Characterization of fucoxanthin and fucoxanthinol esters in the Chinese surf clam, Mactra chinensis

Fucoxanthin and fucoxanthinol fatty acid esters in the clam, Mactra chinensis, were characterized on the basis of 1H NMR and FAB-MS spectra. 1H NMR revealed that the hydroxy group at C-3 in fucoxanthin and fucoxanthinol was acylated. 3'-O-Acylated compounds such as fucoxanthinol 3'-ester or fucoxanthinol 3,3'-diester were not found in the clam. The fatty acids esterified with fucoxanthin and fucoxanthinol were identified as C24:6, C22:5, C22:6, C20:5, C20:0, C20:1, C18:0, C18:1, C16:0, C16:1, and C14:0 by FAB-MS data.     

Radical scavenging and singlet oxygen quenching activity of marine carotenoid fucoxanthin and its metabolites

Antioxidant activity of carotenoids is suggested to be one of the factors for their disease preventing effects. Marine carotenoids fucoxanthin and its two metabolites, fucoxanthinol and halocynthiaxanthin, have been shown to exhibit several biological effects. The antioxidant activities of these three carotenoids were assessed in vitro with respect to radical scavenging and singlet oxygen quenching abilities. The 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity of fucoxanthin and fucoxanthinol was higher than that of halocynthiaxanthin, with the effective concentration for 50% scavenging (EC 50) being 164.60, 153.78, and 826.39 microM, respectively. 2,2'-Azinobis-3-ethylbenzo thizoline-6-sulphonate radical scavenging activity of fucoxanthinol (EC 50, 2.49 microM) was stronger than that of fucoxanthin (EC 50, 8.94 microM). Hydroxyl radical scavenging activity as measured by the chemiluminescence technique showed that the scavenging activity of fucoxanthin was 7.9 times higher than that by fucoxanthinol, 16.3 times higher than that by halocynthiaxanthin, and 13.5 times higher than that by alpha-tocopherol. A similar trend was observed when the hydroxyl radical scavenging was assessed by the electron spin resonance (ESR) technique. ESR analysis of the superoxide radical scavenging activity also showed the superiority of fucoxanthin over the other two carotenoids tested. Singlet oxygen quenching ability of the three carotenoids was lower than that of beta-carotene, with quenching rate constants ( k Q, x10 (10) M (-1) s (-1)) being 1.19, 1.81, 0.80, and 12.78 for fucoxanthin, fucoxanthinol, halocynthiaxanthin, and beta-carotene, respectively. The higher radical scavenging activity of fucoxanthin and fucoxanthinol compared with halocynthiaxanthin is assumed to be due to presence of the allenic bond.     

Antiangiogenic activity of nasunin, an antioxidant anthocyanin, in eggplant peels

Nasunin, delphinidin-3-(p-coumaroylrutinoside)-5-glucoside, an antioxidant anthocyanin isolated from eggplant peels, was demonstrated as an angiogenesis inhibitor. Nasunin at higher 10 microM suppressed microvessel outgrowth in an ex vivo angiogenesis assay using a rat aortic ring. The effect of nasunin was examined in various in vitro angiogenesis models using human umbilical vein endothelial cells (HUVECs). Nasunin suppressed HUVEC proliferation in a dose-dependent manner (50-200 microM); however, it had no significant effect on HUVEC chemotaxis in a Boyden chamber assay and HUVEC tube formation on a reconstituted basement membrane. These results imply that nasunin with both antioxidant and antiangiogenic activities might be useful to prevent angiogenesis-related diseases.     

Effects of DDT and its metabolites on soil algae and enzymatic activity

 The persistence of DDT [1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane] and its metabolites in soil, their toxicity to soil algae, and effects on microbial activities were studied in laboratory microcosms for 45 days. In non-sterile soils, removal of DDD [1,1-dichloro-2,2-bis(p-chlorophenyl)ethane] and DDE [1,1-dichloro-2,2-bis(p-chlorophenyl)ethylene] was less than 3%, while 4–8% of applied DDMU [1-chloro-2,2-bis(p-chlorophenyl)ethylene], DDA [2,2-bis(p-chlorophenyl)acetic acid] and DDT were lost. Added DDOH [2,2-bis(p-chlorophenyl)ethanol] was more labile, as 60% was degraded during the same period. Soil microalgae were not measurably affected by the compounds tested at 10–50 mg kg^–1, but at 100 mg kg^–1 soil, DDD, DBP (p,p′-dichlorobenzophenone) and DDA significantly reduced their growth. Phosphatase activity was not affected by DDT and its metabolites at the concentrations tested (≤50 mg kg^–1), but all compounds inhibited dehydrogenase activity at concentrations of 50 mg kg^–1 soil. The toxic effects of DDT and its metabolites were dose-related. Received: 9 January 1998     

Seasonal variations of bromophenols in brown algae (Padina arborescens,Sargassum siliquastrum,and Lobophora variegata) collected in Hong Kong

Distributions and seasonal variations of the key seafood flavor compounds including 2-bromophenol, 4-bromophenol, 2,4-dibromophenol, 2,6-dibromophenol, and 2,4,6-tribromophenol in three species of brown algae (Padina arborescens, Sargassum siliquastrum, and Lobophora variegata) found in Hong Kong waters were investigated. Bromophenols were extracted by simultaneous steam distillation and solvent extraction apparatus and analyzed by gas chromatography-mass spectrometry. On a dried weight basis, the total bromophenol content (TBC) determined varied widely with seasons (from 40.9 to 7030 ng/g). The TBCs detected were higher in winter and lower in summer. Except for 2-bromophenol, the rest of the bromophenols were detected in all of the algal samples. The TBC of L. variegata was generally the highest among all of the algae collected. Relatively high concentrations of bromophenols in algae supported the fact that marine algae were major producers of bromophenols in the marine environment.     

Nitrocapsanthin and nitrofucoxanthin, respective products of capsanthin and fucoxanthin reaction with peroxynitrite

The in vitro reactivity of capsanthin (1) and fucoxanthin (2) with peroxynitrite was investigated, and the reaction products produced by scavenging with peroxynitrite were analyzed. (14'Z)-Nitrocapsanthin (3) and 12-nitrocapsanthin (4) were isolated from the products of the reaction of capsanthin with peroxynitrite. Similarly, (14Z)-15-nitrofucoxanthin (5), (11Z)-11-nitrofucoxanthin (6), and (14Z,9'Z)-15-nitrofucoxanthin (7) were obtained from the reaction of peroxynitrite reaction with fucoxanthin. Capsanthin and fucoxanthin inhibited the nitration of tyrosine by peroxynitrite. Furthermore, nitrocapsanthins (3 and 4) and nitrofucoxanthins (5 and 6) exhibited an inhibitory effect on Epstein-Barr virus early antigen activation in Raji cells and an antiproliferative effect on human pancreatic carcinoma. Moreover, nitrocapsanthins (3 and 4) inhibited carcinogensis of mouse skin tumors initiated by 7,12-dimethylbenz[a]anthracene (DMBN).     

Petalonia binghamiae extract and its constituent fucoxanthin ameliorate high-fat diet-induced obesity by activating AMP-activated protein kinase

In this study, we investigated the antiobesity properties of Petalonia binghamiae extract (PBE) in mice in which obesity was induced with a high-fat diet (HFD). PBE administration (150 mg/kg/day) for 70 days decreased body weight gain, adipose tissue weight, and the serum triglyceride level in mice fed a HFD. PBE reduced serum levels of glutamic pyruvic transaminase and glutamic oxaloacetic transaminase as well as the accumulation of lipid droplets in the liver. PBE restored the HFD-induced decrease in phosphorylation of AMP-activated protein kinase (AMPK) and acetyl-CoA carboxylase (ACC) in epididymal adipose tissue. PBE increased the phosphorylation of AMPK and ACC and decreased the expression of SREBP1c in mature 3T3-L1 adipocytes. In addition, we further explored the active compound responsible for AMPK activation by PBE in 3T3-L1 adipocytes. Fucoxanthin isolated from PBE increased the phosphorylation of AMPK and ACC with increasing LKB1 phosphorylation in mature 3T3-L1 adipocytes. Taken together, these data suggest that PBE (or fucoxanthin) exert improving effects on HFD-induced obesity by promoting β-oxidation and reducing lipogenesis.     

硼砂对灰棕紫泥土尿素转化与脲酶活性的影响

采用室内培养,连续取样测定培养期内0%、0.5%、1%、1.5%和2%的硼砂对灰棕紫泥土中尿素、铵态氮、硝态氮和脲酶活性变化的影响。结果表明,硼砂能抑制脲酶活性,进而减缓尿素水解速度与铵态氮释放速度,0.5%的硼砂效果较弱,随硼砂用量的增大,效果增强;硼砂对硝态氮的积累影响不大。由于硼本身就是作物必需营养元素,因此作为尿素缓释剂有着良好的应用前景。     

Evaluation of antioxidative activity of extracts from a brown seaweed,Sargassum siliquastrum

Antioxidative activities of the extracts from Sargassum siliquastrum were determined using the inhibition of red blood cell (RBC) hemolysis induced by 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH) radicals, suppression of lipid peroxidation using rat brain homogenate, and scavenging activity of superoxide radicals. The dichloromethane fraction isolated from the methanol crude extract by differential solvent extractions exhibited the strongest antioxidant activity in both RBC hemolysis and lipid peroxidation assays. This fraction was further fractionated into four subfractions F1-F4 by silica gel column chromatography. F1 was found to be most effective in protecting RBC against AAPH radicals and in inhibiting lipid peroxidation. On the basis of thin-layer chromatography and UV and IR spectra analyses, all subfractions contained phenolic compounds. However, there was no correlation between the above antioxidant potency and total phenolic compounds estimated by using the Folin-Ciocalteau method.     

Angiotensin I converting enzyme (ACE) inhibitory activity of hetero-chitooligosaccharides prepared from partially different deacetylated chitosans

Angiotensin I converting enzyme (ACE) inhibitory activity of hetero-chitooligosaccharides (hetero-COSs) prepared from partially different deacetylated chitosans was investigated. Partially deacetylated chitosans, 90, 75, and 50% deacetylated chitosan, were prepared from crab chitin by N-deacetylation with 40% sodium hydroxide solution for durations. In addition, nine kinds of hetero-COSs with relatively high molecular masses (5000-10 000 Da; 90-HMWCOSs, 75-HMWCOSs, and 50-HMWCOSs), medium molecular masses (1000-5000 Da; 90-MMWCOSs, 75-MMWCOSs, and 50-MMWCOSs), and low molecular masses (below 1000 Da; 90-LMWCOSs, 75-LMWCOSs, and 50-LMWCOSs) were prepared using an ultrafiltration membrane bioreactor system. ACE inhibitory activity of hetero-COSs was dependent on the degree of deacetylation of chitosans. 50-MMWCOSs that are COSs hydrolyzed from 50% deacetylated chitosan, the relatively lowest degree of deacetylation, exhibited the highest ACE inhibitory activity, and the IC(50) value was 1.22 +/- 0.13 mg/mL. In addition, the ACE inhibition pattern of the 50-MMWCOSs was investigated by Lineweaver-Burk plots, and the inhibition pattern was found to be competitive.     

Analysis of phenolic compounds in white rice, brown rice, and germinated brown rice

Two hydroxycinnamate sucrose esters, 6'-O-(E)-feruloylsucrose and 6'-O-(E)-sinapoylsucrose, were isolated from methanol extracts of rice bran. Soluble and insoluble phenolic compounds as well as 6'-O-(E)-feruloylsucrose and 6'-O-(E)-sinapoylsucrose from white rice, brown rice, and germinated brown rice were analyzed using HPLC. The results demonstrated that the content of insoluble phenolic compounds was significantly higher than that of soluble phenolics in rice, whereas almost all compounds identified in germinated brown rice and brown rice were more abundant than those in white rice. 6'-O-(E)-Feruloylsucrose (1.09 mg/100 g of flour) and 6'-O-(E)-sinapoylsucrose (0.41 mg/100 g of flour) were found to be the major soluble phenolic compounds in brown rice. During germination, an approximately 70% decrease was observed in the content of the two hydroxycinnamate sucrose esters, whereas free phenolic acid content increased significantly; the ferulic acid content of brown rice (0.32 mg/100 g of flour) increased to 0.48 mg/100 g of flour and became the most abundant phenolic compound in germinated brown rice. The content of sinapinic acid increased to 0.21 mg/100 g of flour, which is nearly 10 times as much as that in brown rice (0.02 mg/100 g of flour). In addition, the total content of insoluble phenolic compounds increased from 18.47 mg/100 g of flour in brown rice to 24.78 mg/100 g of flour in germinated brown rice. These data suggest that appropriate germination of brown rice may be a method to improve health-related benefits.     

Phlorotannins from brown algae (Fucus vesiculosus) inhibited the formation of advanced glycation endproducts by scavenging reactive carbonyls

Accumulation of advanced glycation end products (AGEs) in vivo is associated with aging, diabetes, Alzheimer's disease, renal failure, etc. The objective of this study was to investigate the inhibitory effects of brown algae Fucus vesiculosus phlorotannins on the formation of AGEs. F. vesiculosus phlorotannins were extracted using 70% acetone. The resultant extract was fractionated into dichloromethane, ethyl acetate, butanol, and water fractions. The ethyl acetate fraction was further fractionated into four subfractions (Ethyl-F1 to -F4) using a Sephadex LH-20 column. F. vesiculosus acetone extract or fractions significantly inhibited the formation of AGEs mediated by glucose and methylglyoxal in a concentration-dependent manner. The concentrations of F. vesiculosus extracts required to inhibit 50% of albumin glycation (EC(50)) in the bovine serum albumin (BSA)-methylglyoxal assay were lower than those of aminoguanidine (a drug candidate for diabetic complication), except for F. vesiculosus acetone extract and dichloromethane fraction. In the BSA-glucose assay, F. vesiculosus extracts inhibited BSA glycation more than or as effectively as aminoguanidine, except for Ethyl-F3 and -F4. The ethyl acetate fraction and its four subfractions scavenged more than 50% of methylglyoxal in two hours. The hypothesis whether F. vesiculosus phlorotannins scavenged reactive carbonyls by forming adducts was tested. Phloroglucinol, the constituent unit of phlorotannins, reacted with glyoxal and methylglyoxal. Five phloroglucinol-carbonyl adducts were detected and tentatively identified using HPLC-ESI-MS(n).     

Stability of black tea polyphenol, theaflavin, and identification of theanaphthoquinone as its major radical reaction product

In the current study, we have focused on isolation and detection of major radical oxidation products from theaflavin in order to better understand antioxidation mechanisms of this compound. Theanaphthoquinone was identified as a major oxidation product of theaflavin from two different oxidant model systems: DPPH and peroxidase/hydrogen peroxide. This result indicated that the benzotropolone moiety in theaflavin may play an important role in its antioxidant properties. The stability of theaflavin was studied in varying pH solutions: simulated gastric juice and buffer solutions of pH 5.5, pH 7.4, and pH 8.5. The results indicated that theaflavin is unstable in alkaline conditions, while it was stable in acidic conditions. Theanaphthoquinone was identified as an autoxidation product of theaflavin during its stability study in alkaline conditions.     

稻田中藻类的生长状况及其对肥料氮的固持

为了确定藻类对稻田生态系统中氮素固持与迁移过程的影响程度,本研究采用田间小区和15N微区试验,在水稻生长的不同时期,观测了藻类在稻田中的生长状况,定量测定了藻类的生物量和氮素固持量.结果显示,稻田中的藻类具有明显的群落演替特征,水稻生育初期以球状藻为主,中后期则以丝状藻为主.藻类的生长可分为两个阶段,即生长期和衰亡期,其中,生长期从稻秧移栽初期至拔节期,衰亡期从拔节期至水稻收获.在衰亡期,藻类生物量随时间的变化符合生长曲线模式.藻类生物质干重在2007和2009年水稻生育期的变化范围分别为205～610 kg/hm2和12～353 kg/hm2,而藻类氮素固持量则分别为N 5.4～17.5 kg/hm2和N0.4～11.0 kg/hm2.15N微区试验结果显示,藻类对15N的固持量为N 0.3～6.0 kg/hm2,占氮素总固持量的54％～68％,平均为57％,表明藻类固持的氮素中大部分来源于当季施入的氮肥.     

Dynamics of algal populations and acetylene-reducing activity in five rice soils inoculated with blue-green algae

Summary The dynamics of five inoculated strains of heterocystous blue-green algae (BGA) and indigenous algae were studied for 1 month in 1-m² microplots of five soils previously air-dried or oven-dried. The same soils were then dried and resubmerged for another 2 months to study the effect of controlling algal grazers with neem (Azardirachta indica) seeds on the revival and dynamics of indigenous and inoculated algae. During the month following inoculation, inoculated BGA multiplied to some extend in all soils but never dominated the total algal flora. They rarely dominated the indigeneous heterocystous BGA, and did so only when the growth of N₂-fixing BGA was poor or after the decline of blooms of indigenous strains. Once the soils were dried, two of the five inoculated strains did not reappear. During the 1st month following rewetting, the remaining inoculated strains again exhibited poor growth; however, after 2 months of submergence, inoculated Aulosira fertilissima developed an agronomically significant bloom in neem-treated plots of two soils. Correlations between acetylene-reducing activity and heterocystous BGA populations indicated a major contribution by indigenous BGA and a minor contribution by inoculated BGA to the N₂-fixing activity of the soils during the first experiment and the 1st month of the second experiment. The establishment of inoculated BGA exhibited clear differences among strains but was less affected by the nature of the soil and heat treatment. Neem application might have had a delayed positive effect on the late establishment of inoculated A. fertilissima and favored BGA growth and N₂ fixation by the total algal population.Visiting Scientist at IRRI     

TBHQ对阿维菌素光解的影响及对灰飞虱的增效作用

为延缓阿维菌素的光解,研究了不同浓度抗氧化剂TBHQ对阿维菌素光解的影响,并通过稻茎浸渍法测定其与阿维菌素混用后对灰飞虱的毒力。结果表明:阿维菌素的光解行为符合化学反应一级动力学方程,阿维菌素在太阳光下的光解速率小于紫外光下的光解速率;在TBHQ抑制阿维菌素光解试验中,其与阿维菌素比值为1︰1时,阿维菌素的光解速率最小,光解半衰期为11.81h,比不添加TBHQ在紫外光下的光解半衰期延长167.19%。室内生物测定结果表明,TBHQ与阿维菌素混用后对灰飞虱表现为增效作用;经太阳光和紫外光光照3h后,加入TBHQ的阿维菌素对灰飞虱的毒力均显著高于未添加TBHQ的处理,增效比分别为2.00和2.63。     

Fucoxanthin and fucoxanthinol enhance the amount of docosahexaenoic acid in the liver of KKAy obese/diabetic mice

This study examined the effect of dietary fucoxanthin or fucoxanthinol on the amount of docosahexaenoic acid (DHA) in the liver of KKAy mice, a model for obese/type II diabetes. In the first experiment, mice were fed diets containing crude fucoxanthin or glyceroglycolipid for 4 weeks. Results showed a significant increase in the level of DHA in mice fed 0.53% crude fucoxanthin, from 2.3% in control mice to 5.1% of fatty acid composition of total liver lipids. On the other hand, in mice fed crude glyceroglycolipid, the level of DHA as a proportion of total liver fatty acids remained unchanged. To clarify the enhancement of hepatic DHA, in the second experiment, KKAy mice were fed a diet containing purified fucoxanthin or its deacetylated derivative, fucoxanthinol. Results from a quantitative analysis using an internal standard showed that in mice fed 0.2% fucoxanthin, the amount of hepatic DHA was 2-fold higher than in control mice, whereas DHA levels in the small intestine remained unchanged. Furthermore, 0.2% fucoxanthinol led to 1.8- and 1.2-fold increases in the amount of hepatic DHA and arachidonic acid compared to control mice, respectively. These results indicate for the first time that dietary fucoxanthin and fucoxanthinol enhance the amount of DHA in the liver of KKAy mice.     

Determination of strontium isotope ratio of brown rice for estimating its provenance

The strontium isotope ratio (⁸⁷Sr / ⁸⁶Sr) of brown rice (Oryza sativa L.) was determined by multiple collector inductively coupled plasma mass spectrometry (MC-ICP-MS) in order to evaluate the values of ⁸⁷Sr / ⁸⁶Sr for use in the estimation of the area of rice production. Sample solutions were prepared from 5 g of rice samples using the acid (HNO₃-HClO₄-HF) digestion method. Removal of rubidium from the sample solutions was performed using ion-exchange resin (Dowex 50W X8). The Sr isotope ratios were determined with a precision of < 0.01% (RSD, repetitions = 60) by MC-ICP-MS. Typical analysis time for a single sample was about 15 min, reflecting the high sample throughput. The Sr isotope ratios of the Japanese rice samples ranged from 0.706 to 0.709. The Sr isotope ratios of the Chinese and Vietnamese rice samples (0.710 to 0.711) were slightly higher than those of almost all the Japanese samples. Australian rice showed the highest Sr isotope ratio (0.715 to 0.717) among all the rice samples examined. In contrast, the Sr isotope ratio of Californian rice (0.706) was lower than that of almost all the Japanese samples. The variation in the ⁸⁷Sr / ⁸⁶Sr ratios for the rice samples analyzed in this study clearly demonstrated that the Sr isotope ratios could provide a key information for the estimation of rice provenance.