Current status and challenges for cell-cultured milk technology: a systematic review.

Cellular agriculture is an innovative technology for manufacturing sustainable agricultural products as an alternative to traditional agriculture. While most cellular agriculture is predominantly centered on the production of cultured meat, there is a growing demand for an understanding of the production techniques involved in dairy products within cellular agriculture. This review focuses on the current status of cellular agriculture in the dairy sector and technical challenges for cell-cultured milk production. Cellular agriculture technology in the dairy sector has been classified into fermentation-based and animal cell culture-based cellular agriculture. Currently, various companies synthesize milk components through precision fermentation technology. Nevertheless, several startup companies are pursuing animal cell-based technology, driven by public concerns regarding genetically modified organisms in precision fermentation technology. Hence, this review offers an up-to-date exploration of animal cell-based cellular agriculture to produce milk components, specifically emphasizing the structural, functional, and productive aspects of mammary epithelial cells, providing new information for industry and academia.

The Role of Progesterone in Elf5 Activation and Milk Component Synthesis for Cell-Cultured Milk Production in MAC-T Cells.

Prolactin is essential for mammary gland development and lactation. Progesterone also induces ductal branching and alveolar formation via initial secretory differentiation within the mammary gland. Herein, we aimed to evaluate the role of progesterone as a prolactin substitute for the production of cell-cultured milk components in MAC-T cells. Cells were treated with various hormones such as prolactin (PRL), progesterone (P4), 17ß-estradiol (E2), cortisol (COR), and insulin (INS) for 5 d. MAC-T cells cultured in a P4 differentiation media (2500 ng/mL of P4, 25 ng/mL of E2, 25 ng/mL of COR, and 25 ng/mL of INS) showed similar levels of E74-like factor 5 (Elf5) and milk component synthesis (α-casein, ß-casein, α-lactalbumin, ß-lactoglobulin, and triglycerides) compared to those cultured in a PRL differentiation media (5000 ng/mL of PRL, 500 ng/mL of CORT, and 50 ng/mL of INS). The levels of α-casein and triglycerides in the optimal P4 differentiation media were present at comparable levels to those in the PRL differentiation media. Our results demonstrated that P4 induces the activation of Elf5 and the synthesis of milk components in MAC-T cells, similar to PRL. Therefore, P4 may be used as an effective substitute of PRL for cell-cultured milk production in in vitro frameworks.

Physicochemical properties of Pickering emulsion fabricated with polysaccharides/pea protein isolate complex and its application in plant-based patty.

Incorporation of structured liquid oil within plant-based patties can be achieved through the utilization of food-grade Pickering emulsion (PE). Therefore, the aim of this study was to evaluate the quality characteristics of PE and its application in plant-based patty. The PEs were formulated using sunflower oil (SO), polysaccharides and protein, and the specific ratios employed were as following: methylcellulose (MC) 2 % only (MP0); MC 1.5 % + pea protein isolate (PPI) 0.5 % (MP1); MC 1 % + PPI 1 % (MP2); xanthan gum (XG) 2 % only (XP0); XG 1.5 % + PPI 0.5 % (XP1); XG 1 % + PPI 1 % (XP2). MP0 and MP1 were unstable as PEs, whereas MP2 and XP groups (XP0, XP1, and XP2) exhibited stability as a PE. In addition, MP2 and all XP groups showed increased oil binding capacity, hydrophobic interaction, thermal stability, crystallization, rheological properties, and oxidative stability, compared to MP0 and MP1. In PE-applied plant-based patties, MP2 and all XP groups had significantly lower cooking loss and higher emulsion stability than SO. Particularly, MP2-employed plant-based patties exhibited significantly improved textural and sensory properties. Therefore, our data suggest that PEs with methylcellulose and pea protein isolate could be an effective replacement of plant oil in plant-based meat analogs.

Comparative Evaluation of Polysaccharide Binders on the Quality Characteristics of Plant-Based Patties.

Polysaccharides have been used in the production of plant-based meat analogs to replicate the texture of real meat. However, there has been no study that comprehensively compares the effects of different polysaccharides, and a limited number of polysaccharides have been evaluated. Thus, we aimed to identify the most suitable polysaccharide and concentration for plant-based patties. Plant-based patties were manufactured by blending different concentrations (0%, 1%, and 2%) of six polysaccharides with other ingredients, and the quality characteristics and sensory properties were evaluated. The L* values of plant-based patties reduced during the cooking process resembled the color change of beef patty (BP). In particular, a 2% κ-carrageenan-added patty (Car-2) exhibited the lowest L* value among the plant-based patties, measured at 44.05 (p < 0.05). Texture parameters exhibited high values by adding 2% κ-carrageenan and locust bean gum, which was close to BP. In the sensory evaluation, Car-2 showed higher scores for sensory preferences than other plant-based patties. Based on our data, incorporating 2% κ-carrageenan could offer a feasible way of crafting plant-based meat analogs due to its potential to enhance texture and flavor. Further studies are required to evaluate the suitability of polysaccharides in various types of plant-based meat analogs.

In Vitro Synergistic Antibacterial and Anti-Inflammatory Effects of Nisin and Lactic Acid in Yogurt against Helicobacter pylori and Human Gastric Cells.

Helicobacter pylori is a bacterium that naturally thrives in acidic environments and has the potential to induce various gastrointestinal disorders in humans. The antibiotic therapy utilized for treating H. pylori can lead to undesired side effects, such as dysbiosis in the gut microbiota. The objective of our study was to explore the potential antibacterial effects of nisin and lactic acid (LA) in yogurt against H. pylori. Additionally, we investigated the anti-inflammatory effects of nisin and LA in human gastric (AGS) cells infected with H. pylori. Nisin and LA combination showed the strongest inhibitory activity, with confirmed synergy at 0.375 fractional inhibitory concentration index. Also, post-fermented yogurt with incorporation of nisin exhibited antibacterial effect against H. pylori. The combination of nisin and LA resulted in a significant reduction of mRNA levels of bacterial toxins of H. pylori and pro-inflammatory cytokines in AGS cells infected with H. pylori. Furthermore, this also increased bacterial membrane damage, which led to DNA and protein leakage in H. pylori. Overall, the combination of nisin and LA shows promise as an alternative therapy for H. pylori infection. Additionally, the incorporation of nisin into foods containing LA presents a potential application. Further studies, including animal research, are needed to validate these findings and explore clinical applications.

Optimizing hormonal and amino acid combinations for enhanced cell proliferation and cell cycle progression in bovine mammary epithelial cells.

OBJECTIVE: The number of bovine mammary epithelial cells (BMECs) is closely associated with the quantity of milk production in dairy cows; however, the optimal levels and the combined effects of hormones and essential amino acids (EAAs) on cell proliferation are not completely understood. Thus, the purpose of this study was to determine the optimal combination of individual hormones and EAAs for cell proliferation and related signaling pathways in BMECs. METHODS: Immortalized BMECs (MAC-T) were treated with six hormones (insulin, cortisol, progesterone, estrone, 17ß-estradiol, and epidermal growth factor) and ten EAAs (arginine, histidine, leucine, isoleucine, threonine, tryptophan, lysine, methionine, phenylalanine, and valine) for 24 h. RESULTS: Cells were cultured in a medium containing 10% fetal bovine serum (FBS) as FBS supplemented at a concentration of 10% to 50% showed a comparable increase in cell proliferation rate. The optimized combination of four hormones (insulin, cortisol, progesterone, and 17ß-estradiol) and 20% of a mixture of ten EAAs led to the highest cell proliferation rate, which led to a significant increase in cell cycle progression at the S and G2/M phases, in the protein levels of proliferating cell nuclear antigen and cyclin B1, cell nucleus staining, and in cell numbers. CONCLUSION: The optimal combination of hormones and EAAs increased BMEC proliferation by enhancing cell cycle progression in the S and G/2M phases. Our findings indicate that optimizing hormone and amino acid levels has the potential to enhance milk production, both in cell culture settings by promoting increased cell numbers, and in dairy cows by regulating feed intake.

Effect of sous-vide cooking conditions on the physicochemical, microbiological and microstructural properties of duck breast meat.

OBJECTIVE: Sous-vide cooking offers several advantages for poultry meat, including enhanced tenderness, reduced cooking loss, and improved product yield. However, in duck meat, there are challenges associated with using the sous-vide method. The prolonged cooking time at low temperatures can lead to unstable microbial and oxidative stabilities. Thus, we aimed to assess how varying sous-vide cooking temperatures and durations affect the physicochemical and microbial characteristics of duck breast meat, with the goal of identifying an optimal cooking condition. METHODS: Duck breast meat (Anas platyrhynchos) aged 42 days and with an average weight of 1,400±50 g, underwent cooking under various conditions (ranging from 50°C to 80°C) for either 60 or 180 min. Then, physicochemical, microbial, and microstructural properties of the cooked duck breast meat were assessed. RESULTS: Different cooking conditions affected the quality attributes of the meat. The cooking loss, lightness, yellowness, Hue angle, whiteness, and thiobarbituric acid reactive substance (TBARS) values of the duck breast meat increased with the increase in cooking temperature and time. In contrast, the redness and chroma values decreased with the increase in cooking temperature and time. Cooking of samples higher than 60°C increased the volatile basic nitrogen contents and TBARS. Microbial analysis revealed the presence of Escherichia coli and Coliform only in the samples cooked at 50°C and raw meat. Cooking at lower temperature and shorter time increased the tenderness of the meat. Microstructure analysis showed that the contraction of myofibrils and meat density increased upon increasing the cooking temperature and time. CONCLUSION: Our data indicate that the optimal sous-vide method for duck breast meat was cooking at 60°C for 60 min. This temperature and time conditions showed good texture properties and microbial stability, and low level of TBARS of the duck breast meat.

Cinnamaldehyde-Rich Cinnamon Extract Induces Cell Death in Colon Cancer Cell Lines HCT 116 and HT-29.

Cinnamon is a natural spice with a wide range of pharmacological functions, including anti-microbial, antioxidant, and anti-tumor activities. The aim of this study is to investigate the effects of cinnamaldehyde-rich cinnamon extract (CRCE) on the colorectal cancer cell lines HCT 116 and HT-29. The gas chromatography mass spectrometry analysis of a lipophilic extract of cinnamon revealed the dominance of trans-cinnamaldehyde. Cells treated with CRCE (10-60 µg/mL) showed significantly decreased cell viability in a time- and dose-dependent manner. We also observed that cell proliferation and migration capacity were inhibited in CRCE-treated cells. In addition, a remarkable increase in the number of sub-G1-phase cells was observed with arrest at the G2 phase by CRCE treatment. CRCE also induced mitochondrial stress, and finally, CRCE treatment resulted in activation of apoptotic proteins Caspase-3, -9, and PARP and decreased levels of mu-2-related death-inducing gene protein expression with BH3-interacting domain death agonist (BID) activation.

Apoptosis-Inducing Effects of Short-Chain Fatty Acids-Rich Fermented Pistachio Milk in Human Colon Carcinoma Cells.

Pistachio milk (PM), an extraction product of pistachio, is protein- and fat-dense food. Short-chain fatty acids (SCFAs) are known for inducing cytotoxicity and apoptosis in colon carcinoma cells. This study aimed to find an optimal combination of probiotics that can produce a higher amount of SCFAs in PM. In addition, the anti-cancer effect of fermented PM on human colon carcinoma cells (Caco-2) was determined. The combinations of probiotics were as follows: Streptococcus thermophilus + Lactobacillus bulgaricus (C); C + Lactobacillus acidophilus (C-La); C + Lactobacillus gasseri (C-Lg); C + Bifidobacterium bifidum (C-Bb). The results indicated that fermented PM was produced after a short fermentation time in all the probiotics combinations. C-Bb produced up to 1.5-fold more acetate than the other probiotics combinations did. A significant amount of cytotoxicity, i.e., 78, 56, and 29% cell viability was observed in Caco-2 cells by C-Bb-fermented PM at 1, 2.5 and 5%, respectively. C-Bb-fermented PM (5%) induced early and late apoptosis up to 6-fold. Additionally, Caco-2 cells treated with C-Bb-fermented PM significantly induced the downregulation of α-tubulin and the upregulation of cleaved caspase-3, as well as nuclear condensation and fragmentation. Our data suggest that fermented PM, which is rich in acetate, may have the potential as a functional food possessing anti-colon cancer properties.

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.

Development of ß-Cyclodextrin/Konjac-Based Emulsion Gel for a Pork Backfat Substitute in Emulsion-Type Sausage.

Emulsion gel has been used to replace animal fats in meat products. Konjac is a widely used gelling agent; however, its low emulsion stability limits its use in meat products. This study aimed to examine the quality characteristics of ß-cyclodextrin (CD)-supplemented konjac-based emulsion gel (KEG) (CD-KEG) and its application as a fat substitute in emulsion-type sausages. The supplementation of CD increased hydrogen bonds and hydrophobic interactions with konjac and oil in the gels, respectively. Additionally, CD increased the structural complexity and strength of KEG. Since adding more than 6% of CD to KEG did not increase the gel strength, 6% CD-added KEG was adopted to substitute for pork backfat in manufacturing low-fat emulsion-type sausages. The following formulations of the sausages were prepared: pork backfat 20% (PF20); pork backfat 10% + KEG 10% (KEG10); KEG 20% (KEG20); pork backfat 10% + CD-KEG 10% (CD-KEG10); CD-KEG 20% (CD-KEG20); and pork backfat 5% (PF5). The CD-KEG20 formulation exhibited higher viscosity and viscoelasticity than KEG20, which suggested that CD improves the rheological properties and the thermal stability of meat batter. Additionally, CD-KEG20 showed similar emulsion stability, cooking yield and texture parameters compared with PF20. Therefore, 6% CD-added KEG is a suitable fat substitute for preparing low-fat emulsion-type sausages.

Drying Characteristics and Physicochemical Properties of Semi-Dried Restructured Sausage Depend on Initial Moisture Content.

Semi-dried restructured sausages are restructured meat products with a high nutritional and economic value. However, excessively long drying times can have negative effects on the energy consumption, texture, and sensory properties of semi-dried restructured sausages. The objective of this study was to investigate the effects of different water contents on the drying and physicochemical characteristics of semi-dried restructured sausages. Sausages were prepared with different initial moisture contents (0%-50%) and drying time (0-580 min). The drying characteristics, including the drying rate, effective moisture diffusivity, and water activity of sausage were significantly improved as the initial moisture content was increased. When the initial moisture content of the sausage was 50%, physicochemical properties, such as color, porosity, shear force, and volatile basic nitrogen, were improved the most along with the decreased drying time. Scanning electron microscopy data showed greater porosity and pore size in sausages with the increase of initial moisture content. Collectively, our data suggest that an increase in the initial moisture content of semi-dried restructured sausages improves their drying characteristics and physicochemical properties.

On-Site Deployment of an Air-Liquid-Interphase Device to Assess Health Hazard Potency of Airborne Workplace Contaminants: The Case of 3-D Printers.

Biomonitoring of workers is an approach of evaluating workers' exposure to chemicals and particulate matter by measuring biomarkers of parent chemicals, their metabolites, and reaction products in workers' biospecimens. Prerequisites for biological monitoring in the workplace include permission to enter the workplace, approval of the study plan from the IRB (Institutional Review Board), and obtaining consent from workers. Because of the complex legal process involved in biomonitoring, few studies have been conducted so far on biomonitoring of workers' exposures to nanoparticles and other hazards from emerging materials and advanced nanotechnologies. We have developed a cell-based biomonitoring device that can evaluate acute cytotoxicity and various other effect biomakers, such as inflammation, at realistic workplace exposure. This device is based on air-liquid interphase (ALI) and can be used to evaluate cell toxicity and early effect biomarkers along adverse outcome pathways. Following exposure of A549 lung epithelial cells in ALI to workplace air for 1-2 h, the cells were processed to assess the induction of inflammatory and cell damage biomarkers. Initially, we estimated the deposition rate of nanoparticles in the transwell by exposing the cell-free ALI device to silver nanoparticle aerosols (AgNP 20-30 nm) for 2 h in the laboratory. Then A549 lung epithelial cells cultured on the transwell in the ALI device were exposed to AgNP nanoaerosols for 2 h and evaluated for cytotoxicity and induction of mRNAs of pro-inflammatory cytokines IL-1b, IL-6, and TNF-α. Then the cells in the ALI device were exposed to 3-D printer emissions at the workplace and evaluated for the same matched endpoints. The mRNA levels for IL-1b, IL-6, and TNF-α increased significantly at the end of 2-h exposure of A549 cells to the positive control AgNP aerosols. These mRNAs, as well as LDH and microprotein concentrations, increased even more after 24-h post-exposure incubation (p < 0.05). Cytotoxicity evaluation of 3-D printer emissions at 810 and 957 µg/m3, which was more than 80 times higher than the airborne total suspended particulate concentrations in the workplace air (9-12.5 µg/m3), suggested no significant acute cytotoxicity at the end of 2-h exposure to 3-D-printing emission, as well as at 24-h post-exposure incubation. Hyperspectral microscopic observation showed that 3-D printers emitted particles to be attached to A549 cells after 2-h exposure, and many particles were internalized by A549 cells after 24 h of post-exposure incubation. The mRNA expression of pro-inflammatory cytokine IL-1b and IL-6 increased significantly after 2-h exposure to 3-D printer emissions and after 24-h incubation (only IL-6). In contrast, the expression of TNF-α mRNA decreased significantly after 2 h of exposure to 3-D printers and decreased even more after 24-h post-exposure incubation. These results support the use of cell-based ALI devices for direct assessment of airborne hazards in the workplace, for probing toxicological properties of airborne contaminants using adverse molecular pathways, and for guiding study design for workplace biomonitoring. ALI devices can bridge conventional exposure assessment with cellular toxicity testing platforms for hazard and risk assessment.

Effects of Chitosan and Duck Fat-Based Emulsion Coatings on the Quality Characteristics of Chicken Meat during Storage.

Chicken meat is a popular food commodity that is widely consumed worldwide. However, the shelf-life or quality maintenance of chicken meat is a major concern for industries because of spoilage by microbial growth. The aim of this study was to evaluate the effects of chitosan and duck fat-based emulsion coatings on the quality characteristics and microbial stability of chicken meat during refrigerated storage. The coated chicken meat samples were as follows: control (non-coated), DFC0 (coated with duck fat), DFC0.5 (coated with duck fat and 0.5% chitosan), DFC1 (coated with duck fat and 1% chitosan), DFC2 (coated with duck fat and 2% chitosan), and SOC2 (coated with soybean oil and 2% chitosan). The results showed that the apparent viscosity and coating rate were higher in DFC2 than in other groups. Physicochemical parameters (pH, color, and Warner-Bratzler shear force) were better in DFC2 than those in other groups during 15 days of storage. Moreover, DFC2 delayed lipid oxidation, protein deterioration, and growth of microorganisms during storage. These data suggest that chitosan-supplemented duck fat-based emulsion coating could be used to maintain the quality of raw chicken meat during refrigerated storage.

Effect of Different Brine Injection Levels on the Drying Characteristics and Physicochemical Properties of Beef Jerky.

Meat jerky is a type of meat snack with a long shelf life, light weight, and unique sensory properties. However, meat jerky requires a long manufacturing time, resulting in high energy consumption. In this study, beef jerky was prepared by injecting different concentrations of brine at different hot-air drying times (0-800 min). When the brine injection levels were increased to 30%, the drying characteristics of beef jerky, such as drying time and effective moisture diffusivity, were significantly improved owing to the relatively high water content and the formation of porous structures. The physicochemical properties (e.g. meat color, porosity, shear force, and volatile basic nitrogen) of the beef jerky injected with 30% brine were improved owing to the shortened drying time. Scanning electron microscopy images showed that the beef jerky structure became porous and irregular during the brine injection process. Our novel processing technique for manufacturing beef jerky leads to improved quality characteristics and shortened drying times.

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.

Toxicity Assessment of a Single Dose of Poly(ethylene glycol) Diglycidyl Ether (PEGDE) Administered Subcutaneously in Mice.

Poly(ethylene glycol) diglycidyl ether (PEGDE) is widely used to cross-link polymers, particularly in the pharmaceutical and biomaterial sectors. However, the subcutaneous toxicity of PEGDE has not yet been assessed. PEGDE samples (500-40,000 µg/mouse) were subcutaneously injected into the paraspinal dorsum of BALB/c male mice. Cage-side observations were carried out with measurement of organ weight, body weight variation, and feed intake, as well as histopathological characterization on day 28 post-exposure. Mice that received 40,000 µg of PEGDE showed severe toxic response and had to be euthanized. Subcutaneous injection of PEGDE did not alter feed intake and organ weight; however, the body weight variation of mice injected with 20,000 µg of PEGDE was significantly lower than that of the other groups. Exposure to 10,000 and 20,000 µg of PEGDE induced epidermal ulcer formation and hair loss. The histology of skin tissue in mice administered with 20,000 µg of PEGDE showed re-epithelialized or unhealed wounds. However, the liver, spleen, and kidneys were histologically normal. Collectively, PEGDE, particularly above 10,000 µg/mouse, caused subcutaneous toxicity with ulceration, but no toxicity in the other organs. These results may indicate the optimal concentration of subcutaneously injected PEGDE.

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.

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.