抗流感病毒鸡卵黄免疫球蛋白IgY的制备和性质

用流感病毒免疫产蛋鸡,获得的免疫卵黄经两步盐析和一步凝胶过滤分离得到的IgY经SDS-PAGE分析其纯度在95%以上,纯化的IgY经胃蛋白酶(pH=4 0,37℃)酶解后,得到的片段抗体为Fab。采用竞争抑制ELISA法对IgY和Fab的免疫反应活性进行了对比,表明IgY经胃酶切解后保留了约70%的反应活性。     

Screening of Novel Bioactive Peptides from Goat Casein: In Silico to In Vitro Validation

Food-derived bioactive peptides are of great interest to science and industry due to evolving drivers of food product innovation, including health and wellness. This study aims to draw attention through a critical study on how bioinformatics analysis is employed in the identification of bioactive peptides in the laboratory. An in silico analysis (PeptideRanker, BIOPEP, AHTpin, and mAHTPred) of a list of peptides from goat casein hydrolysate was performed to predict which sequences could potentially be bioactive. To validate the predictions, the in vitro antihypertensive potential of the five peptides with the highest potential was first measured. Then, for three of these, gastrointestinal digestion was simulated in vitro, followed by the analysis of the resulting ACE inhibitory activity as well as antioxidant capacity. We thus observed that the use of new computational biology technologies to predict peptide sequences is an important research tool, but they should not be used alone and complementarity with various in vitro and in vivo assays is essential.     

Evaluation of Antioxidant and Free Radical Scavenging Capacities of Polyphenolics from Pods of Caesalpinia pulcherrima

Thirteen polyphenolics were isolated from fresh pods of Caesalpinia pulcherrima using various methods of column chromatography. The structures of these polyphenolics were elucidated as gallic acid (1), methyl gallate (2), 6-O-galloyl-d-glucoside (3), methyl 6-O-galloyl-β-d-glucoside (4), methyl 3,6-di-O-galloyl-α-d-glucopyranoside (5), gentisic acid 5-O-α-d-(6'-O-galloyl)glucopyranoside (6), guaiacylglycerol 4-O-β-d-(6'-O-galloyl)glucopyranoside (7), 3-methoxy-4-hydroxyphenol 1-O-β-d-(6'-O-galloyl) glucopyranoside (8), (+)-gallocatechin (9), (+)-catechin (10), (+)-gallocatechin 3-O-gallate (11), myricetin 3-rhamnoside (12), and ampelopsin (13). All isolated compounds were tested for their antioxidant activities in the 1,1-diphenyl-2-picrylhydrazyl (DPPH), hydroxyl, and peroxynitrite radicals scavenging assays. Among those compounds, 11, 12, and 2 exhibited the best DPPH-, hydroxyl-, and peroxynitrite radical-scavenging activities, respectively. Compound 7 is a new compound, and possesses better scavenging activities towards DPPH but has equivalent hydroxyl radical scavenging activity when compared to BHT. The paper is the first report on free radical scavenging properties of components of the fresh pods of Caesalpinia pulcherrima. The results obtained from the current study indicate that the free radical scavenging property of fresh pods of Caesalpinia pulcherrima may be one of the mechanisms by which this herbal medicine is effective in several free radical mediated diseases.     

Antioxidant and Antithrombotic Activities of Rapeseed Peptides

The antioxidant and antithrombotic activities of crude rapeseed peptides (CRPs) and peptide fractions (RP25 and RP55) prepared from aqueous enzymatic extraction (AEE) of rapeseed were determined. The reducing power of RP55 and CRPs was higher than that of RP25 at the same concentrations. Rapeseed peptides exhibited marked antioxidant activities. The median effective dose (ED₅₀) values of CRPs, RP25 and RP55 for α,α-diphenyl-β-picrylhydrazyl (DPPH) radical scavenging were 72, 499 and 41 μg/mL, respectively. The ED₅₀ values for RP25 and RP55 for hydroxyl radicals scavenging were 2.53 and 6.79 mg/mL, respectively while the ED₅₀ values of RP55 and CRPs for inhibition of lipid peroxidation in a liposome model system were 4.06 and 4.69 mg/mL, respectively. The inhibitory effect on lipid oxidation of RP55 was similar to that of ascorbic acid at a concentration of 5.0 mg/mL. A good positive correlation existed between the peptide concentration and antioxidant activity. RP55 generally showed more potent antioxidant activities except for hydroxyl radicals scavenging ability than RP25 and CRPs at the same concentrations, which was thought to relate to the significantly higher contents of hydrophobic amino acid, tannin, and the brown color substances in RP55. Rapeseed peptides possessed marked inhibitory activities on the thrombin-catalyzed coagulation of fibrinogen, however, their inhibitory effects were not comparable to that of heparin.     

紫草抗氧化成分的提取及其活性研究

分别用石油醚、三氯甲烷和乙醇依次提取紫草成分 ,用OSI仪测定了三种提取物对猪油氧化稳定性的影响 ,发现石油醚提取成分 (LE - pet)和三氯甲烷提取成分 (LE -chl)有明显的抗氧化活性 ,添加到猪油中后 ,w =0 0 2 %时 ,猪油的氧化稳定性比空白样品提高了约 1倍。对LE -pet和LE -chl进行薄层层析分离鉴定 ,得到三种化合物 :β ,β 二甲基丙烯酰紫草醌 (Ⅰ)、乙酰紫草醌 (Ⅱ)和紫草醌 (Ⅲ)。对它们的抗氧化活性进行测试 ,化合物Ⅱ和Ⅲ表现出明显的抗氧化活性 ,当w(Ⅱ)=0 .0 6 %、w(Ⅲ) =0 0 6 %时 ,猪油的氧化稳定性分别提高了 6 0 4和 3 94倍。化合物Ⅰ、Ⅱ、Ⅲ与VE均有较强的协同作用 ,化合物Ⅰ、Ⅱ、Ⅲ的质量分数为 0 .0 2 %与VE 的质量分数为 0 0 2 %共同使用时 ,其抗氧化协同系数 (SE)分别为 81 4(Ⅰ)、5 2 7(Ⅱ)和 132 6 (Ⅲ )     

Kunitz-Type Peptides from Sea Anemones Protect Neuronal Cells against Parkinson’s Disease Inductors via Inhibition of ROS Production and ATP-Induced P2X7 Receptor Activation

Parkinson’s disease (PD) is a socially significant disease, during the development of which oxidative stress and inflammation play a significant role. Here, we studied the neuroprotective effects of four Kunitz-type peptides from Heteractis crispa and Heteractis magnifica sea anemones against PD inductors. The peptide HCIQ1c9, which was obtained for the first time, inhibited trypsin less than other peptides due to unfavorable interactions of Arg17 with Lys43 in the enzyme. Its activity was reduced by up to 70% over the temperature range of 60–100 °C, while HCIQ2c1, HCIQ4c7, and HMIQ3c1 retained their conformation and stayed active up to 90–100 °C. All studied peptides inhibited paraquat- and rotenone-induced intracellular ROS formation, in particular NO, and scavenged free radicals outside the cells. The peptides did not modulate the TRPV1 channels but they affected the P2X7R, both of which are considered therapeutic targets in Parkinson’s disease. HMIQ3c1 and HCIQ4c7 almost completely inhibited the ATP-induced uptake of YO-PRO-1 dye in Neuro-2a cells through P2X7 ion channels and significantly reduced the stable calcium response in these cells. The complex formation of the peptides with the P2X7R extracellular domain was determined via SPR analysis. Thus, these peptides may be considered promising compounds to protect neuronal cells against PD inductors, which act as ROS production inhibitors and partially act as ATP-induced P2X7R activation inhibitors.     

Quality Changes and Antioxidant Properties of Microencapsulated Kenaf (Hibiscus cannabinus L.) Seed Oil During Accelerated Storage

Quality changes and antioxidant properties of microencapsulated kenaf (Hibiscus cannabinus L.) seed oil (MKSO) were investigated at 6-day intervals during 24 days of storage at 65 °C. DPPH, ABTS, FRAP and BCB assays were used to determine the antioxidant activity (AOA) and their correlations were reported. Total flavonoids content (TFC) and oxidative stability in terms of evolution of thiobarbituric acid reactive substances (TBARS) were also studied. The results showed that there was a significant (p < 0.05) effect of microencapsulation whereby AOA of MKSO showed only a 23.28 % decrease as compared to a 61.51 % decrease in the control bulk oil (KSO) when assessed by FRAP. MKSO showed significantly (p < 0.05) lower reduction of its initial TFC as compared to KSO and total increase in TBARS of MKSO was reported to be significantly (p < 0.05) lower than that of KSO. AOA determined by all assays were well correlated with TFC and also among themselves. Microencapsulation of kenaf seed oil was found to be effective in the stabilisation of natural antioxidants and prolonging the shelf-life.     

Screening for the Active Anti-Inflammatory and Antioxidant Polyphenols of Gaultheria procumbens and Their Application for Standardisation: From Identification through Cellular Studies to Quantitative Determination

Aerial parts, leaves, and stems of Gaultheria procumbens are polyphenol-rich herbal medicines with anti-inflammatory and antioxidant effects. The present study focused on identifying active markers of the G. procumbens extracts in an integrated approach combining phytochemical and biological capacity tests. The target compounds, representing all classes of Gaultheria polyphenols, were pre-selected by LC-ESI-PDA-MS/MS. For unambiguous identification, the key analytes, including a rare procyanidin trimer (cinnamtannin B-1), miquelianin potassium salt, and two new natural products: quercetin and kaempferol 3-O-β-d-xylopyranosyl-(1→2)-β-d-glucuronopyranosides, were isolated by preparative HPLC and investigated by spectroscopy (HR-ESI-MS, UV-vis, CD, 1D- and 2D-NMR), thiolysis, flame photometry, optical rotation experiments, and absolute configuration studies. The significant contribution of the pre-selected compounds to the biological effects of the extracts was confirmed in vitro: the analytes significantly and in a dose-dependent manner down-regulated the pro-oxidant and pro-inflammatory functions of human neutrophils ex vivo (inhibited the release of reactive oxygen species, IL-1β, TNF-α, and neutrophils elastase, ELA-2), inhibited two key pro-inflammatory enzymes (cyclooxygenase, COX-2, and hyaluronidase), and most of them, except gaultherin, exerted potent direct antioxidant activity (ferric reducing antioxidant power and superoxide anion scavenging capacity). Moreover, cellular safety was confirmed for all compounds by flow cytometry. Eventually, as these mechanisms have been connected to the health benefits of G. procumbens, 11 polyphenols were accepted as active markers, and a simple, accurate, reproducible, and fully validated RP-HPLC-PDA method for standardisation of the target extracts was proposed.     

Supercritical Carbon Dioxide Extraction of Flavonoids from Pomelo (Citrus grandis (L.) Osbeck) Peel and Their Antioxidant Activity

Supercritical carbon dioxide (SC-CO₂) extraction of flavonoids from pomelo (Citrus grandis (L.) Osbeck) peel and their antioxidant activity were investigated. Box-Behnken design combined with response surface methodology was employed to maximize the extraction yield of flavonoids. Correlation analysis of the mathematical-regression model indicated that a quadratic polynomial model could be used to optimize the SC-CO₂ extraction of flavonoids. The optimal conditions for obtaining the highest extraction yield of flavonoids from pomelo peel were a temperature of 80 °C, a pressure of 39 MPa and a static extraction time of 49 min in the presence of 85% ethanol as modifier. Under these conditions, the experimental yield was 2.37%, which matched positively with the value predicted by the model. Furthermore, flavonoids obtained by SC-CO₂ extraction showed a higher scavenging activity on hydroxyl, 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) radicals than those obtained by conventional solvent extraction (CSE). Therefore, SC-CO₂ extraction can be considered as a suitable technique for the obtainment of flavonoids from pomelo peel.     

Biological activities of Solanum paludosum Moric. extracts obtained by maceration and supercritical fluid extraction

Solanum paludosum extracts obtained by maceration (SP-EtOH_CRUDE and SP-EtOH_MARC) and supercritical fluid extraction (SP-SFECO₂) were evaluated. Three extracts with distinct chromatographic profiles were obtained: SP-EtOH_CRUDE, with a complex phenolic composition; SP-SFECO₂, containing methoxylated flavonoids; and SP-EtOH_MARC, which bears mainly polar polyphenols. All extracts were strong antioxidants, as measured using the diphenylpicrylhydrazyl (DPPH) method, xanthine/xanthine oxidase/luminol and iron-induced lipid peroxidation systems. Extracts were also evaluated as modifiers of antibiotic activity in strains of Staphylococcus aureus expressing fluoroquinolone, macrolide and tetracycline efflux proteins. Despite their lack of antibacterial activity, all extracts were able to potentiate antibiotics activities by as much as eight fold. Both the maceration and supercritical fluid extraction processes were adequate to obtain extracts with different biological activities according to their patterns of phenolic compounds. The data suggest that S. paludosum could be a valuable source of antioxidants and putative efflux pumps inhibitors.