Magnetic field-driven biochemical landscape of browning abatement in goat milk using spatial-omics uncovers.

Influence of magnetic field (MF) treatment on the glycation of goat milk proteins is yet to be elucidated. Proteomic and metabolomic analyses of brown goat milk samples with and without MF treatment were performed. Assessed glycation degree and structural modification of proteins explained that MF treatment dramatically down-regulated the glycation of brown goat milk protein, possibly due to the aggregation behavior induced by MF treatment, which consumed additional glycation sites as well as altered their accessibility and preference. Integrated datasets uncovered that the energy metabolism-related biological events including carbohydrate metabolism, glycerophospholipid metabolism, TCA cycle may mainly account for the browning abatement mechanism of MF. In addition, MF treatment enhanced both the quality and flavor of brown goat milk. This study suggests the feasibility of MF treatment to reduce glycation in brown goat milk for producing high-quality dairy ingredients and products.

Thymol and trans-cinnamaldehyde induce apoptosis through a metacaspase-dependent mitochondrial pathway in food-spoilage yeast Zygosaccharomyces rouxii.

This work aimed for the first time to provide detailed insights into thymol and trans-cinnamaldehyde's mechanisms of action on the food-spoilage yeast Zygosaccharomyces rouxii and offers evidence in favor of the activation of an apoptosis-like phenotype. The action mechanisms of thymol and trans-cinnamaldehyde were investigated by the measurement of a series of typical apoptotic features using flow cytometer or microplate reader. Moreover, quantitative reverse transcription PCR (QRT-PCR) was performed to investigate the effects of thymol and trans-cinnamaldehyde on the transcription of key regulators of apoptosis in Z. rouxii. The results indicated that the treatment of Z. rouxii with thymol or trans-cinnamaldehyde (minimum inhibitory and subinhibitory concentrations) triggered reactive oxygen species (ROS) accumulation, elevated intracellular Ca2+ level, and depolarized mitochondrial membrane potential (MMP) coupled with hallmarks of apoptosis including mitochondrial cytochrome c (cyt c) release, metacaspase activation, phosphatidylserine (PS) exposure, and DNA fragmentation. Moreover, thymol or trans-cinnamaldehyde treatment upregulated the transcription of proapoptotic regulators including Yca1, Dnm1, Nuc1, Ndi1, and Mmi1 and downregulated the transcription of antiapoptotic regulators of Fis1 and Cdc48, further confirming the apoptosis induced by thymol and trans-cinnamaldehyde in Z. rouxii. In summary, thymol and trans-cinnamaldehyde probably induced apoptosis through a metacaspase-dependent mitochondrial pathway in Z. roxuii. These findings provide theoretical support for the development of safe and efficient agents used in food preservation. PRACTICAL APPLICATION: The results will provide a new idea for the systematic analysis of the antifungal mechanisms of thymol and trans-cinnamaldehyde and also provide a theoretical support for the development and application of natural food preservatives, which is of positive significance for the effective control of the spoilage caused by Z. rouxii in food processing and storage and the protection of consumers' health.

Feasibility study on early identification of freshness decay of fresh-cut kiwifruit during cold chain storage by Fourier transform-near infrared spectroscopy combined with chemometrics.

This work mainly aimed to evaluate the feasibility of Fourier transform-near infrared spectroscopy (FT-NIRS) combined with chemometrics in early identification of freshness decay of fresh-cut kiwifruit during simulated cold chain storage, with organoleptic evaluation as a reference. By linear discriminant analysis, the freshness decay could be identified after only 2 days of cold storage, corresponding to freshness level of 3.41 ± 0.27 N (hardness), 0.70 ± 0.05 g/kg (total acid), 8.62 ± 0.06 g/100 g (reducing sugars), 62.04 ± 1.03 mg/100 g (vitamin C) and 2.05 ± 0.11 log10 CFU/g (total plate count). Organoleptic evaluators could not perceive the freshness decay that occurred after 2 days of cold storage. Moreover, the freshness decay could be well quantitatively predicted by partial least squares regression, with low RMSEp (0.18-05.42) and high R2 (0.90-0.96). FT-NIRS appears to be a promising option for early warning of the freshness decay of fresh-cut kiwifruit during cold chain storage, thereby preventing serious spoilage and ensuring fresh fruits for consumers. PRACTICAL APPLICATION: This work is based on the fact that fresh-cut kiwifruit is prone to freshness decay under unstable cold chain conditions, using FT-NIRS combined with chemometrics to identify the freshness decay early and rapidly, to a certain extent, early warn freshness decay and effectively prevent serious spoilage. The technology can be used for food regulatory agencies to monitor the freshness of fresh-cut kiwifruit, and can also be applied for fruit processing enterprises and dealers to ensure the freshness and high quality of fresh-cut kiwifruit.

Antifungal activities and mechanisms of trans-cinnamaldehyde and thymol against food-spoilage yeast Zygosaccharomyces rouxii.

Due to hydrophobicity, plant essential oil components trans-cinnamaldehyde and thymol exert broad-spectrum antimicrobial activities by interfering with the structures and functions of microbial cell membranes, yet their antifungal properties against food-spoilage yeast Zygosaccharomyces rouxii are still unclear. This work aimed to elucidate their antifungal activities and mechanisms against Z. rouxii. Minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) were measured using broth dilution method to evaluate the antifungal activities. Membrane permeability, potential and integrity, intracellular ATP content, and intracellular pH (pHin ) were determined, and membrane damage was observed by scanning electron microscope (SEM) to investigate the antifungal mechanisms. The MICs of trans-cinnamaldehyde and thymol were 0.3125 and 0.0625 mg/ml, respectively, and the MFCs were 1.25 and 0.125 mg/ml, respectively. Increased membrane permeability, membrane hyperpolarization, decreased membrane integrity, reduced intracellular ATP content, and lowered pHin were found after trans-cinnamaldehyde or thymol treatment at the MIC and MFC levels, and thymol exhibited more severe influence on cell membrane. SEM observation further confirmed that trans-cinnamaldehyde caused cell membrane shrinkage and invagination, while thymol resulted in perforation in cell membrane. These results suggest that trans-cinnamaldehyde and thymol exert anti-Z. rouxii activities by inducing cell membrane damage and subsequent intracellular homeostasis disruption. Cellular contents leakage and intracellular homeostasis disruption probably proceed more rapidly after thymol inducing perforation in the cell surface, thereby thymol exerting better anti-Z. rouxii activity. PRACTICAL APPLICATION: The addition of a small amount of thymol (0.125 mg/ml) is sufficient to effectively inhibit the growth of Z. rouxii in apple juice concentrate. Based on our previous work, the addition of thymol (0.125 mg/ml) in apple juice concentrate did not significantly affect the organoleptic property of the diluted apple juice (5°Brix) used for producing commercial apple juice products. Therefore, thymol is practical for application at the effective concentration (0.125 mg/ml) in apple juice concentrate.

Assessment of different antimicrobials to inhibit the growth of Zygosaccharomyces rouxii cocktail in concentrated apple juice.

Zygosaccharomyces rouxii represents the main spoilage cause of concentrated apple juice, leading to waste of products or recalls. Essential oils components derived from plants have been found to present antimicrobial activities against various microbes. However, few work has been reported about their antimicrobial activities against Z. rouxii in concentrated apple juice. In this work, reparameterized Gompertz equation was used to evaluate the antimicrobial activities of different antimicrobials to inhibit the growth of a Z. rouxii cocktail (6.3 lg colony forming units/mL) composed of six strains isolated from concentrated apple juice and two strains from honey and grape must. The obtained mathematical models presented that thymol, carvacrol and trans-cinnamaldehyde were the promising options to inhibit Z. rouxii in 30 oBrix apple juice, reaching a maximal decrease on yeast growth of around 99.65 ± 0.61%. Whereas other antimicrobials showed lower antimicrobial activities with a maximal growth decrease of ranging from 67.13 ± 3.62% to 13.38 ± 1.16%. Additionally, the sensorial characteristics were not affected when the antimicrobials assayed were applied at the effective concentrations in commercial apple juice product. This work provided a theoretical feasibility that thymol, carvacrol and trans-cinnamaldehyde could be applied as natural preservatives for the control of Z. rouxii-related spoilage in fruit juice industry.

Potential use of electronic tongue coupled with chemometrics analysis for early detection of the spoilage of Zygosaccharomyces rouxii in apple juice.

Apple juice spoilage by Zygosaccharomyces rouxii could scarcely be identified at early stage. It is crucial to recognize the spoilage at early stage to prevent waste of products. In present study, electronic tongue was applied to detect the spoilage of Z. rouxii in apple juice, using taste evaluation by panelists as reference. Combined with linear discriminant analysis, identification of the contaminated juice was fulfilled after 12 h, equivalent to yeast population of less than 2.0 lg colony forming units/mL. At the level, panelists were not capable of discerning the spoilage. Sensors HA, ZZ, BB and BA were relatively more sensitive to the changes in overall taste of apple juice. Moreover, cell number of Z. rouxii could be properly quantified by partial least squares regression models with high determination coefficient of 0.98-0.99. Electronic tongue appears to be a powerful approach to realize early detection of contamination of Z. rouxii.

Protein abundance changes of Zygosaccharomyces rouxii in different sugar concentrations.

Zygosaccharomyces rouxii is a yeast which can cause spoilage in the concentrated juice industries. It exhibits resistance to high sugar concentrations but genome- and proteome-wide studies on Z. rouxii in response to high sugar concentrations have been poorly investigated. Herein, by using a 2-D electrophoresis based workflow, the proteome of a wild strain of Z. rouxii under different sugar concentrations has been analyzed. Proteins were extracted, quantified, and subjected to 2-DE analysis in the pH range 4-7. Differences in growth (lag phase), protein content (13.97-19.23mg/g cell dry weight) and number of resolved spots (196-296) were found between sugar concentrations. ANOVA test showed that 168 spots were different, and 47 spots, corresponding to 40 unique gene products have been identified. These protein species are involved in carbohydrate and energy metabolism, amino acid metabolism, response to stimulus, protein transport and vesicle organization, cell morphogenesis regulation, transcription and translation, nucleotide metabolism, amino-sugar nucleotide-sugar pathways, oxidoreductases balancing, and ribosome biogenesis. The present study provides important information about how Z. rouxii acts to cope with high sugar concentration at molecular levels, which might enhance our global understanding of Z. rouxii's high sugar-tolerance trait.

Accessing spoilage features of osmotolerant yeasts identified from kiwifruit plantation and processing environment in Shaanxi, China.

Osmotolerant yeasts originating from kiwifruit industrial chain can result in spoilage incidences, while little information is available about their species and spoilage features. This work identified possible spoilage osmotolerant yeasts from orchards and a manufacturer (quick-freeze kiwifruit manufacturer) in main producing areas in Shaanxi, China and further characterized their spoilage features. A total of 86 osmotolerant isolates dispersing over 29 species were identified through 26S rDNA sequencing at the D1/D2 domain, among which Hanseniaspora uvarum occurred most frequently and have intimate relationships with kiwifruit. RAPD analysis indicated a high variability of this species from sampling regions. The correlation of genotypes with origins was established except for isolates from Zhouzhi orchards, and the mobility of H. uvarum from orchard to the manufacturer can be speculated and contributed to spoilage sourcing. The manufacturing environment favored the inhabitance of osmotolerant yeasts more than the orchard by giving high positive sample ratio or osmotolerant yeast ratio. The growth curves under various glucose levels were fitted by Grofit R package and the obtained growth parameters indicated phenotypic diversity in the H. uvarum and the rest species. Wickerhamomyces anomalus (OM14) and Candida glabrata (OZ17) were the most glucose tolerant species and availability of high glucose would assist them to produce more gas. The test osmotolerant species were odor altering in kiwifruit concentrate juice. 3-Methyl-1-butanol, phenylethyl alcohol, phenylethyl acetate, 5-hydroxymethylfurfural (5-HMF) and ethyl acetate were the most altered compound identified by GC/MS in the juice. Particularly, W. anomalus produced 4-vinylguaiacol and M. guilliermondii produced 4-ethylguaiacol that would imperil product acceptance. The study determines the target spoilers as well as offering a detailed spoilage features, which will be instructive in implementing preventative measures to increase production safety of kiwifruit.

A novel method to quantify the activity of alcohol acetyltransferase Using a SnO2-based sensor of electronic nose.

Alcohol acetyltransferase (AATFase) extensively catalyzes the reactions of alcohols to acetic esters in microorganisms and plants. In this work, a novel method has been proposed to quantify the activity of AATFase using a SnO2-based sensor of electronic nose, which was determined on the basis of its higher sensitivity to the reducing alcohol than the oxidizing ester. The maximum value of the first-derivative of the signals from the SnO2-based sensor was therein found to be an eigenvalue of isoamyl alcohol concentration. Quadratic polynomial regression perfectly fitted the correlation between the eigenvalue and the isoamyl alcohol concentration. The method was used to determine the AATFase activity in this type of reaction by calculating the conversion rate of isoamyl alcohol. The proposed method has been successfully applied to determine the AATFase activity of a cider yeast strain. Compared with GC-MS, the method shows promises with ideal recovery and low cost.

Early detection of Zygosaccharomyces rouxii--spawned spoilage in apple juice by electronic nose combined with chemometrics.

Spoilage spawned by Zygosaccharomyces rouxii can cause sensory defect in apple juice, which could hardly be perceived in the early stage and therefore would lead to the serious economic loss. Thus, it is essential to detect the contamination in early stage to avoid costly waste of products or recalls. In this work the performance of an electronic nose (e-nose) coupled with chemometric analysis was evaluated for diagnosis of the contamination in apple juice, using test panel evaluation as reference. The feasibility of using e-nose responses to predict the spoilage level of apple juice was also evaluated. Coupled with linear discriminant analysis (LDA), detection of the contamination was achieved after 12h, corresponding to the cell concentration of less than 2.0 log 10 CFU/mL, the level at which the test panelists could not yet identify the contamination, indicating that the signals of e-nose could be utilized as early indicators for the onset of contamination. Loading analysis indicated that sensors 2, 6, 7 and 8 were the most important in the detection of Z. rouxii-contaminated apple juice. Moreover, Z. rouxii counts in unknown samples could be well predicted by the established models using partial least squares (PLS) algorithm with high correlation coefficient (R) of 0.98 (Z. rouxii strain ATCC 2623 and ATCC 8383) and 0.97 (Z. rouxii strain B-WHX-12-53). Based on these results, e-nose appears to be promising for rapid analysis of the odor in apple juice during processing or on the shelf to realize the early detection of potential contamination caused by Z. rouxii strains.

Detection of Zygosaccharomyces rouxii and Candida tropicalis in a High-Sugar Medium by a Metal Oxide Sensor-Based Electronic Nose and Comparison with Test Panel Evaluation.

Osmotolerant yeasts are primarily responsible for spoilage of sugar-rich foods. In this work, an electronic nose (e-nose) was used to diagnose contamination caused by two osmotolerant yeast strains (Zygosaccharomyces rouxii and Candida tropicalis) in a high-sugar medium using test panel evaluation as the reference method. Solid-phase microextraction gas chromatography with mass spectrometry (GC-MS) was used to determine the evolution of the volatile organic compound fingerprint in the contaminated samples during yeast growth. Principal component analysis and linear discriminant analysis revealed that the e-nose could identify contamination after 48 h, corresponding to the total yeast levels of 3.68 (Z. rouxii) and 3.09 (C. tropicalis) log CFU/ml. At these levels, the test panel could not yet diagnose the spoilage, indicating that the e-nose approach was more sensitive than the test panel evaluation. Loading analysis indicated that sensors 8 and 6 were the most important for detection of these two yeasts. Based on the result obtained with the e-nose, the incubation time and total yeast levels could be accurately predicted by established multiple regression models with a correlation of greater than 0.97. In the sensory evaluation, spoilage was diagnosed after 72 h in samples contaminated with C. tropicalis and after 48 to 72 h for samples contaminated with Z. rouxii. GC-MS revealed that compounds such as acetaldehyde, acetone, ethyl acetate, alcohol, and 3-methyl-1-butanol contributed to spoilage detection by the e-nose after 48 h. In the high-sugar medium, the e-nose was more sensitive than the test panel evaluation for detecting contamination with these test yeast strains. This information could be useful for developing instruments and techniques for rapid scanning of sugar-rich foods for contamination with osmotolerant yeasts before such spoilage could be detected by the consumer.

Characterization of Osmotolerant Yeasts and Yeast-Like Molds from Apple Orchards and Apple Juice Processing Plants in China and Investigation of Their Spoilage Potential.

Yeasts and yeast-like fungal isolates were recovered from apple orchards and apple juice processing plants located in the Shaanxi province of China. The strains were evaluated for osmotolerance by growing them in 50% (w/v) glucose. Of the strains tested, 66 were positive for osmotolerance and were subsequently identified by 26S or 5.8S-ITS ribosomal RNA (rRNA) gene sequencing. Physiological tests and RAPD-PCR analysis were performed to reveal the polymorphism of isolates belonging to the same species. Further, the spoilage potential of the 66 isolates was determining by evaluating their growth in 50% to 70% (w/v) glucose and measuring gas generation in 50% (w/v) glucose. Thirteen osmotolerant isolates representing 9 species were obtained from 10 apple orchards and 53 target isolates representing 19 species were recovered from 2 apple juice processing plants. In total, members of 14 genera and 23 species of osmotolerant isolates including yeast-like molds were recovered from all sources. The commonly recovered osmotolerant isolates belonged to Kluyveromyces marxianus, Hanseniaspora uvarum, Saccharomyces cerevisiae, Zygosaccharomyces rouxii, Candida tropicalis, and Pichia kudriavzevii. The polymorphism of isolates belonging to the same species was limited to 1 to 3 biotypes. The majority of species were capable of growing within a range of glucose concentration, similar to sugar concentrations found in apple juice products with a lag phase from 96 to 192 h. Overall, Z. rouxii was particularly the most tolerant to high glucose concentration with the shortest lag phase of 48 h in 70% (w/v) glucose and the fastest gas generation rate in 50% (w/v) glucose.

[Differentiation and Identification of Alicyclobacillus Strains by Fourier Transform Near-Infrared Spectroscopy].

Fourier transform near-infrared spectroscopy (FT-NIR) can reflect the overall molecular composition of microbial cells to identify different types of microorganisms. To establish an accurate, effective method about the differentiation and identification of Alicyclobacillus strains between different species, the present research performed the following studies by FT-NIR: (1) The FT-NIR spectra about seven type stains was clustered for data analysis. After preprocessing, reduction of data was performed by Principal Component Analysis (PCA) and Linear Discriminant Analysis(LDA), exploring the feasibility of differentiation and identification between different species, the result suggested that the PCA model can cluster the seven species of Alicyclobacillus strains correctly and the LDA model I can predict the unknown species with 100% accuracy. It evidenced that the method could identify different species of Alicyclobacillus strains preliminary. (2)In order to improve the robustness and practicability of the model, a total of 41 Alicyclobacillus strains including type and isolated strains were prepared for LDA model II, using the same methods as mentioned before. The result indicated that the LDA model validated by fifteen sample with 86.67% accuracy. It was more perfect and more comprehensive. As a result, the FT-NIR technology combined with chemometrics method can accurately and effectively identify Alicyclobacillus strains between different microbial species.

Young environment reverses the declined activity of aged rat-derived endothelial progenitor cells: involvement of the phosphatidylinositol 3-kinase/Akt signaling pathway.

BACKGROUND: Although age-related impairment of endothelial progenitor cells (EPCs) has been documented in recent studies, the detailed role of aging-induced environment in EPCs remains unclear. METHODS: Two and 20 months old Sprague-Dawley female rats were used in the present study. EPCs isolated from young (YEPCs) and aged (AEPCs) rats were cultured with young or aged serum. EPC migration and proliferation were detected with a modified Boyden chamber and the MTT assay, respectively; EPC differentiation was detected by reverse-transcription polymerase chain reaction or fluorescence-activated cell sorting; Akt and phosphorylated-Akt protein expression was detected with Western blotting. EPC transplantation was performed in the rat carotid artery injury models. RESULTS: Young serum significantly promotes AEPC migration, proliferation, and differentiation and increases phosphatidylinositol 3-kinase (PI3-K) and endothelial nitric oxide synthase activity in AEPCs compared with aged serum; total-Akt and phosphorylated-Akt protein expressions in AEPCs are also significantly upregulated by young serum. Transplanted AEPC numbers at vascular injury sites in the young rat carotid artery injury model significantly increased compared with those in aged models. Those effects could be reasonably attenuated by the PI3-K-specific blocker wortmannin. CONCLUSION: A young environment partly restores the declined AEPC activity and promotes AEPC homing to vascular injury sites; activation of the PI3-K/Akt signaling pathway is at least partly responsible for this process.