Chemical characterization and sensory properties of apple brandies aged with different toasted oak chips and ultra-high-pressure treatments.

Aging is an important processing step of producing high quality apple brandy. In this study, apple brandies aged by traditional method and using three different toasted oak chips combined with or without ultra-high-pressure (UHP) treatment were prepared to compare their differences in chemical characterization and sensory properties. The results indicated that the brandies aged with toasted oak chip increased the levels of total acidity, volatile acidity and phenolic compounds. It also had the desirable color and taste. The brandy aged with toasted oak chip combined with UHP reached the highest levels of total acidity (1.06 g/L), total phenolic content (284.92 mg/L) and aromatic esters (49.37 %). Therefore, the aging with high toasted oak chip combined with UHP treatment could cut the traditional aging time to meet the same quality as traditional aging method. The results are very useful to develop a fast and efficient aging technique for brandy production.

AChE activity self-breathing control mechanisms regulated by H2Sn and GSH: Persulfidation and glutathionylation on sulfhydryl after disulfide bonds cleavage.

Hydrogen sulfide (H2S), or dihydrogen sulfane (H2Sn), acts as a signal molecule through the beneficial mechanism of persulfidation, known as the post-translational transformation of cysteine residues to persulfides. We previously reported that Glutathione (GSH) could regulate enzyme activity through S-desulfurization or glutathionylation of residues to generate protein-SG or protein-SSG, releasing H2S. However, little is known about the mechanisms by which H2Sn and GSH affect the disulfide bonds. In this study, we provide direct evidences that H2Sn and GSH modify the sulfhydryl group on Cys272, which forms disulfide bonds in acetylcholinesterase (AChE), to generate Cys-SSH and Cys-SSG, respectively. Glutathionylation of disulfide is a two-step reaction based on nucleophilic substitution, in which the first CS bond is broken, then the SS bond is broken to release H2S. H2Sn and GSH controlled self-breathing motion in enzyme catalysis by disconnecting specific disulfide bonds and modifying cysteine residues, thereby regulating AChE activity. Here, we elucidated H2Sn and GSH mechanisms on disulfide in the AChE system and proposed a self-breathing control theory induced by H2Sn and GSH. These theoretical findings shed light on the biological functions of H2Sn and GSH on sulfhydryl and disulfide bonds and enrich the theory of enzyme activity regulation.

Effect of heat-treating methods on components, instrumental evaluation of color and taste, and antioxidant properties of sea buckthorn pulp flavonoids.

Sea buckthorn has strong physiological activities, which might be attributed to its flavonoid compounds. The heat treatment could speed up the transformation of the flavonoids, so it would affect the properties of the flavonoids. To explore the effect of the heat-treating methods on the sea buckthorn pulp flavonoids, the flavonoid compositions, color and taste properties, and antioxidant activity, prepared by three heating methods, were investigated. The results showed that the extractable sea buckthorn flavonoids rose after heat treatment; the high temperature with high-pressure treatment had the largest yield (3.66 ± 0.21 mg/ml). A total of 16 flavonoid monomers were identified in the sea buckthorn, the content of isorhamnetin-3-O-rutinoside was the highest in the pulp (589.34 ± 4.72 µg/ml), the concentrations of eight monomers increased after heat-treating, and the treating methods could significantly affect the contents of flavonoid monomers (p < 0.05). The difference in color was positively correlated to the heat-treating time and temperature (p < 0.01); the ΔE value in high pressure with high temperature treatment was the largest (10.59). The heat treatment enhanced bitterness and acidity, while it greatly reduced the saltiness, umami, and sweetness (p < 0.01). The stronger antioxidant capacity of the total flavonoids was accompanied by the higher temperature and longer time of heat treatment. The antioxidant capacity was closely correlated to the contents of flavonoid monomers. Therefore, the heat treatment boosted the content of the flavonoids, enhanced the taste, and improved the antioxidant properties. PRACTICAL APPLICATION: This study can provide suitable a method for improving the extracting efficiency, improving properties and biological activities of sea buckthorn pulp flavonoids, also provide directions for predicting the antioxidant activity by the flavonoid monomer contents.

Modification and validation of the simultaneous detection of 38 pesticide residues method by ultra-high-performance liquid chromatography/tandem mass spectrometry with QuEChERS extraction in different oil crops and products.

RATIONALE: Oil crops and products are important food materials in daily life. Pesticide residues in food could directly and indirectly endanger human health. However, the method for detecting multiple pesticides simultaneously is limited. In this study, an easy and efficient method for the simultaneous determination of 38 pesticides in oil crops and products was established and validated. METHODS: All samples were treated with a modified QuEChERS procedure followed by ultra-high-performance liquid chromatography/tandem mass spectrometry (UHPLC/MS/MS) analysis. Mass spectrometry was performed in positive and negative ion electrospray ionization mode. The mobile phase consisted of 0.1% formic acid in water and 0.1% formic acid in acetonitrile. The column used was a Poroshell 120 EC-C18 and the flow rate was 0.3 mL/min. RESULTS: The method was validated so that the calibration curves for all pesticides had good linearity in the concentration range of 10-1000 µg/L with correlation coefficients (R2 ) above 0.9945. The recovery rates were between 70.1 and 120.0%, with relative standard deviations (RSDs) (n = 6) ≤20.0%. The limits of quantification (LOQs) ranged from 0.5 to 10 µg/kg, limits of detection (LODs) ranged from 2.0 to 30 µg/kg, and the matrix effect (ME) ranged from -18.77 to 19.33%, respectively. CONCLUSIONS: The method proved to be accurate, sensitive, and stable. It can be used for rapid screening and confirmation of 38 pesticide residues in oil crops and products which takes 10 min for sample extraction and clean-up with less requirement of solvents. This study provides a technical basis for regulatory analysis and quality supervision of foods.

Comprehensive Analysis of Lysine Lactylation in Rice (Oryza sativa) Grains.

Protein lysine lactylation is a new post-translational modification (PTM) prevalently found in fungi and mammalian cells that directly stimulates gene transcription and regulates the glycolytic flux. However, lysine lactylation sites and regulations remain largely unexplored, especially in cereal crops. Herein, we report the first global lactylome profile in rice, which effectively identified 638 lysine lactylation sites across 342 proteins in rice grains. Functional annotations demonstrated that lysine lactylation was enriched in proteins associated with central carbon metabolism and protein biosynthesis. We also observed that proteins serving as nutrition reservoirs in rice grains were frequently targeted by lactylation. Homology analyses indicated that lactylation was conserved on both histone and nonhistone proteins across plants, human cells, and fungi. In addition to lactylation, additional types of acylations could co-occur in many proteins at identical lysine residues, indicating potential cross-talks between these modifications. Our study provided a comprehensive profile of protein lysine lactylation in cereal crop grains.

Cyanidin-3-O-glucoside and cisplatin inhibit proliferation and downregulate the PI3K/AKT/mTOR pathway in cervical cancer cells.

Natural compounds have been increasingly investigated as substances enhancing the effect of drugs and reducing drug-related adverse reactions. The objective of this study was to determine how a combination of cisplatin (DDP) with cyanidin-3-O-glucoside (C3G) affected malignancy features of cervical cancer cells. The results demonstrated that the proliferation of HeLa cells treated with 5 µg/ml DDP, 400 µg/ml C3G, or a combination of both (5 µg/ml DDP and 400 µg/ml C3G) was inhibited by 17.43%, 34.98%, and 63.38%, respectively. The IC50 values for DDP and the DDP/C3G combination treatments in HeLa cells were 18.53 and 6.435 µg/ml, respectively. Flow cytometry analysis indicated that treatment with DDP, C3G, or the combination induced G1 cell cycle arrest and apoptosis in HeLa cells. Furthermore, after treatment, cyclin D1 and Bcl-2 levels decreased; Bax, cleaved caspase-3, p53, and TIMP-1 were activated; and the PI3K/AKT/mTOR signaling pathway was modulated. These anticancer effects were enhanced in cells treated with the combination of DDP and C3G compared to those treated with DDP or C3G alone. Our study indicates that C3G increases the antitumor activity of DDP, suggesting a potential strategy to reduce adverse effects associated with chemotherapy in cervical cancer. PRACTICAL APPLICATION: Natural biologically active food ingredients are suggested to have a potential to enhance the effect of chemotherapy in cancer. We believe that our study makes a significant contribution to the literature because it revealed, for the first time, that C3G could increase the antitumor activity of DDP, suggesting a potential strategy to reduce adverse effects associated with chemotherapy in cervical cancer.

Gut Microbiota Modulation by Polyphenols from Aronia melanocarpa of LPS-Induced Liver Diseases in Rats.

Aronia melanocarpa polyphenols (AMPs) can alleviate the degree of liver diseases in rats. However, the mechanism by which this is achieved through gut microbiota modulation remains unclear. Here, a rich-polyphenol extract of A. melanocarpa (AMPs) was used to treat lipopolysaccharide (LPS)-induced liver diseases in rats. To gain insights into the anti-LPS-induced liver disease, liver function index, expression of apoptosis proteins, inflammatory factors, and activation of inflammatory signaling pathways were determined with western blot analysis, immunohistochemistry, and 16S rRNA sequencing or quantitative real-time polymerase chain reaction (qRT-PCR). After AMPs treatment, the gut microbiota composition was modulated, promoting the intestinal barrier function by increasing the expression of intestinal epithelial cell tight junction proteins to reduce the LPS content in serum. The expression levels of inflammatory factors interleukin 6 (IL-6), interleukin 1ß (IL-1ß), tumor necrosis factor α (TNF-α), and related mRNAs were reduced. These results showed that AMPs, as a bioactive substance, could enhance the intestinal barrier function and modulate the gut microbiota of LPS-induced liver diseases.

Effects of high hydrostatic pressure on the binding capacity, interaction, and antioxidant activity of the binding products of cyanidin-3-glucoside and blueberry pectin.

In this study, the effects of high hydrostatic pressure (HP) treatment on the binding capacity, interaction, and antioxidant activity of the binding products of blueberry pectin (BP) and cyanidin-3-glucoside (C3G) were assessed. HP was found to significantly improve the adsorption between C3G and BP. After binding, the C3G concentration was found to be the highest (382.1 ± 13.2 µg/mg for BP) when using a C3G-BP mass ratio of 1:2, a pressure of 400 MPa, and a holding time of 15 min. HP processing decreased particle size and altered the characteristics of C3G-BP complexes. The main binding form of the complexes before HP treatment was pectin-wrapped C3G by hydrogen bond interaction, while HP caused charged groups in pectin to be more exposed and improve the electrostatic interaction between C3G and BP. The antioxidant activity results showed that the presence of BP could protect the ferric-reducing antioxidant power of C3G after HP treatment.

Chemical constituents from acid hydrolyzates of Panax quinquefolius total saponins and their inhibition activity to α-glycosidase and protein tyrosine phosphatase 1B.

Objective: To investigate the hypoglycemic components from the acid hydrolyzates of Panax quinquefolius total saponins, and screen the active compounds by in vitro inhibitory activities to α-glycosidase enzymes and protein tyrosine phosphatase-1B (PTP1B). Methods: The hydrolyzates were chromatographed repeatedly over silica gel column, and the structures of the compounds were determined by means of NMR. The in vitro bioassay was performed through the inhibitory effects on α-glucosidase or/and PTP1B. Results: Eight compounds were isolated, which identified as 20(S)-panaxadiol (1), (20S,24R)-dammarane-20,24-epoxy-3ß,6α,12ß,25-tetraol (2), 20(R)-dammarane-3ß,12ß,20,25-tetraol (3), 20(S)-dammarane-3ß,6α,12ß,20,25-pentol (4), 20(R)-dammarane-3ß,12ß,20,25-tetrahydroxy-3ß-O-ß-d-glucopyranoside (5), ß-sitosterol (6), oleanolic acid (7) and 20(S)-protopanaxadiol (8). Compound 5 was ginseng triterpenoid isolated from the acid hydrolysates of total saponins from P. quinquefolius for the first time. In this paper, the possible in vitro inhibitory activities were investigated. Compound 5 exhibited significantly inhibitory activity against α-glucosidase, and the IC50 value [(0.22 ± 0.21) µmol/L] was about 43-fold lower than positive control. For the PTP1B inhibition assay, compound 5 indicated the strongest inhibitory effect with IC50 of (5.91 ± 0.38) µmol/L, followed by compound 4 with IC50 of (6.21 ± 0.21) µmol/L, which were all showed competitive inhibitory pattern by using a Lineweaver-Burk plot. Conclusion: These results supported the potential application of dammaranes from acid hydrolyzates of P. quinquefolius total saponins can be used as ingredients of ancillary anti-diabetic agent or functional factor.

Stability and structural characteristics of amylopectin nanoparticle-binding anthocyanins in Aronia melanocarpa.

The bioavailability of anthocyanins from Aronia melanocarpa is limited by their high degradation rates and poor stability. One way to protect anthocyanins is to deliver them using amylopectin nanoparticles (APNPs). In this study, we used distilled water as the reaction system, and achieved the most stable formulation of anthocyanins and APNPs using a mass ratio of 1:12, a pH of 3, and a binding time of 60 min, at which the binding rates were 81.52%, 82.67%, and 84.00%, respectively. Moreover, electron microscopy, X-ray diffraction, and Fourier-transform infrared spectroscopy were applied to observe microstructural changes with each combination. The observed stabilities and antioxidant activities indicated that binding played a role in protecting anthocyanins from heat, oxidization, reduction reactions, and metallic ions. Finally, our in vitro-digestion results suggested that the optimized compound can partially avoid degradation and diffusion, wherein the anthocyanin retention rate and FRAP value increased to 17.05% and 24.85%, respectively.

Anthocyanins Extracted from Aronia melanocarpa Protect SH-SY5Y Cells against Amyloid-beta (1-42)-Induced Apoptosis by Regulating Ca2+ Homeostasis and Inhibiting Mitochondrial Dysfunction.

We investigated the cytoprotective effects of anthocyanins in Aronia melanocarpa against apoptosis induced by Aß1-42, a key mediator of AD pathophysiology. We measured intracellular calcium with a colorimetric kit, cellular apoptosis with DAPI, intracellular ROS with the fluorescent marker 2,3-dimethoxy-1,4-naphthoquinone, mitochondrial membrane potential with JC-1, and ATP with a colorimetric kit. Gene transcription and protein expression levels of calmodulin, cytochrome c, caspase-9, cleaved caspase-3, Bcl-2, and Bax were analyzed by RT-PCR and Western blotting. The results showed that pretreatment with anthocyanins significantly inhibited Aß1-42-induced apoptosis, decreased intracellular calcium and ROS, and increased ATP and mitochondrial membrane potential. RT-PCR and Western blotting revealed that anthocyanins upregulated the gene transcription and protein expression of calmodulin and Bcl-2 and downregulated those of cytochrome c, caspase-9, cleaved caspase-3, and Bax. A. melanocarpa anthocyanins protected SH-SY5Y cells against Aß1-42-induced apoptosis by regulating Ca2+ homeostasis and apoptosis-related genes and inhibiting mitochondrial dysfunction.

Cytotoxic triterpenes from the acid hydrolyzate of Gynostemma pentaphyllum saponins.

One new dammarane-type triterpene, gypsapogenin A (1), was isolated from the acid hydrolyzate of total saponins from Gynostemma pentaphyllum (Thunb.) Makino, together with two known compounds, (20S,24S)-3ß,20,21ß,23ß,25-pentahydroxy-21,24-cyclodammarane (2) and (23S)-3ß-hydroxydammar-20,24-dien-21-oic acid 21,23-lactone (3). Its structural elucidations were accomplished mainly on the basis of the interpretation of spectroscopic data, such as IR, HR-TOF-MS, and NMR. The cytotoxic activities were evaluated against HepG2 and A549 human cancer cell lines.

Proteomic Analysis of the Antibacterial Mechanism of Action of Juglone against Staphylococcus aureus.

Juglone is a plant-derived 1,4-naphthoquinone with confirmed antibacterial activity. However, the mechanism of action of juglone against Staphylococcus aureus remains unclear. Possible mechanisms were explored by a proteomic analysis of S. aureus proteins that are inhibited by juglone. Two-dimensional gel electrophoresis revealed that 21 protein spots were differentially expressed between juglone-treated and untreated cells of which 13 were identified. A bioinformatic analysis revealed that proteins participating in protein synthesis, the tricarboxylic acid cycle, as well as DNA and RNA synthesis were inhibited by juglone, thus leading to cell collapse. These findings provide clues regarding the mechanism of action of juglone, which can be effective for treating cases of S. aureus infection.

Antibacterial Activity of Juglone against Staphylococcus aureus: From Apparent to Proteomic.

The proportion of foodborne disease caused by pathogenic microorganisms is rising worldwide, with staphylococcal food poisoning being one of the main causes of this increase. Juglone is a plant-derived 1,4-naphthoquinone with confirmed antibacterial and antitumor activities. However, the specific mechanism underlying its antibacterial activity against Staphylococcus aureus remains unclear. To elucidate the mechanism underlying its antibacterial activity, isobaric tags for relative and absolute quantitation methods of quantitative proteomics were applied for analysis of the 53 proteins that were differentially expressed after treatment with juglone. Combined with verification experiments, such as detection of changes in DNA and RNA content and quantification of oxidative damage, our results suggested that juglone effectively increased the protein expression of oxidoreductase and created a peroxidative environment within the cell, significantly reducing cell wall formation and increasing membrane permeability. We hypothesize that juglone binds to DNA and reduces DNA transcription and replication directly. This is the first study to adopt a proteomic approach to investigate the antibacterial mechanism of juglone.