Characterization of the potential allergenicity of enzymatically hydrolyzed casein in Balb/c mouse model.

Bovine casein is a major allergen present in cow milk to induce anaphylaxis. In this study, the potential allergenicity of enzymatically hydrolyzed casein (HC) was evaluated based on in vitro and in vivo. The results showed that Alcalase and Protamex treatment (AT, PT) reduced the potential allergenicity of CN, with the greatest reductions of 68.25% and 50.75%, respectively. In addition, in vivo results showed that HC effectively alleviated allergic response symptoms of Balb/c mice; a significant tendency toward decreased serum IgG1 and mast cell tryptase levels was observed, accompanied by a decrease of Th2-associated IL-4, IL-5, and IL-13 and an increase of IFN-γ levels in spleen. Moreover, the inflammation of the lung, jejunum, and ileum was remarkably ameliorated. The findings indicated that HC induced a shift toward Th1 response and maintained the Th1/Th2 immune balance. Importantly, our results provide the basis for the production of hypoallergenic dairy products.

Investigation of pectolytic and PR genes expression, quality and phytochemical contents in organic and non-organic table grapes at harvest and during storage.

The demand for organic table grapes is increasing worldwide. However, comprehensive information of quality parameters and phytochemical compounds in organically grown fruit remain unclear. Furthermore, table grapes are perishable and postharvest quality retention and waste prevention is very important. In this study we have compared the differences between organic and non-organic table grapes in terms of phytochemical compounds and quality parameters as well as the changes in the expression levels of pathogen related and lytic genes during storage. Organic fruit showed higher levels of phenolics, flavonoids, caffeic acid, hydrogen peroxide, protein content, antioxidant and anti-stress enzymes and total antioxidant activities at harvest and during storage. Although, the expression levels of polygalactronases, pectin methyl esterase, chitinase and glucanase genes was lower in organically grown table grapes at harvest, but the expression of all these genes was significantly increased during cold storage. After 60 days of cold storage the expression levels of pectin methyl esterase, chitinase and glucanase genes was significantly higher than the conventionally grown grape berries in organic ones. The highest expression of polygalacturonase was recorded in organic samples after 30 days of storage. There was no significant difference between the two types of table grapes for decay extension and tissue deterioration rate. The results of this study indicate that due to higher levels of phytochemicals and antioxidant compounds the organic table grapes have a higher nutritional quality. Furthermore, the increase in PR and pectolytic genes expression levels is enough for decreasing the fruit susceptibility to decay pathogens and enhancing the postharvest life of organic grapes.

Free-Choice Feeding of Whole Grains Improves Meat Quality and Intestinal Development of Pigeon Squabs Compared with Complete Pelleted Feed.

Effects of different feeding strategies on meat quality and intestinal development in pigeon squabs were investigated. 120 pairs of pigeons with two squabs each were assigned to five groups (four free-choice feeding systems and one complete feeding system): T1 (corn, pea, wheat, and pelleted feed), T2 (corn, wheat, and pelleted feed), T3 (corn, pea, and pelleted feed), T4 (corn and pelleted feed), and T5 (complete pelleted feed). Compared with T5, the diet in T4 made the breast meat redder and more yellow (p < 0.05). T2 and T4 resulted in an enhanced total superoxide dismutase activity of meat. Breast muscle in T1 and T2 was determined to have higher contents of nonessential amino acids, glycine, alanine, and glutamic acid (p < 0.05). The contents of the essential amino acids, lysine, threonine, valine, histidine, and arginine were also higher in T1 (p < 0.05). Villus height, surface area, and alkaline phosphatase activity of the duodenum and jejunum in T2 were the highest among the treatments (p < 0.05). In conclusion, free-choice feeding system can improve the meat quality and intestinal development of pigeon squabs, but which combination method of whole grains to use in the production depends on the feeding purpose.

Oral delivery of bi-autoantigens by bacterium-like particles (BLPs) against autoimmune diabetes in NOD mice.

Induction of oral tolerance by vaccination with type 1 diabetes mellitus (T1DM)-associated autoantigens exhibits great potential in preventing and treating this autoimmune disease. However, antigen degradation in the gastrointestinal tract (GIT) limits the delivery efficiency of oral antigens. Previously, bacterium-like particles (BLPs) have been used to deliver a single-chain insulin (SCI-59) analog (BLPs-SCI-59) or the intracellular domain of insulinoma-associated protein 2 (IA-2ic) (BLPs-IA-2ic). Both monovalent BLPs vaccines can suppress T1DM in NOD mice by stimulating the corresponding antigen-specific oral tolerance, respectively. Here, we constructed two bivalent BLPs vaccines which simultaneously deliver SCI-59 and IA-2ic (Bivalent vaccine-mix or Bivalent vaccine-SA), and evaluated whether there is an additive beneficial effect on tolerance induction and suppression of T1DM by treatment with BLPs-delivered bi-autoantigens. Compared to the monovalent BLPs vaccines, oral administration of the Bivalent vaccine-mix could significantly reduce morbidity and mortality in T1DM. Treatment with the bivalent BLPs vaccines (especially Bivalent vaccine-mix) endowed the mice with a stronger ability to regulate blood glucose and protect the integrity and function of pancreatic islets than the monovalent BLPs vaccines treatment. This additive effect of BLPs-delivered bi-autoantigens on T1DM prevention may be related to that SCI-59- and IA-2-specific Th2-like immune responses could be induced, which was more beneficial for the correction of Th1/Th2 imbalance. In addition, more CD4+CD25+Foxp3+ regulatory T cells (Tregs) were induced by treatment with the bivalent BLPs vaccines than did the monovalent BLPs vaccines. Therefore, multiple autoantigens delivered by BLPs maybe a promising strategy to prevent T1DM by efficiently inducing antigen-specific immune tolerance.

Rapid quantitative detection of Vibrio parahaemolyticus via high-fidelity target-based microfluidic identification.

With the rapid development of logistics, a growing number of pathogenic microorganisms has the means to spread worldwide using food as a carrier; thus, there is an urgent need to develop effective detection strategies to ensure food safety. By combining novel markers identified by pan-genome analysis and a digital recombinase-aided amplification (RAA) detection method based on a microfluidic chip, a strategy of high-fidelity target-based microfluidic identification (HFTMI) has been developed. Herein, a proof-of-concept study of HFTMI for rapid pathogen detection of V. parahaemolyticus was investigated. Specific primers designed for the gene group_41170 identified in the pan-genome analysis showed high sensitivity and a broad spectrum for the detection of V. parahaemolyticus. Different power systems were investigated to increase the partition rate on specifically designed chamber-based digital chips. The performance of HFTMI was greatly improved compared with qPCR. Collectively, this novel HFTMI system provides more reliable guidance for food safety testing.

Possible Reaction Mechanisms Involved in Degradation of Patulin by Heat-Assisted Cysteine under Highly Acidic Conditions.

Patulin (PAT) is one of mycotoxins that usually contaminates apple juice, and it is not easily detoxified by cysteine (CYS) at room temperature due to the highly acidic conditions based on the Michael addition reaction. However, it could be effectively degraded by a heating treatment at 120 °C for 30 min in the presence of cysteine. In our study, a total of eight degradation products (DP A-H) were characterized and identified via liquid chromatography quadrupole time-of-flight mass spectrometry (LC-Q-TOF-MS) in a negative ion mode, and their structures and formulas were proposed based on their accurate mass data. The fragmentation patterns of PAT and its degradation products were obtained from the MS/MS analysis. Meanwhile, the possible reaction mechanisms involved in the degradation of PAT were established and explained for the first time. According to the relation between the structure and toxicity of PAT, it could be deduced that the toxic effects of PAT degradation products were potentially much less than those of PAT-self.

Effects of combined enzymatic hydrolysis and fed-batch operation on efficient improvement of ferulic acid and p-coumaric acid production from pretreated corn straws.

In the present work, the effects of combined enzymatic hydrolysis by cellulase and xylanase (CXEH), fed-batch enzymatic hydrolysis (FBEH) operation and kinetics on production of ferulic acid (FA) and p-coumaric acid (pCA) from pretreated corn straws were investigated. The results showed that CXEH could efficiently increase production of FA and pCA. When performed the FBEH operation by feeding 150 mL enzymatic hydrolysis solution (1.5 % enzyme concentration, 5:4 (v/v) ratio of cellulase to xylanase and 2.0 % substrate loading) to 250 mL batch enzymatic hydrolysis solution at 36 h, the maximum production (2178.58 and 2710.17 mg/L) and production rate (590.95 and 727.89 mg/L.h) of FA and pCA were respectively obtained. Moreover, the disruption of fiber tissues, enhancement of crystallinity and accelerated degradation of hemicelluloses and lignocelluloses caused by CXEH contributed to effectively improving production of FA and pCA in corn straws.

Extraction and activity of chemical constituents from Houttuynia cordata Thunb by ultrasonic method.

Polyphenols and flavonoids are phytochemicals that have essential roles in human nutrition. In this regard, the contents of polyphenols and flavonoids in Houttuynia cordata Thunb and their antioxidant activities were evaluated in the current study. Two Houttuynia cordata materials with the same chromosome number and chemical type were used to comprehensively assess the contents of total phenols and flavonoids in different parts of H. cordata. These chemical components were extracted by the ultrasonic method. The results showed that the total phenols and antioxidant capacity of different parts of H. cordata were significantly different. The content of polyphenols in roots and stems was low, the antioxidant capacity was weak, the total phenols in flowers and leaves were high, and the antioxidant capacity was strong. Therefore, different parts of H. cordata had different pharmacological and food effects. The whole herb can be used as Chinese herbal medicine, and its young leaves and roots can be used as vegetables. Flavonoids are the main phenolic components, and total phenols are the main components of antioxidant activity. It can explain a very significant positive correlation between total phenols and flavonoids. Therefore, in the further breeding work of H. cordata, the procedure can be simplified by determining one of the above indexes to predict the varieties with high total phenolic and antioxidant activity.

Oral delivery of the intracellular domain of the insulinoma-associated protein 2 (IA-2ic) by bacterium-like particles (BLPs) prevents type 1 diabetes mellitus in NOD mice.

Antigen-specific immune tolerance, which possesses great potential in preventing or curing type 1 diabetes mellitus (T1DM), can be induced by oral vaccination with T1DM-related autoantigens. However, direct administration of autoantigens via oral route exhibits a low tolerance-inducing effect as a result of the digestion of protein antigens in the gastrointestinal tract (GIT) and therefore, a large dosage of autoantigens may be needed. In this study, bacterium-like particles (BLPs) made from food-grade lactic acid bacteria were used to deliver the intracellular domain of the insulinoma-associated protein 2 (IA-2ic). For this purpose, BLPs-IA-2ic vaccine in which IA-2ic bound to the surface of BLPs was constructed. BLPs enhanced the stability of the delivered IA-2ic based on the stability analysis in vitro. Oral administration of BLPs-IA-2ic significantly reduced T1DM incidence in NOD mice. The mice fed BLPs-IA-2ic exhibited a significant reduction in insulitis and preserved the ability to secrete insulin. Immunologic analysis showed that oral vaccination with BLPs-IA-2ic induced antigen-specific T cell tolerance. The results revealed that the successful induction of immune tolerance was dependent on the immune deviation (in favor of T helper 2 responses) and CD4+CD25+FoxP3+ regulatory T cells. Hence, oral vaccination with BLPs-IA-2ic shows potential for application in preventing T1DM.

Removal of patulin by thiol-compounds: A review.

Patulin (PAT) is a toxic mycotoxin usually contaminated apple juices, which leads to a serious food safety issue in the world. Thiol-compounds are a class of compounds containing the thiol (-SH) group themselves or obtained the -SH group by physical or chemical modification. They have the ability to efficiently remove patulin in apple juices with manifested negligible effects on juice quality. This review investigates the latest development in the removal of patulin using thiol-compounds, including the removal efficiencies and mechanisms of patulin, the factors influencing the removal efficiency of patulin, as well as the toxicities of thiol-compounds and safety of juices after detoxification. This review shows that thiol-compounds are promising materials for the removal or degradation of patulin in the contaminated juices.

Factors influencing the removal of patulin by cysteine.

The removal of patulin in phosphoric acid buffer solution by cysteine was investigated. Cysteine could effectively decrease the patulin concentration at high acidic condition (pH 3.0-5.0) with the help of high temperature greater than 90 °C. Three removal mechanisms of patulin by cysteine under high acidic and high temperature conditions were deduced. Reaction temperature, pH of reactive media, molar ratio between cysteine and patulin, and reaction time were all the obvious factors influencing the removal efficiency of patulin, and the increase of any one factor could significantly improve the removal efficiency of patulin. The removal process of patulin could be simulated by the zero-order kinetic model, logarithmic model, and pseudo-first-order kinetic model, respectively, and the corresponding correlation coefficients (R2) were all greater than 0.90. Presently, this method can only be applied for the removal of patulin in contaminated water from washing fruits in juice processing industry due to the high treatment temperature more than 120 °C and the long detoxification time greater than 1 h.

Thermal Stability and Degradation Kinetics of Patulin in Highly Acidic Conditions: Impact of Cysteine.

The thermal stability and degradation kinetics of patulin (PAT, 10 µmol/L) in pH 3.5 of phosphoric-citric acid buffer solutions in the absence and presence of cysteine (CYS, 30 µmol/L) were investigated at temperatures ranging from 90 to 150 °C. The zero-, first-, and second-order models and the Weibull model were used to fit the degradation process of patulin. Both the first-order kinetic model and Weibull model better described the degradation of patulin in the presence of cysteine while it was complexed to simulate them in the absence of cysteine with various models at different temperatures based on the correlation coefficients (R2 > 0.90). At the same reaction time, cysteine and temperature significantly affected the degradation efficiency of patulin in highly acidic conditions (p < 0.01). The rate constants (kT) for patulin degradation with cysteine (0.0036-0.3200 µg/L·min) were far more than those of treatments without cysteine (0.0012-0.1614 µg/L·min), and the activation energy (Ea = 43.89 kJ/mol) was far less than that of treatment without cysteine (61.74 kJ/mol). Increasing temperature could obviously improve the degradation efficiency of patulin, regardless of the presence of cysteine. Thus, both cysteine and high temperature decreased the stability of patulin in highly acidic conditions and improved its degradation efficiency, which could be applied to guide the detoxification of patulin by cysteine in the juice processing industry.

Nanostructured Titanium Dioxide (TiO2) Reduces Sperm Concentration Involving Disorder of Meiosis and Signal Pathway.

Nano-titanium dioxide (nano-TiO2) has been widely used in food and cosmetic industries, and the medical sector. However, nano-TiO2 is potentially toxic to the reproductive system. Previous research has shown that nano-TiO2 can reduce sperm concentration but do not yet known whether this effect occurs because of dysfunctional meiosis in spermatogenic cells. In the present paper, we demonstrate that Nano-TiO2 can penetrate through the blood-testis barrier of a mouse model and enter the testicular tissue, thus causing damage to the testis and epididymis. This reduced the number of developing sperm; we demonstrated that the mechanism underlying this effect was the inhibition or destruction of meiosis in spermatogenic cells, particularly during meiosis I. We also found that the inhibition of meiosis I caused by nano-TiO2 exposure was related to dysfunctional meiosis and that the abnormal expression of meiosis-related factors. Therefore, our data demonstrate that nano-TiO2 reduces sperm concentration by disrupting meiosis and related signaling pathways.

Degradation and ozonolysis pathway elucidation of deoxynivalenol.

To explore the degradation products and ozonolysis pathway of deoxynivalenol (DON), DON (~50 mg/L) in acetonitrile solution was treated by ozone at a concentration of 10.84 g/m3 and a flow rate of 80 mL/min for the times ranging from 0 to 9 min. The results showed that DON concentration rapidly reduced from 51.11 mg/L to 14.97 mg/L within 9 min of ozone exposure with 98.30% of degradation rate, and the ozonolysis of DON followed the first-order kinetic model. Four ozonolysis products of DON were identified based on the analysis of Liquid chromatography-quadrupole time-of-flight mass spectra (LC-QTOF/MS). Their structures were similar to that of DON, while the double bond at C9-C10, 12,13-epoxide ring, and the hydroxyl group at C3 or C7 of DON were all destroyed by ozone. It is deduced that the toxicity of ozonolysis products significantly reduced based on the relationship between structure and toxicity of DON. The ozonolysis pathway of DON followed the Criegee reaction mechanism of ozone according to the chemical structures, accurate mass and molecular formulas of these products.

Cytotoxicity of Deoxynivalenol after Being Exposed to Gaseous Ozone.

In this study, deoxynivalenol (DON) in aqueous solution was exposed to gaseous ozone for periods ranging from 0 to 20 min. The degradation efficiency and cytotoxicity of DON were investigated after being treated by ozone. The results showed that DON was rapidly degraded from 10.76 ± 0.09 mg/L to 0.22 ± 0.04 mg/L within 15 min (P < 0.05), representing a reduction of 97.95%, and no DON was detected after being exposed to 14.50 mg/L of ozone at a flow rate of 80 mL/min for 20 min. The degradation of DON depended on the ozone exposure time, and followed the first-order kinetic model (R2 = 0.9972). Human hepatic carcinoma (HepG2) and Henrietta Lacks (Hela) cells were used to evaluate the cytotoxicity of DON treated by ozone using the 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay. The half-maximal inhibitory concentrations (IC50) values of DON on HepG2 and Hela cells were 2.10 and 1.33 mg/L after 48 h of exposure, respectively, and showed a dose-dependent manner. The cell vitalities of HepG2 and Hela cells on DON were both evidently improved after being exposed to ozone for 15 min, and there were no significant differences between the negative control and that treated at 20 min of ozone exposure. Gaseous ozone can potentially be used as a new method to detoxify DON in agricultural products.

Oral delivery of single-chain insulin (SCI-59) analog by bacterium-like particles (BLPs) induces oral tolerance and prevents autoimmune diabetes in NOD mice.

Oral tolerance, induced by oral administration of autoantigens, is a promising therapeutic approach to treat type 1 diabetes mellitus (T1DM). However, the degradation of antigens passing through the gastrointestinal tract (GIT) leads to low induction efficiency. Based on our previous study, a single-chain insulin (SCI-59) analog, bound to the surface of lactic acid bacteria (LAB) bacterium-like particles (BLPs), was more stable in the simulated gastric fluid, compared to free SCI-59 and insulin. Based on the analysis of diabetes progression, a significant decrease in the incidence of diabetes was observed in mice fed BLPs-SCI-59. Oral administration of BLPs-SCI-59 can enhance glucose tolerance in NOD mice and this effect may result from the protection of pancreatic islet beta cells, as compared to the free SCI-59 group and BLPs group. Oral administration of BLPs-SCI-59 can significantly reduce insulitis and preserve the ability of insulin secretion in treated mice. Oral vaccination with BLPs-SCI-59 induced SCI-59 specific T cell tolerance in treated mice, which may due to the repair of Th1/Th2 imbalance and increased CD4+CD25+FoxP3+ regulatory T cells (Tregs). These results show that oral vaccination with BLPs-SCI-59 is a promising way to prevent T1DM in NOD mice.

Effects of ozone processing on patulin, phenolic compounds and organic acids in apple juice.

In this study, ozone processing was used to degrade patulin in apple juice, and the ozonolysis efficiency of patulin and its effects on phenolic compounds and organic acids in apple juice were investigated. Ozone processing was performed using a self-developed ozonolysis reactor at the ozone concentration of 12 mg/L and flow rate of 3 L/min for increasing ozonation times ranged from 0 to 30 min. Ozone processing significantly degraded patulin in apple juice, and decreased it from 201.06 to below 50 µg/L within 15 min, with a reduction of 75.36%. While major phenolic compounds and organic acids in apple juice were seriously destroyed by ozone processing compared with the control. Processors should consider the adverse effects of ozone processing on quality of apple juices and further studies are advised to optimize the ozone processing for remaining the phenols and organic acids in apple juices.

Ozone detoxification of patulin in aqueous solution and cytotoxic evaluation using human hepatic carcinoma cells.

Patulin often contaminates fruits and fruit-base products, which seriously threats the health of consumers. In this study, ozone was used to degrade patulin in aqueous solution, and investigated the cytotoxicity of patulin after ozone detoxification on human hepatic carcinoma cells (HepG2) using MTT assay and apoptosis assay. Patulin was rapidly degraded from 24.59 mg/L to 9.85 mg/L within 180 s by 10.60 mg/L of ozone at a flow rate of 90 mL/min, and reduced by 59.94%. The half maximal inhibitory concentration (IC50) of patulin on HepG2 cells was 9.32 µmol/L after 24 h of exposure, and it showed a dose dependent effect. After 90 s of ozone detoxification, the cell viability of HepG2 cells obviously increased from 42.31% to 93.96%, and the total apoptotic cells significantly reduced from 22.24% to 11.18% after 180 s of ozone treatment. The results clearly show the great potential of ozone in degrading patulin in liquid foods.

Enhanced cytotoxic and apoptotic potential in hepatic carcinoma cells of chitosan nanoparticles loaded with ginsenoside compound K.

Ginsenoside compound K (CK) has been shown to exhibit anticancer properties. In this study, chitosan nanoparticles loaded with ginsenoside compound K (CK-NPs) were prepared as a delivery system using a self-assembly technique with amphipathic deoxycholic acid-O carboxymethyl chitosan as the carrier, which improved the water solubility of CK. By evaluating drug loading, entrapment efficiency, and in vitro release behavior, the feasibility of CK-NPs as a drug carrier nanoparticle for the treatment of human hepatic carcinoma cells (HepG2) was investigated. Result revealed that CK and CK-NPs showed a dose-dependent inhibitory effect on HepG2 cells with IC50 values of 23.33 and 16.58 µg/mL, respectively. Furthermore, fluorescence imaging demonstrated that CK-NPs promoted cellular uptake in vitro. Therefore, all results indicated that CK-NPs might be a novel drug delivery system to improve the solubility and enhance the cytotoxic and apoptotic potentials of CK for effective liver cancer chemotherapy.