Relationship between antibacterial activity of (+)-catechin derivatives and their interaction with a model membrane

(+)-Catechin derivatives with different alkyl chain lengths were synthesized from (+)-catechin and various straight chain alkylaldehydes in the presence of methyl mercaptan, and their antibacterial activities against Gram-positive bacteria were evaluated. The antibacterial activity increased markedly with elongation of the alkyl chain lengths of the derivatives and reached a maximum at a chain of four to seven carbons. Subsequently, interaction of the (+)-catechin derivatives with a model membrane using liposome was investigated. The derivatives with a chain of three carbons or more were found to have very strong affinity for the membrane. The injury action of the derivatives against the membrane was examined with liposome in which calcein was enclosed as a fluorescent indicator. The leakage was observed in the derivatives with chain lengths of four carbons or more. Particularly the derivatives with chains longer than five carbons are considered to destroy the liposome membrane judging from the degree of the fluorescent leakage. These results implied that the lipophilicity and disrupting ability of the (+)-catechin derivatives to the liposome membrane participate in their antibacterial activity.     

Reactivity of green tea catechins with formaldehyde

In the reaction of green tea catechins with formaldehyde at room temperature (25°C), tea catechins were found to have reactivity. In particular, (-)-epicatechin gallate and (-)-epigallocatechin gallate, which have a galloyl moiety at the C-3 position, showed higher reactivity than (+)-catechin, (-)-epicatechin, or (-)-epigallocatechin. Reactivity of various kinds of simple phenolic compounds and flavonoids with formaldehyde was also examined. Among these compounds, only phloroglucinol showed reactivity to the same degree as that of nongalloylated catechins. These results suggest that factors for reactivity with formaldehyde at room temperature may be the presence of a phloroglucinolic A-ring structure and the absence of the electron-attractive group such as a carbonyl group in Cring. The comparison of the reactivity of 3-O-acylated catechins with that of 3-O-galloylated catechins indicated that only a galloyl group effectively enhanced reactivity with formaldehyde.     

Food habits of fishes in the mangrove estuary of Urauchi River,Iriomote Island,southern Japan

The food habits of 67 fish species collected from the mangrove estuary of the Urauchi River, Iriomote Island, southern Japan were investigated using gut content analysis. Ontogenetic changes in food preference were recognized in nine species, including mugilids, gerreids, mullids, gobiids and tetraodontids. In most cases, juveniles of these species fed mostly on small crustaceans (e.g. calanoid and cyclopoid copepods and gammaridean amphipods) or detritus. With their subsequent growth, larger prey items (e.g. crabs and polychaetes) became dominant. A cluster analysis based on dietary overlaps showed that the mangrove fish assemblage comprised eight trophic groups (zooplankton, small benthic crustacean, large benthic crustacean, polychaete, fish, detritus, plant and insect feeders). Of these, large and small benthic crustacean feeders, which consumed mainly crabs and gammaridean amphipods, respectively, were the most abundantly represented in terms of species, whereas polychaete and insect feeders were each represented by only two species.     

Anisotropy and corotation of galactic cosmic rays

The intensity of Galactic cosmic rays is nearly isotropic because of the influence of magnetic fields in the Milky Way. Here, we present two-dimensional high-precision anisotropy measurement for energies from a few to several hundred teraelectronvolts (TeV), using the large data sample of the Tibet Air Shower Arrays. Besides revealing finer details of the known anisotropies, a new component of Galactic cosmic ray anisotropy in sidereal time is uncovered around the Cygnus region direction. For cosmic-ray energies up to a few hundred TeV, all components of anisotropies fade away, showing a corotation of Galactic cosmic rays with the local Galactic magnetic environment. These results have broad implications for a comprehensive understanding of cosmic rays, supernovae, magnetic fields, and heliospheric and Galactic dynamic environments.     

Synthesis of (-)-[4-3H]epigallocatechin gallate and its metabolic fate in rats after intravenous administration

Because a great deal of attention has been focused on the metabolism of (-)-epigallocatechin gallate (EGCg), quantitative analysis of this compound is required. For this purpose we developed a method of chemical synthesis of [4-(3)H]EGCg. Synthesized [4-(3)H]EGCg showed 99.5% radiochemical purity and a specific activity of 13 Ci/mmol. To clarify the excretion route of EGCg, the radioactivity levels of bile and urine were quantified after intravenous administration of [4-(3)H]EGCg to bile-duct-cannulated rats. Results showed that the radioactivity of the bile sample excreted within 48 h accounted for 77.0% of the dose, whereas only 2.0% of the dose was recovered in the urine. The excretion ratio of bile to urine was calculated to be about 97:3. These results clearly showed that bile was the major excretion route of EGCg. Time-course analysis of the radioactivity in blood was also performed to estimate the pharmacokinetic parameters following intravenous administration of [4-(3)H]EGCg. In addition, EGCg metabolites excreted in the bile within 4 h after the intravenous dose of [4-(3)H]EGCg were analyzed by HPLC. The results showed that 4',4"-di-O-methyl-EGCg was present in the conjugated form and made up about 14.7% of the administered radioactivity.     

Utilization patterns of estuarine and marine habitats by the halfbeak Zenarchopterus dunckeri at Iriomote Island,southern Japan,evaluated from otolith microchemistry

Estuarine and marine habitat use patterns in the halfbeak Zenarchopterus dunckeri were examined at Iriomote Island, southern Japan, by analyzing otolith Li/Ca and Sr/Ca ratios. The ranges of both Li/Ca and Sr/Ca ratios in juvenile Z. dunckeri from the maximum (30 psu) to minimum (0.5 psu) salinity levels of brackish water estimated from rearing experiments, were compared with those of wild individuals collected from upstream and downstream stations in the Urauchi River estuary. The majority of wild-caught individuals had invariable Li/Ca and Sr/Ca ratios along an otolith transect from the core to the posterior edge, which fell within the otolith Li/Ca and Sr/Ca ranges estimated for estuarine individuals in the rearing experiments, suggesting that such individuals developed within the estuary without migrating to a marine environment at any time, although some downstream-dwelling fish had higher otolith elemental ratios than the predetermined estuarine ranges in the mid transect section. The latter fish may have been accidentally flushed from the estuary into the sea by heavy flood events, subsequently returning to the estuary. The overall results suggested that Z. dunckeri is essentially an estuarine resident, completing its life cycle within an estuarine system.     

Distribution of carbohydrates and related enzyme activities in yacon (polymnia sonchifolia)

Abstract

Yacon (Polymnia sonchifolia) plants originating from Andean highlands are known to accumulate a large amount of oligofructans in their tubers and tuberous roots (Ohyama et al. 1990; Asami et al. 1992). Asami et al. (1992) have reported that tubers and tuberous roots which were harvested in late fall contained about 57 and 66% respectively (on a dry matter basis) of oligofructans. However, the tubers, tuberous roots and shoots of the yacon contained only a small amount of starch and inulin. The tuberous roots of yacon which resemble those of sweet potato in appearance are usually eaten raw and are expected to become commercially valuable as a source of oligofructans.     

Metabolic fate of (-)-[4-(3)H]epigallocatechin gallate in rats after oral administration

After oral administration of [4-(3)H]EGCg to rats, the radioactivity in blood, major tissues, urine, and feces was measured over time. The radioactivity in blood and most tissues remained low for 4 h postdose, began to increase after 8 h, peaked at 24 h, and then decreased. Major urinary excretion of radioactivity occurred in the 8-24 h period, and the cumulative radioactivity excreted by 72 h was 32.1% of the dose. The radioactivity in the feces was 35.2% of the dose within 72 h postdose. In the case of rats pretreated with antibiotics (antibiotic-pretreated rats), the radioactivity levels of the blood and urine were definitely lower than those in rats not pretreated with antibiotics (normal rats). The radioactivity recovered in the antibiotic-pretreated rat urine was estimated to be only (1)/(100) of that in the normal rat urine. These results clearly demonstrated that the radioactivity detected in the blood and urine of normal rats mostly originated from degradation products of EGCg produced by intestinal bacteria. Furthermore, a main metabolite in the normal rats was purified and identified as 5-(5'-hydroxyphenyl)-gamma-valerolactone 3'-O-beta-glucuronide (M-2). In feces of the normal rats, EGC (40.8% of the fecal radioactivity) and 5-(3',5'-dihydroxyphenyl)-gamma-valerolactone (M-1, 16.8%) were detected. These results suggested that M-1 was absorbed in the body after degradation of EGCg by intestinal bacteria, yielding M-1 with EGC as an intermediate. Furthermore, M-2 was thought to be formed from M-1 in the intestinal mucosa and/or liver, then to enter the systemic circulation, and finally to be excreted in the urine. Taking into account all of the above findings, a possible metabolic route of EGCg orally administered to rats is proposed.     

Identification of metabolites of (-)-epicatechin gallate and their metabolic fate in the rat

After intravenous administration of (-)-epicatechin gallate to Wistar male rats, its biliary metabolites were examined. Deconjugated forms of (-)-epicatechin gallate metabolites were prepared by beta-glucuronidase/sulfatase treatment and purified by HPLC. Five compounds were subjected to FAB-MS and NMR analyses. These metabolites were shown to be (-)-epicatechin gallate, 3'-O-methyl-(-)-epicatechin gallate, 4'-O-methyl-(-)-epicatechin gallate, 4' '-O-methyl-(-)-epicatechin gallate, and 3',4' '-di-O-methyl-(-)-epicatechin gallate. After oral administration, five major metabolites excreted in rat urine were purified in their deconjugated forms and their chemical structures identified. They were degradation products from (-)-epicatechin gallate, pyrogallol, 5-(3,4-dihydroxyphenyl)-gamma-valerolactone, 4-hydroxy-5-(3,4-dihydroxyphenyl)valeric acid, 3-(3-hydroxyphenyl)propionic acid, and m-coumaric acid. Time course analysis of the identified (-)-epicatechin gallate metabolites showed that (-)-epicatechin gallate and its conjugate appeared in the plasma with their highest levels 0.5 h after oral administration; their levels rapidly decreased, and then they disappeared by 6 h. The degradation products, mainly in their conjugated forms, emerged at 6 h, peaked at 24 h, and disappeared by 48 h. In urine samples, (-)-epicatechin gallate and its methylated metabolites were hardly detected and the degradation products began to be excreted in the 6-24 h period, peaked in the 24-48 h period, and then began to disappear. The most abundant metabolite in both the plasma and the urine was found to be the conjugated form of pyrogallol. On the basis of these results, a possible metabolic route of (-)-epicatechin gallate orally administered to the rat is proposed.     

Differences in fish assemblage structure between vegetated and unvegetated microhabitats in relation to food abundance patterns in a mangrove creek

In order to clarify the mechanisms determining fish distribution patterns in a mangrove system on Iriomote Island, in southern Japan, fish assemblage structures were determined by visual observation, along with food abundance and environmental factors, in an area of mangrove roots on the banks, and a bare sand area at the center, within downstream, midstream and upstream portions of a branch creek from the Urauchi River. The fish assemblage structures differed significantly between the area types, with the mangrove-root area supporting a more diverse and abundant fish fauna. A canonical correspondence analysis revealed that the relationships between fish distribution and their food abundance differed among trophic groups. Benthic crustacean or plant feeders were positively associated with their prey i.e. crabs and macroalgae—in other words, these trophic groups were abundant in downstream and/or midstream mangrove-root areas in which their prey were also particularly abundant. However, zooplankton feeders did not show such relationships, their abundance being positively associated with fine sediment particles (characteristic of areas with weak water movement). These results suggested that food availability is a major factor determining the distribution patterns of benthic crustacean feeders and plant feeders, whereas for zooplankton feeders other factors, such as sheltering effects against water current and/or predators, may be more significant.