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Acrylamide is a group 2A carcinogen and potential endocrine disruptor that can enter the ecosystem by various routes and has recently become a dangerous pollutant. This widely used chemical can enter the human body via air inhalation, food or water consumption, or skin contact. In this study, we developed a peptide probe for the detection of acrylamide by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) after its micro-tagging with a peptide. Direct detection of acrylamide by MALDI-TOF MS is not feasible due to its poor ionization in the MALDI interface, which hinders its analysis by the technique. After microwave irradiation for 2 min, the formed acrylamide-peptide derivative was detected easily by MALDI-TOF MS without the need for extraction procedures. The procedure does not involve organic solvents and a water-soluble peptide that allows detection of acrylamide in small sample volumes with a limit of detection (LOD) of 0.05 ng/μL. The relative standard deviation (RSD) and relative error (RE) of the measurements were

The development of a portable sensor for on-site sensing of 2, 4, 6-trinitrotoluene (TNT) and other nitro aromatics (NAs) is still needed to strengthen environmental protection and public security. Herein, we report the fabrication of fluorescent few-layered MnO₂ nanosheets (FL-MnO₂ NSs) via N,N-dimethyl formamide-mediated ultrasonication and hydrothermal treatment. Electron and spectroscopy studies demonstrate that the modification of the FL-MnO₂ NSs (named as PEI-FL-MnO₂ NSs) with amine-rich polyethyleneimine (PEI) produced planar, β-phase, well-crystalline, and positively charged nanosheets. This PEI-related surface modification not only improves the quantum yield of the FL-MnO₂ NSs from 7.6 to 13.2% but also provides multiple binding sites with nitro groups of NAs. It was found that the presence of TNT, as an example of NAs, efficiently quenched the fluorescence of the PEI-FL-MnO₂ NSs due to the formation of Meisenheimer complexes through the efficient adsorption process. In comparison to PEI-modified MnO₂ quantum dots, the PEI-FL-MnO₂ NSs exhibited relatively high sensitivity to TNT with a limit of detection of 20 pM. By depositing PEI-FL-MnO₂ NSs on a filter paper, the as-made paper sensor was capable of visualizing 10 nM TNT in drinking water and 10 μM TNT on a postal envelope through the appearance of a quenching spot under a UV lamp. Besides, the adsorbed PEI polymer enables the PEI-FL-MnO₂ NSs to electrostatically interact with citrate-capped metal nanoparticles, producing the composites for catalyzing NaBH₄-mediated reduction of NAs. Since the PEI-FL-MnO₂ NSs possess a large surface area to capture NAs and serve as an electron acceptor for BH₄–, the formed composites efficiently catalyze the reaction of NaBH₄ and NAs as compared to metal nanoparticles and PEI-FL-MnO₂ NSs alone.

Currently, few phosphorescent materials (PMs) possess a long phosphorescence lasting time and have potential for application in chemical sensors. Herein, we disclose that the incorporation of few-layer molybdenum disulfide quantum dots (FL-MoS2 QDs) into poly(vinyl alcohol) (PVA) matrices leads to the emission of bright green phosphorescence with a long lasting time of 3.0 s and a phosphorescence quantum yield of 20%. This enhanced phosphorescence originates from the formation of O–H⋯S hydrogen bonding networks between the rich sulfur sites of the FL-MoS2 QDs and the hydroxyl groups of the PVA molecules, which not only rigidifies the vibration modes of the FL-MoS2 QDs but also provides an oxygen barrier. Further investigations reveal that the FL-MoS2 QD/PVA composites exhibit a longer phosphorescence lasting time than N,S-doped carbon dots, few layer tungsten disulfide quantum dots, Rhodamine 6G, and Rhodamine B in PVA matrices. Since heat efficiently induced the removal of water moisture from PVA matrices, the FL-MoS2 QD/PVA composites could be implemented for phosphorescence turn-on and naked-eye detection of temperature variations ranging from 30 to 70 °C. By contrast, the carbon dot/PVA composites were incapable of sensing environmental temperature due to their weak hydrogen bonding with the hydroxyl groups of PVA matrices. Additionally, this study reveals the potential of the FL-MoS2 QD/PVA composites as an advanced security ink for anti-counterfeiting and encryption applications. The given results could open a new direction for potential application of two-dimensional quantum dots in phosphorescence-based sensors and security inks.

This study fabricated yellow-emitting CDs (Y-CDs) by hydrothermal treatment of citric acid and urea and applied them as a fluorescence turn-on platform for sensitive and selective detection of lipopolysaccharide (LPS) based on the non-shifted AIEE of peptide-stabilized CD aggregates. The designed peptide (named K3) consisting of aggregation-active and LPS-recognition units triggered the aggregation of Y-CDs, switching on their fluorescence through the blue-shifted AIEE process. The formed K3-stabilized Y-CD aggregates (K3-YCDAs) specifically interacted with LPS at neutral pH, demonstrating that the sequence of the decorated peptide was highly connected with their selectivity and sensitivity. The K3-YCDAs provided a fast response time (within 5 min) to detect LPS with a quantification range of 0.5-100.0 nM and a limit of detection (LOD, signal-to-noise ratio of 3) of 300.0 pM. By integrating ultrafiltration membranes as a concentration device with K3-YCDAs as a sensing probe, the LOD for LPS was further reduced to 3.0 pM. The determination of picomolar levels of plasma LPS by the K3-YCDAs coupled to the centrifugation ultrafiltration was demonstrated to fall within the specificity range of clinical interest for sepsis patients. Also, the K3-YCDAs served as a fluorescent probe to selectively image and quantify E. coli cells. The distinct advantages of the K3-YCDAs for LPS include fast response time, wide linear range, low detection limit, and excellent selectivity compared to previously reported sensors.

Background/purpose Eosinophils are recruited to the brain of mice after infection with Angiostrongylus canonensis. Several factors produced by infected mice are well known playing the role to chemoattract eosinophils from the blood into the brain. The purpose of this study is to investigate whether Angiostronylus cantonensis young-adult worms (AcYA) have components which have eosinophilic chemotactic activity. Methods Eosinophil chemotactic activity of AcYA was tested by Boyden blind-well chamber technique. The components of AcYA were analysed by SDS-PAGE and Mass spectrometry. Furthermore, galectin-9 in the cerebrospinal fluid (CSF) of infected mice and galectin-9-like in AcYA were measured by ELISA technic and also were recognized by western blot analysis respectively. Results Excretory-secretory products of AcYA did not show eosinophil chemotactic activity. However, the extracts of AcYA showed protein concentration-dependent eosinophil chemotactic activity and reached the peak at the 24 μg/ml. The eosinophil chemotactic activity was significantly reduced by lactose. The components of AcYA at molecular weights of approximatively 15 kDa and 35 kDa showed several galectins component in Mass spectrometric analysis. Furthermore, galectin-9-like in AcYA was recognized by ELISA and western blot analysis. In parallel with increase of galectin-9 in the CSF, eosinophils were also significantly increased in mouse after infected with A. cantonensis. Conclusion Galectin-9-like in AcYA and galectin-9 in mouse CSF were confirmed demonstrating eosinophil chemotactic activity both in vitro study and in the infection of mouse in this study.

Pulmonary artery hypertension (PAH) pathology involves extracellular matrix (ECM) remodeling in cardiac tissues, thus promoting cardiac fibrosis progression. miR-29a-3p reportedly inhibits lung progression and liver fibrosis by regulating ECM protein expression, however, its role in PAH-induced fibrosis remains unclear. In this study, we aimed to investigate the role of miR-29a-3p in cardiac fibrosis progression in PAH and its influence on ECM protein thrombospondin-2 (THBS2) expression. The diagnostic and prognostic values of miR-29a-3p and THBS2 in PAH were evaluated. The expressions and effects of miR-29a-3p and THBS2 were assessed in cell culture, monocrotaline-induced PAH mouse model, and patients with PAH. The levels of circulating miR-29a-3p and THBS2 in patients and mice with PAH decreased and increased, respectively. miR-29a-3p directly targets THBS2 and regulates THBS2 expression via a direct anti-fibrotic effect on PAH-induced cardiac fibrosis. The circulating levels of miR-29a-3p and THBS2 were correlated with PAH diagnostic parameters, suggesting their independent prognostic value. miR-29a-3p targeted THBS2 expression via a direct anti-fibrotic effect on PAH-induced cardiac fibrosis, indicating miR-29a-3p acts as a messenger with promising therapeutic effects.

Sensors that can specifically and accurately detect glycosaminoglycans are rare. Here, a dual-mode platform for fluorescence intensity and lifetime sensing of plasma heparin and fluorescence imaging of heparan sulfate proteoglycan-expressed cancer cells was developed by stabilizing the intramolecular charge transfer (ICT) state of dansyl acid-labeling AG73 (DA-AG73) peptide with glutathione-capped gold nanoclusters (GSH-AuNCs). DA-AG73 peptides, including an electron-donor dimethylamino group and an electron-withdrawing sulfonamide moiety in the labeled DA molecules, emitted weak fluorescence due to the formation of the twisted ICT excited state. The complexation of heparin with DA-AG73 peptides followed by interacting with the GSH-AuNCs could restrict the rotation of the dimethylamino groups of the labeled DA molecules, triggering the transition from their twisted ICT state to ICT excited state. As a result, the fluorescence intensity and lifetime of the labeled DA molecules in DA-AG73 peptides were gradually enhanced with increasing the heparin concentration. The proposed platform provided excellent selectivity toward heparin and heparan sulfate and exhibited two linear calibration curves for quantifying 20–800 nM and 20–1000 nM heparin in the fluorescence intensity and lifetime modes, respectively. The proposed platform was practically applied for the fluorescence intensity and lifetime determination of plasma heparin and for the selective imaging of heparan sulfate proteoglycan-expressed cells.

High mobility group box 1 (HMGB1) has been demonstrated to promote the migration and invasion of non-small cell lung cancer (NSCLC). However, the mechanism of action of HMGB1 in regulating tumor mobility remains unclear. Therefore, we aimed to investigate whether HMGB1 affects mitochondria distribution and regulates dynamin-related protein 1 (DRP1)-mediated lamellipodia/filopodia formation to promote NSCLC migration. The regulation of mitochondrial membrane tension, dynamics, polarization, fission process, and cytoskeletal rearrangements in lung cancer cells by HMGB1 was analyzed using confocal microscopy. The HMGB1-mediated regulation of DRP1 phosphorylation and colocalization was determined using immunostaining and co-immunoprecipitation assays. The tumorigenic potential of HMGB1 was assessed in vivo and further confirmed using NSCLC patient samples. Our results showed that HMGB1 increased the polarity and mobility of cells (mainly by regulating the cytoskeletal system actin and microtubule dynamics and distribution), promoted the formation of lamellipodia/filopodia, and enhanced the expression and phosphorylation of DRP1 in both the nucleus and cytoplasm. In addition, HMGB1 and DRP1 expressions were positively correlated and exhibited poor prognosis and survival in patients with lung cancer. Collectively, HMGB1 plays a key role in the formation of lamellipodia and filopodia by regulating cytoskeleton dynamics and DRP1 expression to promote lung cancer migration.

Thrombospondin-1 (TSP1) is involved in corneal wound healing caused by chemical injury. Herein, we examined the effects of TSP1 on hypoxia-induced damages and wound-healing activity in human corneal epithelial (HCE) cells. Exosomal protein expression was determined using liquid chromatography-tandem mass spectrometry, and HCE cell migration and motility were examined through wound-healing assay and time-lapse microscopy. Reestablishment of cell junctions by TSP1 was assessed through confocal microscopy and 3D image reconstruction. Our results show that CoCl2 -induced hypoxia promoted HCE cell death by paraptosis. TSP1 protected these cells against paraptosis by attenuating mitochondrial membrane potential depletion, swelling and dilation of endoplasmic reticulum and mitochondria, and mitochondrial fission. Exosomes isolated from HCE cells treated with TSP1 contained wound healing-associated proteins that were taken up by HCE cells to promote tissue remodeling and repair. TSP1 protected HCE cells against hypoxia-induced damages and inhibited paraptosis progression by promoting cell migration, cell-cell adhesion, and extracellular matrix remodeling. These findings indicate that TSP1 ameliorates hypoxia-induced paraptosis in HCE cells and promotes wound healing and remodeling by regulating exosomal protein expression. TSP1 may, therefore, play important roles in the treatment of hypoxia-associated corneal diseases.

Context Small extracellular vesicles (sEVs) have emerged as modulators of the disease microenvironment, thereby supporting disease progression. However, the potential role of EVs and their content to the pathophysiology of endometriosis remain unclear. Objective This work aimed to investigate whether the EVs from eutopic (Eu) and ectopic (Ec) endometrial stromal cells (ESCs) differ with respect to protein composition and role in endometriosis. Methods Human Eu and Ec endometrium–derived ESCs were isolated from samples of the same patients (n = 3). sEVs were isolated from ESCs via ultracentrifugation; these sEVs were characterized by Western blotting, transmission electron microscopy, and nanoparticle tracking analysis and analyzed using mass spectrometry. The potential role of EcESCs-derived sEVs (EcESCs-sEVs) in endometriosis was explored by assaying their effects on cell viability/proliferation, migration, and angiogenesis. Results In total, 105 ESCs-sEV–associated proteins were identified from EcESCs-sEVs and EuESCs-sEVs by mass spectrometry analysis. The protein content differed between EcESCs-sEVs and EuESCs-sEVs, with annexin A2 (ANXA2) being the most prominent difference—present in EcESCs-sEVs but not EuESCs-sEVs. We also found that sEVs-ANXA2 regulates the motility, proliferation, and angiogenesis of ESCs via the extracellularly regulated kinase (ERK)/STAT3 pathway. Notably, treatment of ESCs with sEVs-ANXA2 resulted in increased proliferation and motility, suggesting that sEVs-ANXA2 may be involved in regulating endometriosis. Our data suggest that EcESCs-sEVs-ANXA2 regulates the motility and the angiogenic potential of ESCs, implying a role for sEVs-ANXA2 in the pathogenesis of endometriosis. Conclusion The study of sEVs-ANXA2 from Ec endometriotic cells uncovers a new mechanism of endometriosis progression and will inform the development of novel therapeutic strategies.

Currently, few phosphorescent materials (PMs) possess a long phosphorescence lasting time and have potential for application in chemical sensors. Herein, we disclose that the incorporation of few-layer molybdenum disulfide quantum dots (FL-MoS

Dipeptidyl peptidase 4 (DPP-4) inhibitors are incretin-based medications used as oral antidiabetic agents for the treatment of type 2 diabetes. However, DPP-4 inhibitors produce side effects like acute pancreatitis, upper respiratory tract infection, nasopharyngitis, urinary tract infection, serious allergies, cardiovascular diseases, hemolysis, and retinopathy. Hence, the development of a fast and simple method to detect DPP-4 inhibitors in body fluids is important. In this study, we developed a derivatization-assisted microextraction method to enhance the detection sensitivity for trace levels of a DPP-4 inhibitor, sitagliptin, from a small volume (10 μL) of human plasma by using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). Subjecting the analyte to 100 W microwave irradiation after derivatization using a quinoline alkylating reagent (8-bromomethyl quinilone, BrMQ) shortened the reaction time to ~120 s and allowed the target analyte to be easily extracted to a small volume of the organic layer (20 μL). The analyte was then detected by MALDI-TOF MS using α-cyano-4-hydroxycinnamic acid as the matrix. The relative standard deviation and relative error were below 10% in intra- and inter-day assays. Using sitagliptin-d4 as an internal standard, the limits of quantitation and detection were found to be 0.03 μg/mL and 0.01 μg/mL, respectively. All the derivatization and extraction procedures described herein were of microliter grade. This method could effectively reduce the use of organic chemicals and solvents, thereby proving to be an eco-friendly strategy that will cause no harm to the environment.

Mass spectrometry is a powerful instrument for identifying the structures of target compounds. However, one of the major disadvantages is that mass spectrometry could not directly distinguish between enantiomers. Cysteine, a special amino acid containing a thiol group, can undergo a variety of oxidative modifications. The different redox states of cysteine existing in a peptide or protein may interact differently with the stationary phase in a chromatographic column. In this study, we developed a peptide that can be used as a chiral derivatization reagent for the enantioseparation of drugs using nano-scale liquid chromatography. Ibuprofen, a model analyte to evaluate the chiral separation efficiency of peptide-based derivatizing reagents, is a widely used anti-inflammatory drug. After the peptide derivatizing reagents were micro-coupled with ibuprofen, the solution was injected into a nano-flow column. The separation of the (R) and (S) forms of ibuprofen was performed on the nano-scale, using a C18 75 μm capillary column at a flow rate of 400 nL/min. The results indicated that the peptide AAAACAAAR comprising tri-oxidized cysteine is most suitable for separating (R)- and (S)-ibuprofen at the nano-scale flow rate without using a chiral column. The detection of (R)- and (S)-ibuprofen in only 10 μL human plasma (by pricking the finger) was sufficient with an injection volume of 200 nL. The linear range for detecting the (R) and (S) forms of ibuprofen in plasma was 1–100 μg/mL, the relative standard deviation (RSD) for the precision analyses was below 17 %, and the recovery for the method was 86%–109%. Using the peptide as a chiral derivatization reagent with a suitable cysteine oxidation state, we successfully separated and quantitated the (R) and (S) forms of ibuprofen in commercial tablets and plasma samples. Finally, our computation simulations suggested that the oxidization state of cysteine affects the formation of solute–solvent hydrogen bonds, which might influence the separation efficiency on the C18 column by changing the molecular binding affinity with the column stationary phase.

The combination of superimposition and hybrid target powers using a superimposed high power impulse magnetron sputtering (HiPIMS) power and middle-frequency (MF) power system, together with the hybrid of direct-current (DC) and radio-frequency (RF) powers was adopted in this work to grow CrTiBN hard coatings. The flexibility of tailoring the chemical composition, phase, deposition rate, microstructure and mechanical properties of CrTiBN coatings through the feedback control of reactive N2 gas flow rate and the corresponding Ti target poisoning status by a plasma emission monitoring (PEM) system was successfully illustrated in this work. Both the peak current and peak power density of Ti target increased with increasing poisoning ratio, i.e., decreasing PEM set point. The 1.64 times higher deposition rate of CrTiBN films can be obtained when the Ti target status changes from poisoning (PEM set point 30%) to transition regime (PEM set point 70%). Fine and compact film microstructure, and a very smooth surface are fabricated for all films. Each coating shows a low coefficient of friction and low wear rate ranging from 0.46 to 0.49 and from 2.06 to 5.73 × 10−6 mm3 N−1m−1, respectively. The highest hardness of 34.1 GPa, adequate adhesion and wear resistance can be found for the CrTiBN coating as deposited at PEM set point of 40% and Ti target poisoning ratio of 60%.

Although reef-building corals are threatened by a number of anthropogenic impacts, certain scleractinian-dinoflagellate (genus Symbiodinium) endosymbioses have proven markedly resilient to environmental change. For instance, corals from upwelling habitats of Southern Taiwan withstand both short- and long-term increases in temperature, potentially due to their routine exposure to highly variable temperature regimes in situ. To gain a greater understanding of the proteomic basis for such acclimatization to unstable environmental conditions, specimens of the Indo-Pacific reef-building coral Seriatopora hystrix Dana 1846 were sampled during a period of stable temperature conditions from 1) a site characterized by frequent upwelling events in Southern Taiwan and 2) a nearby, non-upwelling control site in the Taiwan Strait. Two-dimensional gel electrophoresis followed by sequencing of differentially concentrated proteins with mass spectrometry unveiled significantly more proteins involved in the cellular stress response in coral hosts of the upwelling site. Although such stress protein signatures could be indicative of sub-lethal levels of cellular stress, especially given the relatively higher sediment loads characteristic of the upwelling site, these proteins may, in contrast, have been constitutively maintained at high levels in preparation for large fluctuations in temperature and other abiotic parameters (e.g., nutrient levels) brought upon by upwelling events.

The alternative transcription factor σB in Bacillus cereus governs the transcription of a number of genes that confer protection against general stress. This transcription factor is regulated by protein-protein interactions among RsbV, RsbW, σB, RsbY, RsbM and RsbK, all encoded in the sigB cluster. Among these regulatory proteins, RsbV, RsbW and σB comprise a partner-switching mechanism. Under normal conditions, σB remains inactive by associating with anti-sigma factor RsbW, which prevents σB from binding to the core RNA polymerase. During environmental stress, RsbK activates RsbY to hydrolyze phosphorylated RsbV, and the dephosphorylated RsbV then sequesters RsbW to liberate σB from RsbW. Although the σB partner-switching module is thought to be the core mechanism for σB regulation, the actual protein-protein interactions among these three proteins in the cell remain to be investigated. In the current study, we show that RsbW and RsbV form a long-lived complex under transient stress treatment, resulting in high persistent expression of RsbV, RsbW and σB from mid-log phase to stationary phase. Full sequestration of RsbW by excess RsbV and increased RsbW:RsbV complex stability afforded by cellular ADP contribute to the prolonged activation of σB. Interestingly, the high expression levels of RsbV, RsbW and σB were dramatically decreased beginning from the transition stage to the stationary phase. Thus, protein interactions among σB partner-switching components are required for the continued induction of σB during environmental stress in the log phase and significant down-regulation of σB is observed in the stationary phase. Our data show that σB is temporally regulated in B. cereus.

Background Skin cancer is an important environmentally-related health issue. Although sun exposure is closely associated with increasing environmental heat, the effects of environmental heat on the skin, especially in the context of photocarcinogenesis, has not been carefully examined. Objectives This study aimed to explore the effects and interactions of UVB radiation and environmental heat on photocarcinogenesis of the skin using cell and animal models. Methods Cultured keratinocytes and hairless mice were exposed to different treatment conditions including UVB radiation and environmental heat. The effects of treatment on keratinocyte and mice skin were evaluated at indicated time points. Results UVB induced DNA damage was significantly lower in keratinocytes that were pretreated in an environment with slightly elevated temperature followed by UVB treatment (Heat-UVB) as compared to UVB and UVB radiation followed by exposure to equivalent increase in environmental heat (UVB-Heat) groups. Similar phenomenon was observed in terms of keratinocyte viability. In the animal model, it was found that Heat-UVB treated mice showed delayed and reduced tumor formation as compared to the UVB and UVB-Heat treated groups. Quantum simulation analyses demonstrated that the energy required for CPD formation at environment with higher temperature required considerable higher energy as compared to CPD formation at lower temperature. Conclusion Taken together, our results demonstrated that with equivalent UVB exposure, higher temperature environment may protect cells against subsequent UVB-induced DNA damages.

Reactive oxygen species (ROS) can induce oxidative stress and are associated with cell death and chronic diseases in organisms. In the treatment of disease, drugs that induce ROS are associated with many side effects and unpleasant symptoms. Therefore, during the assessment of new drugs and candidate compounds, ROS generation is an issue of concern, because ROS can modify proteins, lipids, and nucleic acids within organisms and alter their biological functions. In this work, we designed a peptide-based probe for the rapid (

Based on the oxidative stress theory, aging derives from the accumulation of oxidized proteins induced by reactive oxygen species (ROS) in the cytoplasm. Hydrogen peroxide (H2O2) elicits ROS that induces skin aging through oxidation of proteins, forming disulfide bridges with cysteine or methionine sulfhydryl groups. Decreased Ca2+ signaling is observed in aged cells, probably secondary to the formation of disulfide bonds among Ca2+ signaling-related proteins. Skin aging processes are modeled by treating keratinocytes with H2O2. In the present study, H2O2 dose-dependently impaired the adenosine triphosphate (ATP)-induced Ca2+ response, which was partially protected via co-treatment with β-mercaptoethanol, resulting in reduced disulfide bond formation in inositol 1, 4, 5-trisphosphate receptors (IP3Rs). Molecular hydrogen (H2) was found to be more effectively protected H2O2-induced IP3R1 dysfunction by reducing disulfide bonds, rather than quenching ROS. In conclusion, skin aging processes may involve ROS-induced protein dysfunction due to disulfide bond formation, and H2 can protect oxidation of this process.

We report citrate-capped platinum nanoparticles (Pt NPs) as oxidase mimetics for effectively catalyzing the oxidation of 3,3′,5,5′-tetramethylbenzidine (TMB), 2,2′-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid), dopamine, and methylene blue in the presence of O 2 . To confirm oxidase-like activity of citrate-capped Pt NPs, their activity toward oxygen reduction reaction was studied using cyclic voltammetry and rotating ring-disk electrode method. The results obtained showed that Pt NP NPs can catalyze the oxidation of organic substrates to the colored product and the reduction of oxygen to water through a four-electron exchange process. Because the aggregation of Pt NPs can inhibit their oxidase-like activity and protamine can recognize heparin, we prepared the protamine-modified Pt NPs through direct adsorption on the surface of citrate-capped Pt NPs. The electrostatic attraction between heparin and protamine-stabilized Pt NPs induced nanoparticle aggregation, inhibiting their catalytic activity. Therefore, the lowest detectable heparin concentrations through UV–vis absorption and by the naked eye were estimated to be 0.3 and 60 nM, respectively. Moreover, the proposed system enabled the determination of the therapeutic heparin concentration in a single drop of blood.

Enterovirus 71 (EV71) infection can cause airway symptoms, brainstem encephalitis, neurogenic shock, and neurogenic pulmonary edema with high morbidity and mortality. There is no proven therapeutic modality. Flos Farfarae is the dried flower bud of Tussilago farfara L. that has been used to manage airway illnesses for thousands of years. It has neuro-protective activity and has been used to manage neuro-inflammatory diseases. However, it is unknown whether Flos Farfarae has activity against EV71-induced neuropathy. The current study used both human foreskin fibroblast (CCFS-1/KMC) and human rhabdomyosarcoma (RD) cell lines to test the hypothesis that a hot water extract of Flos Farfarae could effectively inhibit EV71 infection. The authenticity of Flos Farfarae was confirmed by HPLC-UV fingerprint. Through plaque reduction assays and flow cytometry, Flos Farfarae was found to inhibit EV71 infection ([Formula: see text]). Inhibition of viral replication and protein expression were further confirmed by reverse transcription polymerase chain reaction (RT-PCR) and quantitative RT-PCR (qRT-PCR), and western blot, respectively. The estimated IC[Formula: see text]s were 106.3[Formula: see text][Formula: see text]g/mL in CCFS-1/KMC, and 15.0[Formula: see text][Formula: see text]g/mL in RD cells. Therefore, Flos Farfarae could be beneficial to inhibit EV71 infection by preventing viral replication and structural protein expression.

RATIONALE Understanding drug-drug interactions and predicting the side effects induced by polypharmacy are difficult because there are few suitable platforms that can predict drug-drug interactions and possible side effects. Hence, developing a platform to identify significant protein markers of drug-drug interactions and their associated side effects is necessary to avoid adverse effects. METHODS Human liver cells were treated with ethosuximide in combination with cimetidine, ketotifen, metformin, metronidazole, or phenytoin. After sample preparation and extraction, mitochondrial proteins from liver cells were isolated and digested with trypsin. Then, peptide solutions were detected using a nano ultra-performance liquid chromatographic system combined with tandem mass spectrometry. The Ingenuity Pathway Analysis tool was used to simulate drug-drug interactions and identify protein markers associated with drug-induced adverse effects. RESULTS Several protein markers were identified by the proposed method after liver cells were co-treated with ethosuximide and other drugs. Several of these protein markers have previously been reported in the literature, indicating that the proposed platform is workable. CONCLUSIONS Using the proposed in vitro platform, significant protein markers of drug-drug interactions could be identified by mass spectrometry. This workflow can then help predict indicators of drug-drug interactions and associated adverse effects for increased safety in clinical prescriptions.

X-inactive-specific transcript (XIST), a long non-coding RNA, is essential for the initiation of X-chromosome inactivation. However, little is known about other roles of XIST in the physiological process in eukaryotic cells. In this study, the bioinformatics approaches revealed XIST could be processed into a small non-coding RNA XPi2. The XPi2 RNA was confirmed by a northern blot assay; its expression was gender-independent, suggesting the role of XPi2 was beyond X-chromosome inactivation. The pull-down assay combined with LC-MS-MS identified two XPi2-associated proteins, nucleolin and hnRNP A1, connected to the formation of G-quadruplex. Moreover, the microarray data showed the knockdown of XPi2 down-regulated the KRAS pathway. Consistently, we tested the expression of ten genes, including KRAS, which was correlated with a G-quadruplex formation and found the knockdown of XPi2 caused a dramatic decrease in the transcription level of KRAS among the ten genes. The results of CD/NMR assay also supported the interaction of XPi2 and the polypurine-polypyrimidine element of KRAS. Accordingly, XPi2 may stimulate the KRAS expression by attenuating G-quadruplex formation. Our present work sheds light on the novel role of small RNA XPi2 in modulating the G-quadruplex formation which may play some essential roles in the KRAS- associated carcinogenesis.

Valproic acid (VA) is the drug of choice for managing epilepsy syndrome and is an effective treatment for various seizures. In recent years, new applications of VA have been developed for clinical treatment of other diseases. In the degradation pathway, cytochrome P450 participates in the biotransformation of VA to other metabolites. The resulting compounds then contribute to apoptosis induced by oxidative stress. Hence, clinical applications of VA medication should be selected carefully to avoid potentially severe side effects such as hepatotoxicity, hypersensitivity syndrome reactions, neurological toxicity and teratogenicity. This work established a simple method of using a microwave assisted derivatization procedure for improved detection and monitoring of VA derivatives by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). The calibration curve was 1–100 μM for VA analysis, and good linearity was obtained. In intra- and inter-day analyses, all relative standard deviation (RSD) and relative error (RE) values were below 7.5%.

Electropolymerizable carbazole-functionalized imidazolium and pyridinium-based ionic liquids (ILs) were synthesized and characterized using 1H, 13C, 19F NMR, and ESI-MS. They were electropolymerized onto indium tinoxide (ITO) electrodes via cyclic voltammetric polymerization in several selected room temperature ILs (RTILs) as well as in conventional organic solvents. Two redox couples, corresponding to the monomer and polymer, respectively, of the carbazole-functionalized ILs, were observed during the cyclic voltammetric polymerization. It was found, as common ILs, that the anionic components exhibit crucial effects on the physicochemical properties of the carbazole-functionalized ILs. The anions of which were replaced by the anions of the electrolytes in which the monomers were dissolved. The RTILs used for the electropolymerization thus determined the electrochemical behavior of the monomers, the surface morphologies and the physicochemical properties of the poly(ILs) films. After the modification of ITO electrodes with poly(ILs), the electrochemical performance of the semiconducting electrodes was significantly improved towards uric acid oxidation. The poly(ILs) show adjustable properties (via counter anions), and exhibit a wide range of solubility in organic solvents, and electrochromic and solvatochromic properties.

It is widely accepted that snake venom cardiotoxins (CTXs) target the plasma membranes of cells. In the present study, we investigated the role of Asp residues in the interaction of Naja atra cardiotoxin 1 (CTX1) and cardiotoxin 3 (CTX3) with phospholipid bilayers using chemical modification. CTX1 contains three Asp residues at positions 29, 40, and 57, CTX3 contains two Asp residues at positions 40 and 57. Compared to Asp29 and Asp40, Asp57 was sparingly modified with semi-carbazide, as revealed by matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass and mass/mass analyses. Thus, semi-carbazide-modified CTX1 (SEM-CTX1) mainly contained modified Asp29 and Asp40, while SEM-CTX3 contained modified Asp40. Compared to that of native toxins, trifluoroethanol easily induced structural transition of SEM-CTX1 and SEM-CTX3, suggesting that the structural flexibility of CTXs was constrained by Asp40. Modification of Asp29 and Asp40 markedly promoted the ability of CTX1 to induce permeability of cell membranes and lipid vesicles, CTX3 and SEM-CTX3 showed similar membrane-damaging activity. Modification of Asp residues did not affect the membrane-binding capability of CTXs. Circular dichroism spectra of SEM-CTX3 and CTX3 were similar, while the gross conformation of SEM-CTX1 was distinct from that of CTX1. The interaction of CTX1 with membrane was distinctly changed by Asp modification. Collectively, our data suggest that Asp29 of CTX1 suppresses the optimization of membrane-bound conformation to a fully active state and that the function of Asp40 in the structural constraints of CTX1 and CTX3 is not important for the manifestation of membrane-perturbing activity.

Aggregation-induced emission enhancement (AIEE) of thiolated gold nanoclusters (AuNCs) has emerged as an attractive and alternative strategy to improve their brightness. This study demonstrates Ce(iii)-triggered AIEE of glutathione-capped AuNCs (GSH-AuNCs) through the coordination between two carboxylic groups of GSH and Ce(iii). The cluster size and valence state of GSH-AuNCs are almost identical to those of a Ce(iii)-induced assembly of GSH-AuNCs (named Ce(iii)-GSH-AuNCs). More importantly, the as-prepared Ce(iii)-GSH-AuNCs exhibit a higher quantum yield (up to 13%), longer luminescence lifetime, and shorter maximum luminescence peak than GSH-AuNCs. Additionally, Ce(iii)-GSH-AuNCs possess redox-switchable luminescence, high salt stability, and long-term storage stability. These findings provide clear evidence that the Ce(iii)-triggered aggregation of GSH-AuNCs is a crucial factor to improve the luminescence property of GSH-AuNCs. Intriguingly, the presence of adenosine triphosphate (ATP) switches off the luminescence of Ce(iii)-GSH AuNCs through the significant formation of Ce(iii)-ATP complexes. Furthermore, the ATP-induced luminescence quenching of Ce(iii)-GSH-AuNCs can be paired with the alkaline phosphatase (ALP)-ATP system to design a turn-on luminescent probe for ALP; the limit of detection for ALP is estimated to be 0.03 U L-1. Also, the biocompatibility of Ce(iii)-GSH-AuNCs enables the proposed system to detect ALP in human serum and HeLa cells.

In the clinic, ethosuximide is commonly used to treat generalized absence seizures but has recently been repurposed for other diseases. Because of adverse effects and drug interactions, high-throughput therapeutic drug monitoring of ethosuximide is necessary. Microextraction is a simple, effective, rapid, and low consumption of organic solvents method for sample preparation. In this study, microderivatization-increased detection (MDID)-combined microextraction was used to detect ethosuximide by mass spectrometry. Ethosuximide is a difficult to retain and ionize compound in the C18 nano-flow column and ionization interface, respectively. Hence, we developed a fast method for detecting ethosuximide in human plasma by using the MDID strategy (within 2 min). Chemical microderivatization parameters were studied and optimized to increase the sensitivity of ethosuximide detection at trace levels. The linear range for the analysis of ethosuximide in 10 μL plasma was 5–500 μg/mL with a coefficient of determination (r2) ≥ 0.995. The precision and accuracy of intraday and interday analyses of ethosuximide were below 13.0%. Furthermore, modifications of major proteins in plasma and blood cells, induced by ethosuximide, were identified. The proposed method effectively utilizes microliter samples to detect drug plasma concentrations under suitable microextraction procedures toward the eco-friendly goal of low consumption of organic solvents.

Exosomes are implicated in cancer cell development, migration and invasion. Pigment epithelium-derived factor (PEDF) is a secreted anticancer protein that can regulate lung cancer progression; however, the role of PEDF in non-small cell lung cancer (NSCLC), including metastasis and cancer cell-derived exosome secretion, is unclear. In this study, we analyzed the effects of PEDF on exosome-mediated migration, invasion, and tumorigenicity of cultured NSCLC cells. The results showed that PEDF overexpression significantly reduced NSCLC invasion and migration, while inducing cell aggregation, whereas PEDF knockdown had the opposite effects. Exosomes from NSCLC cells treated with recombinant PEDF had a significantly reduced ability to promote cancer cell motility, migration, and invasion compared to exosomes from untreated cells. Exosomes from PEDF-treated cells contained thrombospondin 1 (THBS1), which inhibited cytoskeletal remodeling and exosome-induced lung cancer cell motility, migration, and invasion. Furthermore, PEDF-overexpressing NSCLC cells formed smaller xenograft tumors with higher THBS1 expression compared to control tumors. Our findings indicate that PEDF decreases the metastatic potential of NSCLC cells through regulation of THBS1 release in cancer cell-derived exosomes, thus uncovering a new mechanism of lung cancer progression.

In this study, a new non-heme iron complex was prepared by mixing FeCl3∙6H2O and 2,4,6-tris(2-pyridylmethylamino)-1,3,5-triazine (TPMATA) in ionic liquid 1-butyl-1-methylpyrrolidinium bis((trifluoromethyl)sulfonyl)amide (IL [BMP+][TFSA−]). The iron complex (TPMATA-FeCl3) was used as a modifier for preparing the TPMATA-FeCl3-modified carbon ionic liquid electrode (TPMATA-FeCl3/CILE). H2O2 was electrocatalytically reduced at the TPMATA-FeCl3/CILE in neutral aqueous solutions via Fenton's mechanism in which hydroxyl radical, •OH, was produced, and the radical was detected by using a fluorescence probe. This electrode showed a sensitivity of 22.1 μA∙mM−1 and a dynamic range of 0.025∼4.69 mM with a regression coefficient R2 of 0.993 (or 0.025∼0.765 mM, R2 = 0.997, sensitivity = 27.6 μA∙mM−1). The detection limit was 0.0125 mM (S/N = 3). This electrode shows the potential to be used as an electrochemical H2O2 sensor or for the production of hydroxyl radicals.

Retinoblastoma (Rb) is the most common primary intraocular malignant tumor of childhood. It is important to develop the strategy for Rb treatment. We have tested a quinolone derivative 2,9-bis[2-(pyrrolidin-1-yl)ethoxy]-6-{4-[2-(pyrrolidin-1-yl)ethoxy]phenyl}-11H-indeno[1,2-c]quinolin-11-one (BPIQ) for its anti-cancer effects against Rb via cultured human Rb cell line Y79. Our results showed that BPIQ significantly inhibits cell growth of Y79. Furthermore, the flow cytometer-based assays and Western blotting showed that BPIQ induces the apoptosis of Y79 via increasing the level of reactive oxygen species (ROS). Besides, the activation of γH2AX, a DNA damage sensor in human Y79 cells was also observed, indicating the potential of BPIQ for causing DNA damage of Rb cells. On the contrary, BPIQ-induced apoptosis of Y79 cells was attenuated significantly by N-acetyl-L-cysteine (NAC), an ROS scavenger. The results of Western blot showed that BPIQ down-regulates the levels of anti-apoptotic proteins Bcl-2, survivin and XIAP while up-regulates the pro-apoptotic proteins Bad, Bax and Bid. Our present study demonstrated the anti-proliferative effect of BPIQ in human Y79 cells. The inhibitory effect of BPIQ on the proliferation of Y79 cells is, at least, partly mediated by the regulation of ROS and DNA damage pathway. In conclusion, BPIQ may provide an alternative option in the chemotherapeutics or chemoprevention on the Rb therapy in the future.