Carotenoid and Tocopherol Profiling in 18 Korean Traditional Green Leafy Vegetables by LC-SIM-MS.

Fruits and vegetables are a vital source of redox-active phytochemicals in the diet. Traditional green leafy vegetables (GLVs) are a rich source of carotenoids, dietary fiber, minerals, phenols, vitamins, and tocopherols and are commonly consumed in rural areas worldwide. In traditional Korean medicine, many GLVs are used to treat various ailments. However, data on the carotenoid and tocopherol content of many traditional GLVs consumed in the Republic of Korea are insufficient. The current work aims to compare the carotenoid and tocopherol profiles of 18 traditional GLVs by utilizing a single ion monitoring LC-MS approach to identify the potential GLVs for commercial cultivation and healthy diet formulations. Among the traditional GLVs investigated, (all-E)-lutein was the most abundant carotenoid, ranging from 44.4% in Glehnia littoralis to 52.1% in Heracleum moellendorffii. It was followed by (all-E)-violaxanthin and (all-E)-ß-carotene. The highest contents of (all-E)-violaxanthin (75.6 µg/g FW), 9-Z-neoxanthin (48.4 µg/g FW), (all-E)-luteoxanthin (10.8 µg/g FW), (all-E)-lutein (174.1 µg/g FW), total xanthophylls (310.5 µg/g FW), (all-E)-ß-carotene (69.6 µg/g FW), and total carotenoids (380.1 µg/g FW) were recorded in Pimpinella brachycarpa. Surprisingly, Taraxacum mongolicum also showed the highest contents of (all-E)-violaxanthin, (all-E)-lutein, and total carotenoids, which were statistically non-significant (p > 0.05, Tukey HSD) with P. brachycarpa. The highest concentration of (all-E)-zeaxanthin (14.4 µg/g FW) was recorded in Solidago virga-aurea. Among the studied herbs, 13.9 (H. moellendorffii)-133.6 µg/g FW (Toona sinensis) of α-tocopherol was recorded. Overall, the results suggest that P. brachycarpa and T. mongolicum are rich sources of carotenoids. On the other hand, T. sinensis is a rich source of α-tocopherol. These GLVs can be utilized in the diet to enhance the intake of health-beneficial carotenoids and α-tocopherol.

Ginseng leaf extract ameliorates the survival of endotoxemic mice by inhibiting the release of high mobility group box 1.

High mobility group box 1 (HMGB1) is a well-defined mediator involved in the pathophysiologic response to endotoxemia and sepsis. However, the mechanisms and therapeutic agents that could prevent its release are not fully elucidated. Here, the present study demonstrates that the ginseng leaf extract (GLE) regulates lipopolysaccharide (LPS)-triggered release of HMGB1 in macrophages and endotoxemic animal model. Treatment of RAW264.7 macrophages with GLE significantly inhibited the release of HMGB1 stimulated by LPS. GLE also suppressed the generation of nitric oxide (NO) and expression of inducible NO synthase (iNOS) in a dose-dependent manner. These effects of GLE were accompanied by inhibition of HMGB1 release stimulated by LPS, indicating a potential mechanism by which GLE regulates HMGB1 release through NO signaling. Furthermore, induction of suppressor of cytokine signaling 1 by GLE-mediated GLE-dependent suppression of HMGB1 release and NO/iNOS induction by inhibiting Janus kinase 2/signal transducer and activator of transcription 1 signal in RAW 264.7 cells exposed to LPS. Finally, administration of the GLE ameliorated the survival rate of LPS-injected endotoxemic mice in a NO-dependent manner. Thus, GLE may block the LPS-stimulated release of HMGB1 by regulating cellular signal networks, thereby providing a therapeutic strategy for endotoxemia as a functional food. PRACTICAL APPLICATIONS: High mobility group box 1 (HMGB1) is released into the extracellular milieu when immune cells are exposed to pathogen-related molecules such as lipopolysaccharide (LPS), in which it acts as a critical mediator of lethality in sepsis and endotoxemia. The extract of ginseng leaf, which is a part that can be easily thrown away, ameliorated the survival rate of endotoxemic mice by inhibiting HMGB1 secretion in a NO-dependent manner. Thus, this study suggests that ginseng leaf can be used as a functional food by resolving the immune responses in the pathology of endotoxemia.

Effects of Bromelain and Double Emulsion on the Physicochemical Properties of Pork Loin.

Our aim was to investigate the effects of bromelain embedded in double emulsion (DE) on physicochemical properties of pork loin. We evaluated DE characteristics such as size, zeta potential, and microscopy after fabrication. We marinated meat with distilled water (DW), 1% (w/v) bromelain solution, blank DE, and 1% (w/v) bromelain loaded in double emulsion (DE E) for 0, 24, 48, and 72 h at 4°C, and prepared raw meat for control. The marinated samples were assessed for color, water holding capacity, cooking loss, moisture content, pH, protein solubility, Warner-Bratzler shear force (WBSF) and gel electrophoresis. The droplet size of 1% (w/v) bromelain embedded in DE was increased compared with blank DE (p<0.05) and values of zeta potential decreased. The increase in lightness and color difference range of the DE-treated group was lower than that of the DW-treated group (p<0.05). Moreover, treatment by immersion in 1% (w/v) DE E resulted in the highest water holding capacity values (p<0.05) and lower cooking loss values than water base treatment (p<0.05). Results for myofibrillar protein solubility and WBSF showed a similar trend. 1% (w/v) DE E showed degradation of myosin heavy chain after 48 h in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Thus, bromelain-loaded DE is useful for controlling and handling enzyme activity in food industry.

Nanoemulsions improve the efficacy of turmeric in palmitate- and high fat diet-induced cellular and animal models.

Turmeric is a well-known functional food exhibiting multiple biological activities in health and disease. However, low aqueous solubility and poor bioavailability limit its therapeutic potential. Herein, we investigated the utility of nanoemulsions as a carrier to improve the efficacy of turmeric. Compared with turmeric extract (TE), 5% TE-loaded nanoemulsion (TE-NE), which contains 20-fold lower curcumin content than TE, achieved similar inhibition of palmitate-induced lipotoxicity in HepG2 cells. Exposure of HepG2 cells to 5% TE-NE also suppressed the palmitate-induced accumulation of lipid vacuoles and reactive oxygen species comparably with TE, and was accompanied by decreased levels of sterol regulatory element-binding protein (SREBP)-1, peroxisome proliferator-activated receptor-γ2 (PPAR-γ2), cleaved caspase-3, and poly (ADP-ribose) polymerase (PARP). Consistent with these effects in HepG2 cells, oral administration of 5% TE-NE to mice fed a high fat diet (HFD) markedly suppressed lipid accumulation in liver, leading to a significant reduction in body weight and adipose tissue weight, equivalent to the effects observed with TE. Compared with TE, 5% TE-NE also equivalently inhibited the levels of SREBP-1, PPAR-γ2, cleaved caspase-3, and PARP in the liver of mice fed a HFD. Furthermore, TE and 5% TE-NE significantly improved serum lipid profiles in a similar manner. These observations indicate that nanoemulsions can improve the efficacy of turmeric, thereby eliciting more potent biological efficacy against palmitate- and high fat diet (HFD)-induced cellular damage.

Identification and Evaluation of Cytotoxicity of Peptide Liposome Incorporated Citron Extracts in an in Vitro System.

Abstract: Citrons have been widely used for medicinal purposes for a long time, but the application of citron in the food industry is still restricted. The extensive advantages of nanotechnology in the food industry have greatly broadened the application of foods. In this study, by employing nanotechnology, we prepared citron-extract nanoparticle with an average size of 174.11 ± 3.89 nm, containing protein peptide and/or liposome. In order to evaluate the toxicity of nanoparticles and to ensure food safety, biological cytotoxicity at the cell and genomic levels was also identified to examine the toxicity of citron extracts by using an in vitro system. Our results demonstrated that the cytotoxicity of citronliposome was dependent on cell type in high concentrations (1 and 5 mg/mL), selectively against primary human cardiac progenitor cells (hCPCs), and human endothelial progenitor cells (hEPCs) in MTT and lactate dehydrogenase (LDH) assays. Interestingly, for the NIH-3T3 and H9C2 cell lines, cell cytotoxicity was observed with slight genotoxicity, especially from citronpeptide extract for both cell lines. Taken together, our study provides cytotoxicity data on nanoengineered citron extracts according to different cell type as is crucial for further applications.

Studies on the Anti-Oxidative Function of trans-Cinnamaldehyde-Included ß-Cyclodextrin Complex.

trans-Cinnamaldehyde (tCIN), an active compound found in cinnamon, is well known for its antioxidant, anticancer, and anti-inflammatory activities. The ß-cyclodextrin (ß-CD) oligomer has been used for a variety of applications in nanotechnology, including pharmaceutical and cosmetic applications. Here, we aimed to evaluate the anti-inflammatory and antioxidant effects of tCIN self-included in ß-CD complexes (CIs) in lipopolysaccharide (LPS)-treated murine RAW 264.7 macrophages. RAW 264.7 macrophages were treated with increasing concentrations of ß-CD, tCIN, or CIs for different times. ß-CD alone did not affect the production of nitric oxide (NO) or reactive oxygen species (ROS). However, both tCIN and CI significantly reduced NO and ROS production. Thus, CIs may have strong anti-inflammatory and antioxidant effects, similar to those of tCIN when used alone.

Subcritical water-hydrolyzed fish collagen ameliorates survival of endotoxemic mice by inhibiting HMGB1 release in a HO-1-dependent manner.

To investigate potential mechanisms underlying the bioactivity of hydrolyzed fish collagen, we examined the anti-inflammatory actions of subcritical water-hydrolyzed fish collagen (SWFC) in lipopolysaccharide (LPS)-triggered inflammation and endotoxemia. SWFC markedly inhibited LPS-stimulated release of high mobility group box 1 (HMGB1) in murine RAW264.7 macrophages, along with decreased cytosolic translocation of HMGB1. Both the protein and mRNA levels of heme oxygenase-1 (HO-1) were significantly upregulated in SWFC-treated RAW 264.7 cells in an Nrf2-dependent manner. In line with these effects of SWFC, both HO-1 siRNA and ZnPPIX (zinc protoporphyrin IX) actually attenuated the effects of SWFC on HMGB1 release stimulated by LPS, indicating a possible mechanism by which SWFC modulates HMGB1 release through HO-1 signaling. Notably, administration of SWFC improved the survival rates of LPS-injected endotoxemic mice, in which the serum level of HMGB1 was significantly reduced. Taken together, these results indicate that the anti-inflammatory activities of SWFC are achieved by inhibiting HMGB1 release induced by LPS in a HO-1-sensitive manner.

The Effect of GCSB-5 a New Herbal Medicine on Changes in Pain Behavior and Neuroglial Activation in a Rat Model of Lumbar Disc Herniation.

OBJECTIVE: Lumbar disc herniation can induce sciatica by mechanical compression and/or chemical irritation. The aim of this study was to compare the effects of GCSB-5 (Shinbaro®) and NSAIDs on pain-related behavior and on the expressions of microglia, astrocytes, CGRP, TRPV1, IL-6, and CX3CL1 in a rat model of lumbar disc herniation. METHODS: 112 male Sprague-Dawley rats underwent implantation of nucleus pulposus to a dorsal root ganglion (DRG). Rats were divided into five groups as follows; a saline group (the vehicle control group) (n=27), a 10 mg/kg aceclofenac group (the aceclofenac group) (n=22), and 100, 300 or 600 mg/kg GCSB-5 groups (the GCSB-5 100, 300, or 600 groups) (n=21 for each group). Rats were tested for mechanical allodynia at 3 days after surgery and at 1 day, 3 days, 7 days, 14 days, 21 days, 28 days, 35 days, 42 days, 49 days, and 56 days after treatment commencement. Immunohistochemical staining of microglia (Iba1), astrocytes (GFAP), CGRP, and TRPV1, and PCR for IL-6 and CX3CL1 were performed on spinal dorsal horns and DRGs at 56 days after medication commencement. RESULTS: After 56 days of GCSB-5 300 administration, mechanical withdrawal thresholds were significantly increased (p<0.05), and immunohisto-chemical expressions of Iba1, GFAP, CGRP, and TRPV1 were reduced than other groups, but this difference was not statistically significant. CONCLUSION: These results indicate GCSB-5 reduces mechanical allodynia and downregulates neuroglial activity and the expressions of CGRP and TRPV1 in the spinal segments of a rat model of lumbar disc herniation.

Efficacy of capsicum oleoresin nanocapsules formulation by the modified emulsion-diffusion method.

In this study, we investigated the effect of high pressure homogenizer on the physico-chemical properties of capsicum oleoresin loaded nanoemulsion (NE) or nanocapsules (NCs) based on the emulsion-diffusion method. According to the application stage of high pressure process at principle emulsion-diffusion method, NCs was prepared by conventional-emulsion-diffusion method (CED), modified-emulsion-microfluidization-diffusion method (MEMD) and modified-emulsion-diffusion-microfluidization method (MEDM). The nanocapsules of MEMD showed homogeneous and the smallest particle size as compared with CED. In addition, MEMD presented the surface tension at the value 36.5 mN/m. The encapsulated capsicum oleoresin was generated the bright color and suppressed the dark red color. Furthermore, MEMD gave the high encapsulation efficiency of capsicum oleoresin around 95% and showed the slow release rate. On the other hand, MEDM presented the non-homogeneous and agglomerate of the particle, low percentage of encapsulation efficiency and the high initial release rate when compared with CED and MEMD methods. According these results, it was supposed that the microfluidization was interesting technique to ameliorate the physical properties and efficiency of NCs. However, it was depending on the appropriate combination of microfluidization based on the emulsion-diffusion method.