DDAH1 Protects against Acetaminophen-Induced Liver Hepatoxicity in Mice.

In many developed countries, acetaminophen (APAP) overdose-induced acute liver injury is a significant therapeutic problem. Dimethylarginine dimethylaminohydrolase 1 (DDAH1) is a critical enzyme for asymmetric dimethylarginine (ADMA) metabolism. Growing evidence suggests that liver dysfunction is associated with increased plasma ADMA levels and reduced hepatic DDAH1 activity/expression. The purpose of this study was to investigate the involvement of DDAH1 in APAP-mediated hepatotoxicity using Ddah1-/- and DDAH1 transgenic mice. After APAP challenge, Ddah1-/- mice developed more severe liver injury than wild type (WT) mice, which was associated with a greater induction of fibrosis, oxidative stress, inflammation, cell apoptosis and phosphorylation of JNK. In contrast, overexpression of DDAH1 attenuated APAP-induced liver injury. RNA-seq analysis showed that DDAH1 affects xenobiotic metabolism and glutathione metabolism pathways in APAP-treated livers. Furthermore, we found that DDAH1 knockdown aggravated APAP-induced cell death, oxidative stress, phosphorylation of JNK and p65, upregulation of CYP2E1 and downregulation of GSTA1 in HepG2 cells. Collectively, our data suggested that DDAH1 has a marked protective effect against APAP-induced liver oxidative stress, inflammation and injury. Strategies to increase hepatic DDAH1 expression/activity may be novel approaches for drug-induced acute liver injury therapy.

Fabrication of a ferritin-casein phosphopeptide-calcium shell-core composite as a novel calcium delivery strategy.

Plant ferritin has a natural cage-like nanospace for carrying bioactive ingredients. By taking advantage of the calcium binding ability of casein phosphopeptide (CPP) and the cage-like conformation of plant ferritin, a ferritin-CPP shell-core complex (FC) was fabricated with the reversible self-assembly character of ferritin induced by a pH 2.0/7.0 transition strategy. The FC-calcium composite (FCC) was further fabricated by binding of the FC with calcium ions. When the same amount of calcium was loaded, the calcium binding capacity of the FCC was 28.13 ± 1.65%, which was significantly higher than that of ferritin and CPP alone. Fluorescence and Fourier transform infrared analysis indicated that the CPP encapsulation and the calcium binding in the FCC influenced the ferritin structure. Transmission electron microscopy (TEM) and dynamic light scattering (DLS) results showed that the spherical morphology and the 12 nm-diameter size were sustained in the FC and FCC. Moreover, the FCC as a transport carrier could increase the precipitation time of calcium phosphate, and the encapsulated calcium could be released in a more sustained manner as compared with ferritin and CPP under simulated in vitro gastrointestinal conditions. This study presents a novel calcium delivery strategy based on the ferritin cage and CPP, which will improve the applicability of ferritin and CPP and enhance the bioavailability of calcium ions.

Physiological and Metabolic Responses of a Novel Dunaliella salina Strain to Myo-inositol1.

Dunaliella salina is well known for its ability to accumulate large amounts of ß-carotene. Myo-inositol (MI) enhances the biomass production of D. salina, but the underlying mechanisms were unclear. The present study showed that the concentration of exogenous MI decreased gradually and reached a constant level at the 4th day of cultivation. MI enhanced the contents of total colored carotenoids and the activity of photosystem II. Metabolic profiles were significantly changed after the addition of exogenous MI, as revealed by multivariate statistical analysis. The metabolites could be categorized into four groups based on the relative levels in different samples. Exogenous MI increased the levels of most detected sugars, amino acids, and total saturated and unsaturated fatty acids. Based on the physiological and metabolic analyses, a hypothetical growth-promoting model that MI promotes the growth of D. salina TG by increasing the levels of key metabolites and possibly enhancing photosynthesis, was proposed. This study provides valuable information for understanding the growth-promoting mechanisms of MI in D. salina from the metabolic perspective.

Metformin protects against PM2.5-induced lung injury and cardiac dysfunction independent of AMP-activated protein kinase α2.

Fine particulate matter (PM2.5) airborne pollution increases the risk of respiratory and cardiovascular diseases. Although metformin is a well-known antidiabetic drug, it also confers protection against a series of diseases through the activation of AMP-activated protein kinase (AMPK). However, whether metformin affects PM2.5-induced adverse health effects has not been investigated. In this study, we exposed wild-type (WT) and AMPKα2-/- mice to PM2.5 every other day via intratracheal instillation for 4 weeks. After PM2.5 exposure, the AMPKα2-/- mice developed more severe lung injury and cardiac dysfunction than were developed in the WT mice; however the administration of metformin was effective in attenuating PM2.5-induced lung injury and cardiac dysfunction in both the WT and AMPKα2-/- mice. In the PM2.5-exposed mice, metformin treatment resulted in reduced systemic and pulmonary inflammation, preserved left ventricular ejection fraction, suppressed induction of pulmonary and myocardial fibrosis and oxidative stress, and increased levels of mitochondrial antioxidant enzymes. Moreover, pretreatment with metformin significantly attenuated PM2.5-induced cell death and oxidative stress in control and AMPKα2-depleted BEAS-2B and H9C2 cells, and was associated with preserved expression of mitochondrial antioxidant enzymes. These data support the notion that metformin protects against PM2.5-induced adverse health effects through a pathway that appears independent of AMPKα2. Our findings suggest that metformin may also be a novel drug for therapies that treat air pollution associated disease.

Photo-Actuation of Liquid Crystalline Elastomer Materials Doped with Visible Absorber Dyes under Quasi-Daylight.

We studied the effect of visible absorber dyes on the photo-actuation performances of liquid crystalline elastomer (LCE) materials under quasi-daylight irradiation. The dye-doped LCE materials were prepared through infiltrating visible absorber dyes into a polysiloxane-based LCE matrix based on its solvent-swollen characteristic. They demonstrated well absorption properties in visible spectrum range and performed strong actuation upon the irradiation from quasi-daylight source, thus indicating that the presence of visible absorber dyes effectively sensitized the LCE materials to light irradiation since the light energy was absorbed by the dyes and then converted into heat to trigger the phase change of LCE matrix. The photo-actuation properties of dye-doped LCE materials with different visible absorber dyes, varied dye contents, and irradiation intensities were investigated. It was shown that the visible absorber dyes with different absorption bands created different photo-actuation performances of LCE materials, the one whose absorption band is near the intensity peak position of quasi-daylight spectrum created the optimum photo-actuation performance. The result disclosed a valuable light utilization way for photo-controlled LCE materials since it revealed that a light-absorbing dye, whose absorption band is in the high intensity region of light spectrum, is capable of effectively utilizing light energy to drive the actuation of LCE materials.

[GPC Fingerprint Chromatograms of Aloe vera Leaf Gel Polysaccharides].

OBJECTIVE: To establish the gel permeation chromatography (GPC) fingerprint chromatograms of polysaccharides in Aloe vera leaf gel from the same habitat (Beijing) and different habitats for evaluating the quality of Aloe vera leaf gel products commercially available and testing common adulterated substances. METHODS: The samples were prepared by water-extraction and alcohol-precipitation method. GPC separation was performed on a Shodex SUGAR KS-805 (300 mm x 8.0 mm, 7 µm) column and a Shodex SUGAR KS-803 (300 mm x 8.0 mm, 6 µm) column at the temperature of 60 degrees C by eluting with 0.1 mol/L NaNO3 (containing 0.2 per thousand NaN) at a flow rate of 0.8 mL/min, the chromatographic effluent was detected by refractive index detector (RID) at the temperature of 50 degrees C. RESULTS: The common pattern of GPC fingerprint chromatograms was established and four common peaks were demarcated. The similarities of samples from the same habitat (Beijing) and different habitats were over 0.9. Taking the GPC fingerprint chromatograms for the qualified model, three commercially available aloe products were evaluated to be made of Aloe vera by the different manufacturing processes and four common adulterated substances of aloe polysaccharides were identified effectively. CONCLUSION: The method is simple and accurate with a good reproducibility, and it can be used for the identification and quality evaluation of Aloe vera leaf gel products.

[Determination of alpha-linolenic acid in perilla oil by reversed-phase high performance liquid chromatography coupled with evaporative light-scattering detector].

A method for the determination of alpha-linolenic acid in perilla oil was developed using the reversed-phase high performance liquid chromatography coupled with evaporative light-scattering detector (RP-HPLC-ELSD). The perilla oil was saponified by 0.5 mol/L KOH-CH3OH solution for 20 min in a 60 degrees C water bath, then acidified by 6 mol/L HCl and finally the dissociative fatty acids, including alpha-linolenic acid, was extracted by anhydrous ether. After the ether was blown out by nitrogen, the residuals were dissolved by 10 mL methanol. The calibration curve was found to be linear over the range of 6.2 - 45.4 microg (r = 0.997 3, n = 5) and the detection limit was 0.11 microg (S/N = 3). The average recovery was 102% and their relative standard deviation (RSD) was 6.3% (n = 5). The content of alpha-linolenic acid in the determined perilla oil was 6.79% which is consistent with the previous report.

Isolation and purification of inflacoumarin A and licochalcone A from licorice by high-speed counter-current chromatography.

High-speed counter-current chromatography (HSCCC) technique in semi-preparative scale has been applied to isolate and purify bioactive flavone compounds from the ethanol extract of Glycyrrhiza inflata Bat., a particular plant species of licorice. HSCCC separation was performed with a two-phase solvent system composed of n-hexane-chloroform-methanol-water (5:6:3:2, v/v) by eluting the lower mobile phase at a flow rate of 1.8 ml/min and a revolution speed of 800 rpm. Purification was performed with a two-phase solvent system composed of n-hexane-chloroform-methanol-water (1.5:6:3:2, v/v) by eluting the lower mobile phase at a flow-rate of 1.5 ml/min and a revolution speed of 800 rpm. Two major flavone peaks: inflacoumarin A and licochalcone A were collected and the respective yields of the peaks amount to 6 mg (8.6%, w/w) and 8 mg (11.4%, w/w) from 70 mg of the crude extract sample. The purities of inflacoumarin A and licochalcone A reached 99.6% and 99.1%, respectively, after a sequential purification run. The structures of inflacoumarin A and licochalcone A were positively confirmed by 1H NMR and 13C NMR, 1H-13C-COSY, UV, FT-IR and electron ionization MS analyses.