RESUMEN
The inertial motion unit (IMU) is an effective tool for monitoring and assessing gait impairment in patients with lumbar disc herniation(LDH). However, the current clinical assessment methods for LDH gait focus on patients' subjective scoring indicators and lack the assessment of kinematic ability; at the same time, individual differences in the motor function degradation of the healthy and affected lower limbs of LDH patients are also ignored. To solve this problem, we propose an LDH gait feature model based on multi-source adaptive Kalman data fusion of acceleration and angular velocity. The gait phase is segmented by using an adaptive Kalman data fusion algorithm to estimate the attitude angle, and obtaining gait events through a zero-velocity update technique and a peak detection algorithm. Two IMUs were used to analyze the gait characteristics of lumbar disc patients and healthy gait people, including 12 gait characteristics such as gait spatiotemporal parameters, kinematic parameters, gait variability and stability. Statistical methods were used to analyze the characteristic model and verify the biological differences between the healthy affected side of LDH and healthy subjects. Finally, feature engineering and machine learning technology were used to identify the gait pattern of inertial movement units in patients with lumbar intervertebral disc disease, and achieved a classification accuracy of 95.50%, providing an effective gait feature set and method for clinical evaluation of LDH.
RESUMEN
Colloidal quantum-dot (QD) lasing is normally achieved in close-packed solid-state films, as a high QD volume fraction is required for stimulated emission to outcompete fast Auger decay of optical-gain-active multiexciton states. Here a new type of liquid optical-gain medium is demonstrated, in which compact compositionally-graded QDs (ccg-QDs) that feature strong suppression of Auger decay are liquefied using a small amount of solvent. Transient absorption measurements of ccg-QD liquid suspensions reveal broad-band optical gain spanning a wide spectral range from 560 (green) to 675 nm (red). The gain magnitude is sufficient to realize a two-color amplified spontaneous emission (ASE) at 637 and 594 nm due to the band-edge (1S) and the excited-state (1P) transition, respectively. Importantly, the ASE regime is achieved using quasicontinuous excitation with nanosecond pulses. Furthermore, the ASE is highly stable under prolonged excitation, which stands in contrast to traditional dyes that exhibit strong degradation under identical excitation conditions. These observations point toward a considerable potential of high-density ccg-QD suspensions as liquid, dye-like optical gain media that feature readily achievable spectral tunability and stable operation under intense photoexcitation.
RESUMEN
Investigating spin crossover (SCO)-fluorescence bifunctional materials and establishing their structure-function relationships are attractive topics in chemistry and materials science. However, it remains challenging to preserve the fluorescence and SCO properties simultaneously in aggregated solid states. Herein, we design an (E)-2,6-bis(1H-pyrazol-1-yl)-4-(4-(1,2,2-triphenylvinyl)styryl)pyridine (tpe-bpp) ligand, which contains coordinated SCO and fluorescence units of an aggregation-induced emission luminogen (AIEgen). The coordination of the tpe-bpp ligand with different FeII salts generated three mononuclear complexes: [Fe(tpe-bpp)2](ClO4)2·5.75CH2Cl2 (1), [Fe(tpe-bpp)2](ClO4)2·CH2Cl2·3CH3OH (2) and [Fe(tpe-bpp)2](BF4)2·CH2Cl2·3CH3OH (3). Single-crystal X-ray diffraction studies showed that they shared a similar [Fe(tpe-bpp)2]2+ complex cation. Their counterions and co-crystallized solvents were different. Magnetic measurements revealed that 1, 2, and 3 exhibited a complete SCO behavior with the transition temperatures T1/2 of 375, 260, and 248 K, respectively. Fluorescence measurements confirmed the existence of the AIE property for both the tpe-bpp ligand and Fe(II) complexes. A monotonic decrease of the photoluminescence (PL) intensity upon increasing the temperature was behavior observed for all three complexes.
RESUMEN
Few studies have carried out soil washing experiments using pot experiments to simulate in situ soil washing operations, particularly for alkaline soils. This study explored the effects of multiple washing operations using pot experiments on the removal efficiencies of potentially toxic metals (PTM) from alkaline farmland soil and the reuse strategy of washed soil for safe agricultural production. The results showed that the removal efficiencies of Cd, Pb, Cu, and Zn after seven washings with a mixed chelator (EDTA, GLDA, and citric acid) were 41.1%, 47.1%, 14.7%, and 26.5%, respectively, which was close to the results of the EDTA treatment. For the alkaline soil studied, the second washing with the mixed chelators most effectively removed PTM owing to the activation of them after the first washing operation. The mixed chelator more effectively increased the proportion of stable fraction of PTM and maintained soil nutrients (e.g., nitrogen content) than EDTA, indicating little disturbance of alkaline soil quality after washing with the mixed chelator. After the amendment of the washed soil, there was no visible difference in the biomass weight of crops from the soils washed with different agents, indicating that the inhibitory effect of both washing agents on plant growth was effectively alleviated. The Cd and Pb contents in Z. mays were below the threshold of Hygienical Standard for Feeds of China (GB 13078-2017) (1 and 30 mg·kg-1). Moreover, after three cropping operations, the available concentrations of PTM in the soil washed with the mixed chelator were lower than those in the soil washed with EDTA, indicating the value and potential of agricultural reuse of alkaline farmland soil washed with the mixed chelator.
Asunto(s)
Isópodos , Metales Pesados , Contaminantes del Suelo , Animales , Suelo , Metales Pesados/análisis , Ácido Edético , Cadmio , Granjas , Plomo , Contaminantes del Suelo/análisis , QuelantesRESUMEN
Orbital mixing is paramount to chemistry as it plays a central role in bond formation. It is also important for technologies such as molecular doping of polymers, where the concept of fractional charge transfer is essentially orbital mixing between dopants and hosts. Likewise, it would be both fundamentally interesting and technologically relevant to investigate orbital mixing in emerging hybrid materials containing both inorganic and organic moieties. Here we report experimental observation of orbital mixing between valence band levels of strongly confined PbS quantum dots (QDs) and lowest unoccupied molecular levels of surface-bound high-electron affinity molecules (F4TCNQ), manifested as both an absorption blue-shift of PbS and the emergence of visible and infrared signatures of the fractional charge-transfer species of F4TCNQ. The degree of mixing can be controlled by varying the QD size or by varying the molecule/QD ratio for a specific QD size and can be quantitatively reproduced by a nondegenerate, two-level perturbation model.
RESUMEN
Colloidal quantum dots (QDs) can photocatalyze diverse organic reactions. However, reported QD-photocatalysts often contain highly-toxic elements Cd or Pb, and have not surpassed prototypical transition-metal complexes in terms of their photoredox power or excited-state energy. Here we report low-toxicity ZnSe/ZnS core/shell QDs as potent visible photocatalysts to drive challenging organic transformations. To overcome the limitation of short excited-state lifetime of the QDs, we functionalize their surfaces with benzophenone ligands which can rapidly extract electrons from photoexcited QDs and sustain long-lived charge-separated states. The benzophenone anions function as potent electron relay to drive dehalogenation of aryl chlorides and additive-free polymerization of acrylates. Alternatively, the QDs are functionalized with biphenyl ligands to store energy in long-lived, energetic triplets, enabling [2+2] homo-cycloaddition of styrene and cycloaddition of carbonyls with alkenes.
Asunto(s)
Puntos Cuánticos , Puntos Cuánticos/toxicidad , Compuestos de Zinc , Sulfuros , BenzofenonasRESUMEN
A relatively new addition to the application portfolio of lead halide perovskites is to photosensitize molecular triplets for a variety of photochemical applications. Here we report visible-light-driven isomerization and cycloaddition of organic molecules sensitized by spectrally-tunable perovskite nanocrystals. We first demonstrate with stilbene as the substrate molecule that photoisomerization can proceed efficiently and rapidly by either directly grafting carboxylated stilbene onto nanocrystal surfaces or using triplet-acceptor ligands as the energy relay. The relay approach is more generally applicable as it does not require anchoring-group functionalization of substrate molecules, allowing us to facilely extend it to isomerization of a series of substituted stilbene molecules and ring-closing isomerization of diarylethene, as well as intermolecular [2+2] cycloaddition of acenaphthylene. This study opens an avenue of energy-transfer photocatalysis using perovskite nanocrystals.
RESUMEN
GLS1 orchestrates glutaminolysis and promotes cell proliferation when glutamine is abundant by regenerating TCA cycle intermediates and supporting redox homeostasis. CB-839, an inhibitor of GLS1, is currently under clinical investigation for a variety of cancer types. Here, we show that GLS1 facilitates apoptosis when glutamine is deprived. Mechanistically, the absence of exogenous glutamine sufficiently reduces glutamate levels to convert dimeric GLS1 to a self-assembled, extremely low-Km filamentous polymer. GLS1 filaments possess an enhanced catalytic activity, which further depletes intracellular glutamine. Functionally, filamentous GLS1-dependent glutamine scarcity leads to inadequate synthesis of asparagine and mitogenome-encoded proteins, resulting in ROS-induced apoptosis that can be rescued by asparagine supplementation. Physiologically, we observed GLS1 filaments in solid tumors and validated the tumor-suppressive role of constitutively active, filamentous GLS1 mutants K320A and S482C in xenograft models. Our results change our understanding of GLS1 in cancer metabolism and suggest the therapeutic potential of promoting GLS1 filament formation.
Asunto(s)
Glutaminasa , Glutamina , Apoptosis , Asparagina/genética , Glutaminasa/genética , Glutaminasa/metabolismo , Glutamina/metabolismo , Humanos , Especies Reactivas de OxígenoRESUMEN
Sensitization of molecular triplets using PbS quantum dots (QDs), followed by efficient triplet fusion, has been developed as a novel route to near-infrared-to-visible photon upconversion. Fundamentally, however, the mechanisms of triplet energy transfer (TET) from PbS QDs to surface-anchored polyacence acceptors remain highly debated. Here we study and side-by-side compare the kinetic pathways of TET from photoexcited PbS QDs to surface-anchored tetracene and pentacene derivatives using broad-band transient absorption spectroscopy spanning multiple decades of timescales. We find that the TET pathways are dictated by charge-transfer energetics at the QD/molecule interface. Charge transfer from QDs to tetracene was strongly endothermic, and hence spectroscopy showed one-step transformation from QD excited states to tetracene triplets in 302 ns. In contrast, hole transfer from QDs to pentacene was thermodynamically favoured and was confirmed by the formation of pentacene cation radicals in 13 ps, which subsequently evolved into pentacene triplets through a 101 ns electron transfer process. These results not only are consistent with a recently-established framework of charge-transfer-mediated TET, but also provide a route to manipulate triplet sensitization using lead-salt QDs for efficient upconversion of near-infrared photons.
RESUMEN
Land surface temperature (LST) is a critical state variable of land surface energy equilibrium and a key indicator of environmental change such as climate change, urban heat island, and freezing-thawing hazard. The high spatial and temporal resolution datasets are urgently needed for a variety of environmental change studies, especially in remote areas with few LST observation stations. MODIS and Landsat satellites have complementary characteristics in terms of spatial and temporal resolution for LST retrieval. To make full use of their respective advantages, this paper developed a pixel-based multi-spatial resolution adaptive fusion modeling framework (called pMSRAFM). As an instance of this framework, the data fusion model for joint retrieval of LST from Landsat-8 and MODIS data was implemented to generate the synthetic LST with Landsat-like spatial resolution and MODIS temporal information. The performance of pMSRAFM was tested and validated in the Heihe River Basin located in China. The results of six experiments showed that the fused LST was high similarity to the direct Landsat-derived LST with structural similarity index (SSIM) of 0.83 and the index of agreement (d) of 0.84. The range of SSIM was 0.65-0.88, the root mean square error (RMSE) yielded a range of 1.6-3.4 °C, and the averaged bias was 0.6 °C. Furthermore, the temporal information of MODIS LST was retained and optimized in the synthetic LST. The RMSE ranged from 0.7 °C to 1.5 °C with an average value of 1.1 °C. When compared with in situ LST observations, the mean absolute error and bias were reduced after fusion with the mean absolute bias of 1.3 °C. The validation results that fused LST possesses the spatial pattern of Landsat-derived LSTs and inherits most of the temporal properties of MODIS LSTs at the same time, so it can provide more accurate and credible information. Consequently, pMSRAFM can be served as a promising and practical fusion framework to prepare a high-quality LST spatiotemporal dataset for various applications in environment studies.
RESUMEN
Galectins are a family of ß-galactoside binding proteins, and insect galectins play a role in immune responses and may also affect Cry toxin activity. In this study, we aimed to further understand the function and molecular mechanism of Aedes aegypti galectin-6 in modulation of Cry11Aa toxicity. A. aegypti galectin-6 was cloned, and the recombinant galectin-6 was expressed and purified. Bioassays indicated that galectin-6 could reduce the toxicity of Cry11Aa, protecting A. aegypti larvae. To determine interactions among galectin-6, Cry11Aa and putative toxin receptors, Octet Red System, western blotting, far-western blotting and ELISA assays were performed. Octet Red System showed that galectin-6 bound to BBMVs of A. aegypti larvae with lower affinity than that of Cry11Aa. Western blotting and far-western blotting analyses demonstrated that galectin-6 bound to A. aegypti ALP1 and APN2 as well as to BBMVs, consistent with the results of ELISA and protein docking simulations. However, galectin-6 did not bind to Cadherin in far-western blotting or ELISA assay, though the protein docking simulations suggested their binding potential. These findings support the conclusion that galectin-6 may block Cry11Aa from binding to ALP1 and APN2 due to structural similarity, which might decrease the mosquitocidal toxicity of Cry11Aa.
Asunto(s)
Aedes , Bacillus thuringiensis , Animales , Proteínas Bacterianas , Endotoxinas , Galectinas , Proteínas Hemolisinas , Proteínas de Insectos , LarvaRESUMEN
BACKGROUND: Bacillus thuringiensis israelensis (Bti) is a widely used mosquitocidal microbial pesticide due to its high toxicity. ATP-binding proteins (ABP) are prevalently detected in insects and are related to reaction against Bti toxins. However, the function of ABP in mosquito biocontrol is little known, especially in Aedes aegypti. Therefore, this study aimed to clarify the function of ABP in Ae. aegypti against Bti toxin. RESULTS: Aedes aegypti ABP (GenBank: XM_001661856.2) was cloned, expressed and purified in this study. Far-western blotting and ELISA were also carried out to confirm the interaction between ABP and Cry11Aa. A bioassay of Cry11Aa was performed both in the presence and absence of ABP, which showed that the mortality of Ae. aegypti is increased with an increase in ABP. CONCLUSIONS: Our results suggest that ABP in Ae. aegypti can modulate the toxicity of Cry11Aa toxin to mosquitoes by binding to Bti toxin. This could not only enrich the mechanism of Bt toxin, but also provide more data for the biocontrol of this transmission vector.
Asunto(s)
Aedes/genética , Bacillus thuringiensis/patogenicidad , Proteínas de Insectos/genética , Proteínas de Insectos/metabolismo , Aedes/microbiología , Animales , Toxinas de Bacillus thuringiensis , Proteínas Bacterianas/metabolismo , Bioensayo , Clonación Molecular , Endotoxinas/metabolismo , Proteínas Hemolisinas/metabolismo , Mosquitos Vectores/microbiología , Control Biológico de Vectores , Unión ProteicaRESUMEN
Aedes aegypti is a crucial vector for human diseases, such as yellow fever, dengue, chikungunya, and Zika viruses. Today, a major challenge throughout the globe is the insufficient availability of antiviral drugs and vaccines against arboviruses, and toxins produced by Bacillus thuringiensis (Bt) are still used as biological agents for mosquito control. The use of Cry toxins to kill insects mainly depends on the interaction between Cry toxins and important toxin receptors, such as alkaline phosphatase (ALP). In this study, we investigated the function of A. aegypti C-type lectin-20 (CTL-20) in the tolerance of Cry toxins. We showed that recombinant CTL-20 protein interacted with both Cry11Aa and ALP1 by the Far-Western blot and ELISA methods, and CTL-20 bound to A. aegypti larval brush border membrane vesicles (BBMVs). Binding affinity of CTL-20 to ALP1 was higher than that of Cry11Aa to ALP1. Furthermore, the survival rate of A. aegypti larvae fed with Cry11Aa toxin mixed with recombinant CTL-20 fusion protein was significantly increased compared with that of the control larvae fed with Cry11Aa mixed with thioredoxin. Our novel results suggest that midgut proteins like CTLs may interfere with interactions between Cry toxins and toxin receptors by binding to both Cry toxins and receptors to alter Cry toxicity.
Asunto(s)
Aedes/efectos de los fármacos , Fosfatasa Alcalina/metabolismo , Proteínas Bacterianas/toxicidad , Endotoxinas/toxicidad , Proteínas Hemolisinas/toxicidad , Proteínas de Insectos/metabolismo , Lectinas Tipo C/metabolismo , Aedes/genética , Aedes/metabolismo , Animales , Toxinas de Bacillus thuringiensis , Tracto Gastrointestinal/metabolismo , Proteínas de Insectos/genética , Lectinas Tipo C/genética , Proteínas Recombinantes/metabolismoRESUMEN
Globally, Aedes aegypti is one of the most dangerous mosquitoes that plays a crucial role as a vector for human diseases, such as yellow fever, dengue, and chikungunya. To identify (1) transcriptomic basis of midgut (2) key genes that are involved in the toxicity process by a comparative transcriptomic analysis between the control and Bacillus thuringiensis (Bt) toxin (LLP29 proteins)-treated groups. Next-generation sequencing technology was used to sequence the midgut transcriptome of A. aegypti. A total of 17130 unigenes, including 574 new unigenes, were identified containing 16358 (95.49%) unigenes that were functionally annotated. According to differentially expressed gene (DEG) analysis, 557 DEGs were annotated, including 226 upregulated and 231 downregulated unigenes in the Bt toxin-treated group. A total of 442 DEGs were functionally annotated; among these, 33 were specific to multidrug resistance, 6 were immune-system-related (Lectin, Defensin, Lysozyme), 28 were related to putative proteases, 7 were lipase-related, 8 were related to phosphatases, and 30 were related to other transporters. In addition, the relative expression of 28 DEGs was further confirmed through quantitative real time polymerase chain reaction. The results provide a transcriptomic basis for the identification and functional authentication of DEGs in A. aegypti.
Asunto(s)
Aedes/genética , Bacillus thuringiensis/fisiología , Perfilación de la Expresión Génica/métodos , Transcriptoma/genética , Aedes/microbiología , Animales , Biología Computacional , Biblioteca de Genes , Secuenciación de Nucleótidos de Alto Rendimiento , Reacción en Cadena en Tiempo Real de la PolimerasaRESUMEN
Soil washing is an effective technology for the remediation of multi-metal contaminated soils. However, bioavailability of residual heavy metals in soils and soil properties could be changed during washing processes. This study investigated the effects of EDTA, FeCl3 and mixed chelators (MC) on bioavailability of residual heavy metals in soils and soil biological properties after soil washing. The results showed that soil washing by chelators successfully decreased the total concentration of heavy metals in soils, while it did not effectively decrease the exchangeable fraction of heavy metals, especially for calcareous contaminated soil. The toxic effects of the washed soils seemed to exhibit higher correlations with the changes in the soil properties such as soil pH and nutrient concentrations. As compared with FeCl3 and EDTA, MC tended to moderately change soil properties (e.g., pH, total N, available N, available P, and exchangeable K, Ca, and Mg). Additionally, MC-washed soil had the least influence on the soil enzymes activities, and had the highest germination and growth of Chinese cabbage. Accordingly, MC is a moderate washing solution in the removal of heavy metals from multi-metal contaminated soils, and had minimal negative effects on soil qualities.
Asunto(s)
Quelantes/química , Ácido Cítrico/química , Ácido Edético/química , Contaminación Ambiental/análisis , Metales Pesados/química , Contaminantes del Suelo/química , Suelo/química , Disponibilidad Biológica , Metales Pesados/análisis , Contaminantes del Suelo/análisisRESUMEN
PURPOSE: MicroRNAs (miRs) are endogenous, noncoding small RNAs that play a key role in regulating biological and pathological processes. The oncogenic properties of miR-199b-5p have been demonstrated in previous studies but the effect of miR-199b-5p on osteosarcoma (OS) has not yet been clarified. This study aimed to investigate the effect of miR-199b-5p on OS and the relationship between this miR and the pathological parameters and prognosis of OS. METHODS: MiR-199b-5p expression in 57 pairs of OS tissues, corresponding adjacent normal tissues and OS cells was measured by quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR).The relationship between miR-199b-5p and the pathological features and prognosis of OS patients was examined. We constructed small interfering (si) RNA to knock down miR-199b-5p expression in OS cell lines MG63 and U2OS. Cell Counting Kit-8 (CCK-8), cell cloning assay and Transwell cell migration and invasion assay were applied for investigating the biological function of miR-199b-5p, respectively. Finally, western blot was used for exploring its underlying mechanism. RESULTS: MiR-199b-5p expression in OS was significantly higher than that of normal tissues. Compared to patients w\sith low expression of miR-199b-5p, patients with high expression level tended to be with younger age, higher incidence of distant metastases and lower overall survival. Compared with interference sequence negative control (si-NC) group, the abilities of proliferation, invasion and metastasis of cells transfected with si-miR-199b-5p were significantly decreased. Western blot analysis indicated that expressions of key proteins related to epithelial to mesenchymal transition (EMT) signaling pathway, including N-cadherin, Vimentin, ß-catenin and matrix metalloproteinase-9 (MMP9), were significantly decreased after transfection with si-miR-199b-5p. Furthermore, we found that miR-199b-5p promoted the progression of OS mainly through regulating HER2. CONCLUSIONS: Upregulated miR-199b-5p is significantly related with stage, distant metastasis and poor prognosis of OS. This MiR may promote progression of OS through regulating HER2.
Asunto(s)
Biomarcadores de Tumor/metabolismo , Neoplasias Óseas/patología , Regulación Neoplásica de la Expresión Génica , MicroARNs/genética , Osteosarcoma/secundario , Receptor ErbB-2/metabolismo , Adulto , Biomarcadores de Tumor/genética , Neoplasias Óseas/genética , Neoplasias Óseas/metabolismo , Estudios de Casos y Controles , Movimiento Celular , Proliferación Celular , Progresión de la Enfermedad , Transición Epitelial-Mesenquimal , Femenino , Estudios de Seguimiento , Humanos , Masculino , Invasividad Neoplásica , Metástasis de la Neoplasia , Osteosarcoma/genética , Osteosarcoma/metabolismo , Pronóstico , Receptor ErbB-2/genética , Transducción de Señal , Tasa de Supervivencia , Adulto JovenRESUMEN
Cry11Aa displays high toxicity to the larvae of several mosquito species, including Aedes, Culex, and Anopheles. To study its binding characterization against Culex quinquefasciatus, Cry11Aa was purified and western blot results showed that Cry11Aa could bind successfully to the brush border membrane vesicles. To identify Cry11Aa-binding proteins in C. quinquefasciatus, a biotin-based protein pull-down experiment was performed and seven Cry11Aa-binding proteins were isolated from the midgut of C. quinquefasciatus larvae. Analysis of liquid chromatography-tandem mass spectrometry showed that one of the Cry11Aa-binding proteins is the ATP-binding domain 1 family member B. To investigate its binding property and effect on the toxicity of Cry11Aa, western blot, far-western blot, enzyme-linked immunosorbent assay, and bioassays of Cry11Aa in the presence and absence of the recombinant ATP-binding protein were performed. Our results showed that the ATP-binding protein interacted with Cry11Aa and increased the toxicity of Cry11Aa against C. quinquefasciatus. Our study suggests that midgut proteins other than the toxin receptors may modulate the toxicity of Cry toxins against mosquitoes.
Asunto(s)
Adenosina Trifosfato/metabolismo , Proteínas Bacterianas/metabolismo , Culex/metabolismo , Endotoxinas/metabolismo , Proteínas Hemolisinas/metabolismo , Proteínas de Insectos/metabolismo , Animales , Toxinas de Bacillus thuringiensis , Proteínas Bacterianas/toxicidad , Culex/química , Culex/efectos de los fármacos , Culex/genética , Endotoxinas/toxicidad , Proteínas Hemolisinas/toxicidad , Proteínas de Insectos/química , Proteínas de Insectos/genética , Larva/efectos de los fármacos , Larva/genética , Larva/crecimiento & desarrollo , Larva/metabolismo , Unión Proteica , Dominios ProteicosRESUMEN
Lassa virus (LASV) is an enveloped RNA virus endemic to West Africa and responsible for severe cases of hemorrhagic fever. Virus entry is mediated by the glycoprotein complex consisting of a stable-signal peptide, a receptor-binding subunit, GP1, and a viral-host membrane fusion subunit, GP2. Several cellular receptors can interact with the GP1 subunit and mediate viral entry, including alpha-dystroglycan (αDG) and lysosome-associated membrane protein 1 (LAMP1). In order to define the regions within GP1 that interact with the cellular receptors, we implemented insertional mutagenesis, carbohydrate shielding, and alanine scanning mutagenesis. Eighty GP constructs were engineered and evaluated for GP1-GP2 processing, surface expression, and the ability to mediate cell-to-cell fusion after low-pH exposure. To examine virus-to-cell entry, 49 constructs were incorporated onto vesicular stomatitis virus (VSV) pseudoparticles and transduction efficiencies were monitored in HAP1 and HAP1-ΔDAG1 cells that differentially produce the αDG cell surface receptor. Seven constructs retained efficient transduction in HAP1-ΔDAG1 cells yet poorly transduced HAP1 cells, suggesting that they are involved in αDG utilization. Residues H141, N146, F147, and Y150 cluster at the predicted central core of the trimeric interface and are important for GP-αDG interaction. Additionally, H92A-H93A, 150HA, 172HA, and 230HA displayed reduced transduction in both HAP1 and HAP1-ΔDAG1 cells, despite efficient cell-to-cell fusion activity. These mutations may interfere with interactions with the endosomal receptor LAMP1 or interfere at another stage in entry that is common to both cell lines. Insight gained from these data can aid in the development of more-effective entry inhibitors by blocking receptor interactions.IMPORTANCE Countries in which Lassa virus is endemic, such as Nigeria, Sierra Leone, Guinea, and Liberia, usually experience a seasonal outbreak of the virus from December to March. Currently, there is neither a preventative vaccine nor a therapeutic available to effectively treat severe Lassa fever. One way to thwart virus infection is to inhibit interaction with cellular receptors. It is known that the GP1 subunit of the Lassa glycoprotein complex plays a critical role in receptor recognition. Our results highlight a region within the Lassa virus GP1 protein that interacts with the cellular receptor alpha-dystroglycan. This information may be used for future development of new Lassa virus antivirals.
Asunto(s)
Distroglicanos/metabolismo , Virus Lassa/genética , Virus Lassa/metabolismo , Receptores Virales/metabolismo , Proteínas del Envoltorio Viral/genética , Proteínas del Envoltorio Viral/metabolismo , Línea Celular , Análisis Mutacional de ADN , Humanos , Proteínas de Membrana de los Lisosomas/metabolismo , Mutagénesis Insercional , Mutagénesis Sitio-Dirigida , Proteínas Mutantes/genética , Proteínas Mutantes/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Transducción Genética , Vesiculovirus/genética , Vesiculovirus/fisiología , Internalización del VirusRESUMEN
We prepared in this work an anchoring porphyrin and a series of hat-porphyrins. The zinc atom of the hat-porphyrins can be coordinated axially with the pyridine moiety of the anchoring porphyrin which is anchored on the titania surface by a carboxyl group. The structures of the assemblies were confirmed using computational calculations, transmission electron microscopy (TEM) and energy dispersive spectrometry (EDS). Solar cell devices of the monomer anchoring porphyrin and its assemblies were fabricated and the photovoltaic performances were measured under standard AM 1.5 sunlight irradiance. We found that the assembly devices showed higher JSC and lower VOC than that of the monomer anchoring porphyrin device. However, the comprehensive influence of JSC and VOC led to an enhancement in the solar-to-electric power-conversion efficiency (PCE) of the assemblies. We also studied the variation of JSC and VOC using electronic absorption and emission spectroscopy, charge extraction measurements, transient photovoltage decay measurements and electrochemical impedance spectroscopy.
RESUMEN
Colitose, also known as 3,6-dideoxy-L-galactose or 3-deoxy-L-fucose, is one of only five naturally occurring 3,6-dideoxyhexoses. Colitose was found in lipopolysaccharide of a number of infectious bacteria, including Escherichia coli O55 & O111 and Vibrio cholera O22 & O139. To date, no colitosyltransferase (ColT) has been characterized, probably due to the inaccessibility of the sugar donor, GDP-colitose. In this study, starting with chemically prepared colitose, 94.6 mg of GDP-colitose was prepared via a facile and efficient one-pot two-enzyme system involving an L-fucokinase/GDP-L-Fuc pyrophosphorylase and an inorganic pyrophosphatase (EcPpA). WbgN, a putative ColT from E. coliO55:H5 was then cloned, overexpressed, purified and biochemically characterized by using GDP-colitose as a sugar donor. Activity assay and structural identification of the synthetic product clearly demonstrated that wbgN encodes an α1,2-ColT. Biophysical study showed that WbgN does not require metal ion, and is highly active at pH 7.5-9.0. In addition, acceptor specificity study indicated that WbgN exclusively recognizes lacto-N-biose (Galß1,3-GlcNAc). Most interestingly, it was found that WbgN exhibits similar activity toward GDP-l-Fuc (kcat/Km= 9.2 min(-1)mM(-1)) as that toward GDP-colitose (kcat/Km= 12 min(-1)mM(-1)). Finally, taking advantage of this, type 1 H-antigen was successfully synthesized in preparative scale.