Rhamnogalacturonan lyase 1 (RGL1), as a suppressor of E3 ubiquitin ligase Arabidopsis thaliana ring zinc finger 1 (AtRZF1), is involved in dehydration response to mediate proline synthesis and pectin rhamnogalacturonan-I composition.

Proline metabolism plays a crucial role in both environmental stress responses and plant growth. However, the specific mechanism by which proline contributes to abiotic stress processes remains to be elucidated. In this study, we utilized atrzf1 (Arabidopsis thaliana ring zinc finger 1) as a parental line for T-DNA tagging mutagenesis and identified a suppressor mutant of atrzf1, designated proline content alterative 31 (pca31). The pca31 mutant suppressed the insensitivity of atrzf1 to dehydration stress during early seedling growth. Using Thermal Asymmetric Interlaced-PCR, we found that the T-DNA of pca31 was inserted into the promoter region of the At2g22620 gene, which encodes the cell wall enzyme rhamnogalacturonan lyase 1 (RGL1). Enzymatic assays indicated that RGL1 exhibited rhamnogalacturonan lyase activity, influencing cell wall pectin composition. The decrease in RGL1 gene expression suppressed the transcriptomic perturbation of the atrzf1 mutant. Silencing of the RGL1 gene in atrzf1 resulted in a sensitive phenotype similar to pca31 under osmotic stress conditions. Treatment with mannitol, salt, hydrogen peroxide, and abscisic acid induced RGL1 expression. Furthermore, we uncovered that RGL1 plays a role in modulating root growth and vascular tissue development. Molecular, physiological, and genetic experiments revealed that the positive modulation of RGL1 during abiotic stress was linked to the AtRZF1 pathway. Taken together, these findings establish that pca31 acts as a suppressor of atrzf1 in abiotic stress responses through proline and cell wall metabolisms.

Enhancement of debitterness, water-solubility, and neuroprotective effects of naringin by transglucosylation.

Naringin found in citrus fruits is a flavanone glycoside with numerous biological activities. However, the bitterness, low water-solubility, and low bioavailability of naringin are the main issues limiting its use in the pharmaceutical and nutraceutical industries. Herein, a glucansucrase from isolated Leuconostoc citreum NY87 was used for trans-α-glucosylattion of naringin by using sucrose as substrate. Two naringin glucosides (O-α-D-glucosyl-(1'''' → 6″) naringin (compound 1) and 4'-O-α-D-glucosyl naringin (compound 2)) were purified and determined their structures by nuclear magnetic resonance. The optimization condition for the synthesis of compound 1 was obtained at 10 mM naringin, 200 mM sucrose, and 337.5 mU/mL at 28 °C for 24 h by response surface methodology method. Compound 1 and compound 2 showed 1896- and 3272 times higher water solubility than naringin. Furthermore, the bitterness via the human bitter taste receptor TAS2R39 displayed that compound 1 was reduced 2.9 times bitterness compared with naringin, while compound 2 did not express bitterness at 1 mM. Both compounds expressed higher neuroprotective effects than naringin on human neuroblastoma SH-SY5Y cells treated with 5 mM scopolamine based on cell viability and cortisol content. Compound 1 reduced acetylcholinesterase activity more than naringin and compound 2. These results indicate that naringin glucosides could be utilized as functional material in the nutraceutical and pharmaceutical industries. KEY POINTS: • A novel O-α-D-glucosyl-(1 → 6) naringin was synthesized using glucansucrase from L. citreum NY87. • Naringin glucosides improved water-solubility and neuroprotective effects on SH-SY5Y cells. • Naringin glucosides showed a decrease in bitterness on bitter taste receptor 39.

Vision-Based Pedestrian's Crossing Risky Behavior Extraction and Analysis for Intelligent Mobility Safety System.

Crosswalks present a major threat to pedestrians, but we lack dense behavioral data to investigate the risks they face. One of the breakthroughs is to analyze potential risky behaviors of the road users (e.g., near-miss collision), which can provide clues to take actions such as deployment of additional safety infrastructures. In order to capture these subtle potential risky situations and behaviors, the use of vision sensors makes it easier to study and analyze potential traffic risks. In this study, we introduce a new approach to obtain the potential risky behaviors of vehicles and pedestrians from CCTV cameras deployed on the roads. This study has three novel contributions: (1) recasting CCTV cameras for surveillance to contribute to the study of the crossing environment; (2) creating one sequential process from partitioning video to extracting their behavioral features; and (3) analyzing the extracted behavioral features and clarifying the interactive moving patterns by the crossing environment. These kinds of data are the foundation for understanding road users' risky behaviors, and further support decision makers for their efficient decisions in improving and making a safer road environment. We validate the feasibility of this model by applying it to video footage collected from crosswalks in various conditions in Osan City, Republic of Korea.

A review of encapsulation of carotenoids using spray drying and freeze drying.

Carotenoids are potent antioxidants, but they are highly unstable and susceptible during processing and storage. Encapsulation technologies protect against degradation and are capable of releasing individual or combination of bioactive substances during processing as well as development of various functional food products. Moreover, encapsulating agents can be used to increase the stability of carotenoids and form a barrier between the core and wall materials. Suitable encapsulating agents, temperature, and drying methods are the most important factors for the encapsulation process. In this report, we reviewed the current status of encapsulation of carotenoids from different fruits, vegetables, spices, seaweeds, microorganisms, and synthetic sources using various types of encapsulating agents through spray drying and freeze drying. We also focused on the degradation kinetics and various factors that affect the stability and bioavailability of encapsulated carotenoids during their processing and storage.

Improvement of branched-chain amino acid production by isolated high-producing protease from Bacillus amyloliquefaciens NY130 on isolated soy/whey proteins and their muscle cell protection.

Branched-chain amino acids (BCAAs) are vital components of human and animal nutrition that contribute to the building blocks of proteins. In this study, 170 protease-producing strains were isolated and screened from soy-fermented foods. Bacillus amyloliquefaciens NY130 was obtained from Cheonggukjang with high production of BCAAs. Optimal production of protease from B. amyloliquefaciens NY130 (protease NY130) was achieved at 42 °C and pH 6.0 for 21 h. It was purified and determined as 27- and 40 kDa. Protease NY130 showed maximum activity at pH 9.0 and 45 °C with Km value of 10.95 mg for ISP and 1.69 mg for WPI. Protease-treated ISP and WPI showed increased sweetness and saltiness via electronic tongue analysis and enhanced the protective effect against oxidative stress in C2C12 myocytes by increasing p-mTOR/mTOR protein expression to 160%. This work possesses potential in producing BCAAs by using protease for utilization in food.

Modulating flavanone compound for reducing the bitterness and improving dietary fiber, physicochemical properties, and anti-adipogenesis of green yuzu powder by enzymatic hydrolysis.

Yuzu (Citrus junos Sieb.) is a peel-edible fruit with a pleasant aroma, but its bitter taste can impact consumer appeal. In this study, an efficient enzymatic method reduced bitterness in green yuzu powder (GYP). Cellulase KN and naringinase from Aspergillus oryzae NYO-2 significantly decreased naringin and neohesperidin content by over 87 %, while increasing total dietary fiber and soluble dietary fiber by up to 10 % and 51 %, respectively. Insoluble dietary fiber decreased by up to 22 %. Cellulose, hemicellulose, lignin, and pectin contents in enzyme-treated YP decreased by 1.15-2.00-fold, respectively. Enzyme-treated GYP exhibited improved physicochemical properties, including enhanced solubility, oil-holding capacity, and water swelling capacities. 3T3-L1 cells treated with cellulase-treated GYP and naringinase-treated GYP showed lower lipid accumulation and higher lipolysis capability than GYP, along with decreased fatty acid synthase contents. These findings suggest that enzyme-treated GYP holds potential as a functional ingredient in the food industry.

Putrescine regulating by stress-responsive MAPK cascade contributes to bacterial pathogen defense in Arabidopsis.

Polyamines in plants are involved in various physiological and developmental processes including abiotic and biotic stress responses. We investigated the expression of ADCs, which are key enzymes in putrescine (Put) biosynthesis, and roles of Put involving defense response in Arabidopsis. The increased expression of ADC1 and ADC2, and the induction of Put were detected in GVG-NtMEK2(DD) transgenic Arabidopsis, whereas, their performance was partially compromised in GVG-NtMEK2(DD)/mpk3 and GVG-NtMEK2(DD)/mpk6 mutant following DEX treatment. The expression of ADC2 was highly induced by Pst DC3000 inoculation, while the transcript levels of ADC1 were slightly up-regulated. Compared to the WT plant, Put content in the adc2 knock-out mutant was reduced after Pst DC3000 inoculation, and showed enhanced susceptibility to pathogen infection. The adc2 mutant exhibited reduced expression of PR-1 after bacterial infection and the growth of the pathogen was about 4-fold more than that in the WT plant. Furthermore, the disease susceptibility of the adc2 mutant was recovered by the addition of exogenous Put. Taken together, these results suggest that Arabidopsis MPK3 and MPK6 play a positive role in the regulation of Put biosynthesis, and that Put contributes to bacterial pathogen defense in Arabidopsis.

Production of Prunin and Naringenin by Using Naringinase from Aspergillus oryzae NYO-2 and Their Neuroprotective Properties and Debitterization.

Naringin is a flavanone glycoside in citrus fruits that has various biological functions. However, its bitterness affects the quality, economic value, and consumer acceptability of citrus products. Deglycosylation of naringin using naringinase decreases its bitterness and enhances its functional properties. In this study, eight microbial strains with naringinase activity were isolated from 33 yuzu-based fermented foods. Among them, naringinase from Aspergillus oryzae NYO-2, having the highest activity, was used to produce prunin and naringenin. Under optimal conditions, 19 mM naringin was converted to 14.06 mM prunin and 1.97 mM naringenin. The bitterness of prunin and naringenin was significantly decreased compared to naringin using the human bitter taste receptor TAS2R39. The neuroprotective effects of prunin and naringenin on human neuroblastoma SH-SY5Y cells treated with scopolamine were greater than that of naringin. These findings can widen the potential applications of deglycosylation of naringin to improve sensory and functional properties.

A combination of commercial and traditional food-source-derived enzymatic treatment acts as a potential tool to produce functional yuzu (Citrus junos) powder.

Enzymatic modifications have been applied in citrus to enhance their physicochemical and biological properties and reduce their bitterness. Notwithstanding, research on the combination of enzyme treatment of yuzu is lacking. In this study, yuzu was treated with a combination of isolated cellulase NY203, pectinase UF, and cellulase KN, and this enzymatic treatment was found to increase monosaccharide, naringenin, and hesperetin levels. In contrast, dietary fiber, cellulose, hemicellulose, lignin, and pectin levels were decreased. Moreover, the enzymes disintegrated the inner and outer surface structures and chemical bonding of yuzu, thus improving its solubility rate, water-holding capacity, oil-adsorption capacity, cholesterol-binding capacity, and water-swelling capacity. Furthermore, NY203 + UF + KN combination treatment reduced the bitterness of treated yuzu by 50 % compared with the control. Additionally, NY203 + UF + KN treatment yielded a 28 % decrease in lipid accumulation and two-fold higher lipolytic activity in 3T3L-1 adipocytes. These findings are potentially beneficial to the food/nutraceutical industries regarding functional yuzu powder production.

Fermented Curcuma longa L. Prevents Alcoholic Fatty Liver Disease in Mice by Regulating CYP2E1, SREBP-1c, and PPAR-α.

We examined the efficacy of fermented Curcuma longa L. (FT) on the development of alcoholic fatty liver in mice and investigated the underlying mechanism. The protective potential of FT against ethanol-induced fatty liver was determined using C57BL/6 male mice allocated into four groups (8 mice/group). Control groups received either distilled water or 5 g/kg body weight (b.w.) per day ethanol for 8 days. Treatment groups were administered either 300 mg/kg b.w. per day of milk thistle or FT before receiving ethanol. FT contained a higher amount of caffeic acid and tetrahydrocurcumin than C. longa. FT pretreatment significantly suppressed the elevated hepatic lipid droplets associated with ethanol ingestion. In comparison with ethanol-treated control, FT pretreated mice showed inhibited cytochrome P4502E1 (CYP2E1), sterol regulatory element-binding protein-1 (SREBP-1c), and acetyl-CoA carboxylase production but elevated AMP-activated protein kinase, peroxisome proliferator-activated receptor-alpha (PPAR-α), and carnitine palmitoyltransferase 1 (CPT-1) levels. Taken together, FT is a promising hepatoprotectant for preventing of alcoholic fatty liver through modulating fatty acid synthesis and oxidation.

Synthesis of ellagic acid glucoside using glucansucrase from Leuconostoc and characterization of this glucoside as a functional neuroprotective agent.

Ellagic acid glucoside was synthesized via transglucosylation using sucrose and glucansucrase derived from Leuconostoc mesenteroides B-512 FMCM. After such enzymatic synthesis, the product was purified by 50% ethyl acetate fraction and C18 column chromatography. Modification of ellagic acid glucoside was verified by LC-MS/MS at m/z 485.1 (M + Na)- and m/z 531.1 (M + 3Na)-. The yield of ellagic acid glucoside was 69% (3.47 mM) by response surface methodology using 150 mM sucrose, 300 mU/mL glucansucrase, and 5 mM ellagic acid. The synthesized ellagic acid glucoside showed improved water solubility, up to 58% higher brain nerve cell (SH-SY5Y) protective effect, threefold higher cortisol reducing effect, and fourfold stronger inhibitory effect on acetylcholinesterase (AChE) than ellagic acid. These results indicate that ellagic acid glucoside could be used as a neuroprotective agent.

Antioxidative and Analgesic Effects of Naringin through Selective Inhibition of Transient Receptor Potential Vanilloid Member 1.

Transient receptor potential vanilloid member 1 (TRPV1) is activated in response to capsaicin, protons, temperature, and free reactive oxygen species (ROS) released from inflammatory molecules after exposure to harmful stimuli. The expression level of TRPV1 is elevated in the dorsal root ganglion, and its activation through capsaicin and ROS mediates neuropathic pain in mice. Its expression is high in peripheral and central nervous systems. Although pain is a response evolved for survival, many studies have been conducted to develop analgesics, but no clear results have been reported. Here, we found that naringin selectively inhibited capsaicin-stimulated inward currents in Xenopus oocytes using a two-electrode voltage clamp. The results of this study showed that naringin has an IC50 value of 33.3 µM on TRPV1. The amino acid residues D471 and N628 of TRPV1 were involved in its binding to naringin. Our study bridged the gap between the pain suppression effect of TRPV1 and the preventive effect of naringin on neuropathic pain and oxidation. Naringin had the same characteristics as a model selective antagonist, which is claimed to be ideal for the development of analgesics targeting TRPV1. Thus, this study suggests the applicability of naringin as a novel analgesic candidate through antioxidative and analgesic effects of naringin.

Physiochemical properties, dietary fibers, and functional characterization of three yuzu cultivars at five harvesting times.

This research focused on physiochemical and nutritional properties and functional characterization of three cultivars of yuzu-Native, Tadanishiki yuzu, and Namhae1-during different seasons. According to the cultivar and harvest time, yuzu cultivars were analyzed for free sugar, dietary fiber, hesperidin, naringin, and flavonoid content as well as antioxidant and antihypertensive activity. During November, Namhae1 exhibited the highest fruit weight, °Brix/acidity ratio, and total dietary fiber content. Tadanishiki contained the highest fructose and sucrose levels, pectin and cellulose contents, and soluble dietary fiber. Tadanishiki also had the highest hesperidin content in October, while the naringin content and antioxidant activity were the greatest in November. Antihypertensive activity was also the strongest for Tadanishiki, which was picked in October and November. These results indicated that Tadanishiki in October or November was the best for consumption or favorable processing because of its excellent product quality and high levels of nutritional and functional compounds.

Aqueous green tea infusion extracted by ultra-sonication method, but not by conventional method, facilitates GLUT4 membrane translocation in adipocytes which potently ameliorates high-fat diet-induced obesity.

Green tea contains bioactive compounds, such as polyphenols, responsible for its health-promoting effects, including antiobesity and antidiabetic effects. We previously reported that ultra-sonication extraction (UE) could efficiently increase the extraction yield of green tea compounds. In the present study, we found that the extract obtained using UE contained higher phenolic and flavonoid contents than that obtained using the conventional method. We therefore considered the extract as a bioactive metabolite-rich functional green tea extract (BMF-GTE), and tested its glucose-lowering effect by generating an adipocyte cell line stably expressing 7myc-GLUT4-GFP. We found that BMF-GTE treatment increased GLUT4 translocation to the plasma membrane. Moreover, BMF-GTE administration attenuated weight gain in mice fed a high-fat diet (HFD). Importantly, HFD-induced glucose tolerance was ameliorated in the mice receiving BMF-GTE. Therefore, we conclude that BMF-GTE worked against obesity and diabetes, at least partially, by enhancing GLUT4 translocation in adipocytes. PRACTICAL APPLICATIONS: As green tea is one of the most consumed beverages worldwide, its health effects have been widely tested. In our previous studies, we found that ultra-sonication extraction (UE) has the potential to increase the aqueous extraction yield of green tea compounds compared to conventional extraction techniques. In this study, we examined the biological effect of bioactive metabolite-rich functional green tea extract (BMF-GTE) obtained using UE; we observed that administering BMF-GTE lowered the body weight and increased insulin sensitivity in mice fed a high-fat diet, potentially by facilitating the membrane translocation of GLUT4 in adipocytes. Therefore, this study suggests that the extract obtained with UE had antiobesity and antidiabetic properties, indicative of a potential application of UE in maximizing the beneficial effects of green tea on human health.

Significant Performance and Stability Improvements of Low-Temperature IGZO TFTs by the Formation of In-F Nanoparticles on an SiO2 Buffer Layer.

We report the performance improvement of low-temperature coplanar indium-gallium-zinc-oxide (IGZO) thin-film transistors (TFTs) with a maximum process temperature of 230 °C. We treated F plasma on the surface of an SiO2 buffer layer before depositing the IGZO semiconductor by reactive sputtering. The field-effect mobility increases from 3.8 to 9.0 cm2 V-1·s-1, and the threshold voltage shift (ΔVth) under positive-bias temperature stress decreases from 3.2 to 0.2 V by F-plasma exposure. High-resolution transmission electron microscopy and atom probe tomography analysis reveal that indium fluoride (In-F) nanoparticles are formed at the IGZO/buffer layer interface. This increases the density of the IGZO and improves the TFT performance as well as its bias stability. The results can be applied to the manufacturing of low-temperature coplanar oxide TFTs for oxide electronics, including information displays.

Mitogen-activated protein kinase cascade in the signaling for polyamine biosynthesis in tobacco.

Expression of NtNEK2(DD), a constitutively active mutant of NtMEK2, activates endogenous salicylic acid-induced protein kinase (SIPK) and wounding-induced protein kinase (WIPK), and leads to several stress/defense responses in tobacco. In this study, we used ACP (annealing control primer)-based differential display reverse transcription-PCR to isolate the downstream effectors mediated by the NtMEK2-SIPK/WIPK cascade. The arginine decarboxylase gene (ADC), which is involved in plant putrescine biosynthesis, was one of nine differentially expressed genes. When compared with NtMEK2(KR) plants, NtMEK2(DD) transgenic plants exhibited a significant increase in ADC and ODC (ornithine decarboxylase) transcript levels, as well as in putrescine and its catabolite, gamma-aminobutyric acid, following SIPK/WIPK activation. Taken together, these results suggest that the NtMEK2-SIPK/WIPK cascade is involved in regulating polyamine synthesis, especially putrescine synthesis, through transcriptional regulation of the biosynthetic genes in tobacco.

Synthesis and characterization of hydroquinone fructoside using Leuconostoc mesenteroides levansucrase.

Hydroquinone (HQ) functions as a skin-whitening agent, but it has the potential to cause dermatitis. We synthesized a HQ fructoside (HQ-Fru) as a potential skin-whitening agent by reacting levansucrase from Leuconostoc mesenteroides with HQ as an acceptor and sucrose as a fructofuranose donor. The product was purified using 1-butanol partition and silica-gel column chromatography. The structure of the purified HQ-Fru was determined by (1)H and (13)C nuclear magnetic resonance, and the molecular ion of the product was observed at m/z 295 (C12 H16 O7 Na)(+). The HQ-Fru was identified as 4-hydroxyphenyl-beta-D: -fructofuranoside. The optimum condition for HQ-Fru synthesis was determined using a response surface method (RSM), and the final optimum condition was 350 mM HQ, 115 mM sucrose, and 0.70 U/ml levansucrase, and the final HQ-Fru produced was 1.09 g/l. HQ-Fru showed anti-oxidation activities and inhibition against tyrosinase. The median inhibition concentration (IC(50)) of 1,1-diphenyl-2-picrylhydrazyl scavenging activity was 5.83 mM, showing higher antioxidant activity compared to beta-arbutin (IC(50) = 6.04 mM). The K ( i ) value of HQ-Fru (1.53 mM) against tyrosinase was smaller than that of beta-arbutin (K ( i ) = 2.8 mM), indicating that it was 1.8-times better as an inhibitor. The inhibition of lipid peroxidation by HQ-Fru was 105.3% that of HQ (100%) and 118.9 times higher than that of beta-arbutin (0.89% of HQ).

Expression pattern, regulation, and biological role of runt domain transcription factor, run, in Caenorhabditis elegans.

The Caenorhabditis elegans run gene encodes a Runt domain factor. Runx1, Runx2, and Runx3 are the three known mammalian homologs of run. Runx1, which plays an essential role in hematopoiesis, has been identified at the breakpoint of chromosome translocations that are responsible for human leukemia. Runx2 plays an essential role in osteogenesis, and inactivation of one allele of Runx2 is responsible for the human disease cleidocranial dysplasia. To understand the role of run in C. elegans, we used transgenic run::GFP reporter constructs and a double-stranded RNA-mediated interference method. The expression of run was detected as early as the bean stage exclusively in the nuclei of seam hypodermal cells and lasted until the L3 stage. At the larval stage, expression of run was additionally detected in intestinal cells. The regulatory elements responsible for the postembryonic hypodermal seam cells and intestinal cells were separately located within a 7.2-kb-long intron region. This is the first report demonstrating that an intron region is essential for stage-specific and cell type-specific expression of a C. elegans gene. RNA interference analysis targeting the run gene resulted in an early larva-lethal phenotype, with apparent malformation of the hypodermis and intestine. These results suggest that run is involved in the development of a functional hypodermis and gut in C. elegans. The highly conserved role of the Runt domain transcription factor in gut development during evolution from nematodes to mammals is discussed.

Transglycosylation of gallic acid by using Leuconostoc glucansucrase and its characterization as a functional cosmetic agent.

Gallic acid glycoside was enzymatically synthesized by using dextransucrase and sucrose from gallic acid. After purification by butanol partitioning and preparative HPLC, gallic acid glucoside was detected at m/z 355 (C13, H16, O10, Na)+ by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. The yield of gallic acid glucoside was found to be 35.7% (114 mM) by response surface methodology using a reaction mixture of 319 mM gallic acid, 355 mM sucrose, and 930 mU/mL dextransucrase. The gallic acid glucoside obtained showed 31% higher anti-lipid peroxidation and stronger inhibition (Ki = 1.23 mM) against tyrosinase than that shown by gallic acid (Ki = 1.98 mM). In UVB-irradiated human fibroblast cells, gallic acid glucoside lowered matrix metalloproteinase-1 levels and increased the collagen content, which was indicative of a stronger anti-aging effect than that of gallic acid or arbutin. These results indicated that gallic acid glucoside is likely a superior cosmetic ingredient with skin-whitening and anti-aging functions.

Synthesis and Functional Characterization of Caffeic Acid Glucoside Using Leuconostoc mesenteroides Dextransucrase.

Caffeic acid was modified via transglucosylation using sucrose and dextransucrase from Leuconostoc mesenteroides B-512FMCM. Following enzymatic modification, a caffeic acid glucoside was isolated by butanol separation, silica gel chromatography, and preparative HPLC. The synthesized caffeic acid glucoside had a molecular mass-to-charge ratio of 365 m/z, and its structure was identified as caffeic acid-3-O-α-d-glucopyranoside. The production of this caffeic acid-3-O-α-d-glucopyranoside at a concentration of 153 mM was optimized using 325 mM caffeic acid, 355 mM sucrose, and 650 mU mL-1 dextransucrase in the synthesis reaction. In comparison with the caffeic acid, the caffeic acid-3-O-α-d-glucopyranoside displayed 3-fold higher water solubility, 1.66-fold higher antilipid peroxidation effect, 15% stronger inhibition of colon cancer cell growth, and 11.5-fold higher browning resistance. These results indicate that this caffeic acid-3-O-α-d-glucopyranoside may be a suitable functional component of food and pharmaceutical products.