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BACKGROUND Polyphenols and organic acids are important constitutes in the cider because they greatly contribute to organoleptic quality. The determination of their changes is important for monitoring the fermentation process for purposes of quality control. In this study, the evolution of polyphenols and organic acids was monitored throughout the cider fermentation process. The samples were taken periodically and the polyphenols and organic acids contents were determined using HPLC methods. RESULTS The contents of polyphenols and organic acids were in constant change. After fermentation, the content of (+)-catechin, (−)-epicatechin, chlorogenic acid, cinnamic acid, p-coumaric acid, gallic acid, caffeic acid, ferulic acid, rutin and phloridzin decreased by different degrees, while protocatechuic acid increased after fermentation. The content of organic acids was also affected by fermentation. Malic acid, lactic acid, quinic acid, pyruvic acid and citric acid showed different levels of increase, but succinic acid content decreased. CONCLUSIONS The contents of polyphenols and organic acids were affected by fermentation. Their changing profiles during fermentation process were dependent on the type of phenolic compounds and organic acids studied. © 2014 Society of Chemical Industry

In this study, we employed a one-step method to prepare selenium nanoparticles (SeNPs) decorated by the water-soluble derivative of Ganoderma lucidum polysaccharides (SPS). The SeNPs-SPS complexes were stable, and the diameter of the SeNPs was homogeneous at around 25 nm. We investigated the anti-inflammatory activity of SeNPs-SPS against murine Raw 264.7 macrophage cells induced by LPS. SeNPs-SPS were found to significantly inhibit LPS-stimulated nitric oxide (NO) production against Raw 264.7 macrophages. RT-PCR results reveal the down-regulation of mRNA gene expressions for pro-inflammatory cytokines, including inducible NO synthase (iNOS), interleukin (IL)-1 and TNF-α in a dose-dependent manner. However, the anti-inflammation cytokine IL-10 was markedly increased. In the NF-κB signal pathway, SeNPs-SPS significantly inhibited the phosphorylation of Iκ-Bα. Similar results were observed for inhibition of the phosphorylation of JNK1/2 and p38 mitogen-activated protein kinase(MAPKs), whereas ERK1/2 MAPK was not apparently affected by SeNPs-SPS. All of these results suggest that SeNPs-SPS complexes have anti-inflammatory potential modulating pro-/anti-inflammation cytokine secretion profiles, and that the mechanism is partially due to inhibition of activations of NF-κB, JNK1/2 and p38 MAPKs.

To identify human umbilical cord mesenchymal stem cells (hUCMSCs) and increase the stability of the in vitro synthesized EGFP mRNA through modification.Immunophenotype of hUCMSCs was examined by flow cytometry. Adipogenic and osteogenic differentiations were determined by oil red O and alkaline phosphatase staining, respectively. In vitro synthesized EGFP mRNA was modified through polyA tailing, capping and adding base analogues, and then transfected into the hUCMSCs. After transfection, the fluorescence intensity was detected by flow cytometry and cell viability was determined by Trypan blue staining.Flow cytometry revealed that the hUCMSCs were positive for CD29, CD44, CD105, and were negative for CD34, CD45 and HLA-DR. They had the capacity of differentiating into adipocytes and osteoblasts. Through the modification the in vitro synthesized mRNA was more stable than unmodified mRNA, and mRNA transfection had lower cytotoxicity than DNA.Modification can highly improve the stability of in vitro synthesized mRNA, which can be translated into protein in hUCMSCs.

Ultrasensitive and simultaneous determination of amino acids and amines (AAs) in media and winemaking products is very important for evaluating the relationship profile between the depletion of AAs and aroma compounds formation. In this work, by combing the advantages of Taguchi's scheme with Pareto graphs and Range analysis pattern, an analytical method with efficiently improving fluorescence intensity and resolutions of AAs while saving the time and resources is developed to simultaneously and ultrasensitively detect 21 AAs in media, wine and beer. The results indicate the detection limit (DL) could be reached at ng L(-1) level (3.34-284.3 ng L(-1)), which is 10(6) higher than that of DL obtained by general method. Furthermore, the linear ranges are also established so that the proposed method could be accurately applied. These results demonstrate the method provides an easy and effective way to ultrasensitively and simultaneously analyse AAs in complex products.

To characterize and classify apple juices according to apple variety and geographical origin on the basis of their polyphenol composition, the polyphenolic profiles of 58 apple juice samples belonging to 5 apple varieties and from 6 regions in Shaanxi province of China were assessed. Fifty-one of the samples were from protected designation of origin (PDO) districts. Polyphenols were determined by high-performance liquid chromatography coupled to photodiode array detection (HPLC-PDA) and to a Q Exactive quadrupole-Orbitrap mass spectrometer. Chemometric techniques including principal component analysis (PCA) and stepwise linear discriminant analysis (SLDA) were carried out on polyphenolic profiles of the samples to develop discrimination models. SLDA achieved satisfactory discriminations of apple juices according to variety and geographical origin, providing respectively 98.3 and 91.2% success rate in terms of prediction ability. This result demonstrated that polyphenols could served as characteristic indices to verify the variety and geographical origin of apple juices.

This study was conducted to investigate the occurrence of aflatoxin M1 (AFM1) in milk products in China using the competitive enzyme-linked immunosorbent assay method and to estimate the dietary exposure to this toxin through a probabilistic approach. Based on the exposure assessment results, a quantitative cancer potency formula developed by the Joint Food and Agriculture Organization and World Health Organization Expert Committee on Food Additives was applied to assess the cancer risk. AFM1 was detected in 48.07% of the milk samples and 4.49% of the yoghurt samples. No samples contained AFM1 above the current regulatory limit in China. The simulated AFM1 intake (90% confidence interval) in various sex-age groups ranged from 0.023 (0.021 to 0.023) ng/kg of body weight per day for 30- to 45-year-old men to 0.382 (0.354 to 0.386) ng/kg of body weight per day for 2- to 4-year-old girls at the 99th percentile. The cancer risk of AFM1 to the general population of China was assessed to be 0.129 cancer cases per year per 10(8) persons at the 99th percentile. These results indicate that the health risk associated with AFM1 in milk in China is relatively low.

A combination of immunomagnetic separation (IMS) and polymerase chain reaction (PCR) was used to detect Alicyclobacillus acidoterrestris (A. acidoterrestris) in apple juice. The optimum technological parameters of the IMS system were investigated. The results indicated that the immunocapture reactions could be finished in 60 min and the quantity of IMPs used for IMS was 2.5 mg/mL. Then the combined IMS-PCR procedure was assessed by detecting A. acidoterrestris in apple juice samples. The agarose gel electrophoresis results of 20 different strains showed that the IMS-PCR procedure presented high specificity to the A. acidoterrestris. The sensitivity of the IMS-PCR was 2×10(1) CFU/mL and the total detection time was 3 to 4 h. Of the 78 naturally contaminated apple juice samples examined, the sensitivity, specificity and accuracy of IMS-PCR compared with the standardized pour plate method were 90.9%, 97.0% and 96.2%, respectively. The results exhibited that the developed IMS-PCR method will be a valuable tool for detecting A. acidoterrestris and improving food quality in juice samples.

A reliable and sensitive liquid chromatography/tandem mass spectrometry (LC-MS/MS) method was developed for the simultaneous determination of aflatoxins (AFB1 , AFB2 , AFG1 , and AFG2 ), ochratoxin A (OTA), deoxynivalenol (DON), zearalenone (ZEA), fumonisin B1 (FB1 ), and T2-toxin in maize. The samples were first extracted using acetonitrile: water: acetic acid (79 : 20 : 1), and then further cleaned-up using OASIS HLB cartridge. Optimum conditions for the extraction and chromatographic separation were investigated. The mean recoveries of mycotoxins in spiked maize ranged from 68.3% to 94.3%. Limits of detection and quantification ranged from 0.01 to 0.64 μg/kg and from 0.03 to 2.12 μg/kg, respectively. The LC-MS/MS method has also been successfully applied to 60 maize samples, which were collected from Shaanxi Province of China. Twenty-four of the total 60 samples (40%) were contaminated with at least 1 of these 9 mycotoxins. Occurrence of mycotoxins were 6.7%, 1.7%, 3.3%, 6.7%, 1.7%, 23.3%, and 3.3% for AFB1 , AFB2 , OTA, ZEA, DON, FB1 , and T2-toxin, respectively. The results demonstrated that the procedure was suitable for the simultaneous determination of these mycotoxins in maize matrix.

The magnetic carboxymethyl chitosan nanoparticles (MNPs-CMC) were developed as effective magnetic affinity adsorbent for Bovine serum albumin and the adsorption reactions were investigated. The obtained experimental data were compared with the adsorption kinetics models and equilibrium isotherms. The experimental kinetic data were modeled using Pseudo-first order, Pseudo-second order, Bangham's equation, Intra-particle diffusion model and Elovich equations. It was found that the adsorption reactions followed the Pseudo-second order kinetics equation. The experimental isotherm data were analyzed using Langmuir, Freundlich, Dubinin–Radushkevich and Temkin equations. By comparing the correlation coefficients determined for each linear transformation of isotherm analysis, it was found that the Langmuir equation was the best fit equilibrium model for the adsorption of BSA. Error functions have been used to determine the alternative single component parameters by nonlinear regression due to the inherent bias in using the correlation coefficient resulting from linearization. It showed that the Langmuir equation resulted in the lowest values for the error function and thus fitted the data better than the other isotherm. Various thermodynamic parameters such as enthalpy (△ H °), entropy (△ S °) and Gibbs free energy (△ G °) were evaluated. MNPs-CMC nanoaprticles were shown to be a promising material for adsorption of BSA from aqueous solutions.

Aims To purify and primarily characterize an anti-Alicyclobacillus bacteriocin produced by Bifidobacterium animalis subsp. animalis CICC 6165, suggested to be named bificin C6165. Methods and Results During purification of the bificin C6165, optimal recovery was achieved with ammonium sulfate precipitation followed by two chromatographic steps. Mass spectrometry analyses revealed a distinctive peak corresponding to a molecular mass of 3395·1 Da. This bacteriocin was heat stable, effective after refrigerated storage and freeze–thaw cycles. The primary mode of action of bificin C6165 is most probably due to pore formation, as indicated by the efflux of K+ from metabolically active cells of Alicyclobacillus acidoterrestris. In the presence of 10 mmol l−1 gadolinium, bificin C6165 did not affect cells of Alicyclobacillus acidoterrestris. This suggests that the mode of action of bificin C6165 relies on a net negatively charged cell surface. Conclusions Bificin C6165 is indeed a novel bacteriocin and it exhibited remarkable potency for Alicyclobacillus control. Significance and Impact of the Study Application of bacteriocins in preservation of fruit juices has seldom been studied. Bificin C6165 may be an alternative method to control juice spoilage by this Alicyclobacillus acidoterrestris and meet increasing consumer demand for nature and artificial chemical additive-free food products.

Apple juice is a complex mixture of volatile and nonvolatile components. To develop discrimination models on the basis of the volatile composition for an efficient classification of apple juices according to apple variety and geographical origin, chromatography volatile profiles of 50 apple juice samples belonging to 6 varieties and from 5 counties of Shaanxi (China) were obtained by headspace solid-phase microextraction coupled with gas chromatography. The volatile profiles were processed as continuous and nonspecific signals through multivariate analysis techniques. Different preprocessing methods were applied to raw chromatographic data. The blind chemometric analysis of the preprocessed chromatographic profiles was carried out. Stepwise linear discriminant analysis (SLDA) revealed satisfactory discriminations of apple juices according to variety and geographical origin, provided respectively 100% and 89.8% success rate in terms of prediction ability. Finally, the discriminant volatile compounds selected by SLDA were identified by gas chromatography-mass spectrometry. The proposed strategy was able to verify the variety and geographical origin of apple juices involving only a reduced number of discriminate retention times selected by the stepwise procedure. This result encourages the similar procedures to be considered in quality control of apple juices. Practical Application: This work presented a method for an efficient discrimination of apple juices according to apple variety and geographical origin using HS-SPME-GC-MS together with chemometric tools. Discrimination models developed could help to achieve greater control over the quality of the juice and to detect possible adulteration of the product.

The polyphenols were extracted from the unripe apple assisted by a highly efficient and simple method of the ultrasound. Response surface methodology was used to investigate the effects of processing parameters, including ultrasound power, extraction time, temperature, and ethanol concentration on total polyphenols yield and polyphenols composition was analyzed by HPLC. Antioxidant activity of the polyphenols was evaluated as 2, 2-diphenyl-1-picrylhydracyl scavenging activity and inhibition activity of lipid peroxidation. The results showed that 10-100 times higher total polyphenols yield was obtained from the unripe apple than those from the reported apple pomace. The optimum extraction conditions were ultrasonic power of 519.39 W, extraction time of 30 min, extraction temperature 50°C, ethanol concentration of 50% gave the total polyphenols yield of 13.26 ± 0.56 mg GAE/g. HPLC analysis indicated that (-)-epicatechin, procyanidin B2, chlorogenic acid, and procyanidin B1 were the predominant polyphenols in unripe apple, which contributed to the higher antioxidant activity to 2, 2-diphenyl-1-picrylhydracyl of unripe apple polyphenols than other apple polyphenols. The extracted polyphenols had higher ability to inhibit lipid peroxidation than butylated hydroxy toluene, which demonstrated that the unripe apple polyphenols have the potential to be used as a substitute of some synthetic antioxidants.

This study aimed to investigate the adsorption of patulin from apple juice, using two types of inactivated yeast powder: laboratory-prepared yeast powder (LYP) and commercial yeast powder (CYP). The effects of incubation time, pH, incubation temperature, adsorbent amount, and initial concentration of patulin and the stability of the yeast-mycotoxin complex were assessed. The results showed that the efficiencies of the two yeast types in adsorbing patulin were similar. The ability of the powders to remove patulin increased with longer incubation times, and patulin concentration was below detectable levels with LYP and CYP at approximately 36 and 30 h, respectively. The highest removal of patulin was achieved at pH 5.0 for both powder types, and there were no significant differences in patulin decrease at different temperatures (4, 29, and 37°C). Additionally, the adsorption percentage of patulin increased significantly with the increase of absorbent amount and decrease of initial concentration of patulin. Stability of the yeast-patulin complex was assessed, and patulin was more stable when washed in phosphate-buffered saline (pH 4.0) than in absolute ethyl alcohol. These results suggest that inactivated yeast powder has potential as a novel and promising adsorbent to bind patulin effectively.

Coenzyme Q10 (CoQ10, ubiquinone), a potent antioxidative dietary supplement, was produced by submerged fermentation usingAgrobacterium tumefaciensinstead of chemical synthesis or solvent extraction.Agrobacterium tumefaciens1.2554 was subjected to mutagenesis using a series of treatments including high hydrostatic pressure (HHP) treatment, UV irradiation, and diethyl sulfate (DES) treatment to obtain mutant strains showing higher CoQ10 production than wild-type strains. A mutant strain PK38 with four genetic markers was isolated: the specific CoQ10 content of the mutant strain increased by 52.83% compared with the original strain. Effects of carbon and nitrogen sources on CoQ10 production with PK38 were studied. Sucrose at concentration of 30 g/l was tested as the best carbon source, and yeast extract at concentration of 30 g/l supplemented with 10 g/l of ammonium sulfate was identified to be the most favorable for CoQ10 production using PK38. Fed-batch culture strategy was then used for increasing production of CoQ10 in 5-l fermentor. Using the exponential feeding fed-batch culture of sucrose, cell growth and CoQ10 formation were significantly improved. With this strategy, the final cell biomass, CoQ10 production, and specific CoQ10 production increased by 126.11, 173.12, and 22.76%, respectively, compared to those of batch culture.

The yield of CoQ10 , an intracellular product extracted from Agrobacterium tumefaciens cells is dependent on the effectiveness of cell lysis post fermentation. Various cell lysis approaches are investigated, including ultrasound, repetitive freezing/thawing, grinding and acid-heat treatment. The acid-heat combination using hydrochloric acid is found the most effective in releasing CoQ 10, followed by lactic, sulfuric, phosphoric and oxalic acids. The most significant processing parameters, namely the ratio of acid solution volume and bacteria weight (A/B ratio), incubation temperature and reaction time, are optimized by using the central composite design with a quadratic regression model built by response surface methodology. The highest CoQ 10 yield at 1.518 mg/g dry cell is attained using hydrochloric acid (3 mol/L) under optimal A/B ratio, temperature and time at 10.8 mL/g, 84.2 °C and 35.3 min, respectively.

Aims Investigation of the response of mesenchymal stem cells (MSCs) to vascular mechanical forces is very important in the field of cardiovascular intervention. Ser/Thr-protein kinase Pim-1 is a novel transducer of cell survival and the cell cycle that promotes signals in the hematopoietic cell system. Current studies aim to foster an understanding of Pim-1 expression and regulation in MSCs in response to different durations and strengths of laminar shear stress (SS) and to investigate the role of Pim-1 in SS-induced cell proliferation. Main methods A parallel-plate flow chamber was used to control the strength and duration of SS. Proliferation was measured with the BrdU cell proliferation assay. The expressions of Pim-1 mRNA and protein were evaluated by reverse transcription-polymerase chain reaction and western blotting, respectively. RNA interference was used to knock down the Pim-1 gene. Key findings The results showed that SS up-regulation of Pim-1 mRNA and protein was time-dependent. Pim-1 induction was SS strength-dependent, and the expression level reached a maximum at 30 dynes/cm2. Inhibitors of p38MAPK and ERK attenuated the SS-induced expression of Pim-1. In addition, SS significantly increased BrdU-uptake, which was effectively blocked by the silencing of Pim-1. Significance These results demonstrated that Pim-1 is expressed in MSCs and plays an important role in the SS-induced proliferation of MSCs.

Coenzyme Q10 (CoQ10), a potent antioxidative dietary supplement, was produced using a photosynthetic bacteria Rhodospirillum rubrum ATCC 25852 by submerged fermentation supplemented with tobacco biomass hydrolysate (TBH) in comparison with media supplemented with hydrolysates prepared with alfalfa (ABH) or spinach (SBH). Growth medium supplemented with 20% (v/v) TBH was found favorable with regard to cell density and CoQ10 concentration. The stimulation effects on cell growth (shortened lag phase, accelerated exponential growth, and elevated final cell concentration) and CoQ10 production (enhanced specific CoQ10 content per unit cell weight) could be attributed to the presence of solanesol, the precursor of CoQ10, in the tobacco biomass. The final yield of CoQ10 reached 20.16 mg/l in the fermentation medium supplemented with 20% TBH.

The purpose of this study was to identify the key factors involved in patulin adsorption by heat-inactivated lactic acid bacteria (LAB) cells. For preventing bacterial contamination, a sterilization process was involved in the adsorption process. The effects of various physical, chemical, and enzymatic pre-treatments, simultaneous treatments, and post-treatments on the patulin adsorption performances of six LAB strains were evaluated. The pre-treated cells were characterized by scanning electron microscopy (SEM). Results showed that the removal of patulin by viable cells was mainly based on adsorption or degradation, depending on the specific strain. The adsorption abilities were widely increased by NaOH and esterification pre-treatments, and reduced by trypsin, lipase, iodate, and periodate pre-treatments. Additionally, the adsorption abilities were almost maintained at pH 2.2-4.0, and enhanced significantly at pH 4.0-6.0. The effects of sodium and magnesium ions on the adsorption abilities at pH 4 were slight and strain-specific. A lower proportion of patulin was released from the strain with higher adsorption ability. Analyses revealed that the physical structure of peptidoglycan was not a principal factor. Vicinal OH and carboxyl groups were not involved in patulin adsorption, while alkaline amino acids, thiol and ester compounds were important for patulin adsorption. Additionally, besides hydrophobic interaction, electrostatic interaction also participated in patulin adsorption, which was enhanced with the increase in pH (4.0-6.0).

An approach based on immunomagnetic separation (IMS) and SYBR Green I real-time PCR (real-time PCR) with species-specific primers and melting curve analysis was proposed as a rapid and effective method for detecting Alicyclobacillus spp. in fruit juices. Specific primers targeting the 16S rDNA sequences of Alicyclobacillus spp. were designed and then confirmed by the amplification of DNA extracted from standard strains and isolates. Spiked samples containing known amounts of target bacteria were used to obtain standard curves; the correlation coefficient was greater than 0.986 and the real-time PCR amplification efficiencies were 98.9%- 101.8%. The detection limit of the testing system was 2.8×101 CFU/mL. The coefficient of variation for intra-assay and inter-assay variability were all within the acceptable limit of 5%. Besides, the performance of the IMS-real-time PCR assay was further investigated by detecting naturally contaminated kiwi fruit juice; the sensitivity, specificity and accuracy were 91.7%, 95.9% and 95.3%, respectively. The established IMS-real-time PCR procedure provides a new method for identification and quantitative detection of Alicyclobacillus spp. in fruit juice.