Lipid components and enzymatic hydrolysis of lipids in muscle of Chinese freshwater fish

The lipid and fatty acid composition of muscle of 10 species of freshwater fish obtained from a market of Shanghai City was examined. Total lipids (TL) ranged over 0.9–4.7% of muscle for all samples. The content of triacylglycerol (TG) in muscle ranged over 0.2–3.4% and that of polar lipids (PL) was 0.5–1.3%. Differences of TL content were dependent on TG contents. The predominant important fatty acids (>10% of the total fatty acids in TL) were 16∶0 and 18∶1n−9 with some 16∶1n−7, 18∶2n−6, and 22∶6n−3. The polyunsaturated fatty acids (PUFA) content was 10.2–43.4%, and especially Chinese sea bass contained above 20% of 22∶6n−3 in the total fatty acids. There were higher levels of PUFA such as 20∶5n−3 and 22∶6n−3 in PL than in neutral lipids. Muscle of the silver carp was stored at 20°C, and changes of lipid classes during storage were examined. Free fatty acids increased, and PL decreased during storage. This phenomenon was inhibited by heating the muscle, suggesting that lipid hydrolysis by phospholipase occurred in silver carp muscle.     

Oxidation of Methyl Linoleate in Oil-in-Water Micro- and Nanoemulsion Systems

Oxidation of methyl linoleate in O/W emulsions having droplets of median diameters ranging from 17 nm to 8.0 μm was carried out at 40°C. The oxidation process was analyzed on the basis of a kinetic equation of the autocatalytic type. The induction period was found to be shorter and the oxidation rate constant lower for emulsions with smaller oil droplets. The stoichiometry between methyl linoleate and oxygen was observed to be independent of both the size of oil droplet and the type of the surfactant and was found to be unity during the early stage of the oxidation. However, more oxgen was consumed in the oxidation of the methyl linoleate in the later half of the oxidation process.     

Analytical Method for Diacylglycerol Kinase ζ Activity in Cells Using Protein Myristoylation and Liquid Chromatography–Tandem Mass Spectrometry

Specific inhibitors of diacylglycerol kinase (DGK) ζ can be promising anticancer medications via the activation of cancer immunity. Although the detection of cellular activities of target enzymes is essential for drug screening in addition to in vitro assays, it is difficult to detect the activity of DGKζ in cells. In the present study, we generated AcGFP-DGKζ cDNA with a consensus N-myristoylation sequence at the 5′ end (Myr-AcGFP-DGKζ) to target DGKζ to membranes. Using liquid chromatography (LC)-tandem mass spectrometry (MS/MS) (LC–MS/MS), we showed that Myr-AcGFP-DGKζ, but not AcGFP-DGKζ without the myristoylation sequence, substantially augmented the levels of several phosphatidic acid (PtdOH) species. In contrast to Myr-AcGFP-DGKζ, its inactive mutant did not exhibit an increase in PtdOH production, indicating that the increase in PtdOH production was DGK activity-dependent. This method will be useful in chemical compound selection for the development of drugs targeting DGKζ and can be applicable to various soluble (nonmembrane bound) lipid-metabolizing enzymes, including other DGK isozymes.     

Photophysical properties and photostability of novel symmetric polycyclicphenazine-type fluorescent dyes and the dye-doped films

Novel symmetric polycyclicphenazine-type fluorescent dyes have been synthesized and characterized in solution and in polymer films. The dyes exhibited two strong absorption bands at around 365-428 nm and 519-567 nm and an intense fluorescence band at around 560-600 nm (Ф = 0.70-0.87) in 1,4-dioxane. The dye-doped polymer films showed excellent wavelength conversion function that efficiently convert ultraviolet and yellow lights into red light (Ф = 0.61-0.92). Moreover, the photostability of the dye-doped PS, PMMA, and PLA films has been investigated.     

Exogenous ANP Treatment Ameliorates Myocardial Insulin Resistance and Protects against Ischemia–Reperfusion Injury in Diet-Induced Obesity

Increasing evidence suggests natriuretic peptides (NPs) coordinate interorgan metabolic crosstalk. We recently reported exogenous ANP treatment ameliorated systemic insulin resistance by inducing adipose tissue browning and attenuating hepatic steatosis in diet-induced obesity (DIO). We herein investigated whether ANP treatment also ameliorates myocardial insulin resistance, leading to cardioprotection during ischemia–reperfusion injury (IRI) in DIO. Mice fed a high-fat diet (HFD) or normal-fat diet for 13 weeks were treated with or without ANP infusion subcutaneously for another 3 weeks. Left ventricular BNP expression was substantially reduced in HFD hearts. Intraperitoneal-insulin-administration-induced Akt phosphorylation was impaired in HFD hearts, which was restored by ANP treatment, suggesting that ANP treatment ameliorated myocardial insulin resistance. After ischemia–reperfusion using the Langendorff model, HFD impaired cardiac functional recovery with a corresponding increased infarct size. However, ANP treatment improved functional recovery and reduced injury while restoring impaired IRI-induced Akt phosphorylation in HFD hearts. Myocardial ultrastructural analyses showed increased peri-mitochondrial lipid droplets with concomitantly decreased ATGL and HSL phosphorylation levels in ANP-treated HFD, suggesting that ANP protects mitochondria from lipid overload by trapping lipids. Accordingly, ANP treatment attenuated mitochondria cristae disruption after IRI in HFD hearts. In summary, exogenous ANP treatment ameliorates myocardial insulin resistance and protects against IRI associated with mitochondrial ultrastructure modifications in DIO. Replenishing biologically active NPs substantially affects HFD hearts in which endogenous NP production is impaired.     

Yerba-mate (Ilex paraguariensis) extract prevents ethanol-induced liver injury in rats

Ethanol metabolism-associated oxidative stress contributes to the pathogenesis of alcoholic liver disease. We examined the effect of a Mate tea extract on ethanol-induced liver injury in vitro and in vivo models. Isolated hepatocytes were incubated with ethanol. An extract of Yerba-Mate tea (EMT) was added to the cultures simultaneously with ethanol. EMT treatment suppressed the ethanol-induced increase in cell death by inhibiting cytochrome p450 2E1 (CYP2E1) activity, which is related to the production of reactive oxygen species. Furthermore, we examined the effects of EMT on serum transaminase activity, and the progression of liver fibrosis in rats treated with ethanol and CCl₄. Rats were fed a diet that included 0.005% or 0.02% EMT or no EMT. For a period of 3 weeks, the animals were provided drinking water containing 5% ethanol and were also treated with carbon tetrachloride (CCl₄) (0.1 ml/kg of body weight). EMT treatment suppressed plasma ALT and AST activities in the ethanol- and CCl₄-treated rats. EMT treatment also decreased CYP2E1 expression and increased ADH expression in the ethanol- and CCl₄-treated rats. EMT treatment fully protected the rats against ethanol- and CCl₄-induced liver injury. These results suggest that EMT may serve as a candidate for preventing ethanol-induced liver injury.     

Increase in Anticancer Drug-Induced Toxicity by Fisetin in Lung Adenocarcinoma A549 Spheroid Cells Mediated by the Reduction of Claudin-2 Expression

Claudin-2 (CLDN2), a component of tight junction, is involved in the reduction of anticancer drug-induced toxicity in spheroids of A549 cells derived from human lung adenocarcinoma. Fisetin, a dietary flavonoid, inhibits cancer cell growth, but its effect on chemosensitivity in spheroids is unknown. Here, we found that fisetin (20 μM) decreases the protein level of CLDN2 to 22.3%. Therefore, the expression mechanisms were investigated by real-time polymerase chain reaction and Western blotting. Spheroids were formed in round-bottom plates, and anticancer drug-induced toxicity was measured by ATP content. Fisetin decreased the phosphorylated-Akt level, and CLDN2 expression was decreased by a phosphatidylinositol 3-kinase (PI3K) inhibitor, suggesting the inhibition of PI3K/Akt signal is involved in the reduction of CLDN2 expression. Hypoxia level, one of the hallmarks of tumor microenvironment, was reduced by fisetin. Although fisetin did not change hypoxia inducible factor-1α level, it decreased the protein level of nuclear factor erythroid 2-related factor 2, a stress response factor, by 25.4% in the spheroids. The toxicity of doxorubicin (20 μM) was enhanced by fisetin from 62.8% to 40.9%, which was rescued by CLDN2 overexpression (51.7%). These results suggest that fisetin can enhance anticancer drug toxicity in A549 spheroids mediated by the reduction of CLDN2 expression.     

Water-filled single-wall carbon nanotubes as molecular nanovalves

It is known that at low temperature, water inside single-wall carbon nanotubes (water-SWNTs) undergoes a structural transition to form tube-like solid structures. The resulting ice NTs are hollow cylinders with diameters comparable to those of typical gas molecules. Hence, the gas-adsorption properties of ice- and water-SWNTs are of interest. Here, we carry out the first systematic investigation into the stability of water-SWNTs in various gas atmospheres below 0.1 MPa by means of electrical resistance, X-ray diffraction, NMR measurements and molecular dynamics calculations. It is found that the resistivity of water-SWNTs exhibits a significant increase in gas atmospheres below a critical temperature Tc, at which a particular type of atmospheric gas molecule enters the SWNTs in an on-off fashion. On the basis of this phenomenon, it is proposed that water-SWNTs can be used to fabricate a new type of molecular nanovalve.     

A High-Methionine Diet for One-Week Induces a High Accumulation of Methionine in the Cerebrospinal Fluid and Confers Bipolar Disorder-like Behavior in Mice

Methionine (Met) is considered the most toxic amino acid in mammals. Here, we investigated biochemical and behavioral impacts of ad libitum one-week feeding of high-Met diets on mice. Adult male mice were fed the standard rodent diet that contained 0.44% Met (1×) or a diet containing 16 graded Met doses (1.2×–13×). High-Met diets for one-week induced a dose-dependent decrease in body weight and an increase in serum Met levels with a 2.55 mM peak (versus basal 53 µM) on the 12×Met diet. Total homocysteine (Hcy) levels were also upregulated while concentrations of other amino acids were almost maintained in serum. Similarly, levels of Met and Hcy (but not the other amino acids) were highly elevated in the cerebrospinal fluids of mice on the 10×Met diet; the Met levels were much higher than Hcy and the others. In a series of behavioral tests, mice on the 10×Met diet displayed increased anxiety and decreased traveled distances in an open-field test, increased activity to escape from water soaking and tail hanging, and normal learning/memory activity in a Y-maze test, which were reflections of negative/positive symptoms and normal cognitive function, respectively. These results indicate that high-Met ad libitum feeding even for a week can induce bipolar disorder-like disease models in mice.