Alanine Substitution to Determine the Effect of LR5 and YR6 Rice Peptide Structure on Antioxidant and Anti-Inflammatory Activity.

The relationship between the structure of peptides LR5 (LHKFR) and YR6 (YGLYPR) and their antioxidant and anti-inflammatory activity remains unclear. Herein, leucine, tyrosine, proline, and phenylalanine at different positions in the peptides were replaced by Alanine (Ala), and two new pentapeptides (AR5 and LAR5) and four hexapeptides (AGR6, YAR6, YLR6, and YGR6) were obtained. The effect of Ala replacement on the hydrophobicity, cytotoxicity, NO inhibition rate, and active oxygen radical scavenging ability of these peptides and their antioxidant and anti-inflammatory abilities were investigated. The results indicated that the hydrophobicity of the peptides was associated with their amino acid composition and their specific sequence. However, hydrophobicity had no significant effect on cytotoxicity. Ala replacement was shown to enhance hydrophobicity and consequently increased the antioxidant and anti-inflammatory activity of the peptides. The molecular docking studies indicated that the amino acid interactions of the peptide with the Keap1 protein influenced the hydrophobicity and thus affected the antioxidant activity of the peptide.

An iron-based metal-organic framework as a novel dispersive solid-phase extraction sorbent for the efficient adsorption of tetrabromobisphenol A from environmental water samples.

For environmental safety, it is important to establish a simple, rapid, and sensitive method for emerging pollutants. Here, a dispersive solid-phase extraction (d-SPE) method based on an iron-based metal-organic framework (Fe-MIL-88-NH2) combined with high-performance liquid chromatography (HPLC) was developed for tetrabromobisphenol A (TBBPA) in water samples. Fe-MIL-88-NH2 was synthesized using a solvothermal method and completely characterized. Fe-MIL-88-NH2 had good water stability and gave a maximum adsorption capacity of 40.97 mg g-1 for TBBPA. The adsorption of TBBPA on Fe-MIL-88-NH2 followed Langmuir adsorption models and a pseudo-second-order kinetic model. The bromine ion and the hydroxyl group of TBBPA could form strong hydrogen bond interactions with the amino protons around the cavity of Fe-MIL-88-NH2, which was in accord with the molecular simulation calculations. Furthermore, several important d-SPE parameters were optimized, such as the amount of materials, extraction time, pH, ionic strength, elution solvent type, and volume. The established method showed good linearity in the concentration range of 0.005-100 µg g-1 (r2 ≥ 0.9996). This method's limits of detection (LOD) and quantification (LOQ) were 0.001 µg g-1 and 0.005 µg g-1, respectively. The recoveries in spiked water samples ranged from 87.5% to 104.9%. The proposed method was applied successfully to detect TBBPA in environmental water samples.

Magnetic relaxation switching assay based on three-dimensional assembly of Fe3O4@ZIF-8 for detection of cadmium ions.

The design and construction of a novel magnetic resonance switch (MRS) sensor for cadmium ion (Cd2+) detection is described. Fe3O4@ZIF-8 was synthesized through seed-mediated growth of dimercaptosuccinic acid-coated Fe3O4. Fe3O4@ZIF-8 with high relaxation value (163.086 mM-1 s-1) and large negative zeta potential (-20.69 mV) exhibited good magnetic relaxation performance and water solubility. The successfully synthesized Fe3O4@ZIF-8 was used to develop an immune recognition-based MOFs-MRS sensor for highly sensitive detection of Cd2+. The proposed MRS detected a wide linear range of Cd2+ concentration from 2 to 200 ng mL-1 with a low limit of detection of 0.65 ng mL-1 (S/N = 3), and displayed high selectivity towards matrix interference. The robust sensing system was effective even in a complex sample matrix, enabling the quantitative analysis of Cd2+ content in rice samples and drinking water samples with good reliability. Recoveries of Cd2+ ranged from 91.50 to 112.05% for spiked drinking water and from 95.86 to 110.45% for spiked rice samples. The versatility of Fe3O4@ZIF-8 with customized relaxation responses could allow the adaptation of magnetic resonance platforms for food safety purposes.

Application of an improved hollow fiber liquid phase microextraction technique coupled to LC-MS/MS to studying migration of fluorescent whitening agents from plastic food contact materials.

In this paper, a new hollow fiber liquid-phase microextraction method was developed to improve the extraction of five fluorescent whitening agents that migrated from plastics food contact materials. Influencing factors, such as the types of membrane, the extraction solvent, the stirring speed, the addition of salt ion, and extraction time, were investigated in detail. Under the optimal conditions, high enrichment factors (71-205) can be obtained with 15 µL extraction solvent. The new method is advantageous; the polypropylene hollow fiber membrane modified by sepiolite nanoparticles had excellent solvent binding force and mass transfer effect compared with the conventional extraction technique. The extracts were analyzed by high performance liquid chromatography-tandem mass spectrometry, the limits of detection were 0.3 or 0.9 ng kg-1 with good correlation coefficients (r2 ≥ 0.9940) for the five fluorescent whitening agents. The intra-day and inter-day recoveries ranged between 82.6% and 112%, with a relative standard deviation of less than 12%. The established method was successfully applied to the analysis of fluorescent whitening agent migration from four types of plastic food contact materials immersed in three food simulants.

Extraction of antioxidant peptides from rice dreg protein hydrolysate via an angling method.

Selective enrichment of the highly active antioxidant peptides is required as the lack of an efficient method leads to long screening processes, hampering the research of antioxidant peptides. A simple synthetic metal-organic framework MIL-53 (Cr) was initially applied to extract specific antioxidant peptides from rice dreg protein hydrolysate. The highest active fraction was further purified by reversed-phase high-performance liquid chromatography. The antioxidant peptides with the highest antioxidant activities were identified as Gly-Asp-Met-Asn-Pro and Leu-Leu-Leu-Arg-Trp by LC-MS. These two peptides were synthesized and also exhibited good scavenging activity on the DPPH free radical, superoxide anion free radical and hydroxyl radical, and good chelating ability on Fe2+. The results confirmed that the angling method was effective for antioxidant peptide enrichment from protein hydrolysates.

MicroRNA expression profile and functional analysis reveal their roles in contact inhibition and its disruption switch of rat vascular smooth muscle cells.

Contact inhibition and its disruption of vascular smooth muscle cells (VSMCs) are important cellular events in vascular diseases. But the underlying molecular mechanisms are unclear. In this study we investigated the roles of microRNAs (miRNAs) in the contact inhibition and its disruption of VSMCs and the molecular mechanisms involved. Rat VSMCs were seeded at 30% or 90% confluence. MiRNA expression profiles in contact-inhibited confluent VSMCs (90% confluence) and non-contact-inhibited low-density VSMCs (30% confluence) were determined. We found that multiple miRNAs were differentially expressed between the two groups. Among them, miR-145 was significantly increased in contact-inhibited VSMCs. Serum could disrupt the contact inhibition as shown by the elicited proliferation of confluent VSMCs. The contact inhibition disruption accompanied with a down-regulation of miR-145. Serum-induced contact inhibition disruption of VSMCs was blocked by overexpression of miR-145. Moreover, downregulation of miR-145 was sufficient to disrupt the contact inhibition of VSMCs. The downregulation of miR-145 in serum-induced contact inhibition disruption was related to the activation PI3-kinase/Akt pathway, which was blocked by the PI3-kinase inhibitor LY294002. KLF5, a target gene of miR-145, was identified to be involved in miR-145-mediated effect on VSMC contact inhibition disruption, as it could be inhibited by knockdown of KLF5. In summary, our results show that multiple miRNAs are differentially expressed in contact-inhibited VSMCs and in non-contact-inhibited VSMCs. Among them, miR-145 is a critical gene in contact inhibition and its disruption of VSMCs. PI3-kinase/Akt/miR-145/KLF5 is a critical signaling pathway in serum-induced contact inhibition disruption. Targeting of miRNAs related to the contact inhibition of VSMCs may represent a novel therapeutic approach for vascular diseases.

[Construction and expression of a prokaryotic expression plasmid of idiotypic vaccine against B cell lymphoma: encoding the fusion genes of single-chain variable fragment and MCP-3].

The aim was to construct a prokaryotic expression plasmid encoding the fusion gene of single-chain variable fragment and monocyte chemotactic protein-3 (MCP-3). The cDNAs of immunoglobulin (Ig) VH and Ig VL were amplified by RT-PCR and assembled into the single-chain variable fragment (scFv) by recombinant PCR method. The cDNAs of Ig VH and Ig VL were connected by a (Gly4Ser)3 linker. Then, the fragments of scFv and MCP-3 were connected with a NDAQAPKS spacer, using recombinant PCR method again. The results indicated that the fusion gene of scFv-MCP-3 were constructed correctly and cloned into the prokaryotic expression plasmid successfully identified by sequencing and restriction endonucleases examination. Finally, the fusion protein was expressed in E coli DH5alpha under induction by arabinose. And the fusion protein was 65 kD and account for 30% of the total protein of the bacteria. In conclusion, a prokaryotic plasmid, encoding the fusion gene of single-chain variable fragment with MCP-3 and expressing idiotype protein vaccination against B cell lymphoma, was constructed correctly.