Comparative effects of pectin and hydrolyzed pectin coating as pre-frying treatments on acrylamide formation in potato chips.

This study aimed to compare the effects of pectin and hydrolyzed pectin coating as pre-frying treatments on acrylamide content and quality characteristics of fried potato chips. The hydrolyzed pectin with molecular weight (Mw) of 8.81 ± 0.49 kDa was obtained through partial degradation of pectin (Mw: 747.57 ± 6.73 kDa) using pectinase. Results showed that both pectin and hydrolyzed pectin coating significantly inhibited acrylamide formation and inhibition rates exceeded 90 %. Hydrolyzed pectin had stronger inhibitory activity against acrylamide formation than pectin, especially when the concentration of hydrolyzed pectin was >2 %, its inhibitory rate exceeded 95 %. Compared to pectin coating, hydrolyzed pectin coating endow fried potato chips with smaller browning, higher crispness, less moisture but higher oil content. Overall, hydrolyzed pectin had better application prospects than pectin in inhibiting acrylamide formation of fried potato chips.

Galactooligosaccharides in infant formulas: Maillard reaction characteristics and influence on formation of advanced glycation end products.

As prebiotics supplemented in infant formulas (IFs), galactooligosaccharides (GOSs) also have many other biological activities; however, their Maillard reaction characteristics are still unclear. We investigated the Maillard reactivity of GOSs and their effects on advanced glycation end product (AGE) formation during IF processing. The results showed that AGE and HMF formation was temperature-dependent and reached the maximum at pH 9.0 in the Maillard reaction system of GOSs and Nα-acetyl-L-lysine. Acidic conditions accelerated HMF formation; however, protein cross-linking was more likely to occur under alkaline conditions. The degree of polymerization (DP) of GOSs had no significant effect on AGEs formation (except pyrraline); however, the greater the DP, the higher the concentration of HMF and pyrraline. Besides, compared with arginine and casein, lysine and whey protein were more prone to Maillard reaction with GOSs. GOSs promoted AGEs formation in a dose-dependent manner during the processing of IFs. These results provide a reliable theoretical basis for application of GOSs in IFs.

Heat treatment of galangin and kaempferol inhibits their benefits to improve barrier function in rat intestinal epithelial cells.

Flavonols are bioactive substances in plant foods. In this study, two flavonols galangin and kaempferol were heated at 100°C for 30 min prior to assessing their effects on barrier function of rat intestinal epithelial (IEC-6) cells. Both heated and unheated flavonols (2.5-20 µmol/L dosages) were nontoxic to the cells up to 48 h post-treatment, and could promote cell viability values to 102.2-141.2% of control. By treatment with 5 µmol/L flavonols for 24 and 48 h, the treated cells time-dependently showed better improved physical and biological barrier functions than the control cells without any flavonol treatment, including higher transepithelial electrical resistance and antibacterial effect but reduced paracellular permeability and bacterial translocation. The results from real-time PCR and western-blot assays indicated that the cells treated with heated and unheated flavonols of 5 µmol/L dosage had up-regulated mRNA (1.13-1.81 folds) and protein (1.15-5.11 folds) expression for zonula occluden-1, occludin, and claudin-1 that are vital to the tight junctions of the cells. Moreover, protein expression of RhoA and ROCK were down-regulated into 0.41-0.98 and 0.40-0.92 folds, respectively, demonstrating a Rho inactivation that led to enhanced cell barrier integrity via the RhoA/ROCK pathway. Overall, galangin was more active than kaempferol to perform three biofunctions like improving cell barrier function, up-regulating tight junctions protein expression, and down-regulating RhoA/ROCK expression. Moreover, the heated flavonols were less effective than the unheated counterparts to perform these biofunctions. It is concluded that this heat treatment of galangin and kaempferol could inhibit their benefits to improve barrier function of IEC-6 cells.

Barrier-promoting efficiency of two bioactive flavonols quercetin and myricetin on rat intestinal epithelial (IEC-6) cells via suppressing Rho activation.

Polyphenols are beneficial to human health because of their bio-activities. In this study, two flavonols quercetin and myricetin with or without heat treatment at 100 °C for 30 min were assessed for their barrier-promoting efficiency in rat intestinal epithelial (IEC-6) cells. The results indicated that the heated and unheated flavonols at dose levels of 2.5-20 µmol L-1 had a nontoxic effect on the cells treated for 24 and 48 h but enhanced the values of cell viability larger than 100% (especially at a dose level of 5 µmol L-1). Moreover, the cells exposed to these flavonols of 5 µmol L-1 for 24 and 48 h had improved barrier integrity compared to the control cells without any flavonol treatment, reflected by enhanced transepithelial electrical resistance and anti-bacterial effect but decreased paracellular permeability and bacterial translocation. Moreover, the results from both mRNA and protein expression verified 1.1-3.4 fold up-regulation of zonula occludens-1, occludin, and claudin-1 that are critical to tight junctions and barrier function of cells. Furthermore, the expression of other two proteins RhoA and ROCK in the treated cells was also down-regulated, demonstrating suppressed Rho activation and consequently barrier promotion via the RhoA/ROCK signaling pathway. Overall quercetin, due to its lower molecular polarity, mostly gave higher barrier-promoting efficiency than myricetin, while the heated flavonols were always less efficient than the unheated counterparts to promote barrier integrity of IEC-6 cells. It is thus highlighted that flavonols can provide barrier-promoting effects on intestinal epithelial cells with a promoting efficiency dependent on flavonol polarity; however, heat treatment especially excessive heat treatment of plant foods might lead to damaged flavonol activity.

Pectin oligosaccharides from hawthorn (Crataegus pinnatifida Bunge. Var. major) inhibit the formation of advanced glycation end products in infant formula milk powder.

Pectin oligosaccharides (POSs) can not only be used as prebiotics to promote the growth of beneficial bacteria in the intestine but also can be used as natural food-borne antiglycation agents to inhibit the formation of advanced glycation end products (AGEs) in vitro, which is related to their structure, including molecular weight and galacturonic acid content. In this study, haw polysaccharides (HPSs) were isolated and purified, and POSs with high antiglycation activity in vitro were prepared. On this basis, the inhibitory effect of POSs on the formation of AGEs in infant formula milk powder was investigated. The results showed that no obvious inhibitory effect of POSs was found on the formation of AGEs in infant formula milk powder under accelerated storage at 25 °C and 45 °C. But, POSs showed a strong inhibitory effect on the formation of furosine, Nε-carboxymethyllysine (CML), Nε-carboxyethyllysine (CEL) and the total AGEs in infant formula milk powder under accelerated storage at 65 °C. In addition, POSs also had a strong inhibitory effect on the formation of lipid oxidation products but did not affect the formation of protein degradation products. Simultaneously, the cytotoxicity experiments using human umbilical vein endothelial cells showed that the infant formula milk powder supplemented with POSs had the lowest cytotoxicity compared to the blank control (BC) and GOS/FOS supplementation milk powder under accelerated storage at 65 °C, which may be related to the inhibitory effect of POSs on the formation of AGEs in infant formula milk powder. Furthermore, the in vitro fermentation experiments showed that the antiglycation process did not affect the prebiotic activity of POSs in infant formula milk powder.

Effect of Cu/Mn-Fortification on In Vitro Activities of the Peptic Hydrolysate of Bovine Lactoferrin against Human Gastric Cancer BGC-823 Cells.

Bovine lactoferrin hydrolysate (BLH) was prepared with pepsin, fortified with Cu2+ (Mn2+) 0.64 and 1.28 (0.28 and 0.56) mg/g protein, and then assessed for their activity against human gastric cancer BGC-823 cells. BLH and the four fortified BLH products dose- and time-dependently had growth inhibition on the cells in both short- and long-time experiments. These samples at dose level of 25 mg/mL could stop cell-cycle progression at the G0/G1-phase, damage mitochondrial membrane, and induce cell apoptosis. In total, the fortified BLH products had higher activities in the cells than BLH alone. Moreover, higher Cu/Mn fortification level brought higher effects, and Mn was more effective than Cu to increase these effects. In the treated cells, the apoptosis-related proteins such as Bad, Bax, p53, cytochrome c, caspase-3, and caspase-9 were up-regulated, while Bcl-2 was down-regulated. Caspase-3 activation was also evidenced using a caspase-3 inhibitor, z-VAD-fmk. Thus, Cu- and especially Mn-fortification of BLH brought health benefits such as increased anti-cancer activity in the BGC-823 cells via activating the apoptosis-related proteins to induce cell apoptosis.

Effect of the Plastein Reaction in Presence of Extrinsic Amino Acids on the Protective Activity of Casein Hydrolysate against Ethanol-Induced Damage in HHL-5 Cells.

Casein hydrolysates (CH) were prepared using papain and modified by the plastein reaction (CH-P) in the presence of extrinsic phenylalanine (CH-P-Phe) or tryptophan (CH-P-Trp). The in vitro protective activity of CH and its modified products against ethanol-induced damage in HHL-5 cells was investigated. The results showed that the modification by the plastein reaction reduced the amino group content of CH. However, the modification by the plastein reaction in the presence of extrinsic amino acids could enhance the antioxidant, proliferative, cell cycle arresting, and anti-apoptosis activity of CH. Biological activities of CH and its modified products in the HHL-5 cells varied depending on the hydrolysate concentration (1, 2, and 3 mg/mL) and treatment time (24, 48, and 72 h). Generally, higher biological activities were found after cell treatment with CH or its modified products at concentration of 2 mg/mL for 48 h compared to other treatments. In addition, CH modified in the presence of tryptophan (CH-P-Trp) showed higher biological activity than that modified in the presence of phenylalanine (CH-P-Phe). Based on the obtained results, it can be concluded that casein hydrolysates with enhanced biological activity and potential health benefits can be produced by papain and the plastein reaction with the incorporation of extrinsic amino acids.

Copper or Manganese Supplementation Endows the Peptic Hydrolysate from Bovine Lactoferrin with Enhanced Activity to Human Gastric Cancer AGS Cells.

A lactoferrin hydrolysate (LFH) was generated from bovine lactoferrin by pepsin, mixed with Cu2+ and Mn2+ at 0.64-1.28 and 0.28-0.56 mg/g protein, respectively; and then their in vitro effects on human gastric cancer AGS cells were assessed. With incubation times of 24 or 48 h, LFH and its Cu2+/Mn2+ mixtures at 10-30 mg/mL in dose-dependent manner inhibited cell growth; and more, these mixtures showed higher activities than LFH alone. Cell treatments of LFH and the mixtures (25 mg/mL) for 24 h could arrest cell cycle at G0/G1-phase, damage mitochondrial membrane integrity, and induce apoptosis, while the mixtures were also more powerful than LFH to exert these three effects. Higher Cu2+/Mn2+ supplementation level resulted in higher growth inhibition, cell cycle arrest, mitochondrial membrane potential disruption, and apoptosis induction; furthermore, Mn2+ was notable for its higher efficacy than Cu2+ to increase these four effects. Western-blot assay results revealed that four apoptosis-related proteins Bad, Bax, cytochrome c, and p53 were up-regulated, and both caspase-3 and caspase-9 also were cleaved and activated; moreover, two autophagy-related proteins LC3-II and cleaved Beclin-1 were down- and up-regulated, respectively. It is thus concluded that Cu2+ and especially Mn2+ could endow supplemented LFH with increased anti-cancer effects in AGS cells, with two proposed events as enhanced apoptosis induction (via activating apoptosis-related proteins) and autophagy inhibition (via activating autophagy-related proteins).

Administration of Lactobacillus paracasei ameliorates type 2 diabetes in mice.

Probiotics have been proposed as an option for the prevention of type 2 diabetes mellitus (T2DM). The objective of this study was to evaluate the hypoglycemic effects of Lactobacillus paracasei on diabetic mice and explore the possible underlying molecular mechanism. The α-glucosidase inhibitory activities of eight L. paracasei strains were assessed in vitro. L. paracasei TD062 with high α-glucosidase inhibitory activity (31.9%) showed an excellent antidiabetic ability and it could survive in simulated gastrointestinal juices. To investigate the beneficial effects of L. paracasei TD062, diabetic mice were treated with the strain at 109, 108 and 107 CFU ml-1. The results indicated that the administration of L. paracasei TD062 could regulate the levels of fasting blood glucose (FBG), postprandial blood glucose (PBG), glucose tolerance, hepatic glycogen and lipid metabolism. In addition, the antioxidant capacity was also improved by oral administration of L. paracasei TD062. And the hypoglycemic effects exhibited dose dependence to some extent. Furthermore, it was revealed that L. paracasei TD062 had a positive effect on the expression levels of genes related to glucose metabolism and the PI3K/Akt pathway. These results demonstrated that L. paracasei TD062 played an important role in preventing the development of T2DM and might be applied as a new type of hypoglycemic agent in functional foods.

Comparative experiments of fibril formation from whey protein concentrate with homogeneous and secondary nuclei.

Two types of special structures, homogeneous and secondary nuclei, form during fibril formation. The structural and functional properties of amyloid fibrils in whey protein concentrate (WPC) with different ratios of added homogeneous nuclei to secondary nuclei were investigated. Thioflavin T fluorescence analysis and kinetic equations indicated that two types of nuclei could accelerate WPC fibrillation compared with WPC self-assembling into amyloid fibrils, thereby reducing the lag time and increasing the number of fibrils. However, there were considerable differences in the nucleation-inducing capability of WPC fibrillation between homogeneous and secondary nuclei. The number of fibrils formed by adding homogeneous nuclei was higher than that obtained with secondary nuclei, the increase in the Th T fluorescence intensity induced by homogeneous nuclei was 1.83-fold much than secondary nuclei. Meanwhile, secondary nuclei yielded a 2.71-fold faster aggregation rate of WPC than homogeneous nuclei, particularly during the first hour of thermal treatment (protein mass ratio of nuclei to WPC 1:1). The gelation time of WPC after secondary nuclei addition was shorter, from 10 h (WPC (2.0/6.5)) to 4 h (WPC + HN) to 2 h (WPC + SN); however, the gel microstructure of WPC after the addition of homogeneous nuclei was denser, yielding a preferred water holding capacity.

Diversity and evolution of Lactobacillus casei group isolated from fermented dairy products in Tibet.

Bacteria in Lactobacillus casei group, including Lactobacillus casei (L. casei), Lactobacillus paracasei (L. paracasei), and Lactobacillus rhamnosus (L. rhamnosus) are important lactic acid bacteria in the production of fermented dairy products and are faced with the controversial nomenclatural status due to their close phylogenetic similarity. To probe the evolution and phylogeny of L. casei group, 100 isolates of lactic acid bacteria originated from naturally fermented dairy products in Tibet of China were subjected to multilocus sequence typing (MLST). The MLST scheme, based on analysis of the housekeeping genes fusA, ileS, lepA, leuS, pyrG, recA and recG, revealed that all the isolates belonged to a group containing the L. paracasei reference strains and were clearly different from the strains of L. casei and L. rhamnosus. Although nucleotide diversity (π) was low for the seven genes (ranging from 0.00341 for fusA to 0.01307 for recG), high genetic diversity represented by 83 sequence types (STs) with a discriminatory index of 0.98 was detected. A network-like structure based on split decomposition analysis, and the high values of the relative effect of recombination and mutation in the diversification of the lineages (r/m = 4.76) and the relative frequency of occurrence of recombination and mutation (ρ/θ = 2.62) indicated that intra-species recombination occurred frequently and homologous recombination played a key role in generating genotypic diversity amongst L. paracasei strains in Tibet. The discovery of 51 new STs and the results of STRUCTURE analysis suggested that the L. casei group in Tibet had an individual and particular population structure in comparison to European isolates. Overall, this research might be the first report about genetic diversity and population structure of Lactobacillus populations isolated from naturally fermented dairy products in Tibet based on MLST scheme.

[Isolation, identification and characterization of an atrazine degrading bacterium].

An atrazine-degrading bacterial strain named L-6 was isolated from the sludge mixture of the sewage treatment plant by cultivating in raw water with limited nutrition and aeration and was domesticated steadily using SBR (Sequencing Batch Reactor) for two months. The degradation rate of atrazine in inorganic liquid culture medium with atrazine as the sole source of nitrogen could reach 89.2% after 96 hours. The cells showed shape of long rod under scanning electron microscope. After extraction of genomic DNA and PCR amplification, the 16S rRNA gene sequences were used for homology analysis and construction of phylogenetic trees. The results suggested that the 16S rRNA gene sequence of L-6 had up to 99% homology with those of many strains of Pseudomonas strains in GenBank database. With physiological and biochemical reactions, the strain L-6 was identified as Pseudomonas sp. Carbon use test indicated that L-6 can utilize glucose, fructose and citric acid sodium as carbon sources, but could not use sucrose, lactose or starch. The optimum degradation conditions were optimized as following:temperature 30 degrees C, initial pH 7-9.