Formation of Epoxy Fatty Acids in Triacylglycerol Standards during Heating.

Epoxy fatty acid formation during heating was estimated using triolein (OOO) and trilinolein (LLL). Epoxy octadecanoic acids were found in heated OOO, while epoxy octadecenoic acids were found in heated LLL. The content of epoxy fatty acids increased with heating time, and trans-epoxy fatty acids were formed significantly more than cis-epoxy fatty acids. A comparison between OOO and LLL indicated that epoxy fatty acid formation was higher in the OOO than that in the LLL. Heating tests in the presence of α- tocopherol suggested that the formation of epoxy fatty acids could be suppressed by antioxidants.

Upregulated Genes in Atrial Fibrillation Blood and the Left Atrium.

INTRODUCTION: Atrial fibrillation (AF) is a common arrhythmia associated with aging. Many known risk factors are associated with AF, but many senior individuals do not develop AF despite having multiple risk factors. This finding suggests that other factors may be involved in AF onset. This study aimed to identify upregulated genes in the peripheral blood and left atrium of patients with AF. These genes may serve as potential biomarkers to predict AF onset risk and its complications. METHODS: Gene expression data was analyzed from blood (n = 3) and left atrial samples (n = 15) of patients with AF and sinus rhythm. We evaluated the significant genes identified using p-value analysis of weighted average difference to confirm their rankings. We created figures for the genes using GeneMANIA and performed a functional analysis using Cytoscape3.10.1. Hub and bottleneck genes were identified based on degree and betweenness centrality. We used RefEx to confirm the organs in which the extracted genes were expressed. Heatmaps and Gene ontology term evaluation were performed to further elucidate the biological functions of the genes. RESULTS: We identified 12 upregulated genes (CAST, ASAH1, MAFB, VCAN, DDIT4, FTL, HEXB, PROS1, BNIP3L, PABPC1, YBX3, and S100A6) in both the blood and left atrium of patients with AF. We analyzed the gene functions using GeneMANIA and Cytoscape. The identified genes were involved in a variety of pathways, including lysosomal function and lipid and sphingolipid catabolism. Next, we investigated whether the 12 identified genes identified were systemically expressed or had high organ specificity. Finally, Reference expression (RefEx) was used to analyze the gene expression levels in various tissues. Four genes; FTL, ASAH1, S100A6, and PABPC1, were highly expressed in the normal heart tissue. Finally, we evaluated the expression levels of the 12 genes in the blood of patients with AF using a heatmap. Our findings suggest that the 12 genes identified in this study, especially the lysosome-related genes (FTL and ASAH1), may be involved in AF pathogenesis. CONCLUSION: Lysosome-related genes may be important to understand the AF pathophysiology and to develop AF-related future studies.

Coordination Structure of Si(II) Ions in Binary Melts (LiF-KF, LiF-NaF, and NaF-KF) and Ternary Melt (LiF-NaF-KF).

The formation of Si(II) by the comproportionation reaction between electrodeposited silicon and Si(IV) ions [Si(IV) + Si0 ⇄ 2Si(II)] in fluoride melts has been discussed in many previous reports on the electrochemical processing of silicon. In this study, the coordination structure of Si(II) ions in LiF-KF, LiF-NaF, NaF-KF, and LiF-NaF-KF melts was investigated by high-temperature Raman spectroscopy and quantum chemical calculation based on density functional theory. The formation of Si(II) ions was induced by mixing metallic silicon powder as the source of Si0, with the melts containing hexafluoro silicate or silicon dioxide as the source of Si(IV) ions. The simulated vibrational modes of the Si(II) species and the experimental Raman bands showed good agreement. It was found that both Si(II) and Si(IV) species were detected in either fluoride melt, demonstrating that the proposed comproportionation reaction between Si0 and Si(IV) occurred. In addition, the interaction between Si(II) and F-, O2-, Li+, Na+, and K+ ions significantly changed depending on the composition of fluoride melts. The coordination structure of the Si(II) ions reported in this study will provide clues for controlling the electrodeposition process of silicon in fluoride melts.

Tunable structure of chimeric isomaltomegalosaccharides with double α-(1 → 4)-glucosyl chains enhances the solubility of water-insoluble bioactive compounds.

Isomaltomegalosaccharides with α-(1 â†’ 4) and α-(1 â†’ 6)-segments solubilize water-insoluble ligands since the former complexes with the ligand and the latter solubilizes the complex. Previously, we enzymatically synthesized isomaltomegalosaccharide with a single α-(1 â†’ 4)-segment at the reducing end (S-IMS) by dextran dextrinase (DDase), but the chain length [average degree of polymerization (DP) ≤ 9] was insufficient for strong encapsulation. We hypothesized that the conjugation of longer α-(1 â†’ 4)-segment afforded the promising function although DDase is incapable to do so. In this study, the cyclodextrin glucanotransferase-catalyzed coupling reaction of α-cyclodextrin to S-IMS synthesized a new α-(1 â†’ 4)-segment at the nonreducing end (N-4S) of S-IMS to form D-IMS [IMS harboring double α-(1 â†’ 4)-segments]. The length of N-4S was modulated by the ratio between α-cyclodextrin and S-IMS, generating N-4Ss with DPs of 7-50. Based on phase-solubility analysis, D-IMS-28.3/13/3 bearing amylose-like helical N-4S with DP of 28.3 displayed a water-soluble complex with aromatic drugs and curcumin. Small-angle X-ray scattering revealed the chain adapted to rigid in solution in which the radius of gyration was estimated to 2.4 nm. Furthermore, D-IMS with short N-4S solubilized flavonoids of less-soluble multifunctional substances. In our research, enzyme-generated functional biomaterials from DDase were developed to maximize the hydrophobic binding efficacy towards water-insoluble bioactive compounds.

Development of a Sequential Fractionation-and-Recovery Method for Multiple Anti-Inflammatory Components Contained in the Dried Red Alga Dulse (Palmaria palmata).

A separation process was established to sequentially fractionate and recover three anti-inflammatory components derived from sugars, phycobiliprotein, and chlorophyll from the hot-air-dried thalli of the red alga dulse (Palmaria palmata). The developed process consisted of three steps, without the use of organic solvents. In Step I, the sugars were separated by disrupting the cell wall of the dried thalli with a polysaccharide-degrading enzyme, and a sugar-rich extract (E1) was obtained by precipitating the other components, which were simultaneously eluted by acid precipitation. In Step II, the residue suspension from Step I was digested with thermolysin to obtain phycobiliprotein-derived peptides (PPs), and a PP-rich extract (E2) was obtained by separating the other extracts using acid precipitation. In Step III, solubilized chlorophyll was obtained by heating the residue, which was acid-precipitated, neutralized, and re-dissolved to concentrate the chlorophyll-related components (Chls)-rich extract (E3). These three extracts suppressed inflammatory-cytokine secretion by lipopolysaccharide (LPS)-stimulated macrophages, confirming that the sequential procedure had no negative effects on the activities of any of the extracts. The E1, E2, and E3 were rich in sugars, PPs, and Chls, respectively, indicating that the anti-inflammatory components were effectively fractionated and recovered through the separation protocol.

Thermal conductivity and conductance of protein in aqueous solution: Effects of geometrical shape.

Considering the importance of elucidating the heat transfer in living cells, we evaluated the thermal conductivity κ and conductance G of hydrated protein through all-atom non-equilibrium molecular dynamics simulation. Extending the computational scheme developed in earlier studies for spherical protein to cylindrical one under the periodic boundary condition, we enabled the theoretical analysis of anisotropic thermal conduction and also discussed the effects of protein size correction on the calculated results. While the present results for myoglobin and green fluorescent protein (GFP) by the spherical model were in fair agreement with previous computational and experimental results, we found that the evaluations for κ and G by the cylindrical model, in particular, those for the longitudinal direction of GFP, were enhanced substantially, but still keeping a consistency with experimental data. We also studied the influence by salt addition of physiological concentration, finding insignificant alteration of thermal conduction of protein in the present case.

Stable Isotope Tracer to Reveal the Interconversion between 3-Monochloro-1,2-propanediol Ester and Glycidyl Ester during the Deodorization Process.

In this study, the effects of the deodorization process on the interconversion between 3-monochloro-1,2-propanediol ester (3-MCPDE) and glycidyl ester (GE) using 3-MCPDE or GE standards containing deuterium-labeled palmitic acid (*P), oleic acid (*O), or linoleic acid (*L) were examined. Deuterium-labeled 3-MCPDE or GE was added to palm oil then deodorized at 250 °C for 20, 40, or 60 min. In the 3-MCPDE-spiked palm oil, the deuterium-labeled 3-MCPDE content decreased with deodorization time. Moreover, GE containing *P or *O was detected, but there was no GE containing *L in the 3-MCPDE-spiked palm oil. In the GE-spiked oil, GE containing *O or *L decreased with deodorization time, but the content of GE containing *P did not change over the time. Furthermore, deuterium-labeled 3-MCPDE was not detected in the GE-spiked oil. These results suggest that 3-MCPDE is converted into GE and that fatty acid species bound to 3-MCPDE or GE may affect their interconversion.

Antioxidative Self-Assembling Nanoparticles Attenuate the Development of Steatohepatitis and Inhibit Hepatocarcinogenesis in Mice.

Oxidative stress (OS) contributes to nonalcoholic steatohepatitis (NASH) and hepatocarcinogenesis. We investigated whether antioxidative self-assembling nanoparticles (SMAPoTN) could reduce the development of NASH and hepatocellular carcinoma (HCC) in p62/Sqstm1 and Nrf2 double knockout (DKO) mice and studied protective mechanisms. We measured disease development in male DKO mice fed a normal chow (NASH model) or a 60% high-fat diet (HFD; HCC model) with or without SMAPoTN administration for 26 weeks. SMAPoTN inhibited liver fibrosis in both groups and prevented HCC development (0% vs. 33%, p < 0.05) in the HFD group. SMAPoTN reduced OS, inflammatory cytokine signaling, and liver fibrosis. RNA-sequencing revealed that SMAPoTN decreased endoplasmic reticulum stress signaling genes in both groups, HCC driver genes, and cancer pathway genes, especially PI3K-AKT in the HFD groups. In the SMAPoTN treatment HFD group, serum lipopolysaccharide levels and liver lipopolysaccharide-binding protein expression were significantly lower compared with those in the nontreatment group. SMAPoTN improved the α-diversity of gut microbiota, and changed the microbiota composition. Oral SMAPoTN administration attenuated NASH development and suppressed hepatocarcinogenesis in DKO mice by improving endoplasmic reticulum stress in the liver and intestinal microbiota. SMAPoTN may be a new therapeutic option for NASH subjects and those with a high HCC risk.

Quantitative Analysis of Lactone Enantiomers in Butter and Margarine through the Combination of Solvent Extraction and Enantioselective Gas Chromatography-Mass Spectrometry.

We quantified the enantiomeric distributions of δ- and γ-lactones in butter, fermented butter, and margarine through the combination of solvent extraction and enantioselective gas chromatography-mass spectrometry. The main lactones in butter and fermented butter comprised (R)-δ-decalactone, (R)-δ-dodecalactone, (R)-δ-tetradecalactone, (R)-δ-hexadecalactone, and (R)-γ-dodecalactone. In contrast, margarine samples consisted of only δ-decalactone and δ-dodecalactone in racemic forms, indicating that synthetic aroma chemicals were added to margarine. After heat treatment, 13 types of lactones were detected in butter and fermented butter. In heated butter and fermented butter, major δ-lactones in the (R)-form were abundant, but only δ-octalactone in the (S)-form was detected. In contrast, γ-dodecalactone (main γ-lactone in the heated samples) was abundant in the (R)-form, whereas other γ-lactones were detected in the racemic form. These results suggested that the major lactones in dairy products are in the (R)-form. Furthermore, the heat treatment affected the enantiomeric distribution of lactones in butter and fermented butter.

Comparison of Lipoprotein Cholesterol Levels in Golden Syrian Hamster Administrated trans-Octadecenoic Acid Positional Isomers.

We previously conducted a study using HepG2 cells to compare the effect on the secreted apolipoprotein B-100 and apolipoprotein A-1 ratio (B-100/A-1) corresponding to the ratio of low-density to high-density lipoprotein cholesterol (LDL/HDL) among 13 types of trans-octadecenoic acid (t-18:1) positional isomers. The results revealed that trans-5-18:1 (t5) significantly increased B-100/A-1. In this study, 1% of t5 in the diet, corresponding to 2.08 energy%, was administrated golden Syrian hamsters for 4 weeks to reveal the effects on lipid profiles, including LDL/HDL, by comparing cis-9-octadecenoic acid (OA, oleic acid), trans-9-octadecenoic acid (EA), trans-11-octadecenoic acid (VA), and trans-9,trans-12- octadecadienoic acid (TT). LDL/HDL was not significantly different among the groups. However, the cholesterol concentration of medium very low-density lipoprotein (VLDL) was significantly lower in the TT diet than in the OA and t5 diets. The cholesterol concentration of small VLDL was significantly lower in the TT diet than in the OA, t5, and EA diets. The cholesterol concentration of large LDL was significantly lower in the TT diet than in the t5 and EA diets. However, no significant difference was detected between the TT and OA diets. In contrast, the cholesterol concentration of very small HDL was significantly higher in the TT diet than in the t5 diet. These results would support that lipid metabolism is affected by the structure of TFA in animals. However, t5-18:1 did not significantly change any lipid profile compared to OA existing in nature, and the previous result from the cell experiment showing that t5 increased B-100/A-1 (LDL/HDL) was not confirmed in this animal experiment.

Heteroconjugates of quercetin with 4'-O-sulfate selectively accumulate in rat plasma due to limited urinary excretion.

Quercetin and methylquercetin are present in a variety of sulfate and glucuronide conjugates in the plasma of quercetin-fed rats and humans. Quercetin conjugates exhibit various physiological activities, depending on the type and position of conjugation. However, little is known regarding the type and position of isomers of quercetin conjugates in the plasma, their accumulation in the liver and kidneys, and their excretion via urine. Using authentic standards of quercetin conjugates and liquid chromatography-tandem mass spectrometry (LC/MS/MS) analysis, we identified and quantified various quercetin conjugates in blood plasma, urine, liver, and kidney tissues 1, 4, and 10 h after orally administering 33.1 µmol kg-1 quercetin glucosides to rats. The profiles of quercetin conjugates were largely different among plasma, urine, liver, and kidneys. Very limited heteroconjugates (7-O-glucuronide-4'-O-sulfate) of quercetin and methylquercetin dominated in the plasma, but these heteroconjugates were much less excreted via urine and did not largely accumulate in the liver and kidneys. Heteroconjugates constituting sulfates other than 4' position sulfate, 7-O-glucuronide-3'-O-sulfate, 4'-O-glucuronide-7-O-sulfate, and 3'-O-glucuronide-7-O-sulfate were major metabolites in urine, but were minimally detected in the plasma. We also found that mono-sulfate conjugates were abundant in the liver and renal tissues. These results suggest that excretion of quercetin conjugates, especially heteroconjugates, into urine is highly selective. The heteroconjugates with 4'-O-sulfate may be scarcely excreted via urine, and thus accumulate in the blood plasma. Further research is necessary to evaluate the physiological effects of heteroconjugates accumulated in the plasma.

Visualization of dietary docosahexaenoic acid in whole-body zebrafish using matrix-assisted laser desorption/ionization mass spectrometry imaging.

Zebrafish models have been developed for several studies involving lipid metabolism and lipid-related diseases. In the present study, the migration of dietary docosahexaenoic acid (DHA) in whole-body zebrafish was estimated by stable-isotope tracer and matrix-assisted laser desorption/ionization mass spectrometry imaging. Administration of 1-13C-2,2-D2-labeled DHA ((+3)DHA) ethyl ester to male zebrafish was conducted to evaluate its accumulation, migration, and distribution in the body. The (+3)DHA content in the body of zebrafish after administering (+3)DHA for 10 and 15 d was significantly higher than that in the control group. (+3)DHA was observed as a constituent of phosphatidylcholine (PC) in the intestine of zebrafish that were administered (+3)DHA for 5 and 10 d. (+3)DHA-containing PC tended to accumulate in the intestines of zebrafish administered (+3)DHA for 1 d, indicating that recombination of (+3)DHA from ethyl ester to PC occurs quickly at intestine. After administration for 15 d, (+3)DHA-containing PC accumulated in the intestine, liver, and muscle of whole-body zebrafish. In contrast, (+3)DHA-containing PC was not detected in the brain. These results showed that dietary DHA is initially constructed into PC as a structural component of intestinal cell membranes and gradually migrates into peripheral tissues such as muscle.

Quantitative Analysis of Manganese in Underwater Steel Samples Using Long-Short Double-Pulse Laser-Induced Breakdown Spectroscopy.

The long-short double-pulse laser-induced breakdown spectroscopy (LS-DP-LIBS) method was applied to qualitative and quantitative analyses of underwater steel samples to improve the detection ability of underwater measurement. The stable plasma intensity and discrete emission lines were detected using LS-DP-LIBS when comparing the measured results of single-pulse LIBS (SP-LIBS) and LS-DP-LIBS. The long pulse produces a cavitation bubble without plasma, and the short pulse induces the plasma of steel samples within the bubble. The detection features of LS-DP-LIBS for underwater steel samples were discussed in different intra-pulse delay time, long-pulse width, and delay time conditions when analyzing the measured spectra, the signal intensity of Fe(I) at 400.524 nm and 402.187 nm, Mn(I) at 404.136 nm, and intensity ratio of Mn(I) 404.136 nm/Fe(I) 402.187 nm. The results indicated that the plasma stability and spectral signal intensity were improved significantly with a long-pulse width of 80 µs in the intra-pulse delay time of 70 µs, which were appropriate for bubble formation and plasma generation. According to the discussion of the delay time effect, the state of generated plasma was almost stable from 650 ns to 850 ns. Manganese (Mn) contents in steel samples were analyzed quantitatively when measuring five steel samples with different Mn contents using LS-DP-LIBS in optimal experimental conditions. A strong linear dependence was observed with R2=0.9842, which demonstrated the feasibility and appropriateness of quantitative analysis for underwater measurement using LS-DP-LIBS.

Simultaneous Treatment of Long-chain Monounsaturated Fatty Acid and n-3 Polyunsaturated Fatty Acid Decreases Lipid and Cholesterol Levels in HepG2 Cell.

The n-3 type polyunsaturated fatty acids (n-3PUFAs), including eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), from fish oil exhibit health benefits such as triacylglycerol- and cholesterol-lowering effects. Some pelagic fishes contain long-chain monounsaturated fatty acids (LC-MUFAs) such as eicosenoic acid (C20:1), which exert health-promoting effects. However, no study has evaluated beneficial effects of n-3PUFA and LC-MUFA combination. Here, we investigated effects of simultaneous treatment with n-3PUFA (EPA and DHA) and LC-MUFA (cis-5-C20:1 and cis-7-C20:1) and found that n-3PUFA and LC-MUFA combination significantly decreased lipid accumulation and reduced total cholesterol in HepG2 cells. Cholesterol level was significantly lower in DHA + cis-7-C20:1 group than in DHA + EPA group. These results suggest the importance of LC-MUFA as a functional molecule in fish oil.

Heart rate recovery is useful for evaluating the recovery of exercise tolerance in patients with heart failure and atrial fibrillation.

This study aimed to examine the factors that contribute to improvement of exercise tolerance in patients with heart failure (HF) and atrial fibrillation (AF) following cardiac rehabilitation. Our hypothesis is that parasympathetic values are important for recovering exercise tolerance in those patients. We included 84 consecutive patients with HF and AF (mean age: 69 ± 15 years, 80% men). All of the patients underwent a cardiopulmonary exercise test and had pre and post 5 month cardiac rehabilitation assessed. After 155 ± 11 days and 44 ± 8 sessions, 73 patients (86%) showed an increase in peak oxygen uptake (VO2) and VO2 at the anaerobic threshold. In univariate linear regression analysis, the % change in heart rate recovery, plasma B-type natriuretic peptide levels, resting heart rate, and the minute ventilation /carbon dioxide output slope were significantly related to that of peak VO2 (p < 0.01, p = 0.03, p = 0.02, p < 0.01, respectively). Stepwise multivariate linear regression analysis showed that the % change in heart rate recovery was independently related to that of peak VO2 (p < 0.05). Our results suggest that heart rate recovery is closely associated with recovery of exercise tolerance in patients with HF and AF after CR.

Simple generation of various α-monofluoroalkyl radicals by organic photoredox catalysis: modular synthesis of ß-monofluoroketones.

A metal-free and operationally simple strategy for the generation of various α-monofluoroalkyl radicals has been developed. A combination of 1,4-bis(diarylamino)naphthalene photocatalyst and sulfoximine-based fluoroalkylating reagents is the key to success. The protocol can be applied to modular synthesis of ß-monofluoroketones through radical monofluoroalkylation of alkenyl acetates.

A Heteropolynuclear Pt-Ag System Having Cycloplatinated Rollover Bipyridyl Units.

The synthesis and photophysical properties of the heteropolynuclear Pt-Ag complex having cyclometalated rollover bipyridine ligands (bpy*) and bridging pyrazolato ligands are reported. The Pt2Ag2 complex was synthesized by two step reactions from a Pt(II) complex precursor having the rollover bpy* ligand, [Pt(bpy*)(dmso)Cl], with 3,5-dimethylpyrazole (Me2pzH) and a subsequent replacement of NH protons in the Me2pzH moieties with the Ag(I) ion. The Pt2Ag2 complex exists as a mixture of U- and Z-shaped isomers in solution, whose structures were clearly determined by single-crystal X-ray structural analyses. NMR studies using the single crystals revealed rapid isomerization of the Pt2Ag2 complexes in solution, although the Pt2Ag2 structures were supported effectively by the multiple metal-metal interactions. Furthermore, the Pt2Ag2 framework can capture a Ag(I) ion during the U-Z isomerization to afford a Pt2Ag3 core with the formation of Pt → Ag dative bonds. The Pt2Ag3 complex showed further aggregation to form a dimer structure in the presence of coordinating solvent via the crystallization process. The formation of Pt → Ag dative bonds significantly changes the emission energy from the Pt2Ag2 complex, while the emission spectra of U- and Z-isomers of Pt2Ag2 complex almost coincide with each other and their emissive properties are very similar. The density functional theory (DFT) and time-dependent DFT (TD-DFT) calculations revealed the effects of additional Ag(I) ion on the photophysical properties of the heteropolynuclear Pt-Ag complexes bearing the rollover bpy* ligands.

Flux Growth and Magnetic Properties of Helimagnetic Hexagonal Ferrite Ba(Fe1-x Sc x )12O19 Single Crystals.

Fabricating large, high-crystalline-quality single-crystal samples of hexagonal ferrite Ba(Fe1-x Sc x )12O19 is the first important step to elucidating its helimagnetic structure and developing it for further applications. In this study, single crystals of Ba(Fe1-x Sc x )12O19 of various Sc concentrations x were successfully grown by the spontaneous crystallization method using Na2O-Fe2O3 flux. We determined the optimal starting composition of reagents for Ba(Fe1-x Sc x )12O19 growth as a function of x. In situ monitoring of the crystal nucleus generation accelerated the success of crystal growth. The obtained crystals comprised black and lamellate structures with a size of 13 mm × 8 mm × 2 mm and a surface of {001} orientation. X-ray diffraction and elemental analysis revealed that the obtained crystals were composed of single-phase Ba(Fe1-x Sc x )12O19 of high crystalline quality. The lattice constants a and c increased linearly with increasing x, thereby following Vegard's law. The temperature dependence of magnetization and the magnetization curves at 77 K of the x = 0.128 crystal exhibited behavior characteristics of helimagnetism. Neutron diffraction measurements of the x = 0.128 crystal exhibited magnetic satellite reflection peaks below 211 K, providing evidence that Ba(Fe1-x Sc x )12O19 behaves as a helimagnetic material.

DNA markers based on retrotransposon insertion polymorphisms can detect short DNA fragments for strawberry cultivar identification.

In this study, DNA markers were developed for discrimination of strawberry (Fragaria × ananassa L.) cultivars based on retrotransposon insertion polymorphisms. We performed a comprehensive genomic search to identify retrotransposon insertion sites and subsequently selected one retrotransposon family, designated CL3, which provided reliable discrimination among strawberry cultivars. Through analyses of 75 strawberry cultivars, we developed eight cultivar-specific markers based on CL3 retrotransposon insertion sites. Used in combination with 10 additional polymorphic markers, we differentiated 35 strawberry cultivars commonly cultivated in Japan. In addition, we demonstrated that the retrotransposon-based markers were effective for PCR detection of DNA extracted from processed food materials, whereas a SSR marker was ineffective. These results indicated that the retrotransposon-based markers are useful for cultivar discrimination for processed food products, such as jams, in which DNA may be fragmented or degraded.