[Mechanism of astragaloside â £ combined with Panax notoginseng saponins in regulating angiogenesis to treat cerebral ischemia based on network pharmacology and experimental verification].

Network pharmacology and animal and cell experiments were employed to explore the mechanism of astragaloside Ⅳ(AST Ⅳ) combined with Panax notoginseng saponins(PNS) in regulating angiogenesis to treat cerebral ischemia. The method of network pharmacology was used to predict the possible mechanisms of AST Ⅳ and PNS in treating cerebral ischemia by mediating angiogenesis. In vivo experiment: SD rats were randomized into sham, model, and AST Ⅳ(10 mg·kg~(-1)) + PNS(25 mg·kg~(-1)) groups, and the model of cerebral ischemia was established with middle cerebral artery occlusion(MCAO) method. AST Ⅳ and PNS were administered by gavage twice a day. the Longa method was employed to measure the neurological deficits. The brain tissue was stained with hematoxylin-eosin(HE) to reveal the pathological damage. Immunohistochemical assay was employed to measure the expression of von Willebrand factor(vWF), and immunofluorescence assay to measure the expression of vascular endothelial growth factor A(VEGFA). Western blot was employed to determine the protein levels of vascular endothelial growth factor receptor 2(VEGFR2), VEGFA, phosphorylated phosphatidylinositol 3-kinase(p-PI3K), and phosphorylated protein kinase B(p-AKT) in the brain tissue. In vitro experiment: the primary generation of rat brain microvascular endothelial cells(rBEMCs) was cultured and identified. The third-generation rBMECs were assigned into control, model, AST Ⅳ(50 µmol·L~(-1)) + PNS(30 µmol·L~(-1)), LY294002(PI3K/AKT signaling pathway inhibitor), 740Y-P(PI3K/AKT signaling pathway agonist), AST Ⅳ + PNS + LY294002, and AST Ⅳ + PNS + 740Y-P groups. Oxygen glucose deprivation/re-oxygenation(OGD/R) was employed to establish the cell model of cerebral ischemia-reperfusion injury. The cell counting kit-8(CCK-8) and scratch assay were employed to examine the survival and migration of rBEMCs, respectively. Matrigel was used to evaluate the tube formation from rBEMCs. The Transwell assay was employed to examine endothelial cell permeability. Western blot was employed to determine the expression of VEGFR2, VEGFA, p-PI3K, and p-AKT in rBEMCs. The results of network pharmacology analysis showed that AST Ⅳ and PNS regulated 21 targets including VEGFA and AKT1 of angiogenesis in cerebral infarction. Most of these 21 targets were involved in the PI3K/AKT signaling pathway. The in vivo experiments showed that compared with the model group, AST Ⅳ + PNS reduced the neurological deficit score(P<0.05) and the cell damage rate in the brain tissue(P<0.05), promoted the expression of vWF and VEGFA(P<0.01) and angiogenesis, and up-regulated the expression of proteins in the PI3K/AKT pathway(P<0.05, P<0.01). The in vitro experiments showed that compared with the model group, the AST Ⅳ + PNS, 740Y-P, AST Ⅳ + PNS + LY294002, and AST Ⅳ + PNS + 740Y-P improved the survival of rBEMCs after OGD/R, enhanced the migration of rBEMCs, increased the tubes formed by rBEMCs, up-regulated the expression of proteins in the PI3K/AKT pathway, and reduced endothelial cell permeability(P<0.05, P<0.01). Compared with the LY294002 group, the AST Ⅳ + PNS + LY294002 group showed increased survival rate, migration rate, and number of tubes, up-regulated expression of proteins in the PI3K/AKT pathway, and decreased endothelial cell permeability(P<0.05,P<0.01). Compared with the AST Ⅳ + PNS and 740Y-P groups, the AST Ⅳ + PNS + 740Y-P group presented increased survival rate, migration rate, and number of tubes and up-regulated expression of proteins in the PI3K/AKT pathway, and reduced endothelial cell permeability(P<0.01). This study indicates that AST Ⅳ and PNS can promote angiogenesis after cerebral ischemia by activating the PI3K/AKT signaling pathway.

Physicochemical, antibacterial and food preservation properties of active packaging films based on chitosan/ε-polylysine-grafted bacterial cellulose.

To address the environmental and food contamination issues caused by plastics and microorganisms, antimicrobial films using natural polymers has attracted enormous attention. In this work, we proposed a green, convenient and fast approach to prepare antimicrobial films from chitosan (CS), bacterial cellulose (BC) and ε-polylysine (ε-PL). The effects of different concentrations of ε-PL (0 %, 0.25 %, 0.5 %, 0.75 %, 1 %, w/v) on the physicochemical properties and antibacterial activity of composite films (CS-DABC-x%PL) were systematically investigated. Furthermore, a comprehensive comparison with purely physically mixed CS-BC-x%PL films provides a deeper understanding of the subject matter. Characterization tests of the films were conducted using scanning electron microscope (SEM), X-ray diffraction (XRD) and thermogravimetric analysis (TGA). The results suggested that the incorporation of 0.5 % ε-PL reduced the water solubility of the composite film by 19.82 %, along with improved the tensile strength and thermal stability by 37.31 % and 28.54 %. As ε-PL concentration increased to 1 %, the antibacterial performance of the films gradually enhanced. Additionally, the CS-DABC-0.5%PL film demonstrated effectiveness in delaying the deterioration of tilapia. These findings imply that this novel green packaging material holds significant potential in food preservation due to its promising antibacterial properties.

Plant Proteins: Methods of Quality Assessment and the Human Health Benefits of Pulses.

As countries increase their standard of living and individual income levels rise, there is a concomitant increase in the demand for animal-based protein. However, there are alternative sources. One of the alternatives available is that of increased direct human consumption of plant proteins. The quality of a dietary protein is an important consideration when discussing the merits of one protein source over another. The three most commonly used methods to express protein quality are the protein efficiency ratio (PER), a weight gain measurement; protein digestibility-corrected amino acid score (PDCAAS); and the digestible indispensable amino acid score (DIAAS). The possibility that alterations in the quality and quantity of protein in the diet could generate specific health outcomes is one being actively researched. Plant-based proteins may have additional beneficial properties for human health when compared to animal protein sources, including reductions in risk factors for cardiovascular disease and contributions to increased satiety. In this paper, the methods for the determination of protein quality and the potential beneficial qualities of plant proteins to human health will be described.

[Heterogeneity of fork length-weight relationship for juvenile Engraulis japonius based on linear mixed-effects models]. / 基于线性混合效应模型的日本鳀幼鱼叉长-ä½“é‡å ³ç³»å¼‚è´¨æ€§.

In order to examine the growth heterogeneity of juvenile Engraulis japonicus, we conducted heterogeneity analysis of fork length-weight relationships of the species based on the samples of juvenile E. japonius from April to June 2019 collected from the coastal waters of Zhejiang Province by using the generalized linear model and 9 linear mixed-effect models. The results showed that the fork length of juvenile E. japonius was between 14-74 mm, with an average of 33 mm. The dominant fork length group was 21-50 mm. The weight range was 0.01-2.96 g, with an average of 0.28 g, and the dominant body weight group was 0.01-0.50 g. According to Akaike information criterion (AIC), the growth-fitting model with both months and regions random effects on the growth parameters a and b of juvenile E. japonius fitted best.The reliable prediction results was testified by the cross-validation. In the best model, the fixed value of growth parameter a was 0.24×10-5 and its estimated value did not fluctuate significantly, while the fixed value of b was 3.246 and the estimated value ranged from 3.206 to 3.272, indicating that juvenile E. japonius was under positive allometric growth. Our results suggested that month and region significantly affected the fork length-weight relationships of juvenile E. japonius.

LED-Induced Carotenoid Synthesis and Related Gene Expression in Brassica Microgreens.

In this study, various ratios of combined red, blue, and amber light-emitting diodes (rbaLEDs) were investigated for their effect on the expression of carotenoid biosynthetic genes and carotenoid accumulation in eight Brassica microgreens. Total and individual (ß-carotene, lutein, α-carotene, neoxanthin, and violaxanthin) carotenoids were increased 20-44 and 10-55%, respectively, under dose-dependent increasing amber-blue light and decreasing red in most microgreens. Lipophilic 2,2-diphenyl-1-picrylhydrazyl and ferric reducing antioxidant power antioxidant activities were significantly increased under higher amber and blue light fractions, while oxygen radical absorbance capacity was generally decreased. Under rbaLED in mizuna (B. rapa) microgreens, the lycopene epsilon cyclase (LYCε) expression was 10-15-fold higher, which resulted in downstream accumulation of α-carotene and lutein. Lycopene beta cyclase (LYCß) was not significantly changed, suggesting that ß-carotene, violaxanthin and neoxanthin were mainly controlled by upstream phytoene synthase and branch-point LYCε. Increased beta-ring carotenoid hydroxylase (CHXß) expression was also consistent with lutein accumulation. This study demonstrated for the first time that amber LED was involved in the regulatory mechanism of carotenoid biosynthesis, thus a potential novel approach to production of antioxidant-rich microgreens.

Proximate Analysis and Nutritional Evaluation of Twenty Canadian Lentils by Principal Component and Cluster Analyses.

Proximate composition and starch nutritional properties of twenty cooked lentils were assessed to identify unique varieties that could be used in value added foods. Significant variations exist among the lentil varieties (p < 0.05) with respect to their energy, fat, protein, carbohydrate, and dietary fiber content, and these are related to lentil type and seed size. Dazil and Greenstar were unique for their high resistant starch content (RS) and lower area under the starch hydrolysis curve (SHAUC) while Proclaim was opposite. SHAUC was positively correlated (p < 0.001) with rapidly digestible starch (RDS) content (r = 0.626) but negatively correlated with RS content (r = -0.635). Principal component analysis showed that the first three principal components accounted for 62.8% of the total variance and the contribution of SHAUC was 33.2%. These results confirm that in vitro SHAUC and a combination of RDS and RS may be predictive of the digestibility profile of cooked lentils.

A novel invertebrate toll-like receptor is involved in TLR mediated signal pathway of thick shell mussel Mytilus coruscus.

Toll-like receptors (TLRs) are the most well studied pattern recognition receptors (PRRs) that play a crucial role in both innate and adaptive immunity in animals. In the present study, a novel toll-like receptor (McTLRj) was identified and characterised in thick shell mussel Mytilus coruscus. McTLRj possessed a signal peptide, a transmembrane domain, leucine-rich repeats and an intracellular Toll/interleukin-1 receptor domain that were conserved in typical TLRs. McTLRj transcripts were constitutively expressed in all of the examined tissues with high expression level in immune-related tissues, and significantly induced in haemocytes upon live Vibrio alginolyticus, lipopolysaccharide, polyinosinic-polycytidylic acid and peptidoglycans challenge. The overexpression of the McTLRj TLR fragment in Drosophila S2 cells could induce the expression of Drosophila attacin A, drosomycin, cecropin A, and metchnikowin expression. The expression of McTLRj was obviously repressed by dsRNA-mediated RNA interference, and downstream TLR pathway factors, such as MyD88a, IRAK4, and TRAF6 were significantly repressed in McTLRj-silenced mussels upon LPS challenge. These results collectively indicated that McTLRj is a TLR family member that may play a potential PRR role in TLR-mediated signalling pathway. This research contributed to the clarification of innate immune response in molluscs.

Computational Study on the Excited-State Decay of 5-Methylcytosine and 5-Hydroxymethylcytosine: The Common Form of DNA Methylation and Its Oxidation Product.

5-Methylcytosine (5mC) is the predominant epigenetic modification of DNA. 5mC and its sequential oxidation product, 5-hydroxymethylcytosine (5hmC), are crucial epigenetic markers which have a profound impact on gene stability, expression, and regulation. In the present work, ab initio electronic structure computations were performed to investigate the excited-state decay pathways for 5mC and 5hmC in both the neutral and protonated forms. Based on the theoretical quantities, four nonradiative decay pathways via conical intersections (CIs) were identified: ring distortion, ring opening, N-H dissociation, and intersystem crossing (ISC) pathways. Additional calculated potential energy surfaces revealed that ring distortion and ISC pathways were the most effective routes for 5mC and 5hmC, respectively. The influence of environmental factors, such as the solution and an acidic environment, was also explored in this study. Our study demonstrated that excited-state decay pathways via CIs are indispensable for the photostability of DNA epigenetic modifications and may be involved in ingenome stability and mammalian development.

Debranching of pea starch using pullulanase and ultrasonication synergistically to enhance slowly digestible and resistant starch.

Pullulanase (P) and ultrasonication (U) were simultaneously applied to debranch pea starch to enhance slowly digestible starch (SDS) and resistant starch (RS) fractions in the debranched pea starch (DPS). A synergistic debranching effect was found under conditions of pullulanase (40 npun/g) and ultrasonication (100% amplitude in pulse mode, 1 min on followed by 9 min off), which produced 73.5% linear glucans, 18% SDS and 26% RS in the resulting DPS-PU after 6 h of debranching. Even when autoclaving the DPS-PU at 118 °C for 30 min, following cooldown, 11% SDS and 25% RS were retained in the DPS-PU, compared with 0% SDS and 12% RS in autoclaved native pea starch. The SDS fraction in autoclaved DPS-PU further increased to 16% while the RS content remained constant during 14 days of cold storage. In summary, DPS-PU is high in linear glucans, low in starch digestibility and has a thermally stable RS fraction.

Theoretical Studies on the Photophysics and Photochemistry of 5-Formylcytosine and 5-Carboxylcytosine: The Oxidative Products of Epigenetic Modification of Cytosine in DNA.

Cytosine methylation and demethylation play crucial roles in understanding the genomic DNA expression regulation. The epigenetic modification of cytosine and its continuous oxidative products are called the "new four bases of DNA" including 5-methylcytosine (5mC), 5-hydroxymethylcytosine (5hmC), 5-formylcytosine (5fC), and 5-carboxylcytosine (5caC). However, compared to the abundant studies on the classical DNA bases, the photophysical and photochemical properties of those new bases have not yet aroused people's excessive attention. In this contribution, a systematic study on the nonradiative decay and photochemical pathways via excited states or conical intersections upon photoexcitation has been explored through high-level computational approaches such as the complete active space self-consistent field method, complete active space with second-order perturbation theory, and density functional theory. Pathways like the ring-distortion deactivation, hydrogen dissociation, hydrogen transfer, and also Norrish type I and II photochemical reactions have been investigated, and it was proposed that intersystem crossing from the S1 state to the T1 state is the most effective route for 5fC. For 5caC, ring-pucking and intramolecular isomerism are effective deactivation ways at both neutral and protonated forms. In the meantime, the influences of two important environmental factors, the solution and acidic environment (i.e., the protonated state), were also considered in this study. From the theoretical perspective, the initial properties of the photostability and photochemical reactivity for 5fC and 5caC have become a crucial aspect to facilitate a further comprehension of their potential role in gene regulation and transcription.

Effect of roasted pea flour/starch and encapsulated pea starch incorporation on the in vitro starch digestibility of pea breads.

Oven or microwave roasting and alginate encapsulation of pea flour and starch to produce novel pea ingredients for enrichment of slowly digestible starch (SDS) and resistant starch (RS) content in pea bread were investigated. Pea flour treated either by oven roasting (160°C, 30min) or by microwave roasting (1.1kW, 6min) effectively retained its low starch digestibility similar to its native form (∼25% SDS; ∼60% RS). When oven roasting was applied to pea starch, SDS content increased triply compared to the fully boiled counterpart. Alginate encapsulation effectively controlled carbohydrate release to simulated gastric, intestinal and colonic fluids, and thus largely enriched the SDS and RS fractions in starch. Pea bread containing up to 37.5% of encapsulated roasted MPS pea starch not only provided high SDS and RS fractions (23.9% SDS and 30.2% RS) compared to a white bread control (0.2% SDS and 2.5% RS), but also provided an acceptable palatability.

Physicochemical properties and in vitro starch digestibility of potato starch/protein blends.

This study aimed to investigate effects of starch-protein interactions on physicochemical properties and in vitro starch digestibility of composite potato starch/protein blends (0, 5, 10, or 15% protein) during processing (cooking, cooling and reheating). The effect on recrystallization and short-range ordering in starch was studied by light microscopy, differential scanning calorimetry and Fourier transform infrared spectroscopy. The results show that protein in the blend proportionally restricted starch granule swelling during cooking and facilitated amylopectin recrystallization during cold-storage. The facilitating effect of protein diminished with increasing blend ratio. Resistant starch content in the processed blends was positively correlated to intensity ratio of 1053/1035cm(-1) in FTIR spectra arising from slow retrogradation of amylopectin (r(2)>0.88, P≤0.05), whose formation was favored by the presence of protein in the blends and further enhanced by cooling of cooked blends. As a conclusion, starch-protein interaction reduced starch digestibility of the processed blends.

Rheological and structural properties of starches from γ-irradiated and stored potatoes.

Starch was extracted from irradiated and stored potato tubers and the properties were compared to CIPC (chlorpropham) treated tubers. The granule properties and dynamic viscoelasticity in temperature ramp and frequency sweep modes were studied while heating the samples. Starch structural characteristics were investigated by high performance anion exchange chromatography (HPAEC) and Fourier transform infrared spectroscopy (FTIR). Gamma-irradiation of potato tubers at a dosage of 0.1kGy induced some degradation of starch molecules, resulting in earlier swelling of starch granules, and greater extents of amylose and total carbohydrate leaching. The early swelling phenomenon was also enhanced with tuber storage time. The retrogradation rate and extent for a concentrated starch gel also increased with tuber storage time whereas γ-irradiation delayed the gel retrogradation. Sprout inhibiting methods could be selected based on the specific processing and texture requirements of the end products.

In vitro antitumor activity and structure characterization of ethanol extracts from wild and cultivated Chaga medicinal mushroom, Inonotus obliquus (Pers.:Fr.) Pilát (Aphyllophoromycetideae).

Inonotus obliquus (Pers.:Fr.) Pilát has been traditionally used as a folk remedy for treatment of cancers, cardiovascular disease and diabetes in Russia, Poland, and most of the Baltic countries, but natural reserves of this fungus have nearly been exhausted. This study was designed to investigate the artificial cultivation of I. obliquus and the antitumor activity of its tissues. The ethanol extract of cultivated sclerotium had the highest cell growth inhibitory rate (74.6%) as determined by an 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. 78% of the bags produced sclerotia and only 6.17 g/bag of sclerotium was obtained. Extracts of the cultivated fruiting body showed 44.2% inhibitory activity against tumor cells. However, the yield was as high as 18.24 g/bag, and 98% of the bags produced fruiting body. The results of gas chromatography-mass spectroscopy (GC-MS) showed that similar compounds were extracted from the wild and cultivated samples. The principal compounds observed were lanosterol, inotodiol, and ergosterol. Their percentages of the mass fraction were 86.1, 59.9, and 71.8% of the total, for the wild sclerotium, cultivated sclerotium, and cultivated fruiting body, respectively. Ergosterol was found to be much higher (27.32%) in cultivated fruiting body. We conclude that cultivated fruiting body of I. obliquus obtained by inoculation of the substrate with spawn mycelium of the fifth generation could serve as an ideal substitute for the wild I. obliquus.

Inhibition of microbial growth and enrichment of gamma-aminobutyric acid during germination of brown rice by electrolyzed oxidizing water.

Electrolyzed oxidizing water (EOW) has been regarded as a potential environmentally friendly broad spectrum microbial decontaminant. EOW with a pH of 3.0 and oxidation reduction potential of 1,079.0 mV were generated by the electrolysis of a dilute NaCl solution (20 mM) in an electrochemical cell. The effects of EOW, 1% NaClO solution, and alkaline electrolyzed water on controlling microbial growth, germination ratio, and enrichment of gamma-aminobutyric acid in germinated brown rice (GBR) were evaluated in this study. Results show that EOW was the most effective at inhibiting microbial growth during germination. Rinsing the rice grains with EOW at 12-h intervals resulted in aerobic plate count reductions of 4.82 log CFU/g, while soaking resulted in bacterial count reductions of 5.38 log CFU/g after 72 h of germination. Moreover, EOW significantly enriched gamma-aminobutyric acid content in GBR (P < 0.05); content was increased 1.6 times in grain rinsed with EOW and 1.8 times in grain soaked in EOW. The findings indicate that EOW is a feasible disinfectant for industrial GBR production.

Phenolic content, composition, antioxidant activity, and their changes during domestic cooking of potatoes.

Potatoes in the diet contribute significantly to antioxidant daily intake worldwide. The influence of different domestic cooking conditions, boiling, microwaving, and baking, on total phenolics (TP), antioxidant capacity, phenolic composition, and tryptophan content was studied using eight commercial potato varieties. The antioxidant capacity was detected by the methods of oxygen radical absorbance capacity assay (ORAC) and the 2,2-diphenyl-1-picrylhydrazyl free radical (DPPH(*)) assay. The phenolic composition and tryptophan content were determined using high-performance liquid chromatography coupled with a diode array detector (HPLC-DAD), whereas phenolics and tryptophan were identified by means of HPLC-mass spectrometry, HPLC-DAD, and authentic standards. Antioxidant capacity was influenced by potato variety and cooking conditions; however, cooked potatoes retained 68-97% ORAC value depending on cooking procedure and variety. Chlorogenic acid and its isomers dominated the phenolic composition of each variety involved in this study. ORAC and TP were highly and positively correlated (r = 0.9119). Norkotah ranked highest in chlorogenic acid content and antioxidant value. Principal component analysis showed different cooking processes did not influence the trend of the antioxidant profile of the eight potato varieties, but specific compounds exert influence on the antioxidant capacity. The results imply that the potato varieties rich in antioxidant components could be good antioxidant sources as activities are not greatly affected by different cooking conditions.