Molecular mechanisms of selenite reduction by Lactiplantibacillus plantarum BSe: An integrated genomic and transcriptomic analysis.

The reduction of selenite [Se(Ⅳ)] by microorganisms is a green and efficient detoxification strategy. We found that Se(Ⅳ) inhibited exopolysaccharide and protein secretion by Lactiplantibacillus plantarum BSe and compromised cell integrity. In this study, L. plantarum BSe reduced Se(Ⅳ) by increasing related enzyme activity and electron transfer. Genomic analysis demonstrated that L. plantarum BSe should be able to reduce Se(Ⅳ). Further transcriptome analysis showed that L. plantarum BSe enhanced its tolerance to Se(Ⅳ) by upregulating the expression of surface proteins and transporters, thus reducing the extracellular Se(Ⅳ) concentration through related enzymatic reactions and siderophore-mediated pathways. Lactiplantibacillus plantarum BSe was able to regulate the expression of related genes involved in quorum sensing and a two-component system and then select appropriate strategies for Se(Ⅳ) transformation in response to varying environmental Se(Ⅳ) concentrations. In addition, azo reductase was linked to the reduction of Se(Ⅳ) for the first time. The present study established a multipath model for the reduction of Se(Ⅳ) by L. plantarum, providing new insights into the biological reduction of Se(Ⅳ) and the biogeochemical cycle of selenium.

Amide-containing neoepitopes: the key factor in the preparation of hapten-specific antibodies and a strategy to overcome.

For a long time, people have suffered from uncertainty, complexity, and a low success rate in generating and screening antibodies against small molecules, which have become the core bottlenecks of immunochemistry. Here, the influence of antigen preparation on antibody generation was investigated at both molecular and submolecular levels. Neoepitopes (amide-containing neoepitopes) formed in the preparation of complete antigens are one of the most important factors limiting the efficiency of generating hapten-specific antibodies, which was verified by different haptens, carrier proteins, and conjugation conditions. Amide-containing neoepitopes present electron-dense structural components on the surface of prepared complete antigens and, therefore, induce the generation of the corresponding antibody with much higher efficiency than target hapten. Crosslinkers should be carefully selected and not overdosed. According to these results, some misconceptions in the conventional anti-hapten antibody production were clarified and corrected. By controlling the content of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) during the synthesis of immunogen to limit the formation of amide-containing neoepitopes, the efficiency of hapten-specific antibody generation could be significantly improved, which verified the correctness of the conclusion and provided an efficient strategy for antibody preparation. The result of the work is of scientific significance in the preparation of high-quality antibodies against small molecules.

Covalent organic nanospheres as a fiber coating for solid-phase microextraction of genotoxic impurities followed by analysis using GC-MS.

Covalent organic nanospheres (CONs) were explored as a fiber coating for solid-phase microextraction of genotoxic impurities (GTIs) from active ingredients (AIs). CONs were synthesized by an easy solution-phase procedure at 25 °C. The obtained nanospheres exhibited a high specific surface area, good thermostability, high acid and alkali resistance, and favorable crystallinity and porosity. Two types of GTIs, alkyl halides (1-iodooctane, 1-chlorobenzene, 1-bromododecane, 1,2-dichlorobenzene, 1-bromooctane, 1-chlorohexane, and 1,8-dibromooctane) and sulfonate esters (methyl p-toluenesulfonate and ethyl p-toluenesulfonate), were chosen as target molecules for assessing the performance of the coating. The prepared coating achieved high enhancement factors (5097-9799) for the selected GTIs. The strong affinity between CONs and GTIs was tentatively attributed to π-π and hydrophobicity interactions, large surface area of the CONs, and size-matching of the materials. Combined with gas chromatography-mass spectrometry (GC-MS), the established analytical method detected the GTIs in capecitabine and imatinib mesylate samples over a wide linear range (0.2-200 ng/g) with a low detection limit (0.04-2.0 ng/g), satisfactory recovery (80.03%-109.5%), and high repeatability (6.20%-14.8%) and reproducibility (6.20%-14.1%). Therefore, the CON-coated fibers are promising alternatives for the sensitive detection of GTIs in AI samples.

Analysis of dietary exposure and risk assessment of pesticide residues in roots and rhizomes of Chinese herbs.

Medicine food homologous (MFH) plants provide therapeutic and health care effects through diet. Thus, a risk assessment system for hazardous ingredient residues is urgently required to ensure their safe use. In this study, the pesticide contamination of six root and rhizome Chinese herbs, Ginseng Radix et Rhizoma, Panacis Quinquefolii Radix, Pseudostellariae Radix, Salviae Miltiorrhizae Radix et Rhizoma, Codonopsis Radix, and Glehniae Radix, and the risks associated with their intake were investigated. A total of 420 MFH plant samples collected from 22 provinces in China were tested, and 61 pesticides were detected in 413 samples. Multiple pesticide residues were detected in each MFH sample, with contents ranging from 0.0002 to 3.010 mg/kg dry weight. Carbendazim (≥47.14%) and propham (≥40%) were the most frequently detected pesticides. Risk assessment determined by hazard quotients indicated that the risks were acceptable, with no short- or long-term adverse health effects. However, considering the high incidence of residues and the detection of unregistered or even prohibited pesticides, strict supervision of soil quality and pesticide application (particularly cadusafos) in MFH plant cultivation are recommended to aid in monitoring MFH plant quality and ensuring diet and drug safety. PRACTICAL APPLICATION: Ensure the diet and drug safety of Chinese herbs.

Covalent organic nanospheres as a fiber coating for solid-phase microextraction of genotoxic impurities followed by analysis using GC-MS / 药物分析学报

Covalent organic nanospheres(CONs)were explored as a fiber coating for solid-phase microextraction of genotoxic impurities(GTIs)from active ingredients(AIs).CONs were synthesized by an easy solution-phase procedure at 25℃.The obtained nanospheres exhibited a high specific surface area,good ther-mostability,high acid and alkali resistance,and favorable crystallinity and porosity.Two types of GTIs,alkyl halides(1-iodooctane,1-chlorobenzene,1-bromododecane,1,2-dichlorobenzene,1-bromooctane,1-chlorohexane,and 1,8-dibromooctane)and sulfonate esters(methyl p-toluenesulfonate and ethyl p-toluenesulfonate),were chosen as target molecules for assessing the performance of the coating.The prepared coating achieved high enhancement factors(5097-9799)for the selected GTIs.The strong affinity between CONs and GTIs was tentatively attributed to T-T and hydrophobicity interactions,large surface area of the CONs,and size-matching of the materials.Combined with gas chromatography-mass spectrometry(GC-MS),the established analytical method detected the GTIs in capecitabine and imatinib mesylate samples over a wide linear range(0.2-200 ng/g)with a low detection limit(0.04-2.0 ng/g),satisfactory recovery(80.03％-109.5％),and high repeatability(6.20％-14.8％)and reproducibility(6.20％-14.1％).Therefore,the CON-coated fibers are promising alternatives for the sensitive detection of GTIs in AI samples.

Development of a Matrix Sublimation Device with Controllable Crystallization Temperature for MALDI Mass Spectrometry Imaging.

The size and distribution of matrix crystals deposited on the surface of a tissue section play a key role in the performance of MALDI mass spectrometry imaging (MALDI-MSI). In this study, uniform distribution and a restricted size of matrix crystals were achieved via a homemade matrix sublimation device with controllable crystallization temperature. The crystallization temperature was stably controlled at a subzero temperature, and homogeneous matrix crystals with diameters <0.2 µm were generated on the sample surface. Typical MALDI-MSI experiments of endogenous and exogenous components in the tissues of strawberries, kidneys, and mussels were conducted to examine the performance of the sublimator. Good reproducibility of MALDI-MSI was achieved, and the quality of ion images was significantly improved. These results demonstrate that the developed sublimator should have potential in matrix deposition for further high resolution MALDI-MSI application.

In-situ exfoliation of graphitic carbon nitride with metal-organic framework via a sonication-assisted approach for dispersive solid-phase extraction of perfluorinated compounds in drinking water samples.

Monitoring the levels of perfluorinated compounds (PFCs) in the environment is of vital importance, owing to their sustained environmental presence, extensive distribution, and associated health risks. The development of cost-effective and efficient sorbents for the establishment of sensitive analytical methods is critical for achieving trace-level detection. In this study, a graphitic carbon nitride (g-C3N4)-based sorbent is synthesized by a facile sonication-assisted method exfoliated by zeolitic imidazolate framework-67 (ZIF-67) in situ. The novel ZIF-67/g-C3N4 composites were systematically characterized by transmission electron microscopy, scanning electron microscopy, atomic force microscopy, X-ray diffraction, Fourier transform infrared spectroscopy, X-ray photoelectron spectroscopy, thermogravimetric analysis, and N2 adsorption-desorption analysis, exhibiting good dispersity and a large surface area. Moreover, molecular dynamics simulations indicated that g-C3N4 structures can be effectively exfoliated by the introduced ZIF-67 molecules. The hybrid material was successfully utilized as a dispersive solid-phase extraction sorbent, and the extraction factors were systematically optimized by response surface methodology. Under optimal conditions, the synthesized sorbent exhibited desirable linear correlations (R2 > 0.99), a low detection limit (0.3-2 ng L-1), and good repeatability (relative standard deviation <15%, n = 6). The developed method was applied for the analysis of natural and spiked water samples. The study demonstrated that the ZIF-67/g-C3N4 composites are promising materials for pollutant adsorption from drinking water samples.

Quick and convenient construction of lambda-cyhalothrin antigen for the generation of specific antibody.

Lambda-cyhalothrin is a pyrethroid widely used in crop, fruit and vegetable production, but has potential health threats to human. Immunoassay is a cheap, rapid and facile method to detect lambda-cyhalothrin, yet wide application of this method still requires improvement in the construction of antigen. In this study, we developed a one-step lambda-cyhalothrin hapten synthesis that transformed the cyanide group in lambda-cyhalothrin to amide. Complete antigen was assembled by coupling the amide with succinic-anhydride-activated carrier proteins, and corresponding polyclonal antibodies were generated using Balb/c mice. Using antibody generated by the method in this paper, the competitive ELISA demonstrated the lowest detection limit of 3.772 µg/L for lambda-cyhalothrin, and no significant cross-reactivity for other pyrethroid pesticides was observed. All the results suggested we have established a more efficient technique of generating lambda-cyhalothrin antibody. Furthermore, since the activated proteins used in this study are highly controllable, we believe these proteins could potentially be the prototype of a series of standardized carrier proteins for the synthesis of complete antigens.

Metabolomic approach for rapid differentiation of Fritillaria bulbs by matrix-assisted laser desorption/ionization mass spectrometry and multivariate statistical analysis.

The bulbs of Fritillaria have been used for centuries as food and medicinal products in many Asian countries. Different Fritillaria species have distinct pharmacological effects despite of their similar appearances. Effective differentiation of Fritillaria species can avoid adulteration and is crucial to its clinical uses. In this paper, a hybrid method of matrix assisted laser desorption/ionization mass spectrometry and multivariate statistical analysis was developed for the rapid and reliable differentiation of Fritillaria species for the first time. Significantly different patterns for five Fritillaria species were obtained by MALDI-MS after instant sample extractions. Different groups of Fritillaria were confidently differentiated via an orthogonal partial least square model. In addition, a metabolomic taxonomy of five Fritillaria species was obtained based on MALDI-MS data.

In situ analysis of oxytetracycline tablets based on matrix-assisted laser desorption/ionization mass spectrometry imaging.

RATIONALE: A thorough understanding of the content and distribution of active ingredients in pharmaceuticals is essential for drug efficacy and safety. Technological advancements in mass spectrometry imaging present an opportunity for methodological innovation by providing qualification and quantification analysis, as well as spatial information, in the same assay, which has great potential for applications in the rapid analysis and quality control of drugs. METHODS: Matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) was employed to directly analyze oxytetracycline tablets in order to map the distribution of the active constituent within the whole tablet. Quantitative analysis was capable of differentiating tablets containing various doses of the active pharmaceutical ingredient. RESULTS: To establish the methodology, detailed factors that influence matrix spraying and spatial resolution during sample preparation and the data acquisition process were optimized systematically. Quantitative analysis could differentiate the tablets containing various doses of the active compound. The proposed method was successfully applied to analyze real commercial tablets. CONCLUSIONS: The developed method could successfully achieve the spatial location of oxytetracycline in actual tablet samples. These results could contribute to pharmaceutical tracing technology, especially the formulation process of tablets, which is helpful for monitoring the quality of pharmaceutical products and guaranteeing drug security.

Dietary exposure and risk assessment of short-chain chlorinated paraffins in supermarket fresh products in Jinan, China.

Dietary intake is the major route for the exposure of residents to short-chain chlorinated paraffins (SCCPs). SCCPs are classified as persistent organic pollutants (POPs) by the Stockholm Convention since May 2017. This study assessed the general population's dietary exposure to SCCPs through supermarket products. Five food categories, which included 40 food species, were collected from five supermarkets in April 2019. The SCCP concentrations in all food matrices varied between 58.6 and 1977 ng g-1 dry weight (dw), with the average and standard deviation (SD) value of 301 ± 379 ng g-1 dw. Generally, the SCCP concentrations in animal-origin food matrices were higher than those in plant-origin food matrices. The C10Cl7 congeners were predominant among the congener groups of SCCPs. The proportion of C10 congeners in the animal-origin food samples (29.8%) was lower than that in the plant-origin food samples (39.7%), and the C13 congeners showed a contrasting result. The estimated daily intake (EDI) of SCCPs through dietary intake for the residents was 3109 ng kg-1 day-1, which is much lower than the standards of European Food Safety Authority (10 mg kg-1 day-1) for SCCPs. Risk assessment based on the supermarket foods indicated that SCCP exposure through dietary intake does not cause adverse effects to human health according to the margin of exposure (MOE).

Core-shell hollow spheres of type C@MoS2 for use in surface-assisted laser desorption/ionization time of flight mass spectrometry of small molecules.

Mesoporous carbon hollow spheres coated with MoS2 (C@MoS2) were synthesized to obtain a material with large specific surface area, fast electron transfer efficiency and good water dispersibility. The composite material was applied as a matrix for the analysis of small molecules by surface-assisted laser desorption/ionization time-of-flight mass spectrometry (SALDI-TOF MS). The use of a core-shell C@MoS2 matrix strongly reduces matrix background interferences and increases signal intensity in the analysis of sulfonamides antibiotics (SAs), cationic dyes, emodin, as well as estrogen and amino acids. The composite material was applied to the SALDI-TOF MS analysis of selected molecules in (spiked) real samples. The ionization mechanism of the core-shell C@MoS2 as a matrix is discussed. The method exhibits low fragmentation interference, excellent ionization efficiency, high reproducibility and satisfactory salt tolerance. Graphical abstractSchematic representation of the method for fabrication of MoS2-coated mesoporous carbon hollow spheres (core-shell C@MoS2). As a new matrix, the nanocomposites were applied to analysis of small molecules by surface-assisted laser desorption/ionization time-of-flight mass spectrometry.

Principles of Inter-Amino-Acid Recognition Revealed by Binding Energies between Homogeneous Oligopeptides.

We have determined the interaction strengths of the common naturally occurring amino acids using a complete binding affinity matrix of 20 × 20 pairs of homo-octapeptides consisting of the 20 common amino acids between stationary and mobile states. We used a bead-based fluorescence assay for these measurements. The results provide a basis for analyzing specificity, polymorphisms, and selectivity of inter-amino-acid interactions. Comparative analyses of the binding energies, i.e., the free energies of association (ΔG A), reveal contributions assignable to both main-chain-related and side-chain-related interactions originating from the chemical structures of these 20 common amino acids. Side-chain-side-chain and side-chain-main-chain interactions are found to be pronounced in an identified set of amino acid pairs that determine the basis of inter-amino-acid recognition.

Anti-tumor activity of nanomicelles encapsulating CXCR4 peptide antagonist E5.

Cancer is the leading cause of death worldwide, and metastasis is the main attribute to cancer death. CXCR4 and its natural ligand CXCL12 have been known to play a critical role in tumorigenesis, angiogenesis and metastasis. Therefore, designing a new CXCR4 antagonist to prevent tumor metastasis will be of great significance. Herein, a novel chemically synthesized peptide (E5) that has an ability to target CXCR4/CXCL12 axis was loaded in micelle glycol-phosphatidylethanolamine (PEG-PE) block copolymer to form micelle-encapsulated E5 (M-E5). We demonstrated that M-E5 exhibited higher affinity for CXCR4-overexpressing MCF-7 and HepG2 tumor cells as compared to free E5, and efficiently inhibited the tumor cells migration. Mechanistic studies implied that PEG-PE micelle can encapsulate E5 and improve E5 targeting efficiency for CXCR4 by accumulating E5 on the tumor cell membrane. Furthermore, through encapsulation of chemotherapeutic drug doxorubicin (Dox) in PEG-PE micelle, we proved that PEG-PE micelle could serve as a co-carrier for both E5 and Dox (M-E5-Dox). M-E5 enhanced the efficiency of Dox by down-regulating the phosphorylation level of Akt, Erk and p38/MAPK proteins. In conclusion, PEG-PE micelle demonstrated a promising delivery system for E5, and M-E5 is expected to be a potential therapeutic agent that will help to improve the clinical benefits in current therapies used for solid tumors.

Aromatic-interaction-mediated inhibition of ß-amyloid assembly structures and cytotoxicity.

Abnormal aggregation of ß-amyloid (Aß) peptide plays an important role in the onset and progress of Alzheimer's disease (AD); hence, targeting Aß aggregation is considered as an effective therapeutic strategy. Here, we studied the aromatic-interaction-mediated inhibitory effect of oligomeric polypeptides (K8Y8, K4Y8, K8W8) on Aß42 fibrillization process. The polypeptides containing lysine as well as representative aromatic amino acids of tryptophan or tyrosine were found to greatly suppress the aggregation as evaluated by thioflavin T assay. Circular dichroism spectra showed that the ß-sheet formation of Aß42 peptides decreased with the polypeptide additives. Molecular docking studies revealed that the oligomeric polypeptides could preferentially bind to Aß42 through π-π stacking between aromatic amino acids and Phe19, together with hydrogen bonding. The cell viability assay confirmed that the toxicity of Aß42 to SH-SY5Y cells was markedly reduced in the presence of polypeptides. This study could be beneficial for developing peptide-based inhibitory agents for amyloidoses. Copyright © 2017 European Peptide Society and John Wiley & Sons, Ltd.

Dual-affinity peptide mediated inter-protein recognition.

We present for the first time an enhanced interaction affinity between an abundant soluble protein (human serum albumin) and a membrane protein (chemokine receptor 4) mediated by a dual-affinity peptide E5.

Self-assembled NIR nanovesicles for long-term photoacoustic imaging in vivo.

We report a supramolecular approach for the preparation of photostable NIR nanovesicles based on a cyanine dye derivative as a photoacoustic (PA) contrast agent for high-performance nano-imaging.

Inhibition of ß-amyloid peptide self-assembly and cytotoxicity by poly(LVFF-co-ß-amino ester).

The ß-amyloid (Aß) peptide plays an important role in the onset and progress of Alzheimer's disease (AD). Therefore, studies on inhibiting fibril formation and thus neurotoxicity of Aß could be beneficial for the prevention and treatment of AD. We report a convenient synthetic approach for one-pot preparation of poly-(ß-amino ester) copolymerized with the GGLVFF peptide, which is based on the frequently used inhibitor LVFF. The copolymer was found to efficiently inhibit the aggregation and fibrillation of Aß42 by using fluorescence assay and atomic force microscopy. Reduced ß-sheet formation of Aß42 peptide after addition of copolymer was observed by circular dichroism. Furthermore, the cell viability assay confirmed that the toxicity of Aß42 for SH-SY5Y cells was markedly reduced in the presence of copolymer. This could be beneficial for AD prevention and treatment and also for the molecular design of inhibitory agents for other amyloidoses.

A novel three-dimensional CdII metal-organic framework based on [Cd6(malonate)6] metallomacrocycles with zeolite SOD (sodalite) topology: poly[ammine-µ3-malonato-cadmium(II)].

A novel Cd(II) metal-organic framework, [Cd(C(3)H(2)O(4))(NH(3))](n), was synthesized by liquid diffusion conducted in the presence of ammonia. The Cd(II) atom has seven-coordinate O(6)N pentagonal-bipyramidal geometry. Six Cd(II) centers are joined by six malonate ligands to form an S(6)-symmetric [Cd(6)(malonate)(6)] metallomacrocycle, which is further extended through a side-on chelating malonate ligand to form a three-dimensional network. Topologically, each Cd(II) center is connected to four others to yield an infinite three-periodic four-coordinated SOD (sodalite) network with point symbol {4(2)·6(4)}. The overall network structure in the crystal is maintained and stabilized by the presence of N-H...O hydrogen bonds.