Nontargeted metabolomics and enzyme inhibitory and antioxidant activities for chemical and biological characterization of jujube (Ziziphus jujuba) extracts.

The chemical and biologically active characterization of jujube samples (fruits, cores, and leaves) were carried out by the integrated nontargeted metabolomics and bioassay. Firstly, collision cross-section values of active compounds in jujubes were determined by ultrahigh-performance liquid chromatography coupled with ion mobility quadrupole time-of-flight mass spectrometry. Then, a multidimensional statistical analysis that contained principal component analysis, partial least squares-discriminant analysis and hierarchical clustering analysis was employed to effectively cluster different tissues and types of jujubes, making identification more scientific. Furthermore, angiotensin-converting enzyme (ACE) and 2, 2-diphenyl-1-picrylhydrazyl (DPPH) were used to evaluate the quality of jujubes from a double activity dimension. The analytical results obtained by using ACE and DPPH to evaluate the quality of jujube were different from multivariate statistics, providing a reference for the application of jujube. Therefore, integrating chemical and biological perspectives to evaluate the quality of jujube provided a more comprehensive evaluation and effective reference for clinical needs.

The Dissolution Behavior of Feldspar Minerals in Various Low-Molecular-Weight Organic Acids.

Feldspar is a high-abundance mineral in the earth's crust, and its natural weathering and dissolution processes are an important phenomenon on the earth's surface. This study focused on the dissolution behavior of silicon (Si) and aluminum (Al) in feldspar minerals (microcline and albite) when exposed to low-molecular-weight organic acids (LMWOAs). Various analytical techniques, including atomic absorption spectrophotometer, X-ray diffraction, scanning electron microscope, and Fourier-transform infrared spectroscopy, were employed to investigate these processes. The results revealed that the concentration of Si and Al released from alkali feldspar increased after treatment with LMWOAs, exhibiting non-stoichiometric dissolution. The Si/Al release ratio from feldspar deviated from the expected value of three. Among the LMWOAs tested, oxalic acid was found to be more effective in dissolving aluminum, while citric acid showed greater efficacy in dissolving silicon. Notably, the composite acid demonstrated the highest capacity for feldspar dissolution, with values of 538 µM (Si) and 287 µM (Al) after treatment for 720 h, respectively. The dissolution data for Si and Al in the organic acid solution was fittingly described by a first-order equation, with high correlation coefficients (R2 ≥ 0.992). The characterization of feldspar powders indicated that the (040) crystal plane of feldspar was particularly susceptible to attack by organic acids. In the presence of these acids, the chemical bonds Si (Al)-O, Si-Si(Al), and O-Si(Al)-O shifted to higher wavenumbers. Additionally, the surface corrosion morphology of feldspar exhibited distinct nanostructures, which became more pronounced with increasing exposure time. It was also observed that the reactivity of feldspar increased over time. These findings provide valuable insights into the natural dissolution process of feldspar and offer a new perspective for the study of this phenomenon.

Preparation of Anorthite/Mullite In Situ and Phase Transformation in Porcelain.

A high sintering temperature is required to acquire excellent performance in the production of porcelain but results in high fuel consumption. To prepare the porcelain with outstanding performance at a lower temperature, a self-produced additive containing calcium (CaK) was added into a three-component system of kaolinite-feldspar-quartz. XRD and SEM were used to characterize the samples. The toughening mechanism and Gibbs free energy were investigated. After introducing the CaK, the bending strength of the porcelain fired at 1513 K increased from 56.32 ± 0.65 MPa to 95.31 ± 0.63 MPa, which was 21.83% higher than that of the porcelain without CaK at an optimal firing temperature of 1603 K. The main crystal phase of the sample comprised mullite and quartz in the raw materials at 1453~1603 K. The anorthite was observed at 1453 K and interlocked with needle-shaped mullite at 1513 K in the porcelain after adding CaK, which resulted in the higher bending strength. Quantitative analysis indicated that the amount of anorthite decreased at 1513 K and disappeared at 1543 K; the amount of mullite increased with temperature. The Gibbs free energy of the reaction (CaO•Al2O3•2SiO2 + 2(Al2O3•2SiO2) → 3Al2O3•2SiO2 + CaO + 4SiO2) at high temperature was negative, which suggested that the formation of mullite (3Al2O3•2SiO2) from anorthite (CaO•Al2O3•2SiO2) was possible. These findings implied that the addition of CaK contributed to the appropriate phase composition and microstructure, and the excellent performance of the porcelain at a lower temperature. In addition, the transformation between anorthite and mullite was possible in the special raw material system. The results are of interest in producing anorthite/mullite ceramics at reduced sintering temperatures and the conversion between anorthite and mullite.

Rapid stacking of amino acids in soybean and Dendrobium officinale by on-capillary sandwich derivatization in capillary electrophoresis.

An online sandwich derivatization and stacking strategy using capillary electrophoresis was developed and successfully applied to the preconcentration of multi-amino acids in two functional food samples. Amino acids were derived with 4-Chloro-7-nitro-1, 2, 3-benzoxadiazole (NBD-Cl) to make the new compounds with chromophore in the stacking process, then the derivatives were stacked by on-column sample preconcentration and detected at 475 nm UV wavelength. The novel sandwich injection sequence was amino acids, NBD-Cl, and amino acids separately. Additionally, the running buffer was 40 mM borax buffer (pH = 9.0). A succession of derivatization and stacking conditions were optimized, including buffer concentration, pressure, NBD-Cl injection time, waiting time, matrix concentration, and sample injection time. In the appropriate range, good linearity values (R2) were obtained for nine amino acids at 0.996-0.999. Intra-day and inter-day precision with a relative standard deviation < 6.23 % (n = 5), the limit of detection in the range of 2.8-25.2 µM, and recovery ranging from 83.2 to 108.2 % were obtained. Besides, the enrichment factors were in range of 8-62 for nine AAs. The stacking approach was successfully applied to soybean and Dendrobium officinale samples, which shows great potential in the determination of free amino acids in samples containing complex matrices.

The Impact of COVID-19 on Motivation, Involvement, and Behavior of Cyclists in Taiwan.

Coronavirus disease (COVID-19) has spread all over the world and has impacted tourism globally, with countries taking various measures such as travel restrictions, border closures, lockdowns, or quarantines to contain the virus. Tourists' motivation has also been affected by COVID-19, but so far, the literature has not yet discussed their concern over COVID-19 as well as the relationships among their motivation, involvement, and behavior intention. Therefore, this study fills the gap in the literature by taking cycling tourism as an example to understand the involvement of tourists concerning COVID-19 and presents the depth and breadth of its effects upon tourism. Due to the challenge of face-to-face, on-site investigation, we employ an online survey for data collection, use exploratory factor analysis to extract the main factors of motivation, involvement, and behavior intention, and set up a structural equation model to examine the relationships among the three factors. The results show that COVID-19 has positively and significantly affected motivation and involvement. Motivation positively and significantly affects involvement, and involvement affects motivation and behavior intention. The main finding herein is that motivation does not affect behavior, but involvement does mediate between the motivation and behavior of cyclists during COVID-19. Therefore, people may perceive the risk of health and wellbeing through such involvement.

Simultaneous capture of hydrophilic and hydrophobic compounds from complex plants by biosurfactant-assisted mechanical amorphous dispersion extraction.

A biosurfactant-assisted mechanical amorphous dispersion extraction (BA-MADE) procedure was established for the simultaneous capture of hydrophilic phenolic acids and hydrophobic tanshinones from Salvia miltiorrhiza. Single-factor experiments and the response surface methodology were used to optimize and analyze the crucial parameters for the method, such as the type and amount of amorphous-dispersion extractants, grinding time, extraction time and solid-to-liquid ratio. The optimized parameter values for the BA-MADE process were 407.02 mg of sodium chenodeoxycholate, a grinding time of 4.87 min, an extraction time of 4.92 min, and a solid-to-liquid ratio of 0.5:10 g/mL. The calibration curves of danshensu, rosmarinic acid, lithospermic acid, salvianolic acid B, salvianolic acid A, dihydrotanshinone I, cryptotanshinone, tanshinone I, and tanshinone II A exhibited good linearity in the range of 1-500 µg/mL (R2 ≥ 0.9990). The limits of detection of nine analytes ranged from 5.46 to 130 ng/mL, the relative standard deviations (RSDs) of intraday and interday precision were less than 1.95 and 3.56%, respectively, and the recoveries of the real sample were in the range of 85-113%, with RSD% below 3.21%. The BA-MADE method was compared with previously reported methods, such as heating reflux extraction, ultrasonic extraction and microwave-assisted micellar extraction, and the results demonstrated that the developed method has significant advantages in the simultaneous extraction of hydrophilic and hydrophobic active components from Salvia miltiorrhiza.

Optimization of mechanically assisted coamorphous dispersion extraction of hydrophobic compounds from plant tea (Citri Reticulatae Pericarpium) using water.

This study aimed to establish a novel mechanically assisted coamorphous dispersion extraction (MADE) method for the extraction of hydrophobic compounds (hesperidin, nobiletin and tangeretin) from Citri Reticulatae Pericarpium using water. The surface morphology, particle size distributions, phase states and functional groups of the coground product surface were characterized by Scanning Electron Microscopy, X-ray diffraction and Fourier transform infrared spectroscopy. The parameters affecting the efficiency of extraction method were optimized by single-factor experiments and response surface methodology. The method showed good linear relationships in the range of 1-500 µg/mL with correlation coefficients (R2) ≥ 0.9990, low limits of detection ranging from 3.0 to 28.3 ng/mL, and acceptable recoveries ranging from 87.0 to 91.0%. Therefore, the proposed MADE method is a promising, efficient and organic solvent-free method for the extraction of hydrophobic compounds from plant tea.

The two-phase amphiphilic preconcentration based on surfactants to enrich phenolic compounds from diluted plant extracts and rat urine by micellar electrokinetic chromatography.

A novel technology by two-phase amphiphilic preconcentration based on surfactants was established for enriching phenolic compounds by micellar electrokinetic chromatography (MEKC). The cationic surfactant cetyltrimethylammonium chloride (CTAC) was combined with the anionic analytes that existed in the sample solution before injection. The boundary was formed between CTAC and sodium dodecyl sulfate (SDS) in the background solution when the sample solution was injected into the capillary, where the analytes bound inside micelles were released due to the stronger electrostatic force between SDS and CTAC. This procedure accelerated the separation of analytes from CTAC and greatly improved the enrichment efficiency. The optimal conditions were obtained after a series of optimizations, and the sensitivity enrichment factors of the four analytes were in the range of 39-93 compared to typical injections in capillary zone electrophoresis. Good linearity for matrix-matched calibrations was established for all analytes with R2 values of 0.9993-0.9997. The limits of detection (S/N = 3) for kaempferol, quercetin, salvianolic acid C, and salvianolic acid B were 0.0166, 0.0292, 0.0215, and 0.0195 µg/ml, respectively. The intracapillary RSDs of the analytes ranged from 0.8% to 1.3% for migration time and from 0.4% to 1.8% for peak areas. The developed method was successfully applied to the determination of phenolic compounds, the main compounds of Salvia miltiorrhiza Bge., and had been validated for the determination of spiked recoveries in rat urine.

Two-step micelle-to-solvent stacking of arsenic species from foods in permanently coated tubing for capillary electrophoresis.

An effective and sensitive two-step stacking of arsenic species by electrokinetic injection and micelle-to-solvent stacking (MSS) using zwitterionic surfactant 3-(N,N-dimethylpalmitylammonio) propane sulfonate was successfully developed in a co-electro-osmotic flow (co-EOF) capillary zone electrophoresis (CZE). The fused silica capillaries were coated with 1% hexadimethrine bromide solution to satisfy the condition of the same EOF direction between the test anions and capillary. The background solution was 100 mM borax buffer (pH 9.2). The proposed method was performed by a CZE system equipped with a diode array detector, and the detection wavelength was monitored at 192 nm. The separation voltage was -20 kV. The group of micelles and target analytes was stacked by field-enhanced sample injection at -10 kV for 180 s. The second stacking step used 60% MeOH to serve as an organic solvent where the analytes were brought by the micelles to the MSS boundary. The enrichment factors of As(Ⅴ), As(Ⅲ), disodium methyl arsonate hexahydrate (MMA), and dimethylarsinic acid (DMA) were 1230, 840, 3820, and 1450, respectively, compared to typical injections in CZE. The limits of detection (S/N = 3) ranged from 0.382 to 0.911 ng mL-1. The intracapillary repeatabilities (%RSD, n = 3) were 0.5-1.0% for migration time and 0.3-0.7% for peak areas. The technology of two-step stacking by MSS in co-EOF CZE together with a simple extraction procedure for As(Ⅴ), As(Ⅲ), MMA and DMA were demonstrated in kelp and rice samples.

Boron nitride nanosheet-assisted matrix solid-phase dispersion microextraction of alkaloids from lotus plumule by high-performance liquid chromatography coupled with ultraviolet detection and ion mobility quadrupole time-of-flight mass spectrometry.

A boron nitride nanosheet (BNNS)-assisted matrix solid-phase dispersion method was established to microextract alkaloids from medicinal plants. The target compounds were identified by high-performance liquid chromatography coupled with ultraviolet detection and ion mobility quadrupole time-of-flight mass spectrometry. During the experimental process, several important parameters, including the type of dispersant, the amount of dispersant, the grinding time, and the type of elution solvent, were optimized. Finally, the BNNSs were chosen as the best dispersant, and their microcosmic morphologies were identified by scanning electron microscopy and transmission electron microscopy. Because of the special property of BNNSs, the cost of this experiment was greatly reduced, especially in elution volume, sample amount (50 mg), and extraction time (2 min). Under the best conditions, 50 mg of sample powder was dispersed with 50 mg of BNNSs, the grinding time was 120 s, the mixed powder was eluted with 200 µL of methanol, and good linearity (r2  > 0.9993) and satisfactory recoveries (80-100%) were obtained. The inter- and intraday precisions were acceptable, with RSDs lower than 2.01 and 4.84%, respectively. The limits of detection ranged from 2.54 to 15.00 ng/mL, and the limits of quantitation were 8.47 to 50.00 ng/mL. The proposed method was successfully applied for the determination of liensinine, isoliensinine, and neferine in lotus plumule.

Cucurbituril and zwitterionic surfactant-based matrix solid-phase dispersion microextraction to simultaneously determine terpenoids from Radix Curcumae.

A rapid, efficient, and environmentally friendly matrix solid-phase dispersion microextraction was established to determine and quantify terpenoids in Radix Curcumae using ultra-high-performance liquid chromatography with a diode array detector. Various parameters affecting the extraction were investigated in detail, such as the grinding time, amount of adsorbent, type and concentration of elution solvent, and pH. The optimization of single-factor and response surface methodology was performed to confirm the best conditions in this procedure. The final optimized conditions were obtained by applying 70 mg of cucurbituril as adsorbent, 149 s as the optimum grinding time, and 228 mM of 3-(N,N-dimethylpalmitylammonio)propanesulfonate aqueous solution (pH 6.5) as the optimal elution solvent. The validated method showed a satisfactory linear range of 0.10-10 µg/mL for curdione and furanodiene, 0.01-10 µg/mL for isocurcumenol and germacrone, and 0.05-10 µg/mL for furanodienone, while the correlation coefficients ranged from 0.9945 to 0.9970. The recoveries of the investigated analytes at two spiked concentration levels (0.1 and 1.0 µg/mL) ranged from 96.53 to 104.60%. In addition, this method displayed acceptable reproducibility (relative standard deviation ≤ 3.66%). The results showed that the newly proposed matrix solid-phase dispersion microextraction method was successfully applied to analyze curdione, isocurcumenol, furanodienone, germacrone and furanodiene in Radix Curcumae samples.

Management of elbow dysfunction associated with giant cell tumor of the distal humerus: achieving oncologic safety and good function by a combined reconstruction.

BACKGROUND: The purpose of our study was to evaluate the functional outcomes and oncologic results of elbow salvage surgery using arthrolysis combined with ligament repair and external fixation for reconstruction of the elbow after tumor excision and autografting. METHODS: We retrospectively reviewed 6 patients with elbow dysfunction associated with giant cell tumor of the distal humerus. All patients were treated with our combined protocol. We assessed the Musculoskeletal Tumor Society system score, range of motion, Mayo Elbow Performance Score, recurrence, and complications for each patient. RESULTS: The mean follow-up period was 48 months (range, 36-60 months). There were no cases of postoperative fracture, infection, elbow dislocation, elbow stiffness, or local recurrence. The average Musculoskeletal Tumor Society score was 28 of 30 points (93%; range, 87%-100%). The Mayo Elbow Performance Score improved from a mean of 61 points to 93 points, with mean flexion of 135° and mean extension of 3°. CONCLUSIONS: Local tumor resection, autografting, and elbow reconstruction by arthrolysis combined with ligament repair and external fixation can be performed with oncologic safety and provide satisfactory functional outcomes with low complication rates.

Management of elbow stiffness after postoperative treatment of terrible triad elbow injury: maintaining mobility and stability using a combined protocol.

PURPOSES: The purpose of this study was to evaluate the results of our protocol that include the restoration of mobility using open release combined with external fixation and stability using ligament repair, to determine the optimal timing of surgery, and to investigate whether resection and replacement of the radial head are associated with different outcomes. METHODS: Twenty-six patients with elbow stiffness after operation of terrible triad injury of the elbow were treated with our protocol. We assessed the optimal timing of the operation by comparing outcomes between the early treatment group and the delayed treatment group. The comparison was performed to investigate whether the results differed between resection and replacement of the radial head. Stability of the elbow, range of motion (ROM), Mayo Elbow Performance Score (MEPS), and complications were assessed for each patient. RESULTS: The mean interval from the initial surgery to the index procedure was 13 months, and the mean follow-up period was 29 months. The MEPS increased from a mean of 65 points to 94 points. Twenty-five of 26 patients achieved stability of the elbow, and all patients achieved functional ROM. There were no significant differences between the two subgroups with respect to ROM and stability of the elbow. CONCLUSION: Our protocol can restore mobility and stability. Resection and replacement of the radial head are both feasible using this protocol. Lastly, the timing of the surgery was not very rigorous, and the surgical delay may be insignificant. LEVEL OF EVIDENCE: Level IV; Case Series; Treatment Study.

[The Properties of an Infrared Hybrid Single Beam Spectrum].

A hybrid single beam spectrum ø(α)=αø(b1)+(1-α)ø(b2)=αø0e(-Kb1)+(1-α)ø0e(-Kb2) is introduced as the combination of two single beam spectra ø(b1) and ø(b2) from the same sample but with different pathlengths (b(1) and b(2)), where α(0＜α＜1) is the hybrid coefficient. The intensity of hybrid spectrum ø(α) can be controlled easily to the desired point by simply choosing an appropriate α. The experimental results showed that hybrid spectrum ø(α) is very nearly identical to ø(b)=ø(0)e-K(b(2)-αb(2)+αb(b)) under appropriate conditions, namely ø(α)≈ø(b) , where ø(b) is the single beam spectrum of the real sample with the pathlength of b(2)-αb(2)+αb(1). Therefore, the desired single beam spectrum ø(b) can be obtained easily by choosing α and we no longer need to prepare IR sample with the thickness of b. Hybrid spectrum method shows valuable potential in application of eliminating background interference.

A tripeptide (NSK) inhibits Japanese encephalitis virus infection in vitro and in vivo.

Japanese encephalitis virus (JEV) is a major pathogen that can cause acute viral encephalitis in both humans and animals. Domain III of the viral envelope protein (EDIII) is involved in binding to host cell receptor(s) to facilitate virus entry. Our previous study showed that the loop3 peptide of EDIII possesses antiviral activity against JEV infection. In this paper, we demonstrate that three residues (NSK) in loop3 are responsible for the antiviral activity of loop3 peptide. In vitro experiments showed that the tripeptide NSK could inhibit JEV infection in both BHK-21 and Neuro-2A cells by inhibiting attachment of JEV to the cells, with IC50 values of 8 µM and 6.5 µM, respectively. In vivo experiments showed that the tripeptide could increase the survival of mice challenged with JEV to 75 % when administrated intracerebrally. Therefore, this tripeptide may serve as the basis for the development of novel antiviral agents against Japanese encephalitis virus infection.

Porcine reproductive and respiratory syndrome virus induces autophagy to promote virus replication.

An increasing number of studies demonstrate that autophagy, an intrinsic mechanism that can degrade cytoplasmic components, is involved in the infection processes of a variety of pathogens. It can be hijacked by various viruses to facilitate their replication. In this study, we found that PRRSV infection significantly increases the number of double- or single-membrane vesicles in the cytoplasm of host cells in ultrastructural analysis. Our results showed the LC3-I was converted into LC3-II after virus infection, suggesting the autophagy machinery was activated. We further used pharmacological agents and shRNAs to confirm that autophagy promoted the replication of PRRSV in host cells. Confocal microscopy analysis showed that PRRSV inhibited the fusion between autophagosomes and lysosomes, suggesting that PRRSV induced incomplete autophagy. This suppression caused the accumulation of autophagosomes which may serve as replication site to enhance PRRSV replication. It has been shown that NSP2 and NSP3 of arterivirus are two components of virus replication complex. We also found in our studies that NSP2 colocalized with LC3 in MARC-145 cells by performing confocal microscopy analysis and continuous density gradient centrifugation. Our studies presented here indicated that autophagy was activated during PRRSV infection and enhanced PRRSV replication in host cells by preventing autophagosome and lysosome fusion.

Role of lipid rafts in porcine reproductive and respiratory syndrome virus infection in MARC-145 cells.

Lipid rafts play an important role in the life cycle of many viruses. Cholesterol is a critical structural component of lipid rafts. Although the porcine reproductive and respiratory syndrome virus (PRRSV) has restricted cell tropism for cells of the monocyte/macrophage lineage, a non-macrophage cell MARC-145 was susceptible to PRRSV because of the expression of virus receptor CD163 on the cell surface, therefore MARC-145 cells is used as model cell for PRRSV studies. In order to determine if cholesterol is involved in PRRSV infection in MARC-145 cells, we used three pharmacological agents: methyl-ß cyclodextrin (MßCD), mevinolin, and filipin complex to deplete cholesterol in MARC-145. Although these agents act by different mechanisms, they all significantly inhibited PRRSV infection. The inhibition could be prevented by addition of exogenous cholesterol. Cell membrane cholesterol depletion after virus infection had no effect on PRRSV production and cholesterol depletion pre-infection did not reduce the virus attachment, suggesting cholesterol is involved in virus entry. Further results showed that cholesterol depletion did not change expression levels of the PRRSV receptor CD163 in MARC-145, had no effect on clathrin-mediated endocytosis, but disturbed lipid-raft-dependent endocytosis. Collectively, these studies suggest that cholesterol is critical for PRRSV entry, which is likely to be mediated by a lipid-raft-dependent pathway.

PhosphoPOINT: a comprehensive human kinase interactome and phospho-protein database.

MOTIVATION: To fully understand how a protein kinase regulates biological processes, it is imperative to first identify its substrate(s) and interacting protein(s). However, of the 518 known human serine/threonine/tyrosine kinases, 35% of these have known substrates, while 14% of the kinases have identified substrate recognition motifs. In contrast, 85% of the kinases have protein-protein interaction (PPI) datasets, raising the possibility that we might reveal potential kinase-substrate pairs from these PPIs. RESULTS: PhosphoPOINT, a comprehensive human kinase interactome and phospho-protein database, is a collection of 4195 phospho-proteins with a total of 15 738 phosphorylation sites. PhosphoPOINT annotates the interactions among kinases, with their down-stream substrates and with interacting (phospho)-proteins to modulate the kinase-substrate pairs. PhosphoPOINT implements various gene expression profiles and Gene Ontology cellular component information to evaluate each kinase and their interacting (phospho)-proteins/substrates. Integration of cSNPs that cause amino acids change with the proteins with the phosphoprotein dataset reveals that 64 phosphorylation sites result in a disease phenotypes when changed; the linked phenotypes include schizophrenia and hypertension. PhosphoPOINT also provides a search function for all phospho-peptides using about 300 known kinase/phosphatase substrate/binding motifs. Altogether, PhosphoPOINT provides robust annotation for kinases, their downstream substrates and their interaction (phospho)-proteins and this should accelerate the functional characterization of kinomemediated signaling. AVAILABILITY: PhosphoPOINT can be freely accessed in http://kinase. bioinformatics.tw/. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.