First paralytic shellfish poison (PSP) infestation of bivalves due to toxic dinoflagellate Alexandrium tamiyavanichii, in the southeast coasts of the Seto Inland Sea, Japan

The mussel Mytilus edulis and the cultured ark shell Anadara broughtonii in the southeast coasts of the Seto Inland Sea were contaminated with paralytic shellfish poison (PSP) following the appearance of the dinoflagellate Alexandrium tamiyavanichii in early December 1999. A. tamiyavanichii plankton collected around the Straits of Naruto on December 3, 1999 showed PSP toxicity, of which 83 mol% was accounted for by GTX2, GTX3 and GTX4. Its specific toxicity was 112.5 fmol/cell, and one MU was equivalent to 7,200 cells. Toxicity values at the beginning of toxification were 4.7 MU/g for the ark shell and 7.3 MU/g for the mussel. In the former, the value remained at almost 4 MU/g, resulting in prohibition of marketing for about two months. In the latter, it sharply decreased to less than 4 MU/g. These bivalves collected during the toxification period were dissected into five tissues, mantle, adductor muscle, hepatopancreas, gills and "others", and submitted to high-performance liquid chromatography (HPLC). The cultured ark shell accumulated GTX2, GTX3 and STX as major components and GTX1, GTX4, GTX5, neoSTX, dcSTX and PX1-3 (C1-C3) as minor ones. The amount of GTX3 decreased with time, while STX tended to increase. At the early stage of PSP toxification, toxins were accumulated in the gills and "others", most of which were quickly detoxified. On the other hand, PSP of the toxified mussel consisted of GTX4 as a main component, and GTX1, GTX2, GTX3, GTX5, STX and PX1-2 (C1-C2) as minor ones. Its toxin composition pattern was similar to that of the ingested causative plankton. Its total toxin decreased soon after disappearance of the dinoflagellate. During the decrease of toxicity, PSP tended to be retained in the hepatopancreas, resulting in accumulation of 50 mol% of total toxin.     

Toxicity and tetramine contents of salivary glands from carnivorous gastropods

Salivary glands from 29 species of marine carnivorous gastropods in nine families were examined for lethal activity against mice and tetramine content. Mouse lethality was assayed by intravenous injection of buffer extracts into mice, and was detected in 14 species. Heat-stability tests confirmed that toxins in four species were thermolabile, while those in eight species were thermostable. Based on the tetramine contents determined by the colorimetric method using methanolic extracts, the thermostable toxins in seven species (Neptunea eulimatalamellosa, N. vinosa, N. arthritica, N. bulbacea, N. intersculpta f. pribiloffensis, N. intersculpta f. frater pilsbry and Hemifusus tuba) were considered to be tetramine contained at high levels (more than 900 micrograms/g salivary gland), but that in one species (Buccinum opisthoplectum) appeared to be a low-molecular-weight compound differing from tetramine. It is interesting that one (Hemifusus tuba) of the seven species containing high amounts of tetramine belongs to the family Melongenidae, although the other six Neptunea species are members of the family Buccinidae, as expected from previous studies.     

Determination of sucralose in foods by HPLC using pre-column derivatization

The development of a sensitive pre-column derivatization high-performance liquid chromatography (HPLC) method for determination of sucralose is reported. Sucralose is converted into a strongly ultraviolet (UV)-absorbing derivative, possessing strong absorption at 260 nm, by treatment with p-nitrobenzoyl chloride (PNBCl). Homogenized samples were dialyzed and washed with a Bond Elut ENV cartridge, then the eluate was evaporated to dryness and the residue was derivatized. Subsequently, the sucralose derivative was purified with hexane-ethyl actate (9:1) in a silica cartridge, and then the sucralose derivative was eluted with acetone. HPLC was performed on a phenyl column, using acetonitrile-water (73:27) as a mobile phase with UV detection (260 nm). The calibration curve was linear in the range of 1 microgram/mL to 50 micrograms/mL of sucralose. The recoveries of sucralose from eight kinds of foods spiked at the levels of 0.20 and 0.05 g/kg of sucralose were more than 76.2% with SD values in the range from 0.90% to 4.31%. The quantitative limit of the developed method was 0.005 g/kg for sucralose in samples.     

Contents of barbaloin-related compounds in aloe drinks and their change during storage

The contents of barbaloin (BA), isoBA, aloin-dimers A, B, C, D and aloe-emodin (AE) in aloe drinks were investigated. BA and isoBA were detected in 30 of the 31 samples at the levels of 120-570 micrograms/mL and 120-580 micrograms/mL, respectively. Aloin-dimers A, B, C and D were detected in 8 of the 31 samples at the levels of 12-38 micrograms/mL, 13-39 micrograms/mL, 11-36 micrograms/mL and 16-69 micrograms/mL, respectively. AE was detected in all of the 31 samples at the levels of 0.03-1.3 micrograms/mL. When aloe drinks were stored for 4 weeks at 5 degrees C after opening the bottle, decrease of BA and isoBA, and increase of AE and aloin-dimers A, B, C and D were observed in most cases. However, in a few aloe drinks, all of BA, isoBA, aloin-dimers A, B, C, D and AE decreased. In these drinks, the existence of aloin-trimer was elucidated by LC/MS analyses. These data suggested that BA in aloe drinks is converted to the dimer and then to the trimer during storage.     

Numerical simulation of an arbitrarily ended asymmetrical slab waveguide by guided-mode extracted integral equations

The scattering phenomenon from an arbitrary-shaped end of a asymmetrical slab waveguide for the cases of TE and TM guided modes is simulated by means of boundary integral equations that are called guided-mode extracted integral equations. The integral equations that we derive can be solved by the conventional boundary-element method. Numerical results are presented for problems of three-layer asymmetrical waveguides with tilted ends. The reflection coefficient, reflected and scattered powers, and radiation patterns are calculated numerically for the cases of incident TE and TM guided modes.     

An investigation of the pH-activity relationships of Cex, a family 10 xylanase from Cellulomonas fimi: xylan inhibition and the influence of nitro-substituted aryl-beta-D-xylobiosides on xylanase activity

The kinetic parameters of Cex, a family 10 xylanase from Cellulomonas fimi, were determined at various pH levels using soluble birchwood xylan (BWX) as a natural polymeric substrate along with three other synthetic aryl-beta-D-xylobioside substrates. Using BWX, a high level of substrate inhibition was observed which increased with decreasing pH. In contrast, typical Michaelis-Menten-type profiles were obtained using the three aryl-beta-D-xylobiosides as substrates. The k(cat) values determined using o-nitrophenyl-beta-D-xylobioside did not change as the pH increased, whereas the k(cat) values obtained with BWX, phenyl-beta-D-xylobioside and p-nitrophenyl-beta-D-xylobioside decreased, suggesting that the presence of an ortho nitro group affects the activity displayed by Cex. These differences were not observed with XynB from Clostridium stercorarium F9, a member of the same family of xylanases as Cex. These results indicate that a careful evaluation is required when employing substituted aryl-beta-D-xylobiosides in the characterization of xylanases.     

Production of eicosapentaenoic acid by a recombinant marine cyanobacterium, Synechococcus sp.

The eicosapentaenoic acid (EPA) synthesis gene cluster from an EPA-producing bacterium, Shewanella sp. SCRC-2738, was cloned into a broad-host range vector, pJRD215, and then introduced into a marine cyanobacterium, Synechococcus sp. NKBG15041c, by conjugation. The transconjugant cyanobacteria produced 3.7±0.2% (2.24±0.13 mg/L) EPA (n-3) and 2.5 ±0.2% (1.49±0.06 mg/L) eicosatetraenoic acid (n-3) of the total fatty acids when the cells were cultured at 23°C at a light intensity of 1,000–1,500 Lux. The EPA and eico-satetraenoic acid contents of the cells were increased to 4.6±0.6% (3.86±1.11 mg/L) and 4.7±0.3% (3.86±0.82 mg/L), and 7.5±0.3% (1.76±0.10 mg/L) and 5.1±0.2% (1.19±0.06 mg/L) when they were cultured at low temperature (18°C) and at lower light intensity (40 Lux), respectively.     

Endothelin A-receptor blockade worsens endotoxin-induced hepatic microcirculatory changes and necrosis

BACKGROUND & AIMS: Endothelin 1 is considered to be an important regulator of sinusoidal blood flow and increases during endotoxemia. The purpose of this study was to investigate the role of endothelin 1 in hepatic microcirculation, oxygen transport, and liver injury during endotoxemia. METHODS: Male Sprague-Dawley rats were continuously infused with 2.5 mL/h of saline, 0.8 mg . kg-1 . h-1 of lipopolysaccharide (LPS), 3 mg . kg-1 . h-1 of BQ-485, an endothelin A-receptor antagonist, or LPS plus BQ-485 for 7 hours. RESULTS: BQ-485 infusion had no significant effect on hepatic microcirculation and liver injury. LPS increased the plasma levels of aspartate aminotransferase (AST) and total bilirubin and decreased the hepatic adenosine triphosphate (ATP) level and bile flow rate. LPS + BQ-485 infusion further increased the plasma levels of AST and total bilirubin and decreased the bile flow rate and the hepatic ATP level. Dual-spot microspectroscopy revealed mild decreases in sinusoidal erythrocyte velocity and oxygen transport in the LPS group and profound decreases in these parameters in the LPS + BQ-485 group. Histological examinations revealed massive necrotic changes in the pericentral regions of the LPS + BQ-485 group. CONCLUSIONS: These results suggest that blockade of endothelin A receptors disturbs hepatic microcirculation and oxygen transport and aggravates the necrotic injury induced by endotoxin.