Anti-inflammatory effects of Allium cepa L. peel extracts via inhibition of JAK-STAT pathway in LPS-stimulated RAW264.7 cells.

ETHNOPHARMACOLOGICAL RELEVANCE: Allium cepa L. (A. cepa) is one of the oldest cultivated plants in the world. A. cepa has been used in traditional folk medicine to treat inflammatory disease in several regions, such as Palestine and Serbia. A. cepa peel has a higher content of flavonoids, such as quercetin, than the edible parts. These flavonoids alleviate inflammatory diseases. However, the anti-inflammatory effects of A. cepa peel extract-obtained using various extraction methods-and their underlying mechanisms require further investigation. AIM OF THE STUDY: Although research to find safe anti-inflammatory substances in various natural products has been actively conducted for many years, it is important to continue identifying potential anti-inflammatory effects in natural materials. The purpose of this study was to investigate the ethnopharmacological properties of the A. cepa peel extract, whose efficacy when obtained through different extraction methods and underlying action mechanisms is not well known. The present study specifically aimed to observe the anti-inflammatory effects of the A. cepa peel extracts obtained using various extraction methods and the related detailed mechanisms of A. cepa peel extracts in lipopolysaccharide (LPS)-induced RAW264.7 cells. MATERIALS AND METHODS: The total flavonoid content of the A. cepa peel extracts was determined the diethylene glycol colorimetric method and measured using a calibration curve prepared using quercetin as a standard solution. The antioxidant activity was evaluated using the ABTS assay, and cytotoxicity was measured using the MTT assay. NO production was measured using Griess reagent. Protein levels were measured by western blotting, and mRNA expression was measured by RT-qPCR. Secreted cytokines were analyzed using ELISA or cytokine arrays. In the GSE160086 dataset, we calculated Z-scores for individual genes of interest and displayed using a heat map. RESULTS: Of the three A. cepa peel extracts obtained using different extraction methods, the A. cepa peel 50% EtOH extract (AP50E) was the most effective at inhibiting LPS-induced nitric oxide (NO) and inducible nitric oxide synthase (iNOS). Furthermore, AP50E significantly reduced the levels of pro-inflammation cytokines interleukin (IL)-1α, IL-1ß, IL-6, and IL-27. Additionally, AP50E directly inhibited the Janus kinase-signaling transducer and activator of transcription (JAK-STAT) pathway. CONCLUSIONS: These results showed that AP50E exhibited an anti-inflammatory effect in LPS-induced RAW264.7 mouse macrophages by directly inhibiting JAK-STAT signaling. Based on these findings, we propose AP50E as a potential candidate for the development of preventive or therapeutic agents against inflammatory diseases.

Divergent substrate specificities and regioselectivities of three lipase isoforms from Cordyceps militaris: Combinatorial advantages for entomopathogenicity and prospects as biocatalysts.

Cordyceps militaris, an entomopathogenic Cordyceps mushroom, is a crucial ethnopharmacological agricultural product with applications in traditional oriental remedies in East Asia. Since lipases are reported to serve as key enzymatic equipment for entomopathogenic fungi during the host infection, the presence of various lipases with different biochemical features in C. militaris was elucidated. Three lipases from C. militaris (CML) of 60-70 kDa were isolated according to protein hydrophobicity; isoform relationships were identified by peptide mapping using liquid chromatography-electrospray ionization-tandem mass spectrometry. The CML isoforms exhibited distinct substrate specificities, which were related to the hydrophobicity of each isoform. Furthermore, the integral stereoselectivity of each lipase towards trioleoylglycerol diverged into two classes (sn-1,3 and sn-2 regioselectivity) that are rare in canonical fungal lipases. Overall, our results demonstrate that C. militaris secretes lipase isoforms with cocktail-like enzyme functions that may contribute to the entomopathogenic life cycle of C. militaris. Each CML isoform has distinct advantages for biocatalyst applications in the food and oleochemical industries.

Characterization and molecular docking study of cathepsin L inhibitory peptides (SnuCalCpIs) from Calotropis procera R. Br.

Propeptides, released from the autocatalytic activation of its zymogen, are potential inhibitors against proteases involved in cancer cell invasion and migration. Our research team previously obtained novel propeptides (SnuCalCpIs) from transcriptome analysis of the medicinal plant Calotropis procera R. Br. and reported them as promising candidates for cancer therapeutics due to their cathepsin L inhibition activity. In the present study, inhibitory activity among SnuCalCpIs was compared with inhibition efficiency and verified by in silico molecular docking analysis. Only SnuCalCpI03 and SnuCalCpI15, expressed in Escherichia coli, showed inhibitory activity against cathepsin L as competitive inhibitors, and the half-maximal inhibitory concentrations (IC50) values of 2.1 nM and 1.6 nM, respectively. They were stable below 70 °C, maintaining more than 90% inhibitory activity over a wide range of pH (2.0-10.0), except at the isoelectric point (pI). The template-based docking simulation models showed that SnuCalCpI02, SnuCalCpI12, and SnuCalCpI16 could not interact with the substrate-binding cleft of cathepsin L even though they possessed the same conserved domain. In contrast, SnuCalCpI03 and SnuCalCpI15 interacted with cathepsin L along the propeptide binding loop and substrate-binding cleft, resulting in obstruction of substrate access to the active site.

Development of chitosan-coated nanoemulsions of two sulfides present in onion (Allium cepa) essential oil and their nematicidal activities against the pine wood nematode, Bursaphelenchus xylophilus.

Pine wood nematode, Bursaphelenchus xylophilus, is a plant parasitic nematode which causes severe damage to several Pinus species. Two natural compounds, dipropyl trisulfide (DPTS) and methyl propyl trisulfide (MPTS), showed strong nematicidal activity against the pine wood nematode, presenting 4.24 and 17.81 µg/mL LC50 values, respectively. However, hydrophobicity and low stability have limited their practical use in the field as nematicides. To overcome these problems, chitosan-coated nanoemulsions of DPTS and MPTS were developed. The optimum chitosan concentration for the delivery system of the two sulfides was 0.5%. Optimized chitosan-coated nanoemulsions of sulfides have a uniform size distribution (mean diameter = 203.7 and 207.7 nm, mean polydispersity index = 0.176 and 0.178) with sufficient colloidal stability (mean zeta potential = +40 and +45 mV). The LC50 values of DPTS and MPTS nanoemulsions coated with 0.5% chitosan against the pine wood nematode were 5.01 and 16.60 µg/mL, respectively. In addition, chitosan coating improved the long-term storage stability and persistence of nematicidal activity of the nanoemulsions. This study indicates that the chitosan-coated nanoemulsion is a suitable formulation for sulfides as novel nematicides against the pine wood nematode for field application.

Association of Coffee Consumption and Its Types According to Addition of Sugar and Creamer with Metabolic Syndrome Incidence in a Korean Population from the Health Examinees (HEXA) Study.

Coffee is widely consumed worldwide, and numerous studies indicate that coffee consumption may potentially affect the development of chronic diseases. Metabolic syndrome (MetS) may constitute a risk factor for chronic diseases. We aimed to prospectively evaluate the association between coffee consumption and MetS incidence. All participants were selected from the Health Examinees study. MetS was defined by the Adult Treatment Panel III criteria of the National Cholesterol Education Program. A multivariate Cox proportional hazards regression model was used to assess the relationship between coffee consumption and MetS incidence. In comparison with non-consumers, male moderate consumers (≤3 cups/day) showed a lower risk for low high-density lipoprotein cholesterol (HDL-C) (≤1 cup/day, hazard ratio (HR): 0.445, 95% confidence interval (CI): 0.254-0.780; 1-3 cups/day, HR: 0.507, 95% CI: 0.299-0.859) and high fasting blood glucose (FPG) (≤1 cup/day, HR: 0.694, 95% CI: 0.538-0.895; 1-3 cups/day, HR: 0.763, 95% CI: 0.598-0.972). Male 3-in-1 coffee (coffee with sugar and creamer) consumers also showed a lower risk for low HDL-C (HR: 0.423, 95% CI: 0.218-0.824) and high FPG (HR: 0.659, 95% CI: 0.497-0.874). These findings indicate a negative association between moderate coffee consumption and low HDL-C and high FPG among Korean male adults.

Inhibitory characteristics of flavonol-3-O-glycosides from Polygonum aviculare L. (common knotgrass) against porcine pancreatic lipase.

Pancreatic lipase (PL) is an enzyme that plays an essential role in the digestion of dietary lipids and is a suitable target for an anti-obesity dietary supplement. The objective of this study was to find a novel source of PL inhibitors from Korean medicinal plants and investigate the PL-inhibitory properties of the active constituents. From among 34 kinds of methanolic crude extracts, Polygonum aviculare L. showed the highest PL-inhibitory activity (63.97 ± 0.05% of inhibition). Solvent fractionation and liquid chromatography/mass spectrometry (LC/MS) analysis identified flavonol-3-O-glycosides, flavonol-3-O-(2″-galloyl)-glycosides, and flavonol aglycones as active constituents. Furthermore, the inhibitory characteristics of the major compounds were investigated in terms of enzyme kinetics and fluorescence quenching. The results suggested that the inhibitory activity of the major compounds is closely related to the tertiary structural change in PL, and that differences in inhibitory activity occurred due to slight discrepancies in their chemical structure.

Protection of Grain Products from Sitophilus oryzae (L.) Contamination by Anti-Insect Pest Repellent Sachet Containing Allyl Mercaptan Microcapsule.

The purpose of this study was to develop an anti-insect pest repellent sachet to prevent Sitophilus oryzae (L.) (Coleoptera: Curculionidae) contamination in grain packaging. The anti-insect pest activities of essential oils (EOs) from garlic (Allium Sativum), ginger (Zingiber Officinalis), black pepper (Piper nigrum), onion (Allium cepa), and fennel (Foeniculum vulgare) as well as major compounds (allyl disulfide, AD; allyl mercaptan, AM) isolated from of garlic and onion (AD and AM) were measured against S. oryzae. The results revealed that garlic EO, onion EO, AD, and AM showed strong fumigant insecticidal activities. Among these, AM showed the highest acetylcholinesterase (AChE) inhibition rate, indicating that the fumigation insecticidal efficacy of AM is related with its AChE inhibition ability. Subsequently, the microcapsules were produced with a high efficiency (80.02%) by using AM as a core material and rice flour as a wall material. Finally, sachet composed of rice flour microcapsule containing 2% AM (RAM) was produced. Repellent assay was performed to measure anti-insect pest ability of the RAM sachet, showed remarkable repelling effect within 48 h both in the presence or absence of attractant. In a release profile of RAM sachet, it was expected to last over 20 mo during the distribution period of brown rice. Moreover, RAM sachet showed no undesirable changes to the sensory properties of the rice both before and after cooking. Taken together, these results suggest that the newly developed RAM sachet could be used as a packaging material to protect grain products from S. oryzae contamination. PRACTICAL APPLICATION: The rice weevil, Sitophilus oryzae (L.) (Coleoptera: Curculionidae), causes damages to stored products and its contamination in grain products has become a major problem in cereal market. To preserve brown rice, an anti-insect pest repellent sachet containing 2% allyl mercaptan was newly developed and it showed remarkable repellent abilities against S. oryzae. It could be used as an active food packaging system to protect grain products from insect pest contamination.

Cysteine Protease Profiles of the Medicinal Plant Calotropis procera R. Br. revealed by de novo transcriptome analysis.

Calotropis procera R. Br., a traditional medicinal plant in India, is a promising source of commercial proteases, because the cysteine proteases from the plant exhibit high thermo-stability, broad pH optima, and plasma-clotting activity. Though several proteases such as Procerain, Procerain B, CpCp-1, CpCp-2, and CpCp-3 have been isolated and characterized, the information of their transcripts is limited to cDNAs encoding their mature peptides. Due to this limitation, in this study, to determine the cDNA sequences encoding full open reading frame of these cysteine proteases, transcripts were sequenced with an Illumina Hiseq2000 sequencer. A total of 171,253,393 clean reads were assembled into 106,093 contigs with an average length of 1,614 bp and an N50 of 2,703 bp, and 70,797 contigs with an average length of 1,565 bp and N50 of 2,082 bp using Trinity and Velvet-Oases software, respectively. Among these contigs, we found 20 unigenes related to papain-like cysteine proteases by BLASTX analysis against a non-redundant NCBI protein database. Our expression analysis revealed that the cysteine protease contains an N-terminal pro-peptide domain (inhibitor region), which is necessary for correct folding and proteolytic activity. It was evident that expression yields using an inducible T7 expression system in Escherichia coli were considerably higher with the pro-peptide domain than without the domain, which could contribute to molecular cloning of the Calotropis procera protease as an active form with correct folding.

Extraction characteristics of subcritical water depending on the number of hydroxyl group in flavonols.

This study compared the efficiencies of using subcritical water, hot water, and organic solvents to extract flavonols from black tea, celery, and ginseng leaf. The effect of key operating conditions was determined by varying the temperature (110-200°C), extraction time (5-15min), and pressure (about 10MPa) and the extracts were analysed quantitatively using HPLC. The yields of myricetin, quercetin, and kaempferol from plants were maximal at extraction temperatures of 170°C, 170°C and 200°C, respectively, and they depend on the number of hydroxyl groups included in the chemical structure of the flavonols, with more of those with fewer hydroxyl (OH) groups attached being extracted at higher temperatures. The results also showed that the yields of flavonols by subcritical water extraction were 2.0- to 22.7- and 1.8- to 23.6-fold higher than those obtained using the ethanol and methanol as traditional extraction methods, respectively.

Enhancing the stability of lipid nanoparticle systems by sonication during the cooling step and controlling the liquid oil content.

Aggregation of unstable particles in water limits the application of lipid nanoparticle (LNP) systems to foods despite the capability to encapsulate lipophilic bioactive components. This study exploits a preparation process that can reduce the aggregation of LNPs. Sonication during the cooling step (postsonication) for 4, 5, or 6 min was applied to increase the covering effect of Tween 20 on the particle. Additionally, LNPs were prepared using fully hydrogenated canola oil (FHCO) blended with 0-30 wt % liquid canola oil (LCO) of the lipid phase. Surfactant surface load data indicate that the postsonication might make nonemulsifying Tween 20 diffuse from the aqueous phase to droplet surfaces, which could decrease the crystallinity index (CI) of LNPs due to the inhibition of lipid crystallization. Moreover, the LCO content in lipid matrix could decrease the CI, which could reduce the formation of hydrophobic patches on the particle surface. Therefore, the postsonication and the LCO addition in the matrix could effectively prevent aggregation among hydrophobic patches. This improved colloidal stability of LNPs was verified by the particle shape in transmission electron microscopy and the gelation test. Consequently, LNPs fabricated using 6 min postsonication and 30 wt % LCO in the lipid exhibited the greatest stability (size, 202.3 nm; CI, 57.5%; Tween 20 surface load, 10.29 mg m(-2)). This study may serve as a basis for further research that aims to develop delivery systems for functional foods.

Development of a novel selective and differential medium for the isolation of Listeria monocytogenes.

A new medium (lecithin and levofloxacin [LL] medium) is described for the isolation of Listeria monocytogenes from food samples. LL medium includes lecithin from soybeans for the detection of phosphatidylinositol-specific phospholipase C (PI-PLC) and phosphatidylcholine-specific phospholipase C (PC-PLC) produced by L. monocytogenes. Levofloxacin is incorporated to inhibit the growth of microorganisms other than L. monocytogenes, especially Bacillus cereus, shown to possess PI-PLC and PC-PLC activities. L. monocyogenes produced white colonies with a halo on LL medium, whereas Listeria innocua appeared as white colonies without a halo. Levofloxacin at 0.20 mg/liter completely inhibited the growth of B. cereus, while the growth of L. monocytogenes was unaffected. In the second phase of the study, the sensitivity and the specificity of LL medium were compared to those of modified Oxford agar (MOX) and two chromogenic media (Brilliance Listeria agar and CHROMagar Listeria), using a total of 250 food samples. From 200 unspiked food samples, the specificity of LL medium (96.0%) was superior to that of MOX (72.0%) and similar to the specificities of Brilliance Listeria agar (96.5%) and CHROMagar Listeria (94.5%). From 50 spiked food samples, LL medium and CHROMagar Listeria represented the highest sensitivities (96.0%), followed by Brilliance Listeria agar (92.0%) and MOX (54.0%). Also, LL medium showed the highest confirmation rate (98.8%), followed by Brilliance Listeria agar (98.7%), CHROMagar Listeria (98.3%), and MOX (52.0%). On the basis of its good specificity and cost effectiveness, LL medium is useful for the isolation of L. monocytogenes from food samples.

Effects of roasting conditions on the physicochemical properties and volatile distribution in perilla oils (Perilla frutescens var. japonica).

UNLABELLED: Perilla seeds have more than 60% of α-linolenic acid, one of omega-3 essential fatty acids. Headspace volatiles and physicochemical properties including color, fluorescence intensity, and the oxidation products in perilla oil (PO) from perilla seeds roasted at different conditions were analyzed. Roasting temperature was 150, 170, 190, and 210°C, and roasting time was 15 and 30 min at each roasting temperature. PO from higher roasting temperature and longer roasting time had lower L* values, higher a*, b*, and chroma values, more brown pigments and fluorescence intensity, and more conjugated dienoic acids. Pyrazines were major volatiles in PO, and furans, sulfur-containing compounds, and hydrocarbons were also detected by a solid phase microextraction gas chromatography/mass spectrometry. In PO, 2,5-Dimethylpyrazine and 2-furancarboxaldehyde were 2 major volatiles. The principal component analysis of volatiles showed the 1st principal component (PC1) and the 2nd principal component (PC2) express 56.64% and 22.72% of the volatile variability in PO, respectively, which can differentiate PO prepared from roasting conditions clearly. Some physicochemical properties especially brown pigment and volatiles were positively correlated with each other in PO. PRACTICAL APPLICATION: Perilla oil (PO) from perilla seeds possesses more than 60% of α-linolenic acid, one of omega-3 fatty acids. Roasting process has been used to extract oil from perilla seeds. Understanding physicochemical properties of PO from diverse roasting conditions are important steps to produce PO in food industry. Roasting process induces darkening of color, increase of fluorescence intensity, and brown pigments in PO. Pyrazines and furans are major headspace volatiles in PO roasted above 170°C. The results of this study can help to produce PO in industrial scales with desired headspace volatiles, colors, and oxidative state.

Optimization, in vitro release and bioavailability of gamma-oryzanol-loaded calcium pectinate microparticles reinforced with chitosan.

Response surface methodology was used to optimize coating conditions, including chitosan concentration (X(1)) and coating time (X(2)), for sustained release of chitosan-coated Ca-pectinate (CP) microparticles containing oryzanol (OZ). The optimized values of X(1) and X(2) were found to be 1.48% and 69.92 min, respectively. These optimized values agreed favorably with the predicted results, indicating the utility of predictive models for the release of OZ in simulated intestinal fluid. In vitro release studies revealed that the chitosan-coated CP microparticles were quite stable under acidic conditions, but swell and disintegrate under alkaline conditions. In vivo release study of OZ, physically entrapped within chitosan-coated CP microcapsules, demonstrated the sustained release of OZ and could be used to improve the bioavailability of OZ following oral administration.

Effects of riboflavin photosensitization on the degradation of bisphenol A (BPA) in model and real-food systems.

Effects of riboflavin photosensitizations on the stability of bisphenol A (BPA), a well-known endocrine disrupting chemical, were studied in model and real-food systems by high-performance liquid chromatography (HPLC). Concentration of BPA was significantly decreased under light exposure (P < 0.05) as the concentration of riboflavin increased while those without riboflavin under light or those with riboflavin in the dark did not change significantly (P > 0.05). Addition of 50, 100, and 200 microM sodium azide significantly increased the stability of BPA in riboflavin photosensitization with concentration dependent manner (P < 0.05), implying that a singlet oxygen or type II pathway played a role in the photodegradation of BPA. Stability of BPA in riboflavin was significantly increased in the presence of high concentration of tert-butanol, a hydroxyl radical quencher, under light storage for 80 min, indicating hydroxyl radicals were involved and contributed to the degradation of BPA, at least in part. Availability of riboflavin photosensitization on the photodegradation of BPA was tested in 2 canned tea beverages with different phenolic contents. BPA was more stable in the beverage sample with higher total phenolic contents and free radical scavenging ability. The photodegradation of BPA in riboflavin photosensitization can be an efficient way to decrease the concentration of BPA from environmental or food systems.