Digestive characteristics of astaxanthin oil in water emulsion stabilized by a casein-caffeic acid-glucose ternary conjugate.

To overcome the barrier of poor oral bioavailability of astaxanthin, a stable oil-in-water emulsion was constructed using casein-caffeic acid-glucose ternary conjugates (CSC) to deliver astaxanthin, and its gastrointestinal behavior was evaluated in vitro with sodium caseinate (CSN) as a control. Results showed that, CSC-stabilized emulsion shower better resistance to the adverse conditions of the gastric environment than CSN-stabilized emulsion, and exhibited lower average particle size and aggregation (4972.33 nm, -5.93 mv) after simulated gastric digestion. Besides, after simulated intestinal digestion, the reducing capacity and astaxanthin transfer efficiency of CSC emulsion into the micellar phase were 686.74 µmol Trolox/100 mL and 26.2 %, which were 2.6 and 4.05-fold higher than that of CSN emulsion. The above results suggest that CSC can be used for better delivery of astaxanthin, which could be useful in designing foods such as functional beverages, pharmaceuticals and nutritional supplements for delivery of lipophilic bioactives.

The effects of mulberry polyphenols on the digestibility and absorption properties of pork myofibrillar protein in vitro.

The objective of the present study was to explore the effect of mulberry polyphenols on the digestibility and absorption properties of myofibrillar protein (MP) in vitro. MP was extracted from the Longissimus et thoracis muscle of 18 different pig carcasses and the MP-mulberry polyphenols complex was prepared. The antioxidant activity of digestive juice, degradation of both MP and polyphenols, and the metabolism of MP and the MP-polyphenols complex by intestinal microbial activity during digestion and fermentation in vitro were compared. The results showed that mulberry polyphenols significantly affect the digestibility of MP and the antioxidant activity of digestive juices during digestion (P < 0.05). After the modification of the polyphenols, the hydrolysis of MP increased from 55.4% to 64.0%, and the molecular weight of protein digestion product significantly decreased (P < 0.05). The scavenging rates of 2, 2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) and 2,2-diphenyl-1-picrylhydrazyl in the final digestive juice were 350.1 µmol Trolox/mg protein and 34.0%, respectively, which were 0.34 and 0.47-fold higher than those of the control (P < 0.05). Furthermore, the release and degradation of phenolic compounds mainly occurred during intestinal digestion, and polyphenols that reached the colon after digestion, through the fermentation of intestinal microorganisms in vitro, enriched Lactobacillus and promoted the production of short-chain fatty acids which has obvious potential to improve intestinal health.

Feeding Aquilaria sinensis Leaves Modulates Lipid Metabolism and Improves the Meat Quality of Goats.

Aquilaria (A.) sinensis is a medicinal plant widely grown in tropical South China. Given the abundant pruning waste of its leaves, the use of A. sinensis leaves is valuable. In this study, goats were fed a diet containing 20% A. sinensis leaves. Compared with the basal diet, feeding A. sinensis leaves to goats did not affect growth performance but considerably reduced the feeding cost. Strikingly, feeding A. sinensis leaves resulted in a significant decrease in the blood cholesterol levels (2.11 vs. 1.49 mmol/L, p = 0.01) along with a significant increase in the high-density lipoprotein levels (1.42 vs. 1.82 mmol/L, p = 0.01). There was also a tendency to lower the content of low-density lipoprotein levels in goats (0.78 vs. 0.45 mmol/L, p = 0.09). Furthermore, metabolomics analysis demonstrated that the reduction in cholesterol levels occurred in both the serum (0.387-fold change) and muscle (0.382-fold change) of goats during A. sinensis leaf feeding. The metabolic responses to feeding A. sinensis leaves suggest that the activation of lipolysis metabolism might happen in goats. These observed changes would be conducive to improving animal health and meat quality, ultimately benefiting human health.

Improved physicochemical stability and bioaccessibility of astaxanthin-loaded oil-in-water emulsions by a casein-caffeic acid-glucose ternary conjugate.

In this study, the influence of casein-caffeic acid-glucose ternary conjugate (CSC) on the physicochemical properties and bioaccessibility of astaxanthin-loaded emulsion was investigated and compared with sodium caseinate (CSN), a synthetic emulsifier commonly used in the food industry. The CSC-stabilized emulsion exhibits droplet characteristics similar to CSN-stabilized emulsion, and can effectively resist the external forces that lead to the phase separation of the emulsion. Although phase separation also occurred at pH 4.0, CSC emulsion had a wider range of pH stability (pH 3.0, 5.0-8.0) and higher salt ion stability than CSN emulsion. Furthermore, CSC-stabilized astaxanthin emulsions showed better astaxanthin protection under different heat treatment conditions and storage temperatures compared with CSN. After 28 days of storage at 4 °C, astaxanthin residues in the CSC-stabilized emulsion reached 92.37 %. The bioaccessibility of astaxanthin in CSC-stabilized emulsion was 26.21 %, much higher than that in CSN (6.47 %). This research study provides a platform for designing astaxanthin-fortified food or beverage systems to achieve better stability and delivery to target sites.

Influence of Gelatin-Chitosan-Glycerol Edible Coating Incorporated with Chlorogenic Acid, Gallic Acid, and Resveratrol on the Preservation of Fresh Beef.

Chlorogenic acid (CA), gallic acid (GA), and resveratrol (RES) were added to a gelatin (GEL)-chitosan (CHI)-glycerol (GLY) edible coating, and their effects on the coating of fresh beef preservation were investigated. The results revealed that CA had the most significant improvement effect on fresh beef preservation. The combination of GEL-CHI-GLY-CA preserved the color of the beef better and delayed the increase of the total volatile base nitrogen, even though its total phenolic content decreased at a faster rate during beef preservation. GA also improved the preservation effect as on the 12th day of storage, the beef samples treated with GEL-CHI-GLY-GA had the lowest thiobarbituric acid reactive substances (0.76 mg Malondialdehyde (MDA)/kg) and total viable count (6.0 log cfu/g). On the whole, though RES showed an improvement on beef preservation, the improvement was not as good as the other two polyphenols. After 12 days of storage, the beef samples treated with GEL-CHI-GLY-RES had a higher pH value (6.25) than the other two polyphenol treatmed groups. Overall, the three polyphenol-added combinations increased the shelf life of beef by approximately 3-6 days compared to the control group (treated GEL-CHI-GLY with distilled water).

Source Tracker Modeling Based on 16S rDNA Sequencing and Analysis of Microbial Contamination Sources for Pasteurized Milk.

Milk is rich in fat, protein, minerals, vitamins, peptides, immunologically active substances, and other nutrients, and it plays an important role in satisfying human nutrition and health. However, dairy product safety incidents caused by microbial contamination have occurred. We found that the total bacterial numbers in the pasteurized product were low and far below the limit requirements of the food safety standards of the European Union, the United States, and China. At the genus level, the primary microbial groups found in milk samples were Acinetobacter, Macrococcus, Pseudomonas, and Lactococcus, while in the equipment rinse water and air samples there was contamination by Stenotrophomonas and Acinetobacter. The Source Tracker model analysis indicated that the microorganisms in the final milk products were significantly related to the contamination in product tanks and raw milk. Therefore, it is the hope that this work can provide guidance to pinpoint contamination problems using the proper quality control sampling at specific stages in the pasteurization process.

Effects of mulberry pomace on physicochemical and textural properties of stirred-type flavored yogurt.

Adding functional ingredients is an important method to develop functional dairy products. Mulberry pomace (MPo), a byproduct of mulberry fruit processing, is rich in phenolic compounds and anthocyanins and can be served as the functional ingredient in functional dairy products. The aim of this work was to prepare a functional flavored yogurt by incorporating MPo into stirred yogurt and to investigate the effects of MPo on the physicochemical and textural properties of the product during cold storage. We supplemented MPo powder up to 3% (wt/wt) in fermented milk, and the changes in color, pH, titratable acidity (TA), total phenol content (TPC), total anthocyanin content (TAC), water-holding capacity, rheological behavior, texture, and microstructure of the functional flavored yogurt were monitored during storage under 4°C for 28 d. The MPo powder brought a pink to dark red color to the yogurt, decreased the lightness (L*) and yellow-blue color (b*) values, increased the red-green color (a*) values, decreased the pH value, and increased the contents of TA, TPC, and TAC in a dose-dependent manner. The addition of MPo at 1%, 2%, and 3% (wt/wt) significantly increased water-holding capacity, consistency, viscosity, and viscosity index, and reduced firmness of yogurt samples. Supplementation of MPo significantly reduced the pore spaces and channels inside the samples and improved microstructure of the functional yogurt. During the 28 d of cold storage, MPo-fortified yogurt samples kept relatively constant color, although their L*, a*, and b* showed a decreasing tendency. The pH of all yogurt samples gradually decreased with increasing of TA. Interestingly, TPC and TAC contents and the texture parameters of MPo-fortified yogurt increased gradually and continuously during the 28 d of cold storage. Mulberry pomace is beneficial to improve the physicochemical and textural properties of yogurt and has the potential as a natural stabilizer to be used in functional yogurt rich in phytochemicals.

The dose-dependent effects of polyphenols and malondialdehyde on the emulsifying and gel properties of myofibrillar protein-mulberry polyphenol complex.

The effects of mulberry polyphenols and malondialdehyde (MDA) on the emulsifying and gel properties of myofibrillar protein (MP) were studied. Emulsibility and gel properties of MP were compared in range of mulberry polyphenol/MDA concentrations by particle size, Zeta potential, antioxidant capacity, gel strength, water-holding capacity (WHC), rheological properties and microstructure. Mulberry polyphenols enhanced the inoxidizability of MP emulsion but decreased its emulsifying property. MDA at intermediate concentrations (5-20 mM) improved the elasticity, strength, and WHC of polyphenols-modified MP emulsion gel, while at high concentration (40 mM) it destroyed the emulsion gel, resulting in "oil leakage". Polyphenol is not conducive to the gelation but weaken the oxidative damage of MDA to protein. The gel structure of MP emulsion collapsed after high dose of polyphenols or MDA treated. Thus, to maintain uniform textural and antioxidant activity of meat product, both polyphenols addition and oxidation intensity should be controlled simultaneously.

The Combination of Two Bioactive Constituents, Lactoferrin and Linolenic Acid, Inhibits Mouse Xenograft Esophageal Tumor Growth by Downregulating Lithocholyltaurine and Inhibiting the JAK2/STAT3-Related Pathway.

The addition of lactoferrin and three unsaturated fatty acids, oleic acid, docosahexaenoic acid (DHA), and linolenic acid, to dairy products was approved in recent years. Research into the biological activities of lactoferrin and these three unsaturated fatty acids has revealed anti-inflammatory, antiviral, antioxidant, antitumor, antiparasitic, and antibiotic effects. However, investigations and comparisons of lactoferrin + oleic acid/DHA/linolenic acid combinations in an esophageal cancer cell model and in xenograft tumor models have not been extensively reported, and the related mechanism of these combinations remains elusive. In the present study, the effects of lactoferrin and the three fatty acids on KYSE450 cell viability, migration, and invasion were investigated to choose the proper doses and effective combination in vitro. A tumor-bearing nude mouse model was established to investigate the role of selected combinations in inhibiting esophageal tumor formation in vivo. Metabonomics detection and data analysis were performed to screen special metabolites and related pathways, which were validated by western blotting. The results demonstrated that lactoferrin, the three unsaturated fatty acids, and their combinations inhibited the viability, migration, and invasion of KYSE450 cells and induced apoptosis and the lactoferrin + linolenic acid combination exhibited the strongest activity in suppressing KYSE450 tumor formation in vivo. The lactoferrin + linolenic acid combination inhibited phosphorylation in the JAK2/STAT3-related pathway by downregulating the special metabolite lithocholyltaurine, thereby suppressing formation of KYSE450 tumors.

Angustoline Inhibited Esophageal Tumors Through Regulating LKB1/AMPK/ELAVL1/LPACT2 Pathway and Phospholipid Remodeling.

Esophageal cancer is a type of gastrointestinal carcinoma and is among the 10 most common causes of cancer death worldwide. However, the specific mechanism and the biomarkers in the proliferation and metastasis of esophageal tumors are still unclear. Therefore, the development of several natural products which could inhibit esophageal tumors deserve attention. In the present study, different sources of cancer cells were used to select the sensitive cell line (esophageal cancer cell KYSE450) and the proper dose of angustoline, which were utilized in the following cell viability, migration and invasion assays. Then the lipidomic detection of clinical samples (tissue and blood plasma) from esophageal cancer patients was performed, to screen out the specific phospholipid metabolites [PC (16:0/18:1) and LPC (16:0)]. Considering lysophosphatidylcholine acyltransferase 2 (LPCAT2) was tightly relative with phospholipids conversion, serine/threonine-protein kinase 11 (LKB1), 5'-monophosphate (AMP)-activated protein kinase (AMPK) and embryonic lethal, and abnormal vision, drosophila-like 1 (ELAVL1) were investigated, to evaluate their expression levels in esophageal tumor tissue and KYSE450 cells. Additionally, KYSE450 tumor bearing mouse model was constructed, the role of angustoline in inhibiting esophageal tumors through regulating LKB1/AMPK/ELAVL1/LPCAT2 pathway was validated, and found that the conversion from LPC (16:0) to PC (16:0/18:1) was blocked by angustoline in some degree. The above results for the first time proved that angustoline suppressed esophageal tumors through activating LKB1/AMPK and inhibiting ELAVL1/LPCAT2, which consequently blocked phospholipid remodeling from LPC (16:0) to PC (16:0/18:1).

Investigation and comparison of the protective activities of three functional proteins-lactoferrin, α-lactalbumin, and ß-lactoglobulin-in cerebral ischemia reperfusion injury.

The objective of this study was to evaluate the protection conferred by lactoferrin, α-lactalbumin, and ß-lactoglobulin in cerebral ischemia reperfusion (I/R) injury. Rat pheochromocytoma (PC12) cells were used to construct an oxygen and glucose deprivation model in vitro, and ICR mice underwent carotid artery "ligation-relaxation" to construct a cerebral I/R injury model in vivo. The levels of toll-like receptor 4 (TLR4) and downstream factors including nuclear factor-κB, tumor necrosis factor-α, and IL-1ß were measured. Metabonomics detection and data mining were conducted to identify the specific metabolic sponsor of the 3 proteins. The results showed that lactoferrin, α-lactalbumin, and ß-lactoglobulin protected neurons from cerebral I/R injury by increasing the level of bopindolol and subsequently inhibiting the TLR4-related pathway to different degrees; ß-lactoglobulin had the strongest activity of the 3 proteins. In summary, this study is the first to investigate and compare the protective effects of lactoferrin, α-lactalbumin, and ß-lactoglobulin in a cerebral stroke model. The results implicate TLR4 as a novel target of the 3 bioactive proteins to prevent cerebral I/R injury.

Maillard reaction products with furan ring, like furosine, cause kidney injury through triggering ferroptosis pathway.

Maillard reaction products (MRPs), produced in the heating of food, might pose adverse effects on human health. As a classical MRP, the toxicity of furosine in kidney tissue was evaluated and the related mechanism was investigated here. We detected the concentration of furosine in mouse serum and organs using ultra-high-performance liquid chromatography, and screened out phosphatidylethanolamine (PE; 18:0/16:1) as the special metabolite in kidney by metabonomics analysis. Furthermore, furosine was verified to induce ferroptosis in kidney cells by quantitative real-time RT-PCR (q-PCR) and western blotting. The affinity between furosine and aldose reductase (AR) was measured by surface plasmon resonance (SPR), indicating AR is one of the targets of furosine. In addition, the furan ring was shown to be the main active group via structure-activity relationship (SAR) studies of furosine and other MRPs, which were toxic to kidney cells by activating ferroptosis pathway.

Modulation activity of heat-treated and untreated lactoferrin on the TLR-4 pathway in anoxia cell model and cerebral ischemia reperfusion mouse model.

This study aimed to investigate the modulation activity of heated and nonheated lactoferrins in an inflammatory pathway in anoxia and reoxygenation cell and cerebral ischemic reperfusion mouse models. Rat pheochromocytoma 12 (PC-12) cells were subjected to oxygen and glucose deprivation in vitro to construct an anoxia and reoxygenation cell model, and Institute for Cancer Research (ICR) mice were given carotid artery "ligation-relaxation" in vivo to construct a cerebral ischemic reperfusion mouse model. The protein levels of toll-like receptor 4 (TLR-4) and downstream inflammatory proteins including nuclear factor-κB (NF-κB), tumor necrosis factor-α (TNF-α), and IL-1ß were detected. Meanwhile, metabonomic detection of overall metabolites of PC-12 cells was performed to screen out the specific changed metabolite affected by lactoferrin at the condition of anoxia and reoxygenation. The results showed that lactoferrin could inhibit the TLR-4-related pathway triggered by anoxia and reoxygenation and ischemic reperfusion. A total of 41 significantly changed metabolites were identified by metabonomic analysis, and glutathione was seen as a metabolite of interest in suppressing TLR-4-related pathway in anoxia and reoxygenation cell models. However, heated lactoferrin lost the ability of attenuating the TLR-4-related pathway. The loss of modulation activity of heated lactoferrin might be due to its protein aggregation, which was evidenced by larger average particle diameter than the unheated lactoferrin. This study is the first to investigate the effect of heat treatment on the modulation activity of lactoferrin in the TLR-4-related pathway in anoxia and reoxygenation cell and cerebral ischemic reperfusion mouse models, and indicate that lactoferrin may serve as a dietary intervention for cerebral ischemia.

Furosine Posed Toxic Effects on Primary Sertoli Cells through Regulating Cep55/NF-κB/PI3K/Akt/FOX01/TNF-α Pathway.

As one of the Maillard reaction products, furosine has been widely reported in a variety of heat-processed foods, while the toxicity of furosine on the reproductive system and related mechanisms are unclear. Here, we constructed an intragastric gavage male mice model (42-day administration, 0.1/0.25/0.5 g furosine/Kg body weight per day) to investigate its effects on mice testicle index, hormones in serum, and mice sperm quality. Besides, the lipid metabonomics analysis was performed to screen out the special metabolites and relatively altered pathways in mice testicle tissue. Mice primary sertoli cells were separated from male mice testicle to validate the role of special metabolites in regulating pathways. We found that furosine affected testicle index, hormones expression level and sperm quality, as well as caused pathological damages in testicle tissue. Phosphatidylethanolamine (PE) (18:0/16:1) was upregulated by furosine both in mice testicle tissue and in primary sertoli cells, meanwhile, PE(18:0/16:1) was proved to activate Cep55/NF-κB/PI3K/Akt/FOX01/TNF-α pathway, and as a functional protein in dairy products, lactoferrin could inhibit expression of this pathway when combined with furosine. In conclusion, for the first time we validated that furosine posed toxic effects on mice sperms and testicle tissue through upregulating PE(18:0/16:1) and activating Cep55/NF-κB/PI3K/Akt/FOX01/TNF-α pathway.

Furosine, a Maillard Reaction Product, Triggers Necroptosis in Hepatocytes by Regulating the RIPK1/RIPK3/MLKL Pathway.

As one of the typical Maillard reaction products, furosine has been widely reported in a variety of heat-processed food. Though furosine was shown to be toxic on organs, its toxicity mechanism is still unclear. The present study aimed to investigate the toxicity mechanism of furosine in liver tissue. An intragastric gavage mice model (42-day administration, 0.1/0.25/0.5 g/kg of furosine per day) and a mice primary hepatocyte model were employed to investigate the toxicity mechanism of furosine on mice liver tissue. A metabonomics analysis of mice liver, serum, and red blood cells (RBC) was performed. The special metabolic mediator of furosine, lysophosphatidylcholine 18:0 (LPC (18:0)) was identified. Then, the effect of the upstream gene phospholipase A2 gamma (PLA2-3) on LPC (18:0), as well as the effect of furosine (100 mg/L) on the receptor-interacting serine/threonine-protein kinase (RIPK)1/RIPK3/mixed lineage kinase domain-like protein (MLKL) pathway and inflammatory factors, was determined in liver tissue and primary hepatocytes. PLA2-3 was found to regulate the level of LPC (18:0) and activate the expression of RIPK1, RIPK3, P-MLKL, and of the inflammatory factors including tumor necrosis factor α (TNF-α) and interleukin (IL-1ß), both in liver tissue and in primary hepatocytes. Upon treatment with furosine, the upstream sensor PLA2-3 activated the RIPK1/RIPK3/MLKL necroptosis pathway and caused inflammation by regulating the expression of LPC (18:0), which further caused liver damage.

l-Proline Alleviates Kidney Injury Caused by AFB1 and AFM1 through Regulating Excessive Apoptosis of Kidney Cells.

The toxicity and related mechanisms of aflatoxin B1 (AFB1) and aflatoxin M1 (AFM1) in the mouse kidney were studied, and the role of l-proline in alleviating kidney damage was investigated. In a 28-day toxicity mouse model, thirty mice were divided into six groups: control (without treatment), l-proline group (10 g/kg body weight (b.w.)), AFB1 group (0.5 mg/kg b.w.), AFM1 (3.5 mg/kg b.w.), AFB1 + l-proline group and AFM1 + l-proline group. Kidney index and biochemical indicators were detected, and pathological staining was observed. Using a human embryonic kidney 293 (HEK 293) cell model, cell apoptosis rate and apoptotic proteins expressions were detected. The results showed that AFB1 and AFM1 activated pathways related with oxidative stress and caused kidney injury; l-proline significantly alleviated abnormal expressions of biochemical parameters and pathological kidney damage, as well as excessive cell apoptosis in the AF-treated models. Moreover, proline dehydrogenase (PRODH) was verified to regulate the levels of l-proline and downstream apoptotic factors (Bax, Bcl-2, and cleaved Caspase-3) compared with the control (p < 0.05). In conclusion, l-proline could protect mouse kidneys from AFB1 and AFM1 through alleviating oxidative damage and decreasing downstream apoptosis, which deserves further research and development.

Lactoferrin Induces the Synthesis of Vitamin B6 and Protects HUVEC Functions by Activating PDXP and the PI3K/AKT/ERK1/2 Pathway.

As a nutritional active protein in foods, multiple studies of the biological activities of lactoferrin had been undertaken, including antioxidant, antiviral, anti-inflammatory, antitumor, antibiosis, and antiparasitic effects, while the mechanism related with its protection of cardiovascular system remained elusive. In the present work, the effect of lactoferrin on the viability of HUVECs (human umbilical vein endothelial cells) was detected to select the proper doses. Moreover, transcriptomics detection and data analysis were performed to screen out the special genes and the related pathways. Meanwhile, the regulation of lactoferrin in the functional factors thromboxane A2 (TXA2) and prostacyclin (PGI2) was detected. Then, the small interfering RNA (SiRNA) fragment of the selected gene pyridoxal phosphatase (PDXP) was transfected into HUVECs to validate its role in protecting HUVECs function. Results showed that lactoferrin inhibited the expression of TXA2 and activated expression of PGI2, as well as activated expression of PDXP, which significantly up-regulated the synthesis of vitamin B6 (VB6) and the phosphoinositide 3-kinase (PI3K)/ serine/threonine-protein kinase (AKT)/ extracellular regulated protein kinases (ERK) 1/2 pathway. For the first time, we revealed that lactoferrin could induce the synthesis of VB6 and protect HUVECs function through activating PDXP gene and the related pathway.

Effect of Heat Treatment on the Antitumor Activity of Lactoferrin in Human Colon Tumor (HT29) Model.

To investigate the effect of heat treatment on the antitumor activity of lactoferrin in colon cancer cells and colon tumors, the HT-29 (human intestinal epithelial tumor cell) cell line was exposed to lactoferrin and various heat treatments. The impacts on cell proliferation, invasion, and migration were observed in vitro, and nude mice bearing HT29 tumors were administered lactoferrin and underwent various heat treatments in vivo. In the HT29 cell proliferation test using transwell and scratch analyses, lactoferrin (20 mg/mL) without or with heat treatment (50 and 70 °C) significantly inhibited cell proliferation, migration, and invasion (compared with the control, p < 0.05), while lactoferrin with heat treatment (100 °C) did not affect these parameters. In vivo, HT29 tumor weight was significantly reduced in the lactoferrin (without heat treatment and with 50 and 70 °C treatment) groups (1.59 ± 0.20, 1.67 ± 0.25, and 2.41 ± 0.42 g, compared with the control, p < 0.05), and there was no significant difference between the control (3.73 ± 0.33 g) and the 100 °C treatment group (3.58 ± 0.29 g). Moreover, 100 °C heat treatment reduced inhibition of the VEGFR2/VEGFA/PI3K/Akt/Erk1/2 angiogenesis pathway by lactoferrin. In summary, HT29 tumors were effectively suppressed by lactoferrin via inhibition of VEGFR2/VEGFA/PI3K/Akt/Erk1/2 pathway, and heat treatment affected the antitumor activity of lactoferrin in a temperature-dependent manner.

Transcriptional and Proteomic Analysis Revealed a Synergistic Effect of Aflatoxin M1 and Ochratoxin A Mycotoxins on the Intestinal Epithelial Integrity of Differentiated Human Caco-2 Cells.

Aflatoxin M1 (AFM1) is a common mycotoxin in dairy milk, and it is typically concurrently present with other mycotoxins that may represent a threat to food safety. However, knowledge of how AFM1, alone or in combination with other mycotoxins, may affect human intestinal epithelial integrity remain to be established. We employed transcriptome and proteome analysis integrated with biological validation to reveal the molecular basis underlining the effect of exposure to AFM1, ochratoxin A (OTA), or both on the intestinal epithelial integrity of differentiated Caco-2 cells. Exposure to 4 µg/mL of OTA was found to disrupt human gut epithelial integrity, whereas 4 µg/mL of AFM1 did not. The integrated transcriptome and proteome analysis of AFM1 and OTA, alone or in combination, indicate the synergistic effect of the two mycotoxins in disrupting intestinal integrity. This effect was mechanistically linked to a broad range of pathways related to intestinal integrity enriched by down-regulated genes and proteins, associated with focal adhesion, adheren junction, and gap junction pathways. Furthermore, the cross-omics analysis of mixed AFM1 and OTA compared to OTA alone suggest that kinase family members, including myosin light-chain kinase, mitogen-activated protein kinases, and protein kinase C, are the potential key regulators in modulating intestinal epithelial integrity. These findings provide novel insight into the synergistic detrimental role of multiple mycotoxins in disrupting intestinal integrity and, therefore, identify potential targets to improve milk safety related to human health.

Susceptibility towards enterotoxigenic Escherichia coli F4ac diarrhea is governed by the MUC13 gene in pigs.

Enterotoxigenic Escherichia coli (ETEC) F4ac is a major determinant of diarrhea and mortality in neonatal and young pigs. Susceptibility to ETEC F4ac is governed by the intestinal receptor specific for the bacterium and is inherited as a monogenic dominant trait. To identify the receptor gene (F4acR), we first mapped the locus to a 7.8-cM region on pig chromosome 13 using a genome scan with 194 microsatellite markers. A further scan with high density markers on chromosome 13 refined the locus to a 5.7-cM interval. Recombination breakpoint analysis defined the locus within a 2.3-Mb region. Further genome-wide mapping using 39,720 informative SNPs revealed that the most significant markers were proximal to the MUC13 gene in the 2.3-Mb region. Association studies in a collection of diverse outbred populations strongly supported that MUC13 is the most likely responsible gene. We characterized the porcine MUC13 gene that encodes two transcripts: MUC13A and MUC13B. Both transcripts have the characteristic PTS regions of mucins that are enriched in distinct tandem repeats. MUC13B is predicated to be heavily O-glycosylated, forming the binding site of the bacterium; while MUC13A does not have the O-glycosylation binding site. Concordantly, 127 independent pigs homozygous for MUC13A across diverse breeds are all resistant to ETEC F4ac, and all 718 susceptible animals from the broad breed panel carry at least one MUC13B allele. Altogether, we conclude that susceptibility towards ETEC F4ac is governed by the MUC13 gene in pigs. The finding has an immediate translation into breeding practice, as it allows us to establish an efficient and accurate diagnostic test for selecting against susceptible animals. Moreover, the finding improves our understanding of mucins that play crucial roles in defense against enteric pathogens. It revealed, for the first time, the direct interaction between MUC13 and enteric bacteria, which is poorly understood in mammals.