Influence of Isolation Temperature on Isolating Diverse Lactic Acid Bacteria from Kimchi and Cultural Characteristics of Psychrotrophs.

Kimchi is a traditional Korean fermented vegetable that is stored and fermented at low temperatures. However, kimchi lactic acid bacteria (LAB) are typically isolated under mesophilic conditions, which may be inappropriate for isolating the diverse LAB. Therefore, this study investigated the suitable conditions for isolating various LAB from kimchi. Here, LAB were isolated from four kimchi samples using MRS, PES, and LBS media and varying isolation temperatures (30, 20, 10, and 5°C). Then, MRS was selected as the suitable medium for LAB isolation. A comparison of culture-dependent and culture-independent approaches indicated that 5°C was not a suitable isolation temperature. Thus, the number and diversity of LAB were determined at 30, 20, and 10°C using 12 additional kimchi samples to elucidate the effect of isolation temperature. With the exception of two samples, most samples did not substantially differ in LAB number. However, Leuconostoc gelidum, Leuconostoc gasicomitatum, Leuconostoc inhae, Dellaglioa algida, Companilactobacillus kimchiensis, Leuconostoc miyukkimchii, Leuconostoc holzapfelii, and Leuconostoc carnosum were isolated only at 10 and 20°C. The growth curves of these isolates, except Leu. holzapfelii and Leu. carnosum, showed poor growth at 30°C. This confirmed their psychrotrophic characteristics. In Weissella koreensis, which was isolated at all isolation temperatures, there was a difference in the fatty acid composition of membranes between strains that could grow well at 30°C and those that could not. These findings can contribute to the isolation of more diverse psychrotrophic strains that were not well isolated under mesophilic temperatures.

Optimizing Conditions in the Acid Tolerance Test for Potential Probiotics Using Response Surface Methodology.

Acid tolerance is an important feature of probiotic development. It is one of the factors underlying the beneficial effects of probiotics in the intestine. However, the methods used by different researchers to test acid tolerance vary, causing confusion in the interpretation of the results. Therefore, in this study, we determine the optimal conditions for the acid tolerance test using response surface methodology. The factors of pH (2.5 to 3.5), exposure time (1 to 2 h), and pepsin (presence or absence) were used as independent variables, and the survival rates of seven strains (Lacticaseibacillus casei KACC 12413, Lactiplantibacillus plantarum KACC 15357, Limosilactobacillus fermentum KACC 11441, Lactiplantibacillus plantarum WCFS1, Lacticaseibacillus rhamnosus GG, Lactiplantibacillus plantarum KCTC 21024, and Lactiplantibacillus plantarum WiKim 0112) known to have probiotic properties were used as dependent variables. The results of the analysis of variance (ANOVA) indicated that the pH value and exposure time in acidic environments significantly affected the acid tolerance test model, and their interaction also had an effect (P < 0.05). Using the ANOVA results, the condition of the acid tolerance test was optimized with a target of an 85% survival rate for each strain. The optimized conditions of the acid tolerance test were as follows: pH 2.92, exposure time of 1.73 h, and presence of pepsin and pH 3, exposure time of 1.98 h, and absence of pepsin. These results can optimize strain selection with rigorous acid tolerance without confusion by unifying the conditions for the acid tolerance test. IMPORTANCE The acid tolerance test, which is the first step in selecting probiotics, is not standardized and can often cause confusion in the interpretation of results. Thus, in the present study, we optimized the conditions for the acid tolerance test using response surface methodology. These optimized conditions can be used to screen for strains with acid tolerance.

Safety assessment of white colony-forming yeasts in kimchi.

White colony-forming yeasts (WCFYs) have been reported to form a white colony on the surface of kimchi, resulting in the deterioration of kimchi sensory quality. However, toxicity of WCFY has rarely been studied. Thus, to evaluate the safety of WCFY (i.e., Kazachstania servazzii, Candia sake, and Pichia kudriavzevii), we conducted cell and animal experiments as well as genomic analysis. In vitro studies indicated that WCFY did not induce cytotoxic responses such as lactate dehydrogenase release, excessive oxidative stress, and mitochondrial damage at concentrations of up to 2.5 × 105 CFU/mL in human intestinal and liver cells. In animal studies using rats (single-dose and 14-day repeated-dose oral toxicity studies), WCFY did not induce death, clinical signs of toxicity, histological alterations of the liver, or increases in the expression of pro-inflammatory cytokines nor cytochrome P450-2E1 in liver tissue at concentrations of up to 5 × 108 CFU/head/day. Genomic analysis revealed that P. kudriavzevii did not harbor genes related to toxicity and antimicrobial resistance. Taken together, our data suggest that exposure to WCFY through kimchi intake did not induce toxic response in the Caco-2, HepG2, and Sprague-Dawley rats. The current work provides evidence for the safety of accidental major WCFY ingestion via kimchi.

Effects of duck fat and κ-carrageenan as replacements for beef fat and pork backfat in frankfurters.

OBJECTIVE: Frankfurters are emulsion-type sausages that are widely consumed worldwide. However, some concerns regarding negative health effects have been raised because of the high fat content and the type of fat. This study aimed to evaluate the effects of duck fat and κ-carrageenan as replacements for beef fat and pork backfat in frankfurters. METHODS: The different formulations for the frankfurters were as follows: 20% beef fat (BF), 20% pork backfat (PBF), 20% duck fat (DF), 20% soybean oil (SO), 20% duck fat/1% κ-carrageenan (DFC), and 20% soybean oil/1% κ-carrageenan (SOC). Physicochemical (fatty acid profile, color, rheological properties, cooking loss, water holding capacity, emulsion stability, and texture profile analysis), oxidative stability and sensory properties of frankfurters were evaluated. RESULTS: Duck fat and κ-carrageenan improved rheological properties of meat batter, and physicochemical properties (emulsion stability, cooking loss, and hardness) of frankfurters. Moreover, duck fat added-frankfurters (DF and DFC) had higher oxidative stability than that of soybean-added frankfurters (SO and SOC) during refrigerated storage for 28 days. In sensory evaluation, flavor, texture, and overall acceptability of DFC were acceptable to untrained panelists. CONCLUSION: Our data suggest that duck fat and κ-carrageenan can replace beef fat and pork backfat in frankfurters. Duck fat and κ-carrageenan contributed to improve the physicochemical properties and oxidative stability while maintaining sensory properties. Therefore, the use of duck fat and κ-carrageenan may be a suitable alternative for replacing beef fat or pork backfat in frankfurters.

Tebuconazole Fungicide Induces Lipid Accumulation and Oxidative Stress in HepG2 Cells.

Tebuconazole (TEB), a triazole fungicide, is frequently applied to agriculture for the increase of food production. Although TEB causes liver toxicity, its effects on cellular lipid accumulation are rarely investigated. Therefore, this study aimed to study the effects of TEB on lipid metabolism and accumulation in HepG2 cells. HepG2 cells were exposed to 0-320 µM TEB for 1-24 h. TEB (20-80 µM, 24 h)-treated cells showed lipid accumulation. Further, TEB (20-80 µM, 1-12 h) increased the nuclear translocation of peroxisome proliferator-activated receptors and the expression of lipid uptake and oxidation-related markers such as cluster of differentiation 36, fatty acid transport protein (FATP) 2, FATP5, and carnitine palmitoyltransferase 1. Oxidative stress levels in TEB-treated cells (20-80 µM, 24 h) were higher, compared to those in the control. TEB (20-80 µM, 24 h) also induced the loss of mitochondrial membrane potential and lower levels of microsomal triglyceride transfer protein in the cells. Thus, TEB can induce lipid accumulation by altering the expression of lipid-metabolizing molecules and can therefore impair lipid metabolism. Our data suggest that human exposure to TEB may be a risk factor for non-alcoholic fatty liver disease.

Comparison of the Probiotic Potential between Lactiplantibacillus plantarum Isolated from Kimchi and Standard Probiotic Strains Isolated from Different Sources.

In the present study, the properties of the Lactiplantibacillus (Lpb.) plantarum WiKim0112 isolated from kimchi were evaluated by comparing its probiotic properties to those of Lpb. plantarum WCFS1 and KACC 11451 isolated from different sources. In both pH 2 and 3, media containing pepsin, Wikim0112, and WCFS1 showed higher cell viability than KACC11451. Viability of all Lpb. plantarum strains in a medium containing pancreatin and bile salt oxgall was significantly decreased compared to the control. WCFS1 showed the highest thermotolerance, followed by Wikim0112 and KACC11451. Wikim0112 showed a similar level of antibacterial activity to WCFS1 and exhibited an overall higher antibacterial activity than KACC11451 against six pathogens. All Lpb. plantatum strains showed high antioxidant activities in SOD, DPPH, and ABTS assays, especially Wikim0112 and WCFS1 exhibited a higher antioxidant activity than KACC11451. All Lpb. plantarum strains showed approximately 60-62% adhesion rates to Caco-2 cells. Moreover, in LPS-stimulated Caco-2 cells, all Lpb. plantarum strains significantly decreased the mRNA expression of pro-inflammatory cytokines (i.e., IL-1ß, IL-6, and TNF-α); Wikim0112 significantly increased the mRNA expression of IL-4 and IFN-γ. Wikim0112 was resistant to streptomycin and vancomycin, whereas WCFS1 and KACC11451 were resistant to four (clindamycin, ciprofloxacin, tetracycline, and vancomycin) and three (ciprofloxacin, tetracycline, and vancomycin) antibiotics, respectively. These results, taken together, indicated that compared to Lpb. plantarum strains isolated from different sources, Wikim0112 showed desirable probiotic properties, suggesting its potential applications in the food and pharmaceutical industries.

Biological activity and processing technologies of edible insects: a review.

The burgeoning global population growth has raised concerns regarding the expected increase in the demand for food, which could be partially tackled by identifying novel food sources. To this end, edible insects have recently attracted research interest. Several technologies for utilizing edible insect-derived proteins have been introduced; however, research into their functional utilization is insufficient. Herein, we reviewed the relevant literature on the importance of insects as food sources, extraction of edible insects, the nutritional value of insects, biological activities of components, and their applications in food industries. We summarized the studies primarily focused on the functional utilization of edible insects, suggesting that for successful incorporation and growth of edible insects in food and pharmaceutical industries, strategies to improve the extraction methods are required to explore the biological activity of edible insects. Furthermore, the awareness of edible insects with a focus on their allergens warrants consideration.

Effects of Flutriafol Fungicide on the Lipid Accumulation in Human Liver Cells and Rat Liver.

Flutriafol (FTF) is a triazole fungicide that can cause liver toxicity through the ingestion of its residues in food and water. However, little is known about the liver toxicity of FTF, particularly nonalcoholic fatty liver disease (NAFLD) in humans. Therefore, the purpose of this study was to investigate whether FTF induces NAFLD in human liver cells and animal liver. HepG2 cells and Sprague Dawley (SD) rats were treated with FTF at doses of 0-640 µM for 24 h and 0-150 mg/kg bw/day for 28 days, respectively. FTF (80, 160, and 320 µM) treatment to cells induced lipid accumulation. FTF (80 and 160 µM)-treated cells had higher levels of cytochrome P450 enzymes and reactive oxygen species and increased mitochondrial membrane potential loss than the control. FTF also increased the mRNA levels of antioxidant enzymes through oxidative stress and nuclear factor erythroid 2-related factor 2 pathways in HepG2 cells. However, a higher level of FTF (320 µM) induced apoptosis. The treatment of SD rats with FTF (2.5-150 mg/kg bw/day) induced fatty infiltration in the liver by impairing liver metabolism and inducing apoptosis. Therefore, our data suggest that human exposure to FTF residues may be a risk factor for liver diseases, such as NAFLD.

In Vitro and In Vivo Study on the Toxic Effects of Propiconazole Fungicide in the Pathogenesis of Liver Fibrosis.

Propiconazole (PCZ) is a hepatotoxic triazole fungicide. There are insufficient data on how PCZ induces liver fibrosis in humans. This study aimed to investigate the effect of PCZ on liver fibrosis and its underlying mechanisms. HepG2 cells and Sprague-Dawley rats were exposed to PCZ at doses of 0-160 µM (3-72 h) and 0.5-50 mg/kg body weight/day (28 days), respectively. PCZ-treated cells activated intracellular oxidative stress via cytochrome P450 and had higher mRNA levels of interleukin-1ß, tumor necrosis factor-α, matrix metalloproteinase (MMP)-2, MMP-9, and transforming growth factor-ß (TGF-ß) than the control. PCZ treatment in cells induced a morphological transition with E-cadherin decrease and vimentin and Snail increase via the oxidative stress and TGF-ß/Smad pathways. PCZ administration in rats induced liver fibrosis through pathological changes, epithelial-mesenchymal transition, and collagen deposition. Thus, our data suggest that exposure of PCZ to humans may be a risk factor for the functional integrity of the liver.

Physicochemical properties and oxidative stability of duck fat-added margarine for reducing the use of fully hydrogenated soybean oil.

Soybean oil (SBO) and fully hydrogenated soybean oil (FHSBO) have been used for margarine production. However, SBO-based margarine requires a considerable amount of trans fatty acid-containing FHSBO due to its low melting point. We aimed to reduce the FHSBO content in margarine by employing duck fat, which has a higher melting point than SBO. Margarines were prepared using different ratios of duck fat and reduced levels of SBO and FHSBO. Physicochemical, sensory, and oxidative properties of the margarines were evaluated. The quality characteristics of margarine improved when duck fat replaced SBO and FHSBO. Furthermore, the lipid oxidation parameters were lower in duck fat-added margarines than the control during storage at 60 °C for 28 days. The margarine containing 80% duck fat showed the best sensory properties. Collectively, duck fat can replace SBO in margarine while reducing the use of FHSBO and maintaining desirable physicochemical properties, oxidative stability, and sensory properties.

Growth Inhibitory Effect of Garlic Powder and Cinnamon Extract on White Colony-Forming Yeast in Kimchi.

White colony-forming yeast (WCFY), also referred to as film forming yeast or spoilage yeast, that appear on the surface of kimchi can deteriorate the sensory properties of kimchi, such as odor and texture. Thus, the aim of this study was to develop a method to inhibit the formation of the white colony in kimchi. First, alterations in kimchi manufacturing and storage conditions, including temperatures, pH, salinity, and anaerobic condition, were investigated to determine if they could inhibit the growth of WCFY (i.e., Kazachstania servazzii, Candida sake, Debaryomyces hansenii, Pichia kudriavzevii, and Hanseniaspora uvarum). Thereafter, the anti yeast activity of freeze-dried garlic powder (FGP) and cinnamon ethanol extract (CEE) was evaluated against WCFY using the agar-well diffusion assay. Following the direct application of FGP and CEE to the surface of the kimchi, the inhibitory effects on white colony were determined. The results showed that WCFY can grow under various manufacturing and storage conditions of kimchi. Regarding the growth inhibitory effect on WCFY, FGP exhibited anti yeast activity against four WCFYs. It did not show anti yeast activity against K. servazzii. However, CEE showed anti yeast activity against K. servazzii. In particular, the mixture of 10% FGP and 1.75% CEE, which was manufactured considering the influence of sensory properties in kimchi, exhibited anti yeast activity against all WCFY. Furthermore, the application of the FGP and CEE mixture supplemented with 0.02% xanthan gum to kimchi to enhance adhesion to the kimchi surface, led to a delay in the formation of a white colony on the surface of the kimchi by an average of 17 d at 10 °C compared to the control group. Collectively, the use of a FGP, CEE, and xanthan gum mixture could be an effective method for the inhibition of white colony formation on the surface of kimchi, extending its shelf life.

In vitro toxicity assessment of crosslinking agents used in hyaluronic acid dermal filler.

Hyaluronic acid (HA) dermal fillers are produced by crosslinking HA with agents, such as 1,4-butanediol diglycidyl ether (BDDE) and poly (ethylene glycol) diglycidyl ether (PEGDE) to acquire desired properties. Thus, the safety evaluation of these crosslinkers is needed at the cellular level. In the present study, cell viability, cytotoxicity, membrane integrity, reactive oxygen species (ROS), mitochondrial membrane potential (MMP), and inflammatory responses were evaluated in the human keratinocyte cell line, HaCaT and human dermal fibroblast cell line, HDF in response to treatment with the crosslinkers. In both the cell lines, BDDE significantly decreased cell viability at 100-1000 ppm, while PEGDE showed a decrease at 500-1000 ppm. In HaCaT cells, BDDE markedly increased cytotoxicity (lactate dehydrogenase release) at 100-1000 ppm, but PEGDE showed an increase at 500-1000 ppm. Cells treated with BDDE (100 ppm) caused alteration in the integrity of cell membrane and shape. In both the cell lines, BDDE-treated cells showed significantly higher ROS levels and MMP loss than PEGDE-treated cells. Also, BDDE-treated cells exhibited higher COX-2 expression at 100 ppm. Expression of inflammatory cytokines (TNF-α, and IL-1 ß) was higher in BDDE-treated cells. Taken together, PEGDE-treated cells showed markedly lower cytotoxicity, ROS production, and inflammatory responses than BDDE-treated cells. Our data suggest that PEGDE is safer than BDDE as a crosslinker in HA dermal fillers.

Evaluation of gels formulated with whey proteins and sodium dodecyl sulfate as a fat replacer in low-fat sausage.

Consumers are increasingly interested in low-fat meat products. Therefore, there is demand for new fat replacers that improve the quality of low-fat meat products. Whey protein isolate (WPI; 10% (w/v)) and sodium dodecyl sulfate (SDS; 0-0.09% (w/v)) were used to produce WPI-SDS gel as a fat replacer of low-fat meat products. Characteristics of WPI-SDS gel were evaluated using SDS-PAGE, FT-IR, viscometer, and texture analyzer. Addition of SDS to WPI increased gelation while reducing aggregation. Addition of 0.06% SDS to WPI-SDS gel has the highest viscosity and hardness, while 0.09% SDS decreased the heat stability of WPI. Quality characteristics including cooking loss, emulsion stability, hardness, and chewiness were significantly improved in WPI-SDS gel-supplemented low-fat sausages. Particularly, the highest hardness and chewiness were obtained in the low-fat sausage added with WPI-SDS gel containing 0.06% SDS. Our results suggest that WPI-SDS gel can be used as a fat replacer in low-fat meat products.

Probiotic Properties of Lactiplantibacillus plantarum LB5 Isolated from Kimchi Based on Nitrate Reducing Capability.

The purpose of this study was to investigate the probiotic properties of lactic acid bacteria isolated from Korean radish water kimchi (dongchimi). A total of 800 isolates of lactic acid bacteria were isolated from kimchi, and the strain having reduction and tolerance capability for nitrate and nitrite was selected and identified as Lactiplantibacillus plantarum LB5 (LPLB5) by 16S rRNA sequencing. LPLB5 showed higher tolerance to acidic pH values (pH 2.5), 0.3% bile salts, and heat treatment (40, 50, and 60 °C). Antibacterial activity showed strong inhibition against four food-borne pathogenic bacteria (E. coli O157:H7 ATCC 35150, Pseudomonas aeruginosa KCCM 12539, Listeria monocytogenes KCCM 40307, and Staphylococcus aureus ATCC 25923). The strain did not show any antibiotic resistance, ß-hemolytic activity, or ability to produce ß-glucuronidase. LPLB5 also exhibited a 30% auto-aggregation ability and 33-60% co-aggregation ability with four pathogenic bacteria (E. coli O157: H7 ATCC 35150, E. coli KCTC 2571, L. monocytogenes ATCC 51776, and S. aureus ATCC 25923). Moreover, the strain showed approximately 40% 2,2-diphenyl-1-picryl-hydrazyl (DPPH) radical- and 10% 2-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) radical-scavenging activity. In cell culture studies, human colon epithelial cells (Caco-2) were treated with LPLB5 (106 and 107 CFU/mL); the bacteria showed more than 70% adherence onto and a 32% invasion rate into the Caco-2 cells. LPLB5 significantly decreased the mRNA expression levels of pro-inflammatory cytokines (interleukin-1 beta (IL-1ß), interleukin 6 (IL-6), and tumor necrosis factor-alpha (TNF-α)) and increased the mRNA expression levels of anti-inflammatory cytokines (interleukin-4 (IL-4), interleukin-10 (IL-10), and interferon-gamma (IFN-γ)) in lipopolysaccharide-stimulated Caco-2 cells. Our data suggest that LPLB5 is safe and possesses probiotic, antioxidant, and anti-inflammatory activities.

Aluminum exposure promotes the metastatic proclivity of human colorectal cancer cells through matrix metalloproteinases and the TGF-ß/Smad signaling pathway.

Human exposure to aluminum (Al) mainly occurs through food intake. However, influences of Al on the gastrointestinal tract have been rarely reported. In particular, the effect of Al on the metastasis and angiogenesis of colorectal cancer cells has not been studied. Thus, we investigated the effect of Al on the metastatic proclivity using the human colorectal cancer cell line, HT-29. Cells were exposed to 1-16 mM AlCl3 for 3-72 h. The effects of AlCl3 on HT-29 cells for migration/invasion/adhesion, and metastasis-associated protein and gene expression were evaluated. AlCl3 promoted cell migration and invasion, whereas it suppressed cell adhesion. AlCl3-exposed cells showed decreased E-cadherin and increased vimentin and Snail. AlCl3 increased transforming growth factor-beta (TGF-ß) mRNA expression and Smad2/3 nuclear translocation. AlCl3-treated cells had a higher mRNA expression of matrix metalloproteinase (MMP)-7 and -9 than the control. Particularly, AlCl3-treated HT-29 cells promoted the angiogenesis of endothelial cells via increasing the secretion of vascular endothelial growth factor. Taken together, AlCl3 can promote the metastatic proclivity of colorectal cancer cells through MMP-7, -9, and TGF-ß/Smad2/3 pathway. Our data suggest that Al exposure of the gastrointestinal tract may be a risk factor for metastasis initiation in colorectal cancer cells.

Effects of Aluminum on the Integrity of the Intestinal Epithelium: An in Vitro and in Vivo Study.

BACKGROUND: Aluminum (Al) is the most abundant and ubiquitous metal in the environment. The main route of human exposure to Al is through food and water intake. Although human exposure to Al is common, the influence of Al on the gastrointestinal tract remains poorly understood. OBJECTIVES: We aimed to further understand the toxic effect of Al and to elucidate the underlying cellular mechanisms in the intestinal barrier. METHODS: The human intestinal epithelial cell line HT-29 and C57BL6 mice were exposed to AlCl3 at 0-16 mM (1-24h) and 5-50mg/kg body weight (13 weeks), respectively. In cell culture experiments, intracellular oxidative stress, inflammatory protein and gene expression, and intestinal epithelial permeability were measured. In animal studies, histological examination, gene expression, and myeloperoxidase (MPO) activity assays were conducted. RESULTS: Cellular oxidative stress level (superoxide production) in AlCl3-treated cells (4 mM, 3h) was approximately 38-fold higher than that of the control. Both protein and mRNA expression of tight junction (TJ) components (occludin and claudin-1) in AlCl3-treated cells (1-4 mM, 24h) was significantly lower than that of the control. Transepithelial electrical resistance (TEER) decreased up to 67% in AlCl3-treated cells (2 mM, 24h) compared with that of the control, which decreased approximately 7%. Al activated extracellular signal-regulated kinase 1/2 and nuclear factor-kappa B (NF-κB), resulting in mRNA expression of matrix metalloproteinase-9, myosin light-chain kinase, and inflammatory cytokines [tumor necrosis factor alpha (TNF-α), interleukin-1ß (IL-1ß), and IL-6] in HT-29 cells. Moreover, oral administration of AlCl3 to mice induced pathological alteration, MPO activation, and inflammatory cytokine (TNF-α, IL-1ß, and IL-6) production in the colon. CONCLUSION: Al induced epithelial barrier dysfunction and inflammation via generation of oxidative stress, down-regulation of the TJ proteins, and production of inflammatory cytokines in HT-29 cells. In addition, Al induced toxicity in the colon by increasing the levels of inflammatory cytokines and MPO activity and induced histological damage in a mouse model. Our data suggest that Al may be a potential risk factor for human intestinal diseases. https://doi.org/10.1289/EHP5701.

Zearalenone Induces Endoplasmic Reticulum Stress and Modulates the Expression of Phase I/II Enzymes in Human Liver Cells.

Zearalenone (ZEN) is a mycotoxin produced by Fusarium species; however, its mechanisms of action in human livers have not been fully elucidated. Thus, we investigated the toxic mechanisms of ZEN in human liver cells. HepG2 cells were treated with ZEN (0-40 µg/mL) for up to 24 h. A significant decrease in cell viability was observed after treatment with 20 and 40 µg/mL of ZEN, including a significant increase in apoptosis and reactive oxygen species production. ZEN increased GRP78 and CHOP, and eIF2α phosphorylation, indicating ER stress; elevated transcription of the autophagy-associated genes, beclin1 and LC3, and translation of LC3; and increased phase I metabolism by increasing PXR and CYP3A4. The protein expression level of CYP3A4 was higher with ZEN treatment up to 20 µg/mL, but remained at the control level after treatment with 40 µg/mL ZEN. In phase II metabolism, Nrf2 activation and UGT1A expression were increased with ZEN treatment up to 20 µg/mL. Treating cells with an ER stress inhibitor alleviated ZEN-induced cell death and autophagy, and inhibited the expression of phase I/II enzymes. Overall, high ZEN concentrations can modulate the expression of phase I/II enzymes via ER stress and reduced protein levels in human liver cells.

Piperlongumine Induces Cell Cycle Arrest via Reactive Oxygen Species Accumulation and IKKß Suppression in Human Breast Cancer Cells.

Piperlongumine (PL), a natural product derived from long pepper (Piper longum L.), is known to exhibit anticancer effects. However, the effect of PL on cell cycle-regulatory proteins in estrogen receptor (ER)-positive breast cancer cells is unclear. Therefore, we investigated whether PL can modulate the growth of ER-positive breast cancer cell line, MCF-7. We found that PL decreased MCF-7 cell proliferation and migration. Flow cytometric analysis demonstrated that PL induced G2/M phase cell cycle arrest. Moreover, PL significantly modulated the mRNA levels of cyclins B1 and D1, cyclin-dependent kinases 1, 4, and 6, and proliferating cell nuclear antigen. PL induced intracellular reactive oxygen species (hydrogen peroxide) accumulation and glutathione depletion. PL-mediated inhibition of IKKß expression decreased nuclear translocation of NF-κB p65. Furthermore, PL significantly increased p21 mRNA levels. In conclusion, our data suggest that PL exerts anticancer effects in ER-positive breast cancer cells by inhibiting cell proliferation and migration via ROS accumulation and IKKß suppression.

Technical Functional Properties of Water- and Salt-soluble Proteins Extracted from Edible Insects.

The amino acid composition, protein quality, and protein functionality of protein solution extracted from three edible insect species were investigated. We used 0.02% ascorbic acid and 0.58 M saline solution to extract water-soluble and salt-soluble proteins from the three insect species. Extracted protein solutions of Tenebrio molitor (TM), Allomyrina dichotoma (AD), and Protaetia brevitarsis seulensis (PB) were divided into six groups, according to species and solubility: WTM, WAD, WPB (water-soluble), and STM, SAD, and SPB (salt-soluble). Defatted TM had the highest protein content, but its protein solubility was the lowest, for both water and saline solutions. Amino acid composition differed by edible insect species and buffer type; SPB had the highest protein quality, followed by WPB. PB had a higher pH than the other species. Color values also differed among species. SPB had abundant high molecular weight proteins, compared with other treatments; and also had the highest foaming capacity, foam stability, and emulsifying capacity. In conclusion, PB is a good source of functional protein compared with the other studied species. Additionally, protein extraction using saline solution is promising as a useful method for improving edible insect protein functionality.

Moringa Extract Attenuates Inflammatory Responses and Increases Gene Expression of Casein in Bovine Mammary Epithelial Cells.

Bovine mastitis is a common inflammatory disease in the udder of dairy cows that causes economic loss to dairy industries. The development of alternative strategies, especially the utilization of natural products, e.g. Moringa oleifera, has gained a lot of interests. The objective of the current study was to investigate the protective effects of moringa extract (ME) in bovine mammary epithelial cells (MAC-T) in in vitro settings. Radical scavenging capacities and anti-inflammatory properties of ME were examined using lipopolysaccharide (LPS)-challenged MAC-T cells. ME showed significant radical scavenging activities. In addition, ME decreased reactive oxygen species produced by LPS in cells. ME also attenuated inflammatory cyclooxygenase-2 expression induced by LPS by down-regulating NF-κB signaling cascade. Moreover, ME ameliorated LPS-induced pro-inflammatory cytokines including tumor necrosis factor-, interleukin-1, and interleukin-6. Furthermore, ME up-regulated mRNA expression levels of heme oxygenase-1, NAD(P)H: quinone oxidoreductase-1, and thioredoxin reductase 1. Importantly, ME promoted differentiated MAC-T cells by increasing mRNA expression levels of α-casein S1, α-casein S2, and ß-casein. In conclusion, ME has beneficial effects in bovine mammary epithelial cells through its anti-inflammatory, antioxidant, and casein production properties. Our study provides evidence that ME could be a good candidate for a feed supplement to decrease inflammatory responses due to bovine mastitis.