Metabolomics-based component profiling of hard and semi-hard natural cheeses with gas chromatography/time-of-flight-mass spectrometry, and its application to sensory predictive modeling

Gas chromatography/time-of-flight mass spectrometry (GC/TOF-MS) was used to analyze hydrophilic low molecular weight components, including amino acids, fatty acids, amines, organic acids, and saccharides, in cheese, and the sensometric application for practical metabolomic studies in the food industry is described. Derivatization of target analytes was conducted prior to the GC/TOF-MS analysis. Data on 13 cheeses, six Cheddar cheeses, six Gouda cheeses and one Parmigiano-Reggiano cheese, were analyzed by multivariate analysis. The uniqueness of the Parmigiano-Reggiano cheese metabolome was revealed. Principal component analysis (PCA) showed no grouping of the Cheddar cheeses and Gouda cheeses according to production method or country of origin. The PCA loading plot confirms that many amino acids contribute positively to PC1, suggesting that PC1 is closely related to degradation of proteins, and that lactic acid contributed positively to PC2, whereas glycerol contributed negatively to PC2, suggesting that factors regarding degradation of carbohydrates and fats were expressed in PC2. Partial least squares (PLS) regression models were constructed to predict the relationship between the metabolite profile and two sensory attributes, "Rich flavor" and "Sour flavor", which were related to maturation. The compounds that play an important role in constructing each sensory prediction model were identified as 12 amino acids and lactose for "Rich flavor", and 4-aminobutyric acid, ornithine, succinic acid, lactic acid, proline and lactose for "Sour flavor". The present study revealed that metabolomics-based component profiling, focusing on hydrophilic low molecular weight components, was able to predict the sensory characteristics related to ripening.     

Selection of acid tolerant bifidobacteria and evidence for a low-pH-inducible acid tolerance response in Bifidobacterium longum

Acidity is an environmental condition commonly encountered by lactic acid bacteria and bifidobacteria in the gastrointestinal tract and fermented foods. In the present study, 22 strains of Bifidobacterium were screened for acid tolerance in artificial gastric juice (AGJ, pH 3.0) and fermented milk. AGJ tolerance was found to be strain-specific, with a pronounced variation among the strains. Several strains with a high survival rate in AGJ that belonged to Bifid. longum, Bifid. breve and Bifid. adolescentis were selected. Among them, only strain BL1 of Bifid longum was found to possess a high survival rate in fermented milk during refrigerated storage. Strain BL1 exhibited a survival rate of more than 25% in AGJ at pH 3.0 for 2 h and maintained a viable cfu level of more than 10(8) per gram of product in fermented milk (pH 4.6) under refrigerated conditions for 2 weeks. The acid tolerance of strain BL1 was found to depend on the final growth pH (<4.5). Rapid loss of acid tolerance was observed when the cells were shifted from acid to neutral conditions by addition of NaOH. Strain BL1 cells were able to maintain much higher intracellular pH under acid conditions, in comparison with those of AGJ sensitive mutant (BL1-S) or cells that lost acid tolerance following pH shifting from acid to neutral conditions. These results suggested that a cytoplasmic pH homeostasis system may function in the acid tolerance response in this strain.     

Antioxidant Effects of Herbal Tea Leaves from Yacon (Smallanthus sonchifolius) on Multiple Free Radical and Reducing Power Assays,Especially on Different Superoxide Anion Radical Generation Systems

Yacon (Smallanthus sonchifolius), a native Andean plant, has been cultivated as a crop and locally used as a traditional folk medicine for the people suffering from diabetes and digestive/renal disorders. However, the medicinal properties of this plant and its processed foods have not been completely established. This study investigates the potent antioxidative effects of herbal tea leaves from yacon in different free radical models and a ferric reducing model. A hot‐water extract exhibited the highest yield of total polyphenol and scavenging effect on 1,1‐diphenyl‐2‐picryl hydrazyl (DPPH) radical among four extracts prepared with hot water, methanol, ethanol, and ethylacetate. In addition, a higher reducing power of the hot‐water extract was similarly demonstrated among these extracts. Varying concentrations of the hot‐water extract resulted in different scavenging activities in four synthetic free radical models: DPPH radical (EC₅₀ 28.1 μg/mL), 2,2′‐azinobis(3‐ethylbenzothiazoline‐6‐sulfonic acid) cation radical (EC₅₀ 23.7 μg/mL), galvinoxyl radical (EC₅₀ 3.06 μg/mL), and chlorpromazine cation radical (EC₅₀ 475 μg/mL). The yacon tea‐leaf extract further demonstrated superoxide anion (O₂^?) radical scavenging effects in the phenazine methosulfate‐NADH‐nitroblue tetrazolium (EC₅₀ 64.5 μg/mL) and xanthine oxidase assay systems (EC₅₀ 20.7 μg/mL). Subsequently, incubating human neutrophilic cells in the presence of the tea‐leaf extract could suppress the cellular O₂^? radical generation (IC₅₀ 65.7 μg/mL) in a phorbol 12‐myristate 13‐acetate‐activated cell model. These results support yacon tea leaves may be a good source of natural antioxidants for preventing O₂^? radical‐mediated disorders.     

Exfoliation of non-swelling muscovite on dodecylammonium chloride intercalation between layers using wet-jet milling

The exfoliation of layered muscovite with non-swelling property has been performed by combining various processes, such as heating, intercalation, and wet-jet milling. The c axis of muscovite was expanded from 2004.0 to 2022.8 pm at 800 °C without the destruction of crystallinity of muscovite. The heating at 800 °C led to the weak attraction force between potassium ions and silicate layers by hydroxylation of muscovite. The muscovite heated at 800 °C progressed the intercalation of dodecylammonium chloride (DDAC) into the layers effectively. Furthermore, the DDAC molecules were inserted to the interlayer of muscovite effectively by suppressing the formation of micelle of DDAC. The sedimentation test of wet-jet milled muscovite slurry showed that the relative packing density of muscovite was decreasing with increasing the amount of the intercalated DDAC. As results, the aspect ratio of muscovite prepared with combining the heating, the intercalation and the wet-jet milling was increased by 253% as compared to the raw muscovite. The aspect ratio was calculated from laser particle size distribution and thickness size distribution which was estimated from field emission-scanning electron microscopic images. The expansion of the interlayer led to the effective exfoliation of muscovite with high aspect ratio.     

Zirconia Coating on Stainless Steel Sheets from Organozirconium Compounds

The stability and the coating characteristics of a variety of organozirconium compounds have been investigated to coat ZrO₂ films on stainless steets for the improvement of heat resistance against oxidation. Zirconium tetraoctylate and zirconium tetrakis(acetylacetonate) were found to be more stable than zirconium alkoxides in an ambient atmosphere. The ZrO₂ films from zirconium alkoxides became white and discontinuous when the thickness of the films exceeded about 200 Å (20 nm). On the other hand, transparent ZrO₂ films were obtained from zirconium tetraoctylate and zirconium tetrakis(acetylacetonate). The IR spectra of the films showed that the heat treatment at 400°C for 5 min produced organic-free ZrO₂ films. The heat resistance of stainless steel sheets against oxidation was improved by the ZrO₂ coating, depending on the film thickness.     

Formation, physical stability and in vitro antimalarial activity of dihydroartemisinin nanosuspensions obtained by co-grinding method

The purpose of this study was to investigate the formation of drug nanoparticles from binary and ternary mixtures, consisting of dihydroartemisinin (DHA), a poorly water-soluble antimalarial drug, with water-soluble polymer and/or surfactant. Binary mixtures of drug/polyvinyl pyrrolidone K30 (PVP K30), binary mixtures of drug/sodium deoxycholate (NaDC), and ternary mixtures of drug/PVP K30/NaDC were prepared at different weight ratios and then ground by vibrating rod mill to obtain ground mixtures. Nanosuspension was successfully formed after dispersing ternary ground mixtures or DHA/NaDC ground mixtures in water. The ternary ground mixtures did not give superior nanosuspension in terms of particle size reduction and recovery of drug nanoparticles, but they provided more physically stable nanosuspensions than DHA/NaDC ground mixtures. The size of drug nanoparticles was decreased with increasing grinding time and lowering amount of PVP K30 and NaDC. About 95% of drug nanoparticles were found in the nanosuspension from ternary ground mixtures. Zeta potential measurement suggested that stable nanosuspension was attributable to adsorption of NaDC and PVP K30 onto surface of drug particles. Atomic force microscopy and transmission electron microscopy with selected area diffraction indicated that DHA in nanosuspension was existed as nanocrystals. The obtained nanosuspensions had higher in vitro antimalarial acitivity against Plasmodium falciparum than microsuspensions. The results suggest that co-grinding of DHA with PVP K30 and NaDC seems to be a promising method to prepare DHA nanosuspension.     

Kinetic studies on lipase-catalyzed acetylation of 2-alkanol with vinyl acetate in organic solvent

Lipase-catalyzed acetylation of 2-alkanol with vinyl acetate has been studied kinetically using Burkholderia cepacia lipase (BCL), enantiomerically pure (R)- and (S)-2-alkanols and different organic solvents. The rate equation was derived by the steady state method for the simplified mechanism. The second order rate constants (k(R) and k(S)) for (R)- and (S)-2-alkanols were evaluated from the slopes of the double reciprocal plots, v(-1) vs. [2-alkanol](-1), where v is the initial rate of the reaction. The log k(R) value increased with the solvent hydrophobicity log P, where P is a partition coefficient of a given solvent between octanol and water. The log k(S) value also increased with log P except the bulky solvents such as 1,4-dioxane and cyclohexane, in which the rates were faster than those expected from the log k(S) vs. log P plot. The slope of log k(S) vs. log P plot was larger than that for (R)-2-alkanol. Thus, log E (E=k(R)/k(S): enantioselectivity) decreased with log P except the bulky solvents. The rate constants and the enantioselectivity were different depending on the structure (carbon number CN) of 2-alkanol. The log E vs. CN plot was minimized at CN=8 and 10 and the log k(S) vs. CN plot maximized at CN=8 and 10. In contrast the log k(R) vs. CN plot showed a different feature from the log E vs. CN plot. These facts suggest that dependence of E on CN is more strongly affected by the reactivity of (S)-2-alkanol than that of (R) isomer in this acetylation.     

Quantum chemical molecular dynamics study of "shrinking" of hot giant fullerenes

We present a detailed analysis of size-down trajectories of the "Shrinking Hot Giant" road of fullerene formation, revealed before by our quantum chemical molecular dynamics simulations. It is found that shrinkage of giant fullerenes occurs in two stages, fall-off and pop-out stage, respectively, and that it is an irreversible process occurring naturally at high temperatures. The driving force behind the energetically unfavorable C2 evaporation is the increase in entropy and pi-conjugation stabilization of the increasingly more spherical fullerene cages. A comparison of the theoretical results with existing experimental data shows the importance of annealing for the synthesis of smaller (C60, C70) fullerenes.     

Stress-induced voiding in stacked tungsten via structure

The stress-induced voiding (SV) in Al-alloy films with stacked tungsten via structures was investigated. Voids were found in interconnections with stacked and borderless vias that had resistance increase after the aging tests. Failure occurs most frequently when the test structures are stored at approximately 250°C. This behavior can be explained by the diffusion creep model similar to SV in a flat line [1]. Finite-element simulations show that tensile stress in Al-lines between upper and lower plugs increases with temperature increase over 175°C. Al grains on W-plugs were found to have high-angle crystalline misorientation in transmission electron microscopy (TEM) observation. The tensile stress and grain misorientation should accelerate the void growth during high temperature storage. O₂ plasma post metal etch treatment is effective to eliminate SV in stacked via structure.     

A novel quarter‐wave retardation film for improving viewing angle properties in time‐sequential stereoscopic 3D‐LCDs

We have successfully developed a quarter‐wave retardation film (QWF) for wide viewing angle 3D liquid crystal displays (3D‐LCDs) that provides high luminance, low crosstalk, low color change, and low head‐tilt‐angle dependency. It was found that the out‐of‐plane retardation (Rth) of the QWF in the LCD needs to be close to 0 nm in order to improve the 3D display properties at an off‐axis position and that the in‐plane retardation (Re) needs to be adjusted from 120 to 130 nm to achieve low color change with head tilting. We adopted a coating process for making our QWF because of its potential for retardation control. 3D‐LCDs with this QWF whose Rth was nearly zero had high performance and allowed off‐axis other than on‐axis.