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The phagocytic ability of macrophages empowers them to enforce innate immunity. RAW264.7, THP-1 and peripheral blood mononuclear cell-derived macrophages display considerable variability with regards to their phagocytic ability. We identify the underlying causes that attenuate the phagocytic abilities of a macrophage. Deformability of the cytoplasm and cortex influences the macrophage's phagocytic ability, and macrophages use the large cell-to-cell variability of their cytoplasmic stiffness to modulate their phagocytic ability. We find that the more-deformable macrophages have a higher phagocytic ability than those that are less deformable. Further, the subcellular spatial variability of cortex stiffness gives rise to more-deformable subdomains on the membrane for pathogen ingestion. We report a previously unknown negative-feedback loop that is triggered by the phagocytic oxidative burst. Macrophages utilize the excess reactive oxygen species to stiffen the cytoplasm, reducing their phagocytic propensity. In organisms, ageing or pathological conditions impair the phagocytic ability of macrophages. Our findings identify the targets that could potentially be utilized for restoring the phagocytic ability of the defunct macrophages.
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Leucócitos Mononucleares , Fagocitose , Citoplasma , Macrófagos , Espécies Reativas de OxigênioRESUMO
Background: The virulence of phytophagous insects is predominantly determined by their ability to evade or suppress host defense for their survival. The rice gall midge (GM, Orseolia oryzae), a monophagous pest of rice, elicits a host defense similar to the one elicited upon pathogen attack. This could be due to the GM feeding behaviour, wherein the GM endosymbionts are transferred to the host plant via oral secretions, and as a result, the host mounts an appropriate defense response(s) (i.e., up-regulation of the salicylic acid pathway) against these endosymbionts. Methods: The current study aimed to analyze the microbiome present at the feeding site of GM maggots to determine the exchange of bacterial species between GM and its host and to elucidate their role in rice-GM interaction using a next-generation sequencing approach. Results: Our results revealed differential representation of the phylum Proteobacteria in the GM-infested and -uninfested rice tissues. Furthermore, analysis of the species diversity of Pseudomonas and Wolbachia supergroups at the feeding sites indicated the exchange of bacterial species between GM and its host upon infestation. Conclusion: As rice-GM microbial associations remain relatively unstudied, these findings not only add to our current understanding of microbe-assisted insect-plant interactions but also provide valuable insights into how these bacteria drive insect-plant coevolution. Moreover, to the best of our knowledge, this is the first report analyzing the microbiome of a host plant (rice) at the feeding site of its insect pest (GM).
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We report a smartphone-paper-based sensor impregnated with cetyltrimethylammonium bromide modified silver nanoparticles (AgNPs/CTAB) for determination of Fe3+ in water and blood plasma samples. The methodology for determination of Fe3+ is based on the change in signal intensity of AgNPs/CTAB fabricated on a paper substrate after the deposition of analyte, using a smartphone followed by processing with ImageJ software. The mechanism of sensing for detection and determination of Fe3+ is based on the discoloration of AgNPs which impregnated the paper substrate. The discoloration is attributed to the electron transfer reaction taking place on the surface of NPs in the presence of CTAB. Fe3+ was determined when the paper was impregnated with 1 mM AgNPs for 5 min of reaction time and the substrate was kept under acidic conditions. The linear range for determination of total iron in terms of Fe3+ was 50-900 µg L-1 with a limit of determination (LOD) of 20 µg L-1 and coefficient of variation (CV) of 3.2%. The good relative recovery of 91.3-95.0% and interference studies showed the selectivity of the method for determination of total iron in water and blood plasma samples. Smartphone-paper-based sensors have advantages of simplicity, rapidity, user-friendliness, low cost, and miniaturization of the method for on-site determination of total iron compared to methods that require sophisticated analytical instruments. Graphical abstract Smartphone-paper-based sensor with cetyltrimethylammonium bromide modified silver nanoparticles for determination of Fe3+ in water and blood plasma samples.
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Ferro/análise , Ferro/sangue , Papel , Smartphone , Água/química , Limite de DetecçãoRESUMO
An optical colorimetric and smartphone-integrated paper device (SIPD) is demonstrated for determination of As (III) in water and soil samples using sucrose modified gold nanoparticles (AuNPs/Suc) as a nanoprobe. The mechanism for determination of As(III) is experimentally validated by performing UV-Vis, transmission electron microscope (TEM), Fourier transforms infra-red spectroscopy (FTIR) and dynamic light scattering (DLS) measurements. The density function theory (DFT) calculations using B3LYP with 6-311G (2d,2p) and LANL2DZ basis sets is used to theoretically prove the mechanism for determination of As(III). In addition, the paper fabricated with AuNPs/SuC was used as a nanoprobe for quantitative determination of As(III) using smartphone and ImageJ software. Calibration plot was linear over 10-800 µgL-1 for colorimetric determination of As(III) with limit of detection (LOD) of 4 µgL-1 acquired when the absorbance ratio obtained at 594 nm/515 nm. The linearity range of 50-3000 µgL-1 with LOD of 20 µgL-1 was determined using smartphone-integrated paper device. AuNPs/Suc is successfully employed for determination of As (III) from contaminated water and soil samples in colorimetry and SIPD. Graphical abstractColorimetric and Smartphone-integrated paper device used for selective detection of arsenic from contaminated water samples using sucrose modified gold nanoparticles (AuNPs/Suc) as a sensing probe.
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Arsênio/química , Colorimetria/métodos , Ouro/química , Nanopartículas Metálicas/química , Smartphone/instrumentação , Sacarose/química , Colorimetria/instrumentação , Humanos , PapelRESUMO
A novel, facile, and low-cost method was developed for determination of cetyltrimethylammonium (CTA+) cationic surfactant in water samples using diffuse reflectance Fourier transform IR (FTIR) spectroscopy and colorimetry. Cetyltrimethylammonium bromide was chosen as a model compound to demonstrate the optimization of the method for determination of CTA+ in water samples. The absorption peak at 3015.96 cm-1 (for CTA+) was enhanced when gold nanoparticles were used as a chemical sensor in diffuse reflectance FTIR spectroscopy, and this absorption peak was used for determination of CTA+. Alternatively, the color change from wine red (525 nm) to blue (740 nm) and the redshift of the localized surface plasmon resonance band in the visible region were used as a sensing probe for determination of CTA+. A linear calibration curve for determination in water samples was obtained in the range from 10 to 100 ng mL-1 with a limit of detection of 3 ng mL-1 by diffuse reflectance FTIR spectroscopy and in the range from 20 to 400 ng mL-1 with a limit of detection of 7 ng mL-1 by colorimetry. The advantageous features of the methods are their simplicity, rapidity, and sensitivity for the determination of CTA+ in water samples. Graphical abstract.
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All-trans retinoic acid (RA) is a key player in many developmental pathways. Most methods used to study its effects in development involve continuous all-trans RA activation by incubation in a solution of all-trans RA or by implanting all-trans RA-soaked beads at desired locations in the embryo. Here we show that the UV-driven photo-isomerization of 13-cis RA to the trans-isomer (and vice versa) can be used to non-invasively and quantitatively control the concentration of all-trans RA in a developing embryo in time and space. This facilitates the global or local perturbation of developmental pathways with a pulse of all-trans RA of known concentration or its inactivation by UV illumination. In zebrafish embryos in which endogenous synthesis of all-trans RA is impaired, incubation for as little as 5 minutes in 1 nM all-trans RA (a pulse) or 5 nM 13-cis RA followed by 1-minute UV illumination is sufficient to rescue the development of the hindbrain if performed no later than bud stage. However, if subsequent to this all-trans RA pulse the embryo is illuminated (no later than bud stage) for 1 minute with UV light (to isomerize, i.e. deactivate, all-trans RA), the rescue of hindbrain development is impaired. This suggests that all-trans RA is sequestered in embryos that have been transiently exposed to it. Using 13-cis RA isomerization with UV light, we further show that local illumination at bud stage of the head region (but not the tail) is sufficient to rescue hindbrain formation in embryos whose all-trans RA synthetic pathway has been impaired.
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Rombencéfalo/embriologia , Rombencéfalo/metabolismo , Tretinoína/metabolismo , Peixe-Zebra/embriologia , Peixe-Zebra/metabolismo , Animais , Isotretinoína/química , Isotretinoína/metabolismo , Rombencéfalo/efeitos da radiação , Tretinoína/química , Raios UltravioletaRESUMO
BACKGROUND: The Asian rice gall midge (Orseolia oryzae) is a destructive insect pest of rice. Gall midge infestation in rice triggers either compatible or incompatible interactions leading to survival or mortality of the feeding maggots, respectively. In incompatible interactions, generation of plant allelochemicals/defense molecules and/or inability of the maggots to continue feeding on the host initiate(s) apoptosis within the maggots. Unraveling these molecular events, triggered within the maggots as a response to feeding on resistant hosts, will enable us to obtain a better understanding of host resistance. The present study points towards the likely involvement of a defender against apoptotic cell death gene (DAD1) in the insect in response to the host defense. RESULTS: The cDNA coding for the DAD1 orthologue in the rice gall midge (OoDAD1) consisted of 339 nucleotides with one intron of 85 bp and two exons of 208 and 131 nucleotides. The deduced amino acid sequence of OoDAD1 showed a high degree of homology (94.6%) with DAD1 orthologue from the Hessian fly (Mayetiola destructor)--a major dipteran pest of wheat. Southern hybridization analysis indicated that OoDAD1 was present as a single copy in the genomes of the Asian rice gall midge biotypes (GMB) 1, 4 and 4 M. In the interactions involving GMB4 with Jaya (susceptible rice host) the expression level of OoDAD1 in feeding maggots gradually increased to 3-fold at 96 hai (hours after infestation) and peaked to 3.5-fold at 96 hai when compared to that at 24 hai. In contrast, expression in maggots feeding on RP2068 (resistant host) showed a steep increase of more than 8-fold at 24 hai and this level was sustained at 48, 72 and 96 hai when compared with the level in maggots feeding on Jaya at 24 hai. Recombinant OoDAD1, expressed in E. coli cells, when injected into rice seedlings induced a hypersensitive response (HR) in the resistant rice host, RP2068, but not in the susceptible rice variety, Jaya. CONCLUSIONS: The results indicate that the expression of OoDAD1 is triggered in the feeding maggots probably due to the host resistance response and therefore, is likely an important molecule in the initial stages of the interaction between the midge and its rice host.
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Apoptose , Dípteros/fisiologia , Resistência à Doença , Comportamento Alimentar , Proteínas de Insetos/genética , Oryza/parasitologia , Tumores de Planta/parasitologia , 3,3'-Diaminobenzidina/metabolismo , Sequência de Aminoácidos , Animais , Southern Blotting , Western Blotting , Dípteros/genética , Regulação da Expressão Gênica , Proteínas de Insetos/química , Proteínas de Insetos/metabolismo , Larva , Dados de Sequência Molecular , Oryza/imunologia , Filogenia , Doenças das Plantas/parasitologia , Proteínas Recombinantes/metabolismo , Alinhamento de SequênciaRESUMO
The present study attempts to obtain an a priori estimate of the absorbed dose received by an individual engaged in the reconnaissance survey in Uranium exploration using a predictive mathematical regression analysis. Other radiation safety parameters such as excess lifetime cancer risk are also calculated. Study reflects that the proper handling of naturally occurring radioactive materials accounts for an absorbed dose significantly less than the prescribed limit.
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Exposição Ocupacional , Monitoramento de Radiação , Urânio , Urânio/análise , Humanos , Índia , Monitoramento de Radiação/métodos , Exposição Ocupacional/análise , Doses de Radiação , Proteção Radiológica/métodos , Medição de Risco/métodos , Exposição à Radiação/análise , Neoplasias Induzidas por Radiação/prevenção & controle , Neoplasias Induzidas por Radiação/etiologia , Neoplasias Induzidas por Radiação/epidemiologiaRESUMO
Bacterial membrane vesicles (MVs) facilitate the long-distance delivery of virulence factors crucial for pathogenicity. The entry and trafficking mechanisms of virulence factors inside host cells are recently emerging; however, whether bacterial MVs can fuse and modulate the physicochemical properties of the host lipid membrane and membrane lipid parameter for fusion remains unknown. In this study, we reconstituted the interaction of bacterial MVs with host cell lipid membranes and quantitatively showed that bacterial MV interaction increases the fluidity, dipole potential, and compressibility of a biologically relevant multicomponent host membrane upon fusion. The presence of cylindrical lipids, such as phosphatidylcholine, and a moderate acyl chain length of C16 help the MV interaction. While significant binding of bacterial MVs to the raft-like lipid membranes with phase-separated regions of the membrane was observed, however, MVs prefer binding to the liquid-disordered regions of the membrane. Furthermore, the elevated levels of cholesterol tend to hinder the interaction of bacterial MVs, as evident from the favorable excess Gibbs free energy of mixing bacterial MVs with host lipid membranes. The findings provide new insights that might have implications for the regulation of host machinery by bacterial pathogens through manipulation of the host membrane properties.
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T cells are crucial for efficient antigen-specific immune responses and thus their migration within the body, to inflamed tissues from circulating blood or to secondary lymphoid organs, plays a very critical role. T cell extravasation in inflamed tissues depends on chemotactic cues and interaction between endothelial adhesion molecules and cellular integrins. A migrating T cell is expected to sense diverse external and membrane-intrinsic mechano-physical cues, but molecular mechanisms of such mechanosensing in cell migration are not established. We explored if the professional mechanosensor Piezo1 plays any role during integrin-dependent chemotaxis of human T cells. We found that deficiency of Piezo1 in human T cells interfered with integrin-dependent cellular motility on ICAM-1-coated surface. Piezo1 recruitment at the leading edge of moving T cells is dependent on and follows focal adhesion formation at the leading edge and local increase in membrane tension upon chemokine receptor activation. Piezo1 recruitment and activation, followed by calcium influx and calpain activation, in turn, are crucial for the integrin LFA1 (CD11a/CD18) recruitment at the leading edge of the chemotactic human T cells. Thus, we find that Piezo1 activation in response to local mechanical cues constitutes a membrane-intrinsic component of the 'outside-in' signaling in human T cells, migrating in response to chemokines, that mediates integrin recruitment to the leading edge.
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Quimiocinas , Canais Iônicos , Linfócitos T , Humanos , Adesão Celular , Movimento Celular , Quimiotaxia , Antígeno-1 Associado à Função Linfocitária , Canais Iônicos/metabolismoRESUMO
The increasing use of pesticides in the agriculture fields strengthen the crop production to meet the needs of increasing population. The residues in water and food materials cause several health hazards. Herein, nitrogen-doped carbon quantum dot (N-CQDs) is designed for determination of methiocarb pesticide in vegetables by fluorescent paper sensor and compared the results with fluorimetry. The fluorescent paper-based detection is performed by recording the change in fluorescence of N-CQDs with introduction of methiocarb using smartphone and ImageJ software. Good linear range was acquired for analysis of methiocarb from 10 to 1000 µgL-1 with a low detection limit (LOD) of 3.5 µgL-1 in fluorimetry; and 700-10,000 µgL-1 with a LOD of 500 µgL-1 in fluorescent paper sensor. A better recovery from 92.0 to 95.4% illustrating the selectivity of both methods for analysis of methiocarb in vegetables. Thus, the advantage of using N-CQDs as a fluorescent sensor for analysis of methiocarb in vegetables is instrument free, portable and user-friendly.
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Metiocarb , Praguicidas , Pontos Quânticos , Corantes Fluorescentes/química , Verduras , Pontos Quânticos/química , Nitrogênio/química , Carbono/química , Smartphone , Fluorometria , Espectrometria de Fluorescência/métodosRESUMO
The Asian rice gall midge (Orseolia oryzae) is a major pest responsible for immense loss in rice productivity. Currently, very little knowledge exists with regard to this insect at the molecular level. The present study was initiated with the aim of developing molecular resources as well as identifying alterations at the transcriptome level in the gall midge maggots that are in a compatible (SH) or in an incompatible interaction (RH) with their rice host. Roche 454 pyrosequencing strategy was used to develop both transcriptomics and genomics resources that led to the identification of 79,028 and 85,395 EST sequences from gall midge biotype 4 (GMB4) maggots feeding on a susceptible and resistant rice variety, TN1 (SH) and Suraksha (RH), respectively. Comparative transcriptome analysis of the maggots in SH and RH revealed over-representation of transcripts from proteolysis and protein phosphorylation in maggots from RH. In contrast, over-representation of transcripts for translation, regulation of transcription and transcripts involved in electron transport chain were observed in maggots from SH. This investigation, besides unveiling various mechanisms underlying insect-plant interactions, will also lead to a better understanding of strategies adopted by insects in general, and the Asian rice gall midge in particular, to overcome host defense.
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Dípteros/genética , Perfilação da Expressão Gênica , Animais , Dípteros/crescimento & desenvolvimento , Etiquetas de Sequências Expressas , Sequenciamento de Nucleotídeos em Larga Escala , Interações Hospedeiro-Parasita , Larva/genética , Larva/metabolismo , Redes e Vias Metabólicas , Peptídeo Hidrolases/metabolismo , Fosforilação , Proteínas Quinases/metabolismo , Proteólise , Análise de Sequência de DNARESUMO
The brown planthopper (BPH) is a monophagous sap-sucking pest of rice that causes immense yield loss. The rapid build-up of pesticide resistance combined with the ability of BPH populations to quickly overcome host plant resistance has rendered conventional control strategies ineffective. One of the likely ways in which BPH adapts to novel environments is by undergoing rapid shifts in its microbiome composition. To elucidate the rapid adaptation to novel environments and the contributions of Pseudomonas toward insect survival, we performed Pseudomonas-specific 16S rRNA gut-microbiome profiling of BPH populations. Results revealed the differential occurrence of Pseudomonas species in BPH populations with changing climates and geographical locations. Further, the observed variation in Pseudomonas species composition and abundance correlated with BPH survivability. Collectively, this study, while adding to our current understanding of symbiont-mediated insect adaptation, also demonstrated a complex interplay between insect physiology and microbiome dynamics, which likely confers BPH its rapid adaptive capacity.
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Herein, a user-friendly and portable smartphone-integrated printed-paper sensor was designed with Cu@Ag nanoparticles (NPs) for on-site monitoring of dimethoate pesticide in food samples, and the results obtained are compared with those obtained by UV-vis spectrophotometry. The working principle for identification of dimethoate pesticide is the change of yellow color NPs to reddish-yellow with associated bathochromic shift of absorption peak when pesticide introduced onto the fabricated paper or glass vial containing the NPs. A smartphone-color detector App and colorimetry were used for quantitative analysis of dimethoate in food samples. Linearity range for analysis of dimethoate using paper sensor and colorimetry were 100-2000 µgL-1 and 50-2500 µgL-1 with detection limit of 30 and 16 µgL-1, respectively. The advantages of using smartphone-integrated paper devices are rapid, instrument-free detection and economic in terms of consumption of lower amounts of NPs solution compared to other NPs-based colorimetric methods.
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Nanopartículas Metálicas , Praguicidas , Colorimetria/métodos , Dimetoato , Prata , SmartphoneRESUMO
Microsatellite loci were isolated from the genomic DNA of the Asian rice gall midge, Orseolia oryzae (Wood-Mason) using a hybridization capture approach. A total of 90 non-redundant primer pairs, representing unique loci, were designed. These simple sequence repeat (SSR) markers represented di (72%), tri (15.3%), and complex repeats (12.7%). Three biotypes of gall midge (20 individuals for each biotype) were screened using these SSRs. The results revealed that 15 loci were hyper variable and showed polymorphism among different biotypes of this pest. The number of alleles ranged from two to 11 and expected heterozygosity was above 0.5. Inheritance studies with three markers (observed to be polymorphic between sexes) revealed sex linked inheritance of two SSRs (Oosat55 and Oosat59) and autosomal inheritance of one marker (Oosat43). These markers will prove to be a useful tool to devise strategies for integrated pest management and in the study of biotype evolution in this important rice pest.
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Dípteros/genética , Dípteros/patogenicidade , Repetições de Microssatélites/genética , Oryza/parasitologia , Animais , Genoma de Inseto/genética , Virulência/genéticaRESUMO
The Asian rice gall midge, Orseolia oryzae (Wood-Mason), is a serious pest of rice. Investigations into the gall midge-rice interaction will unveil the underlying molecular mechanisms which, in turn, can be used as a tool to assist in developing suitable integrated pest management strategies. The insect gut is known to be involved in various physiological and biological processes including digestion, detoxification and interaction with the host. We have cloned and identified two genes, OoprotI and OoprotII, homologous to serine proteases with the conserved His(87), Asp(136) and Ser(241) residues. OoProtI shared 52.26% identity with mosquito-type trypsin from Hessian fly whereas OoProtII showed 52.49% identity to complement component activated C1s from the Hessian fly. Quantitative real time PCR analysis revealed that both the genes were significantly upregulated in larvae feeding on resistant cultivar than in those feeding on susceptible cultivar. These results provide an opportunity to understand the gut physiology of the insect under compatible or incompatible interactions with the host. Phylogenetic analysis grouped these genes in the clade containing proteases of phytophagous insects away from hematophagous insects.
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Dípteros/genética , Regulação da Expressão Gênica , Proteínas de Insetos/genética , Oryza , Serina Proteases/genética , Animais , Dípteros/enzimologia , Proteínas de Insetos/metabolismo , Dados de Sequência Molecular , Filogenia , Alinhamento de Sequência , Análise de Sequência de Proteína , Serina Proteases/metabolismoRESUMO
Autophagy is an extremely essential cellular process aimed to clear redundant and damaged materials, namely organelles, protein aggregates, invading pathogens, etc. through the formation of autophagosomes which are ultimately targeted to lysosomal degradation. In this study, we demonstrated that mTOR dependent classical autophagy is ubiquitously triggered in differentiating monocytes. Moreover, autophagy plays a decisive role in sustaining the process of monocyte to macrophage differentiation. We have delved deeper into understanding the underlying mechanistic complexities that trigger autophagy during differentiation. Intrigued by the significant difference between the protein profiles of monocytes and macrophages, we investigated to learn that autophagy directs monocyte differentiation via protein degradation. Further, we delineated the complex cross-talk between autophagy and cell-cycle arrest in differentiating monocytes. This study also inspects the contribution of adhesion on various steps of autophagy and its ultimate impact on monocyte differentiation. Our study reveals new mechanistic insights into the process of autophagy associated with monocyte differentiation and would undoubtedly help to understand the intricacies of the process better for the effective design of therapeutics as autophagy and autophagy-related processes have enormous importance in human patho-physiology.
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Autofagia , Monócitos , Autofagia/fisiologia , Diferenciação Celular , Humanos , Macrófagos/metabolismo , Monócitos/metabolismo , NF-kappa B , Serina-Treonina Quinases TOR/metabolismoRESUMO
We have implemented a noninvasive optical method for the fast control of protein activity in a live zebrafish embryo. It relies on releasing a protein fused to a modified estrogen receptor ligand binding domain from its complex with cytoplasmic chaperones, upon the local photoactivation of a nonendogenous caged inducer. Molecular dynamics simulations were used to design cyclofen-OH, a photochemically stable inducer of the receptor specific for 4-hydroxy-tamoxifen (ER(T2)). Cyclofen-OH was easily synthesized in two steps with good yields. At submicromolar concentrations, it activates proteins fused to the ER(T2) receptor. This was shown in cultured cells and in zebrafish embryos through emission properties and subcellular localization of properly engineered fluorescent proteins. Cyclofen-OH was successfully caged with various photolabile protecting groups. One particular caged compound was efficient in photoinducing the nuclear translocation of fluorescent proteins either globally (with 365 nm UV illumination) or locally (with a focused UV laser or with two-photon illumination at 750 nm). The present method for photocontrol of protein activity could be used more generally to investigate important physiological processes (e.g., in embryogenesis, organ regeneration and carcinogenesis) with high spatiotemporal resolution.
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Receptores de Estrogênio/metabolismo , Proteínas Recombinantes de Fusão/metabolismo , Peixe-Zebra/genética , Animais , Linhagem Celular , Chlorocebus aethiops , Ciclofenil/química , Embrião não Mamífero/metabolismo , Proteínas de Fluorescência Verde/análise , Proteínas de Fluorescência Verde/metabolismo , Chaperonas Moleculares/metabolismo , Simulação de Dinâmica Molecular , Processos Fotoquímicos , Fótons , Receptores de Estrogênio/genética , Proteínas Recombinantes de Fusão/análise , Tamoxifeno/análogos & derivados , Tamoxifeno/química , Tamoxifeno/farmacologia , Raios Ultravioleta , Peixe-Zebra/embriologia , Peixe-Zebra/metabolismoRESUMO
The molecular bases of aphid virulence to aphid crop plant resistance genes are poorly understood. The Russian wheat aphid, Diuraphis noxia, (Kurdjumov), and the greenbug, Schizaphis graminum (Rondani), are global pest of cereal crops. Each species damages barley, oat, rye and wheat, but S. graminum includes fescue, maize, rice and sorghum in its host range. This study was conducted to compare and contrast the transcriptomes of S. graminum biotype I and D. noxia biotype 1 when each ingested phloem from leaves of varieties of bread wheat, Triticum aestivum L., containing no aphid resistance (Dn0), resistance to D. noxia biotype 1 (Dn4), or resistance to both D. noxia biotype 1 and S. graminum biotype I (Dn7, wheat genotype 94M370). Gene ontology enrichments, k-means analysis and KEGG pathway analysis indicated that 94M370 plants containing the Dn7 D. noxia resistance gene from rye had stronger effects on the global transcriptional profiles of S. graminum and D. noxia relative to those fed Dn4 plants. S. graminum responds to ingestion of phloem sap from 94M370 plants by expression of unigenes coding for proteins involved in DNA and RNA repair, and delayed tissue and structural development. In contrast, D. noxia displays a completely different transcriptome after ingesting phloem sap from Dn4 or 94M370 plants, consisting of unigenes involved primarily in detoxification, nutrient acquisition and structural development. These variations in transcriptional responses of D. noxia and S. graminum suggest that the underlying evolutionary mechanism(s) of virulence in these aphids are likely species specific, even in cases of cross resistance.
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Afídeos/fisiologia , Resistência à Doença , Perfilação da Expressão Gênica/métodos , Hordeum/genética , Proteínas de Insetos/genética , Ração Animal , Animais , Afídeos/classificação , Afídeos/genética , Regulação da Expressão Gênica , Ontologia Genética , Herbivoria , Sequenciamento de Nucleotídeos em Larga Escala , Folhas de Planta/genética , Proteínas de Plantas/genética , Análise de Sequência de RNA , Especificidade da EspécieRESUMO
Epigenetic mechanisms have not been characterized in ticks despite their importance as vectors of human and animal diseases worldwide. Our investigation identifies and functionally characterizes the orthologue of S-adenosylmethionine (SAM) binding methyltransferase enzyme, disruptor of telomeric silencing 1-like (DOT1L) in Ornithodoros moubata (OmDOT1L), a soft tick vector for the relapsing fever pathogen Borrelia duttonii and the African swine fever virus. The OmDOT1L tertiary structure was predicted and compared to the Homo sapiens DOT1L which had been co-crystalized with SGC0946, a DOT1L-specific inhibitor. The amino acid residues crucial for SAM and SGC0946 binding conserved in most DOT1L sequences available, are also conserved in OmDOT1L. Quantitative PCR of Omdot1l during O. moubata life stages showed that transcripts were significantly upregulated in first-stage nymphs. O. moubata larvae exposed to SGC0946 displayed high mortality during molting to first-stage nymphs. Furthermore, a significant decrease in weight was observed in second-stage nymphs fed on recombinant OmDOT1L-immunized rabbits. In contrast, artificial blood feeding supplemented with SGC0946 did not affect survival and reproductive performance of adult female ticks. We concluded that OmDOT1L plays an essential role in the regulation of larval molting and the feeding of O. moubata second-stage nymphs.