Characterization of alfalfa mosaic virus capsid protein using Cryo-EM.

VLPs are virus-like particles that comprise viral capsid proteins that can self-assemble and mimic the shape and size of real viral particles; however, because they do not contain genetic material they cannot infect host cells. VLPs have great potential as safe drug/vehicle candidates; therefore, they are gaining popularity in the field of preventive medicine and therapeutics. Indeed, extensive studies are underway to examine their role as carriers for immunization and as vehicles for delivery of therapeutic agents. Here, we examined the possibility of developing VLP-utilizing technology based on an efficient VLP production process and high-resolution structural analysis. Nicotiana benthamiana was used as an expression platform to produce the coat protein of the alfalfa mosaic virus (AMV-CP). About 250 mg/kg of rAMV-CP was produced from Nicotiana benthamiana leaves. Structural analysis revealed that the oligomeric status of rAMV-CP changed according to the composition and pH of the buffer. Size exclusion chromatography and electron microscopy analysis confirmed the optimal conditions for rAMV-CP VLP formation, and a 2.4 Å resolution structure was confirmed by cryo-EM analysis. Based on the efficient protein production, VLP manufacturing technology, and high-resolution structure presented herein, we suggest that rAMV-CP VLP is a useful platform for development of various new drugs.

Oat (Avena sativa) Extract against Oxidative Stress-Induced Apoptosis in Human Keratinocytes.

Oat (Avena sativa) is well known for its various health benefits. The protective effect of oat extract against oxidative stress-induced apoptosis in human keratinocytes HaCaT was determined. First, extracts of two varieties of oat, Daeyang and Choyang, were analyzed for fat-soluble antioxidants such as α-tocotrienol, γ-oryzanols, lutein and zeaxanthin using an UPLC system and for antioxidant activity using a DPPH assay. Specifically, an 80% ethanol extract of Daeyang oat (Avena sativa cv. Daeyang), which had high amounts of antioxidants and potent radical scavenging activity, was further evaluated for protective effect against oxidative stress-induced cell death, intracellular reactive oxygen species levels, the phosphorylation of DNA damage mediating genes such as H2AX, checkpoint kinase 1 and 2, and p53 and the activation of apoptotic genes such as cleaved caspase-3 and 7 and poly (ADP-ribose) polymerase in HaCaT cells. The Daeyang and Choyang oat 80% ethanol extracts had 26.9 and 24.1 mg/100 g γ-oryzanols, 7.69 and 8.38 mg/100 g α-tocotrienol, 1.25 and 0.34 mg/100 g of lutein and 1.20 and 0.17 mg/100 g of zeaxanthin, respectively. The oat 80% ethanol extract treatment (Avena sativa cv. Daeyang) had a protective effect on oxidative stress-induced cell death in HaCaT cells. In addition, the oat 80% ethanol extracts led to a significant decrease in the intracellular ROS level at a concentration of 50-200 µg/mL, the attenuation of DNA damage mediating genes and the inhibition of apoptotic caspase activities in a dose dependent manner (50-200 µg/mL). Thus, the current study indicates that an oat (Avena sativa cv. Daeyang) extract rich in antioxidants, such as polyphenols, avenanthramides, γ-oryzanols, tocotrienols and carotenoids, has a protective role against oxidative stress-induced keratinocyte injuries and that oat may a useful source for oxidative stress-associated skin damage.

Enhanced Antioxidant Activity of Bioactives in Colored Grains by Nano-Carriers in Human Lens Epithelial Cells.

The use of phytochemicals for preventing chronic diseases associated with oxidative stress such as cataracts is hindered by their low bioavailability. The effects of nano-carriers on the antioxidant activities of extracts of black rice with giant embryo (BRGEx) and soybeans (SBx) have been determined in human lens epithelial B3 cells. Scanning (SEM) and transmission electron microscopy (TEM) demonstrated that rGO (reduced graphene oxide) has a flat surface unlike GO (graphene oxide), which has a distinctive wrinkled structure with defects. UPLC analysis revealed 41.9 µg/100 g of γ-oryzanols in water extract of BRGE, and 111.8 µg /100 g of lutein, 757.7 µg/100 g of γ-tocotrienol, 4071.4 µg/100 g of γ-tocopherol in 40% ethanol extract of soybeans, respectively. Even though a low concentration of BRGEx alone did not show any antioxidant activity in B3 cells, co-treatment of BRGEx with rGO together substantially reduced hydrogen peroxide and methylglyoxal-induced DNA damage, as determined by phosphorylated γH2AX. In addition, SBx with rGO also attenuated DNA damage. Furthermore, intracellular reactive oxygen species were significantly decreased by combining extracts of these colored grains with rGO. These results suggest a potential application of nanocarriers for enhancing the bioavailability of phytochemicals.

Development of Nano Soy Milk through Sensory Attributes and Consumer Acceptability.

Nanotechnology is currently applied in food processing and packaging in the food industry. Nano encapsulation techniques could improve sensory perception and nutrient absorption. The purpose of this study was to identify the sensory characteristics and consumer acceptability of three types of commercial and two types of laboratory-developed soy milk. A total of 20 sensory attributes of the five different soy milk samples, including appearance, smell (odor), taste, flavor, and mouthfeel (texture), were developed. The soy milk samples were evaluated by 100 consumers based on their overall acceptance, appearance, color, smell (odor), taste, flavor, mouthfeel (texture), goso flavor (nuttiness), sweetness, repeated use, and recommendation. One-way analysis of variance (ANOVA), principal component analysis (PCA), and partial least square regression (PLSR) were used to perform the statistical analyses. The SM_D sample generally showed the highest scores for overall liking, flavor, taste, mouthfeel, sweetness, repeated consumption, and recommendation among all the consumer samples tested. Consumers preferred sweet, goso (nuttiness), roasted soybean, and cooked soybean (nuttiness) attributes but not grayness, raw soybean flavor, or mouthfeel. Sweetness was closely related to goso (nuttiness) odor and roasted soybean odor and flavor based on partial least square regression (PLSR) analysis. Determination of the sensory attributes and consumer acceptance of soymilk provides insight into consumer needs and desires along with basic data to facilitate the expansion of the consumer market.

Black soybeans protect human keratinocytes from oxidative stress-induced cell death.

Black soybeans are functional foods containing a variety of bioactives such as isoflavones, carotenoids, tocopherols, phenolic acid as well as anthocyanins. Here, we examined whether Cheongja#3 black soybean extract has a protective effect on oxidative stress-induced cell death in human keratinocytes HaCaT. First, we identified fat-soluble bioactives in three varieties of soybean extracts (Saedanbaek, Daechan, and Cheongja#3). In particular, black soybean Cheongja#3 had high amounts of lutein than other varieties. We demonstrated that Cheongja#3 extract reduced intracellular reactive oxygen species levels in HaCaT cells. Furthermore, Cheongja#3 protected cells from hydrogen peroxide (H2O2)-induced oxidative stress and triggered cell death determined by cell viabilities and apoptotic caspase activities. Next, we identified the underlying mechanism is due to increased Nrf2 antioxidant system by Cheongja#3, thus increasing the expression of heme oxygenases (HO)-1. These results indicated that Cheongja#3 soybean extract has protective role against oxidative stress by upregulating the Nrf-2 antioxidant system in human keratinocyte HaCaT cells.

Dietary Anthocyanins against Obesity and Inflammation.

Chronic low-grade inflammation plays a pivotal role in the pathogenesis of obesity, due to its associated chronic diseases such as type II diabetes, cardiovascular diseases, pulmonary diseases and cancer. Thus, targeting inflammation is an attractive strategy to counter the burden of obesity-induced health problems. Recently, food-derived bioactive compounds have been spotlighted as a regulator against various chronic diseases due to their low toxicity, as opposed to drugs that induce severe side effects. Here we describe the beneficial effects of dietary anthocyanins on obesity-induced metabolic disorders and inflammation. Red cabbage microgreen, blueberry, blackcurrant, mulberry, cherry, black elderberry, black soybean, chokeberry and jaboticaba peel contain a variety of anthocyanins including cyanidins, delphinidins, malvidins, pelargonidins, peonidins and petunidins, and have been reported to alter both metabolic markers and inflammatory markers in cells, animals, and humans. This review discusses the interplay between inflammation and obesity, and their subsequent regulation via the use of dietary anthocyanins, suggesting an alternative dietary strategy to ameliorate obesity and obesity associated chronic diseases.