Antioxidant activity of Magnolol, honokiol, and related phenolic compounds

The antioxidant activity of 10 Japanese and Chinese crude drugs (Kampo drugs) was determined in vitro. Extract of Magnolia cortex, which had the highest antioxidant activity, contained phenolic compounds magnolol and honokiol. However, inhibitory effects of these compounds on lipid oxidation were weaker than that of α-tocopherol as measured by thiobarbituric acid assay. The structure-activity relationship of phenolic compounds showed that antioxidant activities were in the order 4-allyl-2,6-dimethoxyphenol ≥ p,p′-biphenol > eugenol > 2-allyl-6-methylphenol > honokiol > magnolol > caffeic acid > p-ethylphenol > guaiacol. As expected, these results showed that an electron donor and/or bulky groups at the ortho- or para-position of the phenol were required for inhibition of lipid oxidation. Electron spin resonance spin trapping experiments showed that phenol compounds with an allyl substituent on their aromatic rings directly scavenged superoxide (O ₂ ⁻ ), and that only eugenol trapped hydroxyl radicals. These findings suggest that phenolic compounds that contain allyl groups may be effective antioxidants because of the scavenging ability of O ₂ ⁻ or hydroxyl radical, whereas other phenols, without an allyl moiety such as α-tocopherol, may play a role in the termination of free radical chain reactions.     

Preparation of Fatty Acid Methyl Esters by Selective Methanolysis of Polar Glycerolipids

KOH in aqueous methanol catalyzes selective methanolysis of polar glycerolipids with O-ester-linked acyl residues, while triacylglycerols and sterol esters are inert in the solution. Based on these findings, a convenient and reliable method was developed for the preparation of fatty acid methyl esters (FAMEs) from polar glycerolipids in lipid mixtures without prior isolation. Methanolysis of polar glycerolipids was completed within 2.5 min by vortexing or 20 min by shaking with 0.7 M KOH/70% (v/v) methanol in the presence of hexane at 30 °C. The yields of FAMEs obtained by the present method were greater than 95%. The method was applied successfully to gas chromatographic analysis of the fatty acid compositions of polar glycerolipids in seed oil and blood. No obvious differences were found between the fatty acid compositions determined by the present method and those determined by conventional methods, including lipid extraction with chloroform/methanol followed by isolation of polar lipids by chromatography. The fatty acid composition of polar glycerolipids, including phospholipids, can be determined readily in many crude samples.     

Isolation of Delta12 and omega3-fatty acid desaturase genes from the yeast Kluyveromyces lactis and their heterologous expression to produce linoleic and alpha-linolenic acids in Saccharomyces cerevisiae

Two clones with homology to known fatty acid desaturase genes were isolated from the yeast Kluyveromyces lactis. The first gene, which we designate KlFAD2, consists of 411 amino acids with an overall identity of 73.0% to FAD2 from Saccharomyces kluyveri. It exhibited Delta12 fatty acid desaturase activity when expressed in S. cerevisiae under the control of ADH1 promoter and produced endogenous linoleic acid. The second clone, which we designate KlFAD3, consists of 415 amino acids with an overall identity of 79.3% to FAD3 from S. kluyveri. It exhibited omega3 fatty acid desaturase activity in S. cerevisiae when expressed under the control of ADH1 promoter in the presence of the exogenous substrate linoleic acid and produced alpha-linolenic acid. Co-expression of KlFAD2 and KlFAD3 resulted in the endogenous production of both linoleic and alpha-linolenic acids. The yield of alpha-linolenic acid reached 0.8% of total fatty acids and its production was not increased by adding exogenous oleic acid; alpha-linolenic acid reached 8.7% when exogenous linoleic acid was available.     

Titanium Fiber Plates for Bone Tissue Repair

Titanium plates are widely used in clinical settings because of their high bone affinity. However, owing to their high elastic modulus, these plates are not suitable for bone repair since their proximity to the bone surface for prolonged periods can cause stress shielding, leading to bone embrittlement. In contrast, titanium fiber plates prepared by molding titanium fibers into plates by simultaneously applying compression and shear stress at normal room temperature can have an elastic modulus similar to that of bone cortex, and stress shielding will not occur even when the plate lies flush against the bone's surface. Titanium fibers can form a porous structure suitable for cell adhesion and as a bone repair scaffold. A titanium fiber plate is combined with osteoblasts and shown that the titanium fiber plate is better able to facilitate bone tissue repair than the conventional titanium plate when implanted in rat bone defects. Capable of being used in close contact with bone for a long time, and even capable of promoting bone repair, titanium fiber plates have a wide range of applications, and are expected to make great contributions to clinical management of increasing bone diseases, including bone fracture repair and bone regenerative medicine.     

Characterization and grouping of aquatic fulvic acids isolated from clear-water rivers and lakes in Japan

Characteristics of aquatic fulvic acids (FAs) from 10 clear waters in Japan (around the temperate zone) were revealed by several analytical techniques—high performance size exclusion chromatography (HPSEC), elemental analysis, liquid-state ¹³C NMR spectroscopy, isotopic analyses (δ¹³C and δ¹⁵N), and compared with those of International Humic Substances Society (IHSS) standard samples including FAs from brown waters (Suwannee, Pony, and Nordic FAs). Generally clear-water FAs were different from brown-water FAs in chemical properties. Weight-average molecular weights (Mw) of the clear-water FAs were similar to each other, whereas their elemental compositions and carbon species distribution were different. The clear-water FAs all exhibited a high proportion of alkyl carbons, which may be attributed to microbial activity. δ¹³C and δ¹⁵N values of the FAs indicated that there would be a huge gap between origin and chemical structure of clear-water FA. Results of the chemical structural analyses described above were not always linked to those of the isotopic analyses (δ¹³C and δ¹⁵N). Multivariate statistical analysis, i.e. cluster and principal component analysis was applied to reveal differences or similarities in a more objective manner. The FAs were always classified into two clear-water groups and one brown-water group. Aryl-C and O-Alkyl-C contents were important for the grouping. We speculate that the grouping might depend on the differences of aquatic microbial activity caused by the differences of residence time of water.     

Effects of low or medium-pressure UV irradiation on the release of intracellular microcystin

UV irradiation could be an alternative growth inhibition treatment against toxic Microcystis blooms in lakes. This study examined the effect of UV irradiation on the release of toxic intracellular microcystin. Conventional algicidal treatment (e.g. copper sulfate) was known to cause rapid release of intracellular microcystin and additional problems, but no studies have examined the case of UV treatment. A pure culture of Microcystis aeruginosa PCC 7806 was exposed to monochromatic low-pressure or polychromatic medium-pressure UV lamps. Irradiated pure culture suspension was subsequently incubated for 14 days under white light fluorescent lamps. During incubation, profiles of the number of cells, intracellular and extracellular microcystin concentration were determined. After the UV exposure, the number of cells gradually diminished; the net log cell reduction after 6 days of incubation was 1.6 log or 2.0 log for 600 mJ/cm2 of LP or MP UV irradiation, respectively. There were three findings for UV irradiation effects on the release of intracellular microcystin. First, because UV exposure can inhibit increases in the number of cells for about 6 days, it inhibited the increase of microcystin concentrations in water. Second, intracellular microcystin was gradually released into the surrounding water through a gradual reduction in the number of cells, preventing a rapid increase of microcystin concentration in water. Microcystin concentration in water was not significantly higher in UV-irradiated samples than non-UV-irradiated sample. Third, UV irradiation was able to degrade intracellular microcystin; it was reduced from 24.6 to 7.06 or 7.16 fg/cell by 600 mJ/cm2 of LP or MP UV irradiation, respectively. This contributed to reduce increases in microcystin concentrations in water. UV treatment can inhibit Microcystis growth and reduce intracellular microcystin content without rapid release of intracellular microcystin.     

Correlation of fat content and dioxins, total mercury and methyl mercury levels in tuna

In this study, we analyzed the concentrations of mercury and dioxins in tuna with various fat contents (akami; the leaner meat, Chutoro; the belly area of the tuna along the side of the fish between the akami and the otoro. Otoro; the fattiest portion of the tuna) in wild and farmed bluefin tuna and farmed southern bluefin tuna. In the three kinds of tuna, average dioxins concentrations in Akami, chutoro and otoro were 1.7, 4.7 and 9.6 pg TEQ/g, respectively. The dioxins concentration in all three regions of tuna was in direct proportion to the fat content. In the farmed bluefin tuna, the dioxins concentration was almost the same as that of the wild tuna, but differed from that of the farmed southern bluefin tuna. Average total mercury concentration based on wet weight in akami was 0.42 μg/g, being higher than the values of 0.36 μg/g of chutoro and 0.31 μg/g of otoro, and in inverse proportion to the fat content. In all three regions, the total mercury concentration of the wild bluefin tuna was equal to that of the farmed tuna. The total mercury concentration in the latter was two to three times higher than that of the farmed southern bluefin tuna. If the Japanese intake is one fin of tuna (80 g) a day, the daily intake levels of dioxins and methyl mercury can be estimated as 0.48-37 pg TEQ/kg bw and 0.21-0.90 μg/kg bw, respectively.     

Urban pedestrian mobility for mobile wireless network simulation

In order to perform precise evaluation of MANET applications in the real world, realistic mobility models are needed in wireless network simulation. In this paper, we propose a new method to create urban pedestrian flows (UPF) mobility scenarios, which reproduce the walking behavior of pedestrians in urban areas. From given densities of pedestrians observed at several points, our method derives a UPF mobility scenario that reproduces the walking behavior of pedestrians consistent with the observed densities, using linear programming techniques. We have developed a network simulator MobiREAL to design and evaluate MANET protocols and applications with this realistic mobility model. MobiREAL provides various functions and tools including a mobility model to describe the behavior of individual nodes, a GUI to assist with automatic generation of UPF mobility scenarios and a visualization tool. We have conducted some experiments using the MobiREAL simulator. Through the experiments, we have investigated the influence of node mobility on the performance of MANET protocols and have shown the usefulness of our method and the MobiREAL simulator.