Sensitive and on-Site Detection of Staphylococcus aureus Based on CRISPR/Cas 13a-Assisted Chemiluminescence Resonance Energy Transfer.

Developing a specific, sensitive, rapid, and on-site method for detecting pathogenic bacteria in food samples is critical to ensuring public safety. This article demonstrates a CRISPR/Cas13a system and a chemiluminescence resonance energy transfer (CRET) (CRISPR/Cas 13a-assisted CRET)-based strategy for sensitive and on-site detection of pathogenic bacteria in real samples. Once the hybrid double strand of aptamerS. aureus-cRNA recognizes the target model bacteria of Staphylococcus aureus (S. aureus), the released cRNA would bind with CRISPR/Cas 13a to form a complex of cRNA-CRISPR/Cas 13a, which could cleave the RNA molecule in the detecting probe of horseradish peroxidase (HRP) modified-gold nanoparticles (AuNPs) linked by RNA (AuNPs-RNA-HRP), resulting in an enhanced chemiluminescence signal due to the CRET "OFF" phenomenon after introducing the chemiluminescence substrate of luminol. The CRISPR/Cas 13a-assisted CRET strategy successfully detected S. aureus in drinking water and milk with detection limits of 20 and 30 cfu/mL, respectively, within the recovery of 90.07-105.50%. Furthermore, after integrating with an immunochromatographic test strip (ICTS), the CRISPR/Cas 13a-assisted CRET strategy achieved the on-site detection of as low as 102 cfu/mL of S. aureus in drinking water and milk via a smartphone, which is about 10 times lower than that in the previously reported AuNPs-based colorimetric ICTS, demonstrating a convenient and sensitive detection method for S. aureus in real samples.

Brain Accumulation and Toxicity Profiles of Silica Nanoparticles: The Influence of Size and Exposure Route.

Nanoparticles (NPs) can make their way to the brain and cause in situ damage, which is a concern for nanomaterial application and airborne particulate matter exposure. Our recent study indicated that respiratory exposure to silica nanoparticles (SiO2 NPs) caused unexpected cardiovascular toxic effects. However, the toxicities of SiO2 NPs in other organs have warranted further investigation. To confirm the accumulation of SiO2 NPs in the brain, we introduced SiO2 NPs with different diameters into mice via intranasal instillation (INI) and intravenous injection (IVI) in parallel. We found that SiO2 NPs may target the brain through both olfactory and systemic routes, but the size of SiO2 NPs and delivery routes both significantly affected their brain accumulation. Surprisingly, while equivalent SiO2 NPs were found in the brain regions, brain lesions were distinctly much higher in INI than in the IVI group. Mechanistically, we showed that SiO2 NPs introduced via INI induced brain apoptosis and autophagy, while the SiO2 NPs introduced via IVI only induced autophagy in the brain.

High-performance colorimetric immunoassay for determination of chloramphenicol using metal-organic framework-based hybrid composites with increased peroxidase activity.

Metal-organic frameworks (MOFs) as carriers for high-capacity loading of HRP-IgG and gold nanoparticles are introduced, to prepare MOF hybrids with enhanced peroxidase activity. The prepared MOF hybrids were employed to establish an indirect competitive colorimetric immunoassay for chloramphenicol (CAP) detection, in which the limit of detection for CAP is 0.006 µg·L-1, only one-fifth of that of the conventional ELISA using the same antibodies and antigens. The linear range was 0.008-0.108 µg·L-1, and the recovery of spiked milk samples varied in the range 76.0-106.0% through three independent experiments. Our proposed colorimetric immunoassay using the MOF hybrid immunoprobe provides a novel platform for ultra-sensitive determination of CAP residues, and it also could be used as a signal amplification model for the high-performance colorimetric immunoassay in food safety monitoring.

Impact of Protein Corona on Noncovalent Molecule-Gold Nanoparticle-Based Sensing.

Gold nanoparticle (AuNP)-based sensors have been extensively applied for sensing or imaging. It is known that a protein shell named protein corona (PC) formed around the nanomaterials could not only block the desired function of nanomaterials but also affect their behavior, which is a hot and important issue needing consideration. Therefore, we hypothesize that the formation of PC around AuNPs could inevitably affect the AuNP-based target assay. In this work, the effects of PC on the detection results in sensors based on AuNPs were studied. Three types of noncovalent molecule-AuNP sensors including AuNP-dichlorofluorescein, AuNP-aptamer, and AuNP-antibody-DNA were constructed, and several typical proteins (bovine serum albumin, fibrinogen, hemoglobin, and ß-lactoglobulin), milk, and fetal bovine serum were selected as models for the formation of PCs. This study shows that the PC could cause the loss of detection signals (up to 80%) and result in positive deviation of the measuring value compared with the true value. Moreover, the loss of detection signals could also increase the limits of detection (almost 10 times), decreasing the sensitivity of the three types of sensors, as proposed in this work compared to that without PC. Moreover, the polyethylene glycol backfilling strategy could not resolve the negative effects of PC on noncovalent molecule-AuNP sensors. The impacts of PC on detection results from noncovalent molecule-AuNP sensors would cause misdiagnosis or wasted production, which needs careful reconsideration of the AuNP-based detection in application fields like clinic diagnosis, food safety control, and so forth.

Intracellular Pathogen Detection Based on Dual-Recognition Units Constructed Fluorescence Resonance Energy Transfer Nanoprobe.

The intracellular invasion and survival of a pathogen like Staphylococcus aureus (S. aureus) within host cells enable them to resist antibiotic treatment and colonize long-term in the host, which leads to a series of clinical issues. Rapid and specific detection of intracellular bacteria is important in diagnosis of infection and guiding antibiotic administration. Herein, this work reports a simple one-step fluorescence resonance energy transfer (FRET) platform-based strategy to achieve specific and rapid detection of S. aureus in specimens of phagocytic cells. The aptamer modified quantum dots (Aptamer-QDs) and antibiotic molecule of Teicoplanin functionalized-gold nanoparticles (Teico-AuNPs) dual-recognition units to S. aureus are employed as energy donor and acceptor, respectively. Based on the "off" to "on" signal readout mode, when in the presence of target S. aureus, the donor and acceptor are close to each other and bring high FRET efficiency, which is suitable for analysis of intracellular S. aureus. After it was incubated with the sample for 2 h, the as-prepared FRET sensor showed selectivity to the target S. aureus, and the changed fluorescence signal shows an obvious variation with increasing concentration of S. aureus in pure buffer. When the FRET strategy was further applied to assay intracellular S. aureus, there was an obvious fluorescence signal change obtained both by spectrum analysis and visual fluorescence microscope observation when the average number of S. aureus in one host cell (NS. aureus/cell) was as low as 1, which can be attributed to the high fluorescence quenching efficiency of about 41.3%. It could be envisioned that this FRET nanoprobe with high fluorescence quenching efficiency may provide a simple approach for the facile, selective, and rapid diagnosis of an intracellular bacterial infection.

Detection of chloramphenicol with an aptamer-based colorimetric assay: critical evaluation of specific and unspecific binding of analyte molecules.

A chloramphenicol (CAP)-binding aptamer of 80 nucleotides (nt) was reported in 2011. In 2014, it was truncated to 40 nt and has since been used by most researchers, although a careful binding study is still lacking. In this work, binding assays using isothermal titration calorimetry and various DNA-staining dyes were performed. By comparing the truncated aptamer with three control sequences, no specific binding of CAP was observed in each case. The secondary structures of the original and truncated aptamers were analyzed, and it was shown that the likelihood of the truncated aptamer to retain the same binding mechanism as the original sequence is low. We further examined gold nanoparticle (AuNP)-based label-free colorimetric assays. By quantifying the extinction ratio at 620 nm over that at 520 nm, a similar color response was observed regardless of the sequence of DNA, suggesting the color change mainly reflected other events such as the adsorption of CAP by the AuNPs, instead of aptamer binding to CAP. Salt-induced aggregation experiments suggested direct adsorption of CAP on AuNPs. CAP only weakly inhibited DNA adsorption by AuNPs but did not displace pre-adsorbed DNA. Therefore, CAP adsorption by AuNPs needs to be considered when designing related sensors, for example, by using non-aptamer sequences as controls. This work calls for careful confirmation of aptamer binding and control experiments for designing aptamer and AuNP-based biosensors.

Cloning, expression, purification and characterization of a bispecific single-chain diabody against fluoroquinolones and sulfonamides in Escherichia coli.

A recombinant bispecific single-chain diabody (scDb), recognizing fluoroquinolones (FQs) and sulfonamides (SAs), was successfully constructed with two single-chain variable fragment antibodies (scFvs). The scDb gene was cloned into the expression vector pJB33, and 6×His-tagged scDb was expressed as soluble bodies in Escherichia coli RV308 host, then purified by one step affinity chromatography of immobilized metal ion affinity chromatography (IMAC). SDS-PAGE and Western blotting analysis of the purified scDb indicated that the prepared scDb was successfully expressed as a ∼60 kDa and the final purity of the scDb protein was up to 95% with yields of approximately 6 mg/L of bacterial culture. The scDb was further characterized by indirect competitive enzyme linked immunosorbent assay (icELISA), showing that the affinity and specificity of scDb were fully retained from the two parental scFvs, capable of simultaneously binding FQs and SAs. The 50% inhibition concentration (IC50) values of the optimized immunoassay were 0.45 ng mL(-1) for FQs and 0.75 ng mL(-1) for SAs, respectively. The scDb exhibited high affinity to 20 FQs and 14 SAs. Taken together, these findings suggested that the prepared scDb could be used to develop future novel immunoassay for simultaneous determination of 20 FQs and 14 SAs.

Development of a highly sensitive chemiluminescence enzyme immunoassay using enhanced luminol as substrate.

In this study, a high sensitivity chemiluminescence enzyme immunoassay (CLEIA) based on novel enhancers was developed. Under optimal conditions, we developed an enhanced chemiluminescence reaction (ECR) catalyzed by horseradish peroxidase (HRP-C) in the presence of 3-(10'-phenothiazinyl) propane-1-sulfonate (SPTZ) and 4-morpholinopyridine (MORP) as enhancers. The limit of detection of the newly prepared chemiluminescent cocktail for HRP was 0.33 pg/well, which is lower than that of commercial Super Signal substrate. The results showed that this novel chemiluminescent cocktail can significantly increase the light output of HRP-catalyzed ECR, which can be translated into a corresponding improvement in sensitivity. Similar improvements were observed in CLEIA for the determination of chloramphenicol in milk. In addition, the ECR of N-azoles as secondary enhancer was also presented.

Development of a rapid chemiluminescent ciELISA for simultaneous determination of florfenicol and its metabolite florfenicol amine in animal meat products.

BACKGROUND: A rapid one-step chemiluminescent competitive indirect enzyme-linked immunosorbent assay (CL-ciELISA) for florfenicol (FF) and its major metabolite florfenicol amine (FFA) residues in animal meat products has been developed. RESULTS: The 50% binding inhibition (IC50) values of the method were 0.195 µg kg⁻¹ for FFA and 0.24 µg kg⁻¹ for FF under optimum conditions. The cross-reactive rates for FF and FFA were 100.0% and 81.2%, respectively. FF and FFA were easily extracted from animal meat product with an FF/FFA extraction buffer, obtaining recoveries of 81.8-92.0% (FF) and 77.2-100% (FFA). The whole one-step CL-ciELISA test can be accomplished within 40 min in theory. The detection limits (LODs) of the assay were 0.98 µg kg⁻¹ for FF and 0.80 µg kg⁻¹ for FFA in animal meat samples. Finally, field animal meat samples were analyzed with the CL-ciELISA method, and the results correlated well with those obtained using traditional ELISA and a previously reported liquid chromatographic-tandem mass spectrometric method. CONCLUSION: The combined results confirmed the utility of this faster one-step CL-ciELISA for simultaneous trace analysis of FF and FFA. To date, this is the most rapid developed ELISA and CL-ELISA method for detection of FF and FFA.

Chiral silver nanoparticles with surface-anchored L(D)-Cys exhibit dissimilar biological characteristics in vitro but not in vivo.

This work investigated the influence of surface chirality on cellular internalization, cytotoxicity, and tissue distribution of silver nanoparticles (AgNPs). D-cysteine and L-cysteine are chiral forms of the amino acid cysteine. These enantiomers exhibit distinct spatial arrangements, with D-cysteine having a different configuration from L-cysteine. This structural dissimilarity can lead to variations in how these forms interact with biological systems, potentially impacting their cytotoxic responses. Four distinct types of AgNPs were synthesized, each possessing a unique surface coating: pristine AgNPs (pAgNPs), L-cysteine coated AgNPs (AgNPs@L-Cys), D-cysteine coated AgNPs (AgNPs@D-Cys), and racemic AgNPs coated with both L-Cys and D-Cys (AgNPs@L/D-Cys). We found chiral-dependent cytotoxicity of AgNPs on J774A.1 cells. Specifically, AgNPs@L-Cys exhibited the highest toxicity, and AgNPs@D-Cys exhibited the lowest toxicity. Meanwhile, the cellular uptake of the AgNPs correlated nicely with their cytotoxicity, with AgNPs@L-Cys being internalized to the greatest extent while AgNPs@D-Cys displays the least internalization. Scavenger receptors and clathrin predominantly mediate the cellular internalization of these AgNPs. Strikingly, the dissimilar cellular internalization and cytotoxicity of AgNPs with different chirality were eliminated upon protein corona coverage. Notably, following intravenous injection in mice, these four types of AgNPs showed similar patterns among various organs due to the inevitable protein adsorption in the bloodstream. These findings underscored the pivotal role of surface chirality in governing the biological interactions and toxicity of AgNPs.

Chemiluminescence competitive indirect enzyme immunoassay for 20 fluoroquinolone residues in fish and shrimp based on a single-chain variable fragment.

A chemiluminescent competitive indirect enzyme-linked immunosorbent assay, based on a mutant single-chain variable fragment (scFv), was developed to detect a broad range of fluoroquinolones (FQs) in fish and shrimp matrices. In this study, the best scFvC4A9H1_mut2 was adopted, which showed 10-fold improved affinity to sarafloxacin (SAR), difloxacin (DIF), and trovafloxacin (TRO), while the affinity to other FQs was fully inherited from wild-type scFvC4A9H1. In the optimized generic test, scFvC4A9H1_mut2 in combination with norfloxacin-ovalbumin conjugate and horseradish peroxidase-labeled anti-c-myc 9E10 antibody showed 50 % binding inhibition (IC50) at 0.12 µg kg(-1) for norfloxacin in buffer. Screening for the class of FQ antibiotics is accomplished using a simple, rapid extraction carried out with ethanol/acetic acid (99:1, v/v). This common extraction was able to detect 20 FQ residues such as s ciprofloxacin (CIP), danofloxacin, DIF, enoxacin, enrofloxacin (ENR), fleroxacin, amifloxacin, flumequine, levofloxacin, lomefloxacin hydrochloride, marbofloxacin, norfloxacin (NOR), ofloxacin, orbifloxacin, pazufloxacin, pefloxacin-d5 (PEF), prulifloxacin, SAR, sparfloxacin, and TRO in fish and shrimp. The limit of detection (LOD) for NOR was 0.2 µg kg(-1) and the LODs for CIP and ENR were all <0.2 µg kg(-1). Values of LODs inferred from the cross-reactivity data will range from approximately 0.23 µg kg(-1) for PEF to 2.1 µg kg(-1) for TRO. Field fish and shrimp samples were analyzed and compared to the results obtained from liquid chromatography tandem mass spectrometric method. All five instances (from 0.25 to 15.6 µg kg(-1)) in which FQs were present at concentrations near or above the assay LOD were identified as positive by the newly developed assay, demonstrating the usefulness of this assay as a screening tool.

A colorimetric aptasensor for detecting ochratoxin A based on label-free aptamer and gold nanozyme.

In this study, the aptamer of ochratoxin A (OTA) increased the negative charge density on the surface of gold nanoparticles (AuNPs) and promoted the release of hydroxyl radicals and Au3+ to enhance the peroxidase-like activity of the AuNPs. The OTA bound only to the aptamer and did not adsorb non-specifically to the AuNPs. Based on these two conclusions, a label-free colorimetric aptasensor was successfully developed, enabling the precise detection of OTA within the concentration range of 10-600 nM, with a remarkably low detection limit of 6.20 nM. The colorimetric aptasensor was applied to detect OTA in oats, corn, soybeans, rice, and glutinous rice.

Controllable and complete conversion of agarose into oligosaccharides and monosaccharides by microwave-assisted hydrothermal and enzymatic hydrolysis and antibacterial activity of agaro-oligosaccharides.

Hydrolysis of agar or agarose can yield two types of oligosaccharides: agaro-oligosaccharides (AOS) and neoagaro-oligosaccharides (NAOS). These oligosaccharides have various biological activities and promising applications in the future food industry and pharmaceuticals. In this study, we prepared AOS from agarose by microwave-assisted hydrothermal hydrolysis and then used a commercial ß-galactosidase to treat AOS for producing NAOS. A complete conversion from agarose to AOS or NAOS can be achieved by microwave hydrothermal treatment and one-step enzyme reaction, and the production process was completely green. In addition, we combined ß-galactosidase and α-neoagarobiose hydrolase from Saccharophagus degradans 2-40 (SdNABH) to treat AOS, and AOS was completely converted into monosaccharides. Then the results of the inhibitory activity of AOS on the growth of Streptococcus mutans showed that AOS might be a good potential sugar substitute for dental caries prevention. This study provides an efficient approach for the production of multiple mixed degrees of polymerization (DP) of pure AOS and NAOS without requiring acid catalyst and agarases while simplifying the production processes and reducing costs.

Structure and physicochemical properties of a new variety of purple rice (Oryza sativa L. indica) starch.

The investigation of the structure and physicochemical properties of starch extracted from a new variety of purple rice was the aim of this study. Starch extracted from a new variety of purple rice named Tianzi No. 1 (PRS) is different in structure and physicochemical properties compared with waxy rice starch (WRS), japonica rice starch (JRS), and indica rice starch (IRS). PRS granules were diversified in shape, and the birefringence of starch particles were clearly observed. Fourier-transform infrared (FT-IR) spectroscopy exhibited the degree of double helix and low short-range order structure of PRS differed from IRS, JRS and WRS. X-ray diffraction analysis shows that PRS presented a typical A-type XRD pattern and possessed lower crystallinity. Based on rheological experiment results, PRS had the highest apparent viscosity, storage modulus (G') and loss modulus (G″). According to textural experiments, PRS gels had higher textural paraments before and after retrogradation. Supplementary Information: The online version contains supplementary material available at 10.1007/s10068-022-01205-w.

Metabolites and metagenomic analysis reveals the quality of Pu-erh "tea head".

Tea head, a derivative product of Pu-erh tea, are tight tea lumps formed during pile-fermentation. The aim of this study was to reveal the differences of quality-related metabolites and microbial communities between ripened Pu-erh tea (PE-21) and tea heads (CT-21). Compared with PE-21, CT-21 showed a more mellow and smooth taste with slight bitterness and astringency, and can withstand multiple infusions. Metabolites analysis indicated CT-21 had more abundant water-soluble substances (47.39%) and showed significant differences with PE-21 in the main compositions of amino acids, catechins and saccharides which contributed to the viscosity of tea liquor, mellow taste and the tight tea lumps formation. Microbial communities and COG annotation analysis revealed CT-21 had lower abundance of Bacteria (84.05%), and higher abundance of Eukaryota (15.10%), carbohydrate transport and metabolism (8.28%) and glycoside hydrolases (37.36%) compared with PE-21. The different microbial communities may cause metabolites changes, forming distinct flavor of Pu-erh.

Iron oxide nanoparticles oxidize transformed RAW 264.7 macrophages into foam cells: Impact of pulmonary surfactant component dipalmitoylphosphatidylcholine.

Iron oxide nanoparticles (IONPs) are one of the most important components in airborne particulate matter that originally generated from traffic emission, iron ore mining, coal combustion and melting of engine fragments. Once IONPs entered respiratory tract and deposit in the alveoli, they may interact with pulmonary surfactant (PS) that distributed in the alveolar lining. Thereafter, it is necessary to investigate the interaction of inhaled IONPs and PS, which helps the understanding of health risk of respiratory health induced by IONPs. Using dipalmitoyl phosphatidylcholine (DPPC), the major components of PS, as a lipid model, we explored the interaction of DPPC with typical IONPs, Fe3O4 NPs and amino-functionalized analogue (Fe3O4-NH2 NPs). DPPC was readily adsorbed on the surface of both IONPs. Although DPPC corona depressed the cellular uptake of IONPs, IONPs@DPPC complexes caused higher cytotoxicity toward RAW 264.7 macrophages, compared to pristine IONPs. Mechanistic studies have shown that IONPs react with intracellular hydrogen peroxide, which promotes the Fenton reaction, to generate hydroxyl radicals. Iron ions could oxidize lipids to form lipid peroxides, and lipid hydroperoxides will decompose to generate hydroxyl radicals, which further promote cellular oxidative stress, lipid accumulation, foam cell formation, and the release of inflammatory factors. These findings demonstrated the phenomenon of coronal component oxidation, which contributed to IONPs-induced cytotoxicity. This study offered a brand-new toxicological mechanism of IONPs at the molecular level, which is helpful for further understanding the adverse effects of IONPs.

Preparation, characterization and evaluation of capsaicin-loaded indica rice starch nanoparticles.

Capsaicin (CAP) is an alkaloid with multiple physiological effects, but its application is difficult. In this research, indica rice starch nanoparticles (IRSNPs) based nanocarrier was used to load CAP to obtain capsaicin-loaded indica rice starch nanoparticles (CAP-IRSNPs). The microstructure, characteristics and in vitro release behaviors of CAP-IRSNPs were analyzed. CAP-IRSNPs presented average particle sizes of 617.84 ± 6.38 nm, encapsulation efficiency of 70.05 ± 1.78% and loading capacity of 13.41 ± 0.18%. Fourier-transform infrared spectroscopy confirmed that CAP-IRSNPs might be formed by hydrogen-bonding action. Differential scanning calorimetry and X-ray diffraction showed that IRSNPs influenced the crystallization and melting temperatures of CAP. In in vitro release study, CAP-IRSNPs exhibited a sustained release. The CAP concentration, CAP diffusion from matrix and matrix erosion might be the potentially possible mechanisms for capsaicin release from CAP-IRSNPs. These new results concluded that IRSNPs may be a promising nanocarrier for CAP or other hydrophobic bioactive ingredients.

A survey of ß-lactamase and 16S rRNA methylase genes among fluoroquinolone-resistant Escherichia coli isolates and their horizontal transmission in Shandong, China.

The prevalence of ß-lactamase, 16S rRNA methylase genes, and plasmid-mediated fluoroquinolone-resistance (PMQR) determinants (qnrC and qnrD) was determined by polymerase chain reaction in fluoroquinolone-resistant Escherichia coli isolated from a chicken farm, a pig farm, and a hospital in Shandong, China in 2007. The bla(TEM) and bla(CTX-M) were the most prevalent ß-lactamase genes in isolates from chickens (88.4%, 175/198 and 81.3%, 161/198) and hospitalized patients (87.8%, 122/139 and 69.1%, 96/139). The bla(TEM) was the most prevalent ß-lactamase gene observed in isolates from pigs (98.5%, 135/137). The gene bla(CMY-2) was also predominant among isolates from chickens (20.2%, 40/198). The bla(LAP-1) gene was first detected in one strain from chickens and humans (pig farm workers) in China. Only one strain from hospitalized patients was found to possess bla(SHV). The rmtB was the most prevalent 16S rRNA methylase gene detected in isolates from chickens (19.7%, 39/198) and hospitalized patients (15.8%, 22/139). To our knowledge, this is the first report of the detection of the qnrD gene in E. coli from chickens and pigs in China. The qnrC and bla(KPC) genes were not detected in any of the isolates. Results of southern hybridization revealed that PMQR determinants, ß-lactamases, and 16S rRNA methylase genes were located on the same plasmid in E. coli strains derived from patients. Also, PMQR determinants and ß-lactamase genes were localized on the same plasmid in an E. coli strain of animal origin. Results of conjugation experiments revealed that all of these plasmid-based resistance genes can be transferred by conjugation through horizontal transmission.

Effects of sucrose on pasting, thermal, rheological and textural properties of native and alcohol-alkali-treated waxy rice starch.

In this work, the physicochemical properties of native waxy rice starch (WRS) and alcohol-alkali-treated waxy rice starch (AAT-WRS) were studied in the presence of sucrose. The results indicated that the addition of sucrose improved the transparency and freeze-thaw stability of WRS pastes and AAT-WRS pastes. Differential scanning calorimetry showed that the gelatinization temperatures of WRS increased with increased sucrose concentration, but the gelatinization enthalpy increased at low concentration of sucrose and decreased at high concentration. Rheological measurements indicated that sucrose addition had no significant effect on the pseudoplastic shear-thinning behaviors of WRS pastes and AAT-WRS pastes, but changed the apparent viscosity. Dynamic moduli (G' and G″) values of WRS pastes and AAT-WRS pastes with or without sucrose showed frequency dependency and sucrose addition dependency. The elastic behavior was dominant over viscous in the WRS-sucrose mixed pastes, while the AAT-WRS-sucrose mixed pastes was the opposite. The textural paraments of WRS and AAT-WRS before or after retrogradation increased with the increasing concentration of sucrose. These results suggested that sucrose potentially changed the physicochemical properties of WRS and AAT-WRS.

A tandem activation of NLRP3 inflammasome induced by copper oxide nanoparticles and dissolved copper ion in J774A.1 macrophage.

For the first time, we reported that CuONPs exposure induced interleukin (IL)-1ß-mediated inflammation via NOD-, LRR- and pyrin domain-containing protein 3 (NLRP3) inflammasome in J774A.1 macrophage. Mechanistically, CuONPs activated NLRP3 inflammasome is a two-fold process. Firstly, CuONPs challenge caused lysosomal damage, along with the release of cathepsin B, which directly mediated the activation of NLRP3 inflammasomes. Interestingly, after the deposition in lysosomes, CuONPs may release copper ion due to the acidic environment of lysosomes. Consequently, the released copper ions significantly induced cellular oxidative stress and further mediated the activation of NLRP3 inflammasomes. Moreover, CuONPs exposure could prime J774A.1 macrophage to express pro-IL-1ß through myeloid differentiation factor 88 (MyD88)-dependent Toll-like receptor 4 (TLR4) signal pathway subsequently activating nuclear transcription factor kappa B (NF-κB).