RESUMEN
Actin- and microtubule (MT)-based transport systems are essential for intracellular transport. During influenza A virus (IAV) infection, MTs provide long tracks for virus trafficking toward the nucleus. However, the role of the actin cytoskeleton in IAV entry and especially the transit process is still ambiguous. Here, by using quantum dot-based single-virus tracking, it was revealed that the actin cytoskeleton was crucial for the virus entry via clathrin-mediated endocytosis (CME). After entry via CME, the virus reached MTs through three different pathways: the virus (1) was driven by myosin VI to move along actin filaments to reach MTs (AF); (2) was propelled by actin tails assembled by an Arp2/3-dependent mechanism to reach MTs (AT); and (3) directly reached MTs without experiencing actin-related movement (NA). Therefore, the NA pathway was the main one and the fastest for the virus to reach MTs. The AT pathway was activated only when plenty of viruses entered the cell. The viruses transported by the AF and AT pathways shared similar moving velocities, durations, and displacements. This study comprehensively visualized the role of the actin cytoskeleton in IAV entry and transport, revealing different pathways for IAV to reach MTs after entry. The results are of great significance for globally understanding IAV infection and the cellular endocytic transport pathway.
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Endocitosis , Virus de la Influenza A , Microtúbulos , Puntos Cuánticos , Puntos Cuánticos/química , Microtúbulos/metabolismo , Microtúbulos/virología , Humanos , Virus de la Influenza A/fisiología , Internalización del Virus , Animales , Perros , Células de Riñón Canino Madin Darby , Citoesqueleto de Actina/metabolismoRESUMEN
Viral diseases are among the main threats to public health. Understanding the factors affecting viral invasion is important for antiviral research. Until now, it was known that most viruses have very low plaque-forming unit (PFU)-to-particle ratios. However, further investigation is required to determine the underlying factors. Here, using quantitative single-particle analysis methods, the invasion of Semliki Forest virus (SFV), Japanese encephalitis virus (JEV), and influenza A virus (IAV) containing attachment to the cell surface, entry into the cell, transport towards the cell interior, and fusion with endosomes to release nucleocapsids were quantitatively analysed in parallel. It was found that for SFV with an PFU-to-particle ratio of approximately 1:2, an entry efficiency of approximately 31% limited infection. For JEV, whose PFU-to-particle ratio was approximately 1:310, an attachment efficiency of approximately 27% and an entry efficiency of 10% were the main factors limiting its infection. Meanwhile, for IAV with PFU-to-particle ratios of 1:8100, 5% attachment efficiency, 9% entry efficiency, and 53% fusion efficiency significantly limited its infection. These results suggest that viruses with different infectivities have different limited steps in the invasion process. Moreover, there are significant differences in attachment efficiencies among viruses, emphasizing the pivotal role of attachment in viral invasion. The influence of the virus purification method on virus invasion was also investigated. This study, for the first time, reports the efficiencies of different stages of virus invasion, leading to a better understanding of virus invasion and providing a protocol to quantitatively analyse the virus invasion efficiency.
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Virus de la Influenza A , Virus de los Bosques Semliki , Internalización del Virus , Virus de la Influenza A/fisiología , Animales , Virus de los Bosques Semliki/fisiología , Humanos , Virus de la Encefalitis Japonesa (Especie)/fisiología , Línea Celular , Acoplamiento Viral , Endosomas/virologíaRESUMEN
BACKGROUND: Ranitidine induced tumor adverse events remains a contradictory clinical question, due to the limited evidence of tumor risk associated with ranitidine in the real world. The purpose of this study was to evaluate the association of ranitidine with all types of tumors through the FAERS database and to provide a reference for clinical use. RESEARCH DESIGN AND METHODS: Cancer cases associated with ranitidine in the FAERS database from the first quarter of 2004 to the fourth quarter of 2023 were extracted to analyze demographic characteristics, and a disproportion analysis was performed. RESULT: A total of 662,998 ranitidine-related cancer cases were screened, and the 50-59 and 60-69 groups accounted for the largest proportion. In PT signal detection, ranitidine was associated with 98 PT, including penal cancer stage II, gastric cancer stage II, et al. In terms of outcome events, adverse events were higher in men (20.65%) than in women (18.47%). CONCLUSIONS: Ranitidine may induce various tumor-related adverse reactions, especially in long-term users and elderly patients. For these patients, tumor screening should be strengthened, and long-term use of ranitidine should be avoided. Since this study cannot prove causality, further evidence is needed for prospective studies with a larger sample size.
RESUMEN
Influenza A virus (IAV) is internalized into its host cells by endocytosis, which involves many cellular proteins and molecules. In this study, we focus on the function of calcium ion (Ca2+) in IAV endocytosis. We have found that IAV infection is accompanied by the increase in concentration of cytosolic Ca2+, which is mainly attributed to the influx of extracellular Ca2+. When Ca2+ influx is abolished, IAV internalization will be markedly suppressed, but the virus attachment to its host cells will be unaffected. Extracellular Ca2+ influx is essential to the clathrin-mediated endocytosis (CME) of IAVs but dispensable to the clathrin-independent endocytosis of the virus and is dispensable to the CME of transferrin or low-density lipoprotein as a control. Ca2+ influx might participate in the dynamin-promoted membrane fission in the CME of IAVs. Our study highlights that IAVs enter host cells via extracellular Ca2+ influx-involved clathrin- and dynamin-dependent endocytosis, which will facilitate better understanding of IAV infection and development of anti-influenza drugs.
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Materiales Biocompatibles/química , Calcio/metabolismo , Clatrina/metabolismo , Virus de la Influenza A/metabolismo , Células de Riñón Canino Madin Darby/metabolismo , Animales , Perros , Endocitosis , Células de Riñón Canino Madin Darby/virología , Ensayo de Materiales , Tamaño de la PartículaRESUMEN
AIMS: miRNAs are regarded as potential biomarkers correlated with the development and progression of many diseases. However, it is a challenge to construct a sensitive method to detect them without using time-consuming radioactive labeling or complex amplification strategies. METHODS: A facile resonance light scattering (RLS) system was developed for the detection of miRNA employing magnetic nanoparticles (MNPs) as RLS probes. MNPs were coated with streptavidin. DNA probes were modified on the surface of MNPs based on the specific interaction of streptavidin and biotin forming MNPs@DNA probes. MNPs@DNA probes dispersed in homogeneous media causing low RLS signal. RESULTS & CONCLUSION: miRNA hybridized with DNA probes resulting in the aggregation of MNPs and inducing the enhancement of RLS intensity. miRNAs were determined successfully with limit of detection at 0.9 picomole per liter (pM). The potential clinical application of the present biosensor was also demonstrated by measuring miRNAs in human normal and cancer cells, and human serum samples.
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Bioensayo/métodos , Técnicas Biosensibles/métodos , Nanopartículas de Magnetita/química , MicroARNs/análisis , Células A549 , Biotina/química , Sondas de ADN/química , Dispersión Dinámica de Luz , Humanos , Células MCF-7 , MicroARNs/genética , Microscopía Electrónica de Transmisión , Estreptavidina/químicaRESUMEN
An aptamer based method is described for the determination of 8-hydroxy-2'-deoxyguanosine (8-OHdG) using resonance light scattering (RLS). Magnetic nanoparticles (MNPs) were employed as RLS probes. The probe DNA was placed on the surface of MNPs, which produces a rather low RLS signal. If, however, probe DNA hybridizes with the aptamer against 8-OHdG, a sandwich structure will be formed. This results in a significant enhancement of RLS intensity. The aptamer was used as the recognition element to capture 8-OHdG. 8-OHdG has a stronger affinity for the aptamer than probe DNA, and the conformation of the aptamer therefore switches from a double-stranded to a G-quadruplex structure. As a result, MNPs labeled with probe DNA are released, and RLS intensity decreases. The method allows 8-OHdG to be detected with a linear response in the 32 pM - 12.0 nM concentration range and an 11 pM limit of detection (at 3.29SB/m, according to the recent recommendation of IUPAC). The MNPs can be reused 5 times by applying an external magnetic field for collection. The method was successfully applied to analyze human urine samples for its content of 8-OHdG. It was also found that the levels of 8-OHdG noticeably increased with the increase of the Air Quality Index. Conceivably, the method is a viable tool to investigate the relationship between 8-OHdG levels and the effect of air pollution. Graphical abstract A reusable sensing strategy was constructed to detect urinary 8-OHdG based on "turn-off" resonance light scattering. The LOD was as low as 11 pM. This study showed some preliminary data for the association between oxidative stress and air pollution.
RESUMEN
The design of molecular nanodevices attracted great interest in these years. Herein, a reusable, sensitive and specific aptasensor was constructed based on an extension-contraction movement of DNA interconversion for the application of human thrombin detection. The present biosensor was based on resonance light scattering (RLS) using magnetic nanoparticles (MNPs) as the RLS probe. MNPs coated with streptavidin can combine with biotin labeled thrombin aptamers. The combined nanoparticles composite is monodispersed in aqueous medium. When thrombin was added a sandwich structure can form on the surface of MNPs, which induced MNPs aggregation. RLS signal was therefore enhanced, and there is a linear relationship between RLS increment and thrombin concentration in the range of 60pM-6.0nM with a limit of detection at 3.5pM (3.29SB/m, according to the recent recommendation of IUPAC). The present aptasensor can be repeatedly used for at least 6 cycling times by heat to transfer G-quadruplex conformation to single strand of DNA sequence and release thrombin. MNPs can be captured by applying the external magnetic field. Furthermore, the proposed biosensor was successfully applied to detect thrombin in human plasma.
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Aptámeros de Nucleótidos/química , Técnicas Biosensibles/métodos , Nanopartículas de Magnetita/química , Trombina/análisis , Biotina/química , Dispersión Dinámica de Luz/métodos , Humanos , Límite de Detección , Nanopartículas de Magnetita/ultraestructura , Estreptavidina/químicaRESUMEN
A novel and simple fluorescence Off-On system is proposed for selective pyrophosphate (PPi) sensing in an aqueous solution. The method is constructed based on the strong blue emission of carbon dots (CDs) owing to its outstanding photoluminescence and easy synthesis, which has shown exciting potential in analytical and biological field. The fluorescence of CDs can be remarkably quenched by some transition metal ions such as Cu(2+), Ni(2+), Mn(2+) and Co(2+) due to the coordination reaction between metal ions and the carboxylic groups on the surface of CDs. When PPi was introduced to CDs-metal ion system the fluorescence of CDs was recovered regularly. The increment of fluorescence intensity was proportional with the concentration of PPi in the range of 1-200 µM and correspondingly the limit of detection was calculated as 0.32 µM according to the recommendation of IUPAC as 3.29 S B/m. The possible mechanism was discussed for the detection of PPi and the quenching reaction between CDs and metal ions. Furthermore, the proposed system was successfully used to monitor the content of PPi in water samples from artificial wetland.
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Hexavalent chromium is one of the most toxic heavy metals in aqueous solutions. It has been well documented that the brown seaweed can be used as a promising biosorbent for the sequestration of this heavy metal from wastewater. However, the uptake of Cr(VI) is reportedly a rather slower process; the sorption equilibrium can only be established after a few days, much slower than a few hours for the trivalent chromium ion. In this study, we developed a novel technology of electrochemically assisted biosorption (ECAB) system for the enhancement of the treatment efficiency. It was found through our study that the removal efficiencies of Cr(VI) and total chromium were greatly enhanced by 48.1% and 51.3%, respectively, with the application of -1.0 V in the ECAB system. The conversion of Cr(III) due to the electroreduction of Cr(VI) and the higher pH due to the cathodic H(2) evolution created a favorable condition for the uptake of chromium onto the modified seaweed (MSW). The reduction and adsorption of Cr(VI) by MSW was proved to play a minor role in the removal. Both direct electroreduction and indirect electroreduction by atomic H* contributed to the reduction of Cr(VI).
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Cromo/química , Técnicas Electroquímicas/métodos , Sargassum/química , Purificación del Agua/métodos , Oxidación-ReducciónRESUMEN
The anodic oxidation of aqueous solutions containing dimethyl phthalate (DMP) up to 125 mg/L with sodium sulfate (Na2SO4) as supporting electrolyte within the pH range 2.0-10.0 was studied using a one-compartment batch reactor employing a boron-doped diamond (BDD) as anode. Electrolyses were carried out at constant current density (1.5-4.5 mA/cm2). Complete mineralization was always achieved owing to the great concentration of hydroxyl radical (*OH) generated at the BDD surface. The effects of pH, apparent current density and initial DMP concentration on the degradation rate of DMP, the specific charge required for its total mineralization and mineralization current efficiency were investigated systematically. The mineralization rate of DMP was found to be pH-independent and to increase with increasing applied current density. Results indicated that this electrochemical process was subjected, at least partially, to the mass transfer of organics onto the BDD surface. Kinetic analysis of the temporal change of DMP concentration during electrolysis determined by High Performance Liquid Chromatography (HPLC) revealed that DMP decay under all tested conditions followed a pseudo first-order reaction. Aromatic intermediates and generated carboxylic acids were identified by Gas Chromatography-Mass Spectrometry (GC-MS) and a general pathway for the electrochemical incineration of DMP on BDD was proposed.
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Boro/química , Diamante/química , Electroquímica/métodos , Electrodos , Incineración/métodos , Ácidos Ftálicos/química , Cromatografía Líquida de Alta Presión , Cromatografía de Gases y Espectrometría de Masas , Concentración de Iones de Hidrógeno , Oxidación-ReducciónRESUMEN
Hydrotalcite-supported Pd/Sn catalysts with different Mg/Al atomic ratios (2, 3, 4, and 5) for nitrate adsorption and hydrogenation reduction were successfully synthesized by a coprecipitation method. The results showed that different atomic ratios of Mg/Al resulted in different interlayer spacings and zeta-potentials of the catalysts, which thus influenced its adsorption capacity. With the increase of Mg/Al atomic ratio, the interlayer spacing rose and zeta-potential decreased. The adsorption properties of the catalysts were mainly affected by interlayer spacing when Mg/Al atomic ratios increased from 2 to 4. However, when Mg/Al atomic ratios further increased from 4 to 5, there was a negative impact on the adsorption properties of zeta-potential. Also, the adsorption capacity of the catalysts for nitrate followed the order: Mg/Al=2
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A novel photoanode was prepared by immobilizing TiO(2) film onto activated carbon fibers (TiO(2)/ACF) using liquid phase deposition (LPD) to study the electro-photocatalytic (EPC) degradation of organic compounds exemplified by an azo-dye, namely, Acid Orange II (AOII). Results demonstrated that by applying a 0.5 V bias (vs. SCE) across the TiO(2)/ACF electrode, the AOII degradation rate was increased significantly compared to that of photocatalytic (PC) oxidation. The application of an electric field promotes the separation of photogenerated electrons and holes as confirmed by electrochemical impedance spectroscopy (EIS) measurements. The structural and surface morphology of the TiO(2)/ACF electrode was characterized by field emission scanning electron microscopy (FE-SEM) and X-ray diffraction (XRD). SEM images showed that TiO(2) was deposited on almost every carbon fiber with an average thickness of about 200 nm with the inner space between neighboring fibers being maintained unfilled. The morphological features of the photo-anode facilitated the passage of solution as well as UV light through the felt-form electrode and created a three-dimensional environment favorable to EPC oxidation. Both the large outer surface area of the 3D electrode and the good organic adsorption capacity of the ACF support promoted high contact efficiency between AOII and TiO(2) surface. Anatase was the major crystalline TiO(2) deposited. UV-vis spectrophotometry, TOC (total organic carbon) analysis, and HPLC technique were used to monitor the concentration change of AOII and intermediates as to gain insight into the EPC degradation of AOII using the TiO(2)/ACF electrode.
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Compuestos Azo/química , Carbono/química , Naftalenos/química , Titanio/química , Contaminantes Químicos del Agua/química , Purificación del Agua/métodos , Adsorción , Fibra de Carbono , Impedancia Eléctrica , Técnicas Electroquímicas , Electrodos , Microscopía Electrónica de Rastreo , Oxidación-Reducción , Procesos Fotoquímicos , Difracción de Rayos XRESUMEN
A Microcystis aeruginosa which produced high content of microcystin-LR (MC-LR) but no microcystin-RR (MC-RR) was isolated from Dianchi Lake in China. In the molecular structure of MC-LR, glutamic acid, aspartic acid, leucine, alanine and arginine are the constitutional components which are abundant in natural water. In this paper, effects of six amino acids at their natural concentrations on the growth of the M. aeruginosa and the microcystin (MC) production were studied in batch culture. M. aeruginosa could assimilate alanine, leucine and arginine as sole nitrogen sources for growth and MC production. However, glutamic acid, aspartic acid and lysine could not be assimilated quickly, although they could pass the cell membrane and enter into the cell rapidly. Our experiment demonstrated that the possible reason of such phenomenon was that different amino acids had different effects on the process of metabolism through the free dissolved amino acids within the cells.
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Aminoácidos/química , Biotecnología/métodos , Microcystis/metabolismo , Alanina/química , Arginina/química , Ácido Aspártico/química , Cianobacterias/metabolismo , Ácido Glutámico/química , Leucina/química , Toxinas Marinas , Microcistinas/química , Microscopía Electrónica de Rastreo , Nitrógeno/química , Factores de TiempoRESUMEN
A combined bioelectrochemical and sulfur autotrophic denitrification system (CBSAD) was evaluated to treat a groundwater with nitrate contamination (20.9-22.0mgNO(3)(-)-N/L). The reactor was operated continuously for several months with groundwater to maximize treatment efficiency under different hydraulic retention times (HRT) and electric currents. The denitrification rate of sulfur autotrophic part followed a half-order kinetics model. Moreover, the removal efficiency of bioelectrochemical part depended on the electric current. The reactor could be operated efficiently at the HRT ranged from 4.2 to 2.1h (corresponding nitrogen volume-loading rates varied from 0.12 to 0.24 kg N/m(3)d; and optimum current ranged from 30 to 1000 mA), and the NO(3)(-)-N removal rate ranged from 95% to 100% without NO(3)(-)-N accumulation. The pH of effluent was satisfactorily adjusted by bioelectrochemical part, and the sulfate concentration of effluent was lower than 250 mg/L, meeting the drinking water standard of China EPA.
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Bacterias Anaerobias/metabolismo , Reactores Biológicos/microbiología , Electroquímica/instrumentación , Nitratos/metabolismo , Azufre/metabolismo , Contaminantes Químicos del Agua/metabolismo , Purificación del Agua/instrumentación , Procesos Autotróficos/fisiología , Simulación por Computador , Diseño de Equipo , Análisis de Falla de Equipo , Modelos Biológicos , Nitratos/aislamiento & purificación , Azufre/aislamiento & purificación , Integración de SistemasRESUMEN
Effects of nitrogen-limitation and phosphorus-limitation on microcystin (MC) content and energy charge (EC) of the Microcystis aeruginosa were investigated in batch cultures and semi-continuous cultures. In batch cultures, nitrogen-limitation retarded the MC synthesis and phosphorus-limitation had little effects on MC production. The EC remained constant in nitrogen-limited cultures, while it decreased largely when phosphorus was extinct in phosphorus-limited culture. In the semi-continuous cultures, MC production in nitrogen- and phosphorus-limited cultures increased with the increase of dilution rate; however, MC content in phosphorus-limited cultures was more than that in nitrogen-limited cultures. The EC in nitrogen-limited cultures remained constant and in phosphorus-limited cultures increased with the increase of dilution rate. In phosphorus-limited semi-continuous cultures, a direct relationship between EC and MC content was demonstrated. No correlation was observed in nitrogen-limited semi-continuous cultures. Based on the above analysis, a mechanism of nitrogen and phosphorus effect on the MC synthesis was suggested, that the MC synthesis was determined by the combination of necessary enzymes and precursors and EC.
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Microcystis/metabolismo , Nitrógeno/metabolismo , Fósforo/metabolismo , Técnicas Bacteriológicas , Metabolismo Energético , Toxinas Marinas , Microcistinas/metabolismoRESUMEN
The present study investigated the cyanobacteria and one family of their toxins-microcystins (MCs) in Guanting Reservoir of Beijing, China. The dominant species in the cyanobacteria found in August and September of 2006 was Microcystis, which accounted 99% of total algal cells. The specific species of the Microcystis in the cyanobacteria included Microcystis ichthyobalbe, Microcystis novacekii, Microcystis botrys and Microcystis aeruginosa which had different ratios in different sites. The qualitative analysis by HPLC showed that two microcystins were contained in cyanobacteria and one microcystin was in water of the reservoir. The major microcystins were microcystin-RR (MC-RR) and microcystin-LR (MC-LR), but only MC-LR was detected in water. The quantitative analysis by HPLC indicated that the maximum concentrations of MC-RR and MC-LR contained in cyanobacteria were 0.74 and 0.41 mg/g dry weight, respectively. The maximum microcystin concentration in water was 1.15 microg/L and others were below 1 microg/L.
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Toxinas Bacterianas/análisis , Microcistinas/análisis , Microcystis/aislamiento & purificación , Contaminantes del Agua/análisis , Abastecimiento de Agua/análisis , China , Recuento de Colonia Microbiana , Monitoreo del Ambiente , Toxinas Marinas , Microcystis/química , Microcystis/clasificación , Nitrógeno/análisis , Fósforo/análisis , Microbiología del AguaRESUMEN
Angiotensin (ANG) II receptors have been reported in the nonpigmented ciliary epithelium (NPE) of the eye. In cultured NPE, we found ANG II caused a dose-dependent rise of cytoplasmic sodium. The sodium increase was inhibited by the AT(1)-AT(2) receptor antagonist saralasin (IC(50) = 3.7 nM) and the AT(1) antagonist losartan (IC(50) = 0.6 nM) but not by the AT(2) antagonist PD-123319. ANG II also caused a dose-dependent increase in the rate of ouabain-sensitive (86)Rb uptake. The ANG II-induced cell sodium increase and (86)Rb uptake increase were reduced by dimethylamiloride (DMA; 10 microM). On the basis of this finding, we propose that Na(+)/H(+) exchange is stimulated by ANG II. Simultaneously, ANG II appears to inhibit H(+)-ATPase-mediated proton export. Thus Ang II (10 nM) did not alter the baseline cytoplasmic pH (pH(i)) but reduced pH(i) in cells that were also exposed to 10 microM DMA. Consistent with the notion of H(+)-ATPase inhibition in ANG II-treated NPE, bafilomycin A(1) (100 nM) (BAF) and ANG II were both observed to suppress the pH(i) increase that occurs upon exposure to a mixture of epinephrine (1 microM) and acetylcholine (10 microM) and the pH(i) increase elicited by depolarization. In ATP hydrolysis measurements, H(+)-ATPase activity (bafilomycin A(1)-sensitive ATP hydrolysis) was reduced significantly in cells that had been pretreated 10 min with 10 nM ANG II. In summary, these studies suggest that ANG II causes H(+)-ATPase inhibition and an increase of cell sodium due to activation of Na(+)/H(+) exchange.