilação da Expressão Gênica , Receptores Odorantes/genética , Receptores Odorantes/metabolismo , Aminoácidos/metabolismo , Proteínas de Insetos/metabolismo , Antenas de Artrópodes/metabolismo , Transcriptoma
4.
Environ Res ; 237(Pt 2): 116939, 2023 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-37611781

RESUMO

The UV/sulfite system is a promising source of •SO4- and/or •OH, but its application is largely limited by the use of UV light due to its high cost and high energy consumption. Graphite carbon nitride (g-C3N5), as a new photocatalytic material, has better visible light absorption capacity and narrower band gap than g-C3N4, which is expected to activate sulfite under visible light to solve this problem. Herein, a novel FeS2/CN heterojunction material based on g-C3N5 was constructed by hydrothermal in-situ synthesis method and successfully activated sulfite, which was confirmed by tetracycline degradation experiments in water. Under optimized conditions, the degradation rate of TC in 1 h reached 96%. The experimental results revealed that the FeS2/CN heterostructure enhances the absorption of visible light and inhibits the recombination of carriers, enabling more electrons and holes to be utilized. Holes play a major role in the degradation reaction, promote the sulfite chain reaction, and effectively regulate the cycle of Fe2+ and Fe3+ in the solution. Iron ion leaching is negligible and the degradation reaction remains stable at pH 5-9.

5.
Front Zool ; 19(1): 33, 2022 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-36517816

RESUMO

BACKGROUND: The cotton mealybug Phenacoccus solenopsis Tinsley (Hemiptera: Pseudococcidae) is one of the most devastating sap-sucking pests of cultivated plants. The success of P. solenopsis is attributable to its ecological resilience and insecticide resistance, making its control extremely difficult and expensive. Thus, alternative safe approaches are needed to prevent the pest population from reaching the economic threshold. One of these novel approaches is based on the fact that chemical communication via the olfactory system drives critical behaviors required for the survival and development of the species. This knowledge can be useful for controlling insect pests using traps based on semiochemicals. The antennae of insects are an invaluable model for studying the fundamentals of odor perception. Several efforts have been made to investigate the histological and ultrastructural organization of the olfactory organs, such as the antennae and maxillary palps, in many insect species. However, studies on the antennal sensory structures of Phenacoccus species are lacking. Furthermore, although enormous progress has been made in understanding the antennal structures of many mealybug species, the olfactory sensilla in the antennae of P. solenopsis have not yet been described. In this study, we describe, for the first time, the morphology and distribution of the antennal sensilla in male and female P. solenopsis using scanning electron microscopy. RESULTS: Our results revealed that the entire antennae length and the number of flagellar segments were different between the sexes. Eight morphological types of sensilla were identified on male antennae: trichoid sensilla, chaetic sensilla (three subtypes), basiconic sensilla (two subtypes), and campaniform sensilla (two subtypes). Six morphological types of sensilla were found on female antennae. Sensilla chaetica of subtype 2 and campaniform sensilla of subtype 1 were distributed only on male antennae, suggesting that these sensilla are involved in the recognition of female sex pheromones. The subtype 1 of sensilla chaetica was significantly more abundant on female antennae than on male ones, while subtype 3 was only located on the terminal flagellar segment of the antenna in both sexes. CONCLUSIONS: This study provides insightful information for future electrophysiological and behavioral studies on chemical communication in insects, particularly the cotton mealybug, P. solenopsis that could help in developing new strategies for controlling this economically important insect species.

6.
PLoS Genet ; 15(10): e1008419, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31609971

RESUMO

Microorganisms sense environmental fluctuations in nutrients and light, coordinating their growth and development accordingly. Despite their critical roles in fungi, only a few G-protein coupled receptors (GPCRs) have been characterized. The Aspergillus nidulans genome encodes 86 putative GPCRs. Here, we characterise a carbon starvation-induced GPCR-mediated glucose sensing mechanism in A. nidulans. This includes two class V (gprH and gprI) and one class VII (gprM) GPCRs, which in response to glucose promote cAMP signalling, germination and hyphal growth, while negatively regulating sexual development in a light-dependent manner. We demonstrate that GprH regulates sexual development via influencing VeA activity, a key light-dependent regulator of fungal morphogenesis and secondary metabolism. We show that GprH and GprM are light-independent negative regulators of sterigmatocystin biosynthesis. Additionally, we reveal the epistatic interactions between the three GPCRs in regulating sexual development and sterigmatocystin production. In conclusion, GprH, GprM and GprI constitute a novel carbon starvation-induced glucose sensing mechanism that functions upstream of cAMP-PKA signalling to regulate fungal development and mycotoxin production.


Assuntos
Adaptação Fisiológica/efeitos da radiação , Aspergillus nidulans/fisiologia , Proteínas Fúngicas/metabolismo , Luz , Receptores Acoplados a Proteínas G/metabolismo , Carbono/metabolismo , Perfilação da Expressão Gênica , Regulação Fúngica da Expressão Gênica/efeitos da radiação , Glucose/metabolismo , Morfogênese , Esporos Fúngicos/crescimento & desenvolvimento , Esporos Fúngicos/efeitos da radiação , Esterigmatocistina/biossíntese
7.
Ecotoxicol Environ Saf ; 245: 114101, 2022 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-36155334

RESUMO

Recent studies have indicated that the plant volatile methyl benzoate (MB) exhibits significant insecticidal bioactivity against several common insects. However, the potential environmental hazards of MB and its safety to non-target organisms is poorly understood. In the present study, these characteristics were investigated through laboratory experiments and field investigations. The results revealed that MB was highly toxic to the agricultural pest, fall armyworm Spodoptera frugiperda. Compared with the commercial pesticide lambda-cyhalothrin, the toxicities of MB against S. frugiperda larvae and adults were comparable and 3.41 times higher, respectively. Behavioral bioassays showed that the percentage repellency of MB to S. frugiperda larvae was 56.72 %, and MB induced 69.40 % oviposition deterrence rate in S. frugiperda female adults. Furthermore, in terms of median lethal concentration (LC50) and median lethal doses (LD50), MB exhibited non-toxic effects on non-target animals with 3-d LC50 of > 1 % to natural predators (Coccinella septempunctata and Harmonia axyridis), 3-d LD50 of 467.86 µg/bee to the bumblebee Bombus terrestris, 14-d LC50 of 971.09 mg/kg to the earthworm Eisenia fetida, and 4-d LC50 of 47.30 mg/L to the zebrafish Brachydanio rerio. The accumulation of MB in the soil and earthworms was found to be extremely limited. Our comparative study clearly demonstrated that MB is effective as a selective botanical pesticide against S. frugiperda and it is safe to use in the tested environment, with no toxic effects on non-target animals and natural predators.


Assuntos
Besouros , Inseticidas , Oligoquetos , Animais , Benzoatos , Feminino , Inseticidas/toxicidade , Larva , Solo , Spodoptera , Peixe-Zebra
8.
Lasers Med Sci ; 37(3): 1605-1614, 2022 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-34480664

RESUMO

It had been suggested, after facilitating the hatching process, improved pregnancy outcomes could be attained in embryos with thick and hard zona. This study aimed to determine the effect of zona thinning on pregnancy outcomes in poor-quality frozen-thawed blastocysts. This retrospective study included 230 women (≤ 40 years) who underwent frozen embryo transfer of poor-quality blastocysts (scored < 3BB). In total, 105 patients were in the assisted hatching group in which the zona was thinned by laser before transfer and 125 patients were in the control group in which the blastocysts were non-manipulated. Patients' demographics, cycle characteristics, and pregnancy outcomes were compared between the assisted hatching group and the control group. Further, regression analysis was applied to test the correlation between assisted hatching and live birth. All parameters in the patients' demographic characteristics and the cycle's characteristics were not significantly different between two groups. As for pregnancy outcomes, the second trimester pregnancy loss was significantly higher in the assisted hatching group (P = 0.035). Other pregnancy outcomes, including implantation rate, clinical pregnancy rate, biochemical miscarriage rate, the first trimester pregnancy loss, ongoing pregnancy rate, and live birth rate were comparable between two groups. The logistic regression analysis demonstrated no association between live birth and assisted hatching (univariate, OR = 0.787, P > 0.05; multivariate, OR = 0.652, P > 0.05), and the area under the receiver operating characteristic curve of the regression model was almost 0.7. It suggested that zona thinning may not be supposed to perform on poor-quality, frozen-thawed blastocysts. The indications of assisted hatching were still needed to further investigate.


Assuntos
Criopreservação , Resultado da Gravidez , Blastocisto , Transferência Embrionária , Feminino , Humanos , Lasers , Gravidez , Estudos Retrospectivos
9.
Plant Cell Environ ; 44(3): 948-963, 2021 03.
Artigo em Inglês | MEDLINE | ID: mdl-33099790

RESUMO

Plant defence homoterpenes can be used to attract pest natural enemies. However, the biosynthetic pathway of homoterpenes is still unknown in rice, and the practical application of such indirect defence systems suffers from inherent limitations due to their low emissions from plants. Here, we demonstrated that the protein OsCYP92C21 is responsible for homoterpene biosynthesis in rice. We also revealed that the ability of rice to produce homoterpenes is dependent on the subcellular precursor pools. By increasing the precursor pools through specifically subcellular targeting expression, genetic transformation and genetic introgression, we significantly enhanced homoterpene biosynthesis in rice. The final introgressed GM rice plants exhibited higher homoterpene emissions than the wild type rice and the highest homoterpene emission reported so far for such GM plants even without the induction of herbivore attack. As a result, these GM rice plants demonstrated strong attractiveness to the parasitic wasp Cotesia chilonis. This study discovered the homoterpene biosynthesis pathway in rice, and lays the foundation for the utilisation of plant indirect defence mechanism in the "push-pull" strategy of integrated pest management through increasing precursor pools in the subcellular compartments and overexpressing homoterpene synthase by genetic transformation.


Assuntos
Alquil e Aril Transferases/metabolismo , Oryza/metabolismo , Defesa das Plantas contra Herbivoria , Proteínas de Plantas/metabolismo , Compostos Orgânicos Voláteis/metabolismo , Animais , Proteína 9 Associada à CRISPR , Sistemas CRISPR-Cas , Edição de Genes , Técnicas de Inativação de Genes , Oryza/genética , Plantas Geneticamente Modificadas , Reação em Cadeia da Polimerase em Tempo Real , Terpenos/metabolismo , Vespas
10.
Opt Lett ; 46(18): 4614-4617, 2021 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-34525061

RESUMO

Optical trapping of single nanoparticles in vacuum has various applications in both precise measurements and fundamental physics. However, to date, the number and size of randomly loaded nanoparticles in an optical trap is difficult to determine unless in vacuum. In this Letter, an efficient method for nanoparticle size estimation in an optical tweezer system before the evacuation of air was proposed and demonstrated experimentally, using scattering light from levitated particles. The particle radii deduced from the scattering light power in our proposal and from the kinetic theory of particles in gas match well (with the differences of less than 10%). For sample particles with radii ranging within 50-100 nm, we also provide a preselection rule based on this method, where over half of the trapped particles are verified as single particles. Such a particle analysis method is applicable also for the size estimation of levitated diamond particles, gold particles, and other plasmonic particles and can be applied to discovering novel scattering effects.

11.
BMC Genomics ; 21(1): 242, 2020 Mar 18.
Artigo em Inglês | MEDLINE | ID: mdl-32183717

RESUMO

BACKGROUND: The fall webworm Hyphantria cunea is an invasive and polyphagous defoliator pest that feeds on nearly any type of deciduous tree worldwide. The silk web of H. cunea aids its aggregating behavior, provides thermal regulation and is regarded as one of causes for its rapid spread. In addition, both chemosensory and detoxification genes are vital for host adaptation in insects. RESULTS: Here, a high-quality genome of H. cunea was obtained. Silk-web-related genes were identified from the genome, and successful silencing of the silk protein gene HcunFib-H resulted in a significant decrease in silk web shelter production. The CAFE analysis showed that some chemosensory and detoxification gene families, such as CSPs, CCEs, GSTs and UGTs, were expanded. A transcriptome analysis using the newly sequenced H. cunea genome showed that most chemosensory genes were specifically expressed in the antennae, while most detoxification genes were highly expressed during the feeding peak. Moreover, we found that many nutrient-related genes and one detoxification gene, HcunP450 (CYP306A1), were under significant positive selection, suggesting a crucial role of these genes in host adaptation in H. cunea. At the metagenomic level, several microbial communities in H. cunea gut and their metabolic pathways might be beneficial to H. cunea for nutrient metabolism and detoxification, and might also contribute to its host adaptation. CONCLUSIONS: These findings explain the host and environmental adaptations of H. cunea at the genetic level and provide partial evidence for the cause of its rapid invasion and potential gene targets for innovative pest management strategies.


Assuntos
Adaptação Fisiológica/genética , Espécies Introduzidas , Mariposas/classificação , Mariposas/genética , Animais , Sequência de Bases , Perfilação da Expressão Gênica , Genoma , Filogenia
12.
Anal Chem ; 92(9): 6538-6547, 2020 05 05.
Artigo em Inglês | MEDLINE | ID: mdl-32255342

RESUMO

In this work, we designed and fabricated Pd/Au bimetallic thin film electrodes with isolated Pd nanoparticles via underpotential deposition of copper on a gold substrate followed by in situ redox replace reaction in a Pd salt solution. The Pd/Au electrode was characterized by AFM and XPS as well as multiple electrochemical techniques including CV and electrochemical quartz crystal microbalance (EQCM) in sulfuric acid and phosphate buffer electrolytes. Results show that the reduction reactions of the analytes (i.e., H2O2 and 3-nitrotyrosine (3-NT)) at the Pd/Au thin film surfaces affect the nature and reactivity of Pd/Au surface electrochemistry including the adsorbed/absorbed hydrogen and/or the premonolayer palladium oxide redox processes at Pd. The EQCM experiment supports the arrangement of small size Pd nanoparticles in the Pd thin film in the presence of gold exhibits unusual properties, acting as a new physicochemical dimension between the electrode and target H2O2 and 3-NT molecules. The Pd/Au thin film was demonstrated as an extremely sensitive and selective probe for detection of common ROS and RNS (i.e., H2O2 and 3-NT). The integration of two different metallic species, Pd and Au, into a surface structure on nanoscale by exploiting their unique surface electrochemistry establishes an innovative analytical method for highly sensitive and selective detection of H2O2 and 3-NT simultaneously. This method has a general scope for detecting a broad range of redox active and nonredox active species simultaneously, which opens up new opportunities to develop new electrocatalytic materials and innovative sensing approaches.

13.
Mol Ecol ; 29(19): 3795-3808, 2020 10.
Artigo em Inglês | MEDLINE | ID: mdl-32681685

RESUMO

Migratory insects are capable of actively sustaining powered flight for several hours. This extraordinary phenomenon requires a highly efficient transport system to cope with the energetic demands placed on the flight muscles. Here, we provide evidence that the role of the hydrophobic ligand binding of odorant binding proteins (OBPs) extends beyond their typical function in the olfactory system to support insect flight activity via lipid interactions. Transcriptomic and candidate gene analyses show that two phylogenetically clustered OBPs (OBP3/OBP6) are consistently over-expressed in adult moths of the migrant Old-World bollworm, Helicoverpa armigera, displaying sustained flight performance in flight activity bioassays. Tissue-specific over-expression of OBP6 was observed in the antennae, wings and thorax in long-fliers of H. armigera. Transgenic Drosophila flies over-expressing an H. armigera transcript of OBP6 (HarmOBP6) in the flight muscle attained higher flight speeds on a modified tethered flight system. Quantification of lipid molecules using mass spectrometry showed a depletion of triacylglyerol and phospholipids in flown moths. Protein homology models built from the crystal structure of a fatty acid carrier protein identified the binding site of OBP3 and OBP6 for hydrophobic ligand binding with both proteins exhibiting a stronger average binding affinity with triacylglycerols and phospholipids compared with other groups of ligands. We propose that HarmOBP3 and HarmOBP6 contribute to the flight capacity of a globally invasive and highly migratory noctuid moth, and in doing so, extend the function of this group of proteins beyond their typical role as chemosensory proteins in insects.


Assuntos
Mariposas , Receptores Odorantes , Animais , Proteínas de Transporte/genética , Proteínas de Insetos/genética , Proteínas de Insetos/metabolismo , Mariposas/genética , Odorantes , Receptores Odorantes/genética , Transcriptoma
14.
Genome ; 63(1): 1-12, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31533014

RESUMO

The larvae of Holotrichia parallela, a destructive belowground herbivore, causes tremendous damages to maize plants. However, little is known if there are any defense mechanisms in maize roots to defend themselves against this herbivore. In the current research, we carried out RNA-sequencing to investigate the changes in gene transcription level in maize roots after H. parallela larvae infestation. A total of 644 up-regulated genes and 474 down-regulated genes was found. In addition, Gene ontology (GO) annotation analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed. Weighted gene co-expression network analysis (WGCNA) indicated that peroxidase genes may be the hub genes that regulate maize defenses to H. parallela larvae attack. We also found 105 transcription factors, 44 hormone-related genes, and 62 secondary metabolism-related genes within differentially expressed genes (DEGs). Furthermore, the expression profiles of 12 DEGs from the transcriptome analysis were confirmed by quantitative real-time PCR experiments. This transcriptome analysis provides insights into the molecular mechanisms of the underground defense in maize roots to H. parallela larvae attack and will help to select target genes of maize for defense against belowground herbivory.


Assuntos
Besouros/fisiologia , Herbivoria/genética , Zea mays/genética , Animais , Besouros/crescimento & desenvolvimento , Regulação da Expressão Gênica de Plantas , Ontologia Genética , Genes de Plantas , Larva/fisiologia , Reguladores de Crescimento de Plantas/fisiologia , Raízes de Plantas/genética , Raízes de Plantas/metabolismo , RNA-Seq , Reação em Cadeia da Polimerase em Tempo Real , Metabolismo Secundário/genética , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Transcriptoma , Zea mays/metabolismo
15.
J Surg Res ; 249: 145-155, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-31958599

RESUMO

BACKGROUND: Emerging evidence has linked autophagy to skin wound healing; however, the underlying cellular and molecular mechanisms remain poorly understood. The present study was designed to determine the role of autophagy in endothelial cell (EC)-mediated skin wound healing in mice. METHODS: Autophagy-related gene (Atg7) in mouse ECs was inactivated by the Cre-loxP system under the control of an EC-specific VE-Cadherin (Cdh5) promoter (Atg7EC-/- mice). Full-thickness skin wounds were created on the dorsum of wild-type (WT), Cdh5-Cre+, floxed Atg7 (Atg7F/F), and Atg7EC-/- mice. Autophagic activity was determined by autophagic flux assay in the primary culture of ECs isolated from these mice. The wound re-epithelialization and angiogenesis was examined by histological analyses. The angiogenic activity of ECs was evaluated by tube formation assay in vitro. EC proliferation was examined by a cell count CCK-8 kit. EC-originated intercellular communication with dermal fibroblasts and keratinocytes was assessed by measuring the effect of EC conditional medium on the growth of keratinocytes and fibroblasts. The levels of VEGF, EGF, bFGF in EC conditional medium were measured by ELISA. RESULTS: Autophagy deficiency in ECs markedly enhanced the re-epithelialization and the wound closure during skin wound healing. However, it has minimal impact on angiogenesis in the wounded skin. Notably, autophagy deficiency in ECs did not affect their proliferation and migration or angiogenic activity per se but enhanced the EC conditional medium-induced proliferation and migration of keratinocytes and fibroblasts. CONCLUSIONS: These results demonstrate for the first time an inhibitory role of autophagy in the EC-originated paracrine regulation of skin wound healing.


Assuntos
Proteína 7 Relacionada à Autofagia/genética , Autofagia/genética , Células Endoteliais/imunologia , Ferida Cirúrgica/imunologia , Cicatrização/imunologia , Animais , Autofagia/imunologia , Comunicação Celular/genética , Comunicação Celular/imunologia , Movimento Celular/genética , Proliferação de Células/genética , Células Cultivadas , Meios de Cultivo Condicionados/metabolismo , Modelos Animais de Doenças , Endotélio Vascular/citologia , Feminino , Fibroblastos , Humanos , Queratinócitos , Masculino , Camundongos , Camundongos Knockout , Miocárdio/citologia , Neovascularização Fisiológica/genética , Neovascularização Fisiológica/imunologia , Comunicação Parácrina/genética , Comunicação Parácrina/imunologia , Cultura Primária de Células , Pele/irrigação sanguínea , Pele/lesões
16.
Arch Insect Biochem Physiol ; 101(3): e21557, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31062883

RESUMO

The European grapevine moth, Lobesia botrana (Denis & Schiffermüller), is a serious pest in vineyards in North and South America. Mating disruption techniques have been used to control and monitor L. botrana on the basis of its sexual communication. This needs a well-tuned olfactory system, in which it is believed that pheromone-binding proteins (PBPs) are key players that transport pheromones in the antennae of moths. In this study, the selectivity of a PBP, named as LbotPBP1, was tested by fluorescence binding assays against 11 sex pheromone components and 6 host plant volatiles. In addition, its binding mechanism was predicted on the basis of structural analyses by molecular docking and complex and steered molecular dynamics (SMD). Our results indicate that LbotPBP1 binds selectively to sex pheromone components over certain host plant volatiles, according to both in vitro and in silico tests. Thus, chain length (14 carbon atoms) and functional groups (i.e., alcohol and ester) appear to be key features for stable binding. Likewise, residues such as Phe12, Phe36, and Phe118 could participate in unspecific binding processes, whilst Ser9, Ser56, and Trp114 could participate in the specific recognition and stabilization of sex pheromones instead of host plant volatiles. Moreover, our SMD approach supported 11-dodecenyl acetate as the best ligand for LbotPBP1. Overall, the dynamics simulations, contact frequency analysis and SMD shed light on the binding mechanism of LbotPBP1 and could overcome the imprecision of molecular docking, supporting the in vitro binding assays. Finally, the role of LbotPBP1 in the chemical ecology of L. botrana is discussed.


Assuntos
Proteínas de Transporte/genética , Proteínas de Insetos/genética , Mariposas/genética , Atrativos Sexuais/metabolismo , Animais , Proteínas de Transporte/metabolismo , Proteínas de Insetos/metabolismo , Masculino , Simulação de Acoplamento Molecular , Mariposas/metabolismo , Ligação Proteica
17.
Plant Cell Environ ; 41(1): 111-120, 2018 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-28370092

RESUMO

Volatile terpenoids play a key role in plant defence against herbivory by attracting parasitic wasps. We identified seven terpene synthase genes from lima bean, Phaseolus lunatus L. following treatment with either the elicitor alamethicin or spider mites, Tetranychus cinnabarinus. Four of the genes (Pltps2, Pltps3, Pltps4 and Pltps5) were up-regulated with their derived proteins phylogenetically clustered in the TPS-g subfamily and PlTPS3 positioned at the base of this cluster. Recombinant PlTPS3 was able to convert geranyl diphosphate and farnesyl diphosphate to linalool and (E)-nerolidol, the latter being precursor of the homoterpene (E)-4,8-dimethyl-1,3,7-nonatriene (DMNT). Recombinant PlTPS4 showed a different substrate specificity and produced linalool and (E)-nerolidol, as well as (E,E)-geranyllinalool from geranylgeranyl diphosphate. Transgenic rice expressing Pltps3 emitted significantly more (S)-linalool and DMNT than wild-type plants, whereas transgenic rice expressing Pltps4 produced (S)-linalool, DMNT and (E,E)-4,8,12-trimethyl-1,3,7,11-tridecatetraene (TMTT). In laboratory bioassays, female Cotesia chilonis, the natural enemy of the striped rice stemborer, Chilo suppressalis, were significantly attracted to the transgenic plants and their volatiles. We further confirmed this with synthetic blends mimicking natural rice volatile composition. Our study demonstrates that the transformation of rice to produce volatile terpenoids has the potential to enhance plant indirect defence through natural enemy recruitment.


Assuntos
Alquil e Aril Transferases/metabolismo , Fabaceae/enzimologia , Regulação da Expressão Gênica de Plantas , Oryza/genética , Oryza/parasitologia , Alquil e Aril Transferases/genética , Animais , Comportamento Animal , Feminino , Cromatografia Gasosa-Espectrometria de Massas , Genes de Plantas , Parasitos/fisiologia , Filogenia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas , Compostos Orgânicos Voláteis/metabolismo , Vespas/fisiologia
18.
J Chem Ecol ; 43(2): 207-214, 2017 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-28070757

RESUMO

Pheromone binding proteins (PBPs) are thought to play key roles in insect sex pheromone recognition; however, there is little in vivo evidence to support this viewpoint in comparison to abundant biochemical data in vitro. In the present study, two noctuid PBP genes HarmPBP1 and HarmPBP2 of the serious agricultural pest, Helicoverpa armigera were selected to be knocked down by RNA interference, and then the changes in electrophysiological and behavioral responses of male mutants to their major sex pheromone component (Z)-11-hexadecenal (Z11-16:Ald) were recorded. There were no significant electrophysiological or behavioral changes of tested male moths in response to Z11-16:Ald when either single PBP gene was knocked down. However, decreased sensitivity of male moths in response to Z11-16:Ald was observed when both HarmPBP1 and HarmPBP2 genes were silenced. These results reveal that both HarmPBP1 and HarmPBP2 are required for the recognition of the main sex pheromone component Z11-16:Ald in H. armigera. Furthermore, these findings may help clarify physiological roles of moth PBPs in the sex pheromone recognition pathway, which in turn could facilitate pest control by exploring sex pheromone blocking agents.


Assuntos
Comportamento Animal/fisiologia , Proteínas de Insetos/metabolismo , Cetonas/farmacologia , Mariposas , Interferência de RNA , Atrativos Sexuais/metabolismo , Animais , Antenas de Artrópodes/efeitos dos fármacos , Antenas de Artrópodes/fisiologia , Comportamento Animal/efeitos dos fármacos , Fenômenos Eletrofisiológicos , Técnicas de Silenciamento de Genes , Controle de Insetos , Proteínas de Insetos/genética , Cetonas/metabolismo , Masculino , Mariposas/genética , Mariposas/metabolismo , Mariposas/fisiologia , Ligação Proteica , Atrativos Sexuais/genética , Comportamento Sexual Animal/efeitos dos fármacos
20.
Mol Biol Evol ; 32(1): 63-80, 2015 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-25234705

RESUMO

Copy number variation (CNV) makes a major contribution to overall genetic variation and is suspected to play an important role in adaptation. However, aside from a few model species, the extent of CNV in natural populations has seldom been investigated. Here, we report on CNV in the pea aphid Acyrthosiphon pisum, a powerful system for studying the genetic architecture of host-plant adaptation and speciation thanks to multiple host races forming a continuum of genetic divergence. Recent studies have highlighted the potential importance of chemosensory genes, including the gustatory and olfactory receptor gene families (Gr and Or, respectively), in the process of host race formation. We used targeted resequencing to achieve a very high depth of coverage, and thereby revealed the extent of CNV of 434 genes, including 150 chemosensory genes, in 104 individuals distributed across eight host races of the pea aphid. We found that CNV was widespread in our global sample, with a significantly higher occurrence in multigene families, especially in Ors. We also observed a decrease in the gene probability of being completely duplicated or deleted (CDD) with increase in coding sequence length. Genes with CDD variants were usually more polymorphic for copy number, especially in the P450 gene family where toxin resistance may be related to gene dosage. We found that Gr were overrepresented among genes discriminating host races, as were CDD genes and pseudogenes. Our observations shed new light on CNV dynamics and are consistent with CNV playing a role in both local adaptation and speciation.


Assuntos
Afídeos/classificação , Afídeos/genética , Variações do Número de Cópias de DNA , Fabaceae/fisiologia , Proteínas de Insetos/genética , Simbiose , Adaptação Biológica , Animais , Afídeos/fisiologia , Biologia Computacional/métodos , Evolução Molecular , Fabaceae/classificação , Especiação Genética , Variação Genética , Genoma de Inseto , Sequenciamento de Nucleotídeos em Larga Escala , Filogenia , Receptores Odorantes/genética , Análise de Sequência de DNA
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