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Single-particle tracking has enabled real-time, in situ quantitative studies of complex systems. However, inferring dynamic state changes from noisy and undersampling trajectories encounters challenges. Here, we introduce a data-driven method for extracting features of subtrajectories with historical experience learning (Deep-SEES), where a single-particle tracking analysis pipeline based on a self-supervised architecture automatically searches for the latent space, allowing effective segmentation of the underlying states from noisy trajectories without prior knowledge on the particle dynamics. We validated our method on a variety of noisy simulated and experimental data. Our results showed that the method can faithfully capture both stable states and their dynamic switch. In highly random systems, our method outperformed commonly used unsupervised methods in inferring motion states, which is important for understanding nanoparticles interacting with living cell membranes, active enzymes, and liquid-liquid phase separation. Self-generating latent features of trajectories could potentially improve the understanding, estimation, and prediction of many complex systems.
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Nanopartículas , Imagem Individual de Molécula , Movimento (Física) , Membrana CelularRESUMO
Catalytic enzymes exhibiting enhanced motion have drawn extensive attention over the past decade; nevertheless, little is known about the effect on the environment induced by enzymes. Herein, we studied the active urease system by simultaneously monitoring the diffusion of single anisotropic gold nanorods (AuNRs) with high speed dark-field imaging. We found both the translational and the rotational diffusion coefficients of AuNRs were enhanced but with inconsistent degrees, indicating the catalytic reaction had a minor effect on the physiochemical properties of the environment according to the Stokes-Einstein equation. With the increase of substrate concentration, the diffusion of AuNRs showed increased spatial but decreased temporal heterogeneity. Additionally, high speed imaging revealed AuNRs could experience intermittent ballistic motion for tens of milliseconds. These results imply inhomogeneous distribution of enzymes in free solution induced by active enzymatic reactions.
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Nanopartículas , Nanotubos , Anisotropia , Difusão , Ouro/química , Nanotubos/químicaRESUMO
BACKGROUND: Previous studies have confirmed that exposure to fine particulate matter (PM2.5) is associated with respiratory disease mortality. However, due to the differences in PM2.5 concentration, composition and population susceptibility within different regions, the estimates of the association between PM2.5 concentration and mortality are different. Moreover, few studies have examined the potential hazard of excessive PM2.5 exposure in terms of respiratory disease mortality. METHODS: Daily recorded data on meteorological indices, environmental pollutants, and causes of death data in Changsha from January 2015 to December 2018 were obtained. The potential relationship between PM2.5 concentrations and respiratory disease mortality was determined using distributed lag nonlinear model (DLNM), which includes the relative risk (RR) and cumulative relative risk (CRR) of the lagged effect. The synergistic effects of other air pollutants were also considered. RESULTS: A total of 8,825 cases of respiratory disease mortality occurred in Changsha between 2015 and 2018. The acute effect of PM2.5 concentration was associated with an increased risk of respiratory disease mortality. Regarding the lag specific effect, a 10 µg/m3 increase in PM2.5 concentration on respiratory disease mortality was statistically significant at lag day 0 and lag day 7 with a relative risk of 1.019 (95% CI 1.007- 1.031) and 1.013(95%CI: 1.002-1.024). As for the cumulative lag effect, a 4-day moving average of PM2.5 concentrations was significantly associated with a cumulative relative risk of 1.027 (95%CI: 1.011-1.031). The single-day lag effect and cumulative 4-day lag effect for male individuals were more significant than those observed in females. The effect of PM2.5 concentrations and respiratory disease mortality remained statistically significant in the multi-pollutant models (SO2, NO2, and O3). A higher risk was observed in the cold season than in the warm season. CONCLUSIONS: Our findings show a potential association between exposure to PM2.5 concentration and respiratory disease mortality in Changsha, with male individuals observed to have particularly higher risk.
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Poluentes Atmosféricos , Poluição do Ar , Poluentes Ambientais , Transtornos Respiratórios , Doenças Respiratórias , Feminino , Masculino , Humanos , Material Particulado/efeitos adversos , Material Particulado/análise , Poluição do Ar/efeitos adversos , Poluentes Atmosféricos/efeitos adversos , Poluentes Atmosféricos/análise , China/epidemiologia , Exposição Ambiental/efeitos adversosRESUMO
A high-speed darkfield microscope has been developed to monitor the rapid rotation of single gold nanorods (AuNRs) and used to study the spatiotemporal heterogeneity of chemical reactions in free solution. A wide range of viscosities from 237â cP to 0.8â cP could be detected conveniently. We studied H2 O2 decomposition reactions that were catalyzed by AuNRs coated with Pt nanodots (AuNR@PtNDs) and observed two different rotational states. The two states and their transitions are related to the production and the amalgamation of O2 nanobubbles on the nanorod surface depending on H2 O2 concentration. In addition, the local fluidic environment of pure water was found to be non-uniform in time and space. This technique could be applied to study other chemical and biochemical reactions in solution.
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BACKGROUND: Serine protease inhibitors (SPIs) have been found in all living organisms and play significant roles in digestion, development and innate immunity. In this study, we present a genome-wide identification and expression profiling of SPI genes in the diamondback moth, Plutella xylostella (L.), a major pest of cruciferous crops with global distribution and broad resistance to different types of insecticides. RESULTS: A total of 61 potential SPI genes were identified in the P. xylostella genome, and these SPIs were classified into serpins, canonical inhibitors, and alpha-2-macroglobulins based on their modes of action. Sequence alignments showed that amino acid residues in the hinge region of known inhibitory serpins from other insect species were conserved in most P. xylostella serpins, suggesting that these P. xylostella serpins may be functionally active. Phylogenetic analysis confirmed that P. xylostella inhibitory serpins were clustered with known inhibitory serpins from six other insect species. More interestingly, nine serpins were highly similar to the orthologues in Manduca sexta which have been demonstrated to participate in regulating the prophenoloxidase activation cascade, an important innate immune response in insects. Of the 61 P.xylostella SPI genes, 33 were canonical SPIs containing seven types of inhibitor domains, including Kunitz, Kazal, TIL, amfpi, Antistasin, WAP and Pacifastin. Moreover, some SPIs contained additional non-inhibitor domains, including spondin_N, reeler, and other modules, which may be involved in protein-protein interactions. Gene expression profiling showed gene-differential, stage- and sex-specific expression patterns of SPIs, suggesting that SPIs may be involved in multiple physiological processes in P. xylostella. CONCLUSIONS: This is the most comprehensive investigation so far on SPI genes in P. xylostella. The characterized features and expression patterns of P. xylostella SPIs indicate that the SPI family genes may be involved in innate immunity of this species. Our findings provide valuable information for uncovering further biological roles of SPI genes in P. xylostella.
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Perfilação da Expressão Gênica , Regulação da Expressão Gênica/efeitos dos fármacos , Mariposas/efeitos dos fármacos , Mariposas/genética , Inibidores de Serina Proteinase/farmacologia , Transcriptoma , Sequência de Aminoácidos , Animais , Análise por Conglomerados , Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Insetos/química , Proteínas de Insetos/genética , Estágios do Ciclo de Vida/genética , Mariposas/classificação , Mariposas/crescimento & desenvolvimento , Filogenia , Serpinas/química , Serpinas/genéticaRESUMO
Using the linear approach to design a controller is still prevalent. The state feedback control (SFC) is applied in this paper to improve the dynamic response of permanent magnet synchronous machine (PMSM) speed regulation systems. First, a third-order augmented system is constructed for the reason that a higher-order system has better disturbance rejection. It can be found through analysis and comparison that the order of the proposed speed controller is increased. The parameters of SFC are selected by utilizing the linear quadratic regulator (LQR), and the influence of matrix Q on dynamic performance is detailed through the Bode diagram. Additionally, considering parametric uncertainties and unmodeled dynamics, a disturbance observer (DOB) using the Luenberger observer is designed to further boost anti-disturbance performance. Finally, plenty of experimental results verify the effectiveness of the proposed methods.
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This paper proposes a composite sliding mode control (SMC) to optimize the tracking performance and the anti-disturbance performance of permanent magnet synchronous machine (PMSM) speed regulation systems. The differential term in the control law can magnify the measurement noise, resulting in more discontinuity. To filter out the high frequency noise and make the control law smoother, the first-order differentiator (FOD) is employed to estimate the speed error and its derivative. Since the feedforward compensation can improve the robustness of the system, a disturbance observer (DOB) based on the sliding mode observer (SMO) is designed to reinforce the dynamic performance under disturbance variation. Under the effect of the feedforward compensation, chattering can be further weakened by decreasing the switching gain appropriately. Finally, the effectiveness of the proposed methods is confirmed by various experimental results.
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Liquid-liquid phase separation (LLPS) underlies the formation mechanism of membraneless biomolecular condensates locally to perform important physiological functions such as selective autophagy, but little is known about the relationship between their dynamic structural organization and biophysical properties. Here, a dark-field microscopy based single plasmonic nanoparticle tracking (DFSPT) technique was introduced to simultaneously monitor the diffusion dynamics of multiple gold nanorod (AuNR) probes in a protein LLPS system and to quantitatively characterize the spatiotemporal heterogeneity of the LLPS condensates during their phase transformation. Based on spatially and temporally resolved analysis of the diffusional behavior of the AuNRs, structure and material properties of p62 condensates, such as the viscoelasticity, the compartmentalization, and the recruitment of protein-covered nanoparticles into the large droplet, have been observed. Moreover, the nonsmooth droplet interface, its solidification after further phase transition or maturation, and the size effect of the inner vacuoles have also been revealed. Our method can be potentially applied to in vitro investigation of different reconstituted membrane-free biomolecular condensates and in vivo study of their dynamic evolution.
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Nanopartículas , Proteínas , Transição de Fase , Análise Espaço-Temporal , Estudos de Tempo e MovimentoRESUMO
Since the formation of protein corona inevitably leads to an increase in the particle size, it is important to develop technologies enabling in situ monitoring of the size change of nanoparticles. Traditional diffusion-based methods for particle size measurement focused on the translational diffusion coefficient; however, the detection sensitivity can be improved by determining the rotational diffusion coefficient, which has a cubic dependence on the particle radius. Here, using an optically anisotropic gold nanorod as the rotational probe and using high-speed dark-field microscopy, we can extract the rotational diffusion coefficient of a single nanorod and monitor the size change induced by the formation of protein corona in situ in real time. We successfully determined the thermodynamic parameters for the interactions between AuNRs with BSA and fibrinogen, and also studied corona formation in complex media and with AuNRs with different surface chemistry. This work would provide new avenues for the study of interactions between nanomedicines and proteins.
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Ouro/química , Nanopartículas Metálicas/química , Microscopia , Nanotubos/química , Coroa de Proteína/análise , Soroalbumina Bovina/análise , Animais , Anisotropia , BovinosRESUMO
Invasive species may change the life history strategies, distribution, genetic configuration and trophic interactions of native species. The diamondback moth, Plutella xylostella L., is an invasive herbivore attacking cultivated and wild brassica plants worldwide. Here we present phylogeographic analyses of P. xylostella and one of its major parasitoids, Cotesia vestalis, using mitochondrial markers, revealing the genetic diversity and evolutionary history of these two species. We find evidence that C. vestalis originated in Southwest China, then adapted to P. xylostella as a new host by ecological sorting as P. xylostella expanded its geographic range into this region. Associated with the expansion of P. xylostella, Wolbachia symbionts were introduced into local populations of the parasitoid through horizontal transfer from its newly associated host. Insights into the evolutionary history and phylogeographic system of the herbivore and its parasitoid provide an important basis for better understanding the impacts of biological invasion on genetic configuration of local species.
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Brassica/parasitologia , Herbivoria/fisiologia , Himenópteros/classificação , Mariposas/classificação , Adaptação Fisiológica , Animais , China , DNA Mitocondrial , Variação Genética , Himenópteros/fisiologia , Espécies Introduzidas , Mariposas/fisiologia , Filogeografia , SimbioseRESUMO
The diamondback moth, Plutella xylostella (L.), is a worldwide agricultural pest that has developed resistance to multiple classes of insecticides. Genetics-based approaches show promise as alternative pest management approaches but require functional studies to identify suitable gene targets. Here we use the CRISPR/Cas9 system to target a gene, abdominal-A, which has an important role in determining the identity and functionality of abdominal segments. We report that P. xylostella abdominal-A (Pxabd-A) has two structurally-similar splice isoforms (A and B) that differ only in the length of exon II, with 15 additional nucleotides in isoform A. Pxabd-A transcripts were detected in all developmental stages, and particularly in pupae and adults. CRISPR/Cas9-based mutagenesis of Pxabd-A exon I produced 91% chimeric mutants following injection of 448 eggs. Phenotypes with abnormal prolegs and malformed segments were visible in hatched larvae and unhatched embryos, and various defects were inherited by the next generation (G1). Genotyping of mutants demonstrated several mutations at the Pxabd-A genomic locus. The results indicate that a series of insertions and deletions were induced in the Pxabd-A locus, not only in G0 survivors but also in G1 individuals, and this provides a foundation for genome editing. Our study demonstrates the utility of the CRISPR/Cas9 system for targeting genes in an agricultural pest and therefore provides a foundation the development of novel pest management tools.
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Sistemas CRISPR-Cas , Proteínas de Drosophila/genética , Mariposas/genética , Proteínas Nucleares/genética , Fatores de Transcrição/genética , Abdome/crescimento & desenvolvimento , Abdome/fisiologia , Animais , Sequência de Bases , Proteínas de Drosophila/metabolismo , Feminino , Genes Homeobox , Larva/genética , Larva/crescimento & desenvolvimento , Larva/metabolismo , Masculino , Mariposas/crescimento & desenvolvimento , Mariposas/metabolismo , Proteínas Nucleares/metabolismo , Óvulo/crescimento & desenvolvimento , Óvulo/metabolismo , Fatores de Transcrição/metabolismoRESUMO
Objective@#To learn the status and influencing factors of the purchase of supplementary insurance for adverse events following immunization ( AEFI ) by parents in Changsha, so as to provide basis for the development of compensatory strategies.@*Methods@#Stratified random sampling method was used to select the parents who lived in Changsha for more than six months and had children under seven years old as subjects. A questionnaire survey was conducted to collect the information about demographic features, awareness of AEFI and the purchase of supplementary insurance. Logistic regression model was used to analyze the influencing factors for purchasing supplementary insurance. @*Results@#Among 712 respondents ( response rate, 94.93% ) , 354 ( 49.72% ) purchased supplementary insurance. The results of multivariate logistic regression analysis showed that the parents aged 36-71 years ( OR=0.325, 95%CI: 0.144-0.732 ) were less likely to purchase supplementary insurance; the parents who were aware of supplementary insurance ( OR=3.622, 95%CI: 2.218-5.913 ) and compensation range ( OR=1.332, 95%CI: 1.164-1.524 ) , and who scored higher in the knowledge and attitude of AEFI ( OR=1.137, 95%CI: 1.049-1.231 ) were more likely to purchase supplementary insurance.@*Conclusion @#About 49.72% of the parents purchased of supplementary insurance. Age, awareness of supplementary insurance and compensation range,as well as knowledge and attitude of AEFI were associated with the purchase of supplementary insurance.