Electrospray ionization mass spectrometry of cyclodextrin complexes with amino acids in incubated solutions and in eluates of gel permeation chromatography

1:1 complexes of beta-cyclodextrin (CD) with three amino acids (Gly, Phe and Trp) have been detected as ions in the gas phase using infusion positive and negative ion electrospray ionization mass spectrometry (ESI-MS). In contrast with the positive ion ESI mass spectra of simple aqueous solutions, the aggregates and adducts usually formed in the ESI process did not appear in the positive ion ESI spectra of solutions buffered with ammonium acetate (NH4Ac), even at higher analyte concentrations, These studies suggest that addition of buffer and/or use of a low analyte concentration should be used to overcome formation of aggregates and metal ion adducts in such mass spectrometry studies. Also, the deprotonated complexes are dissociated by collision induced dissociation (CID) to form an abundant product ion, the deprotonated CD, requiring transfer of a proton to the amino acid carboxyl group, To understand formation of complexes in the gas phase, gel permeation chromatography (GPC) was used to separate free amino acids (AAs) from complexes in an incubated solution. The ESI mass spectra of the GPC fractions show the presence of 1:1 complexes of both CD-aromatic amino acids and CD-aliphatic amino acids. Compared with CD-aliphatic amino acid complexes, CD-aromatic amino acid complexes appear to be destabilized in the gas phase, possibly because the hydrophobic interaction which binds the aromatic group of amino acids in the CD cavity in solution may become repulsive when solvent evaporates from the droplets during the electrospray process, whereas those complex ions formed as proton bound dimers are stabilized by electrostatic forces, the major binding force for such complexes in the gas phase. In addition, the GPC technique coupled with off-line ESI-MS can rapidly separate CD complexes by size, and provides some information on the character of the complexes in solution. (C) 1998 John Wiley & Sons, Ltd.

Synthesis and structure of [NH4](4)H[(PMo8V4V2O42)-V-IV-O-V].24H(2)O

[NH4](4)H[(PMo8V4V2O42)-V-IV-O-V] . 24H(2)O has been hydrothermally synthesized from ammonium vanadate and sodium molybdate in aqueous solution by adding phosphorous acid, and its structure determined by single crystal X-ray analysis. The heteropoly molybdovanadophosphate anion is a bicapped 'pseudo-Keggin' polyanion, two vanadium atoms are distributed at two 'capping' metal atom positions and other four vanadium atoms randomly distributed at eight metal atom positions. (C) 1998 Elsevier Science Ltd. All rights reserved.

Probe beam deflection combined with cyclic voltammetry studies of C-60-modified electrode film

C-60 films, prepared by solution casting, were studied by means of in situ probe beam deflection (PBD) combined with cyclic voltammetry (CV). PBD is a powerful technique for investigation of phenomena at the electrode/electrolyte interface in acetonitrile with quaternary ammonium and alkali metal salts as supporting electrolytes. In tetra-n-butylammonium (TBA(+)) salt solution, a stable CV can be obtained during the first two reduction/reoxidation waves. On reduction, injection of cations to maintain charge balance and dissolution of small amount of C-60(-) (TEA(+)) and/or C-60(2-) (TBA(+))(2) are detected. During the reoxidation process ejection of cations and injection of anions occur simultaneously, especially for the second reoxidation wave. In the case where TBABr is the supporting electrolyte, the accompanied behavior is more complicated than in TBABF(4), TBAClO(4), and TBAPF(6) solutions. A small pair of prewaves in CV are proposed due to oxidation/reduction of C-60 domains but not dissolution/redeposition of C-60 film. Extending the potential scan range to the third reduction wave, no apparent corresponding reoxidation wave is related to the third reduction wave, the electroactivity of the film disappears rapidly and dissolution of C-60 film is observed. In tetraethylammonium (TEA(+)) and NAClO(4) solutions, the electrochemistry of the C-60 films is unstable, and potential scans lead to dissolution of flaking of the film.

WATER-SOLUBLE AND AMPHIPHILIC POLYMERS .11. GRAFT-POLYMERIZATION OF ACRYLIC-ACID ON POLY(VINYL ALCOHOL)

The graft polymerization of acrylic acid(AA) on poly(vinyl alcohol) (PVAL) has been investigated by using either potassium persulfate (KPS) or ceric ammonium nitrate(CAN) as an initiator. In the case of KPS initiation, the formation of the graft polymer always lags behind the homopolymer formation. The graft polymer is separated by acetone, and the increase of reaction temperature favors the homopolymer formation at the early stage. In the case of CAN initiation, graft polymers with a high PAA content can hardly be obtained when the polymerization is performed under nitrogen and at < 0.06 mol/L HNO3 concentration. It has been found that incorporation of a small amount of oxygen in a protective nitrogen gas accelerates markedly the graft polymerization, and that the resulting graft polymers can not be separated by acetone precipitation technique in most cases. The Dalian nitrogen(containing 0.7% oxygen) is a good protective gas for CAN-initiated PVAL-AA graft polymerization.

STUDY OF THE CHLORINATION REACTION OF Li2CO3 WITH NH4Cl

The chlorination reaction of Li2CO3 with NH4Cl has been studied in detail by a series of thermal analysis methods. When NH4Cl/Li2CO3 mole ratio equals 4, Li2CO3 can be transformed into LiCl quantitatively in a stream of Ar gas flow. All residual NH4Cl is decomposed completely at 400 degrees C and carried away from the reaction cell by Ar gas.Analysis by X-Ray diffraction and Ion Chromatography show that there are almost no NH4Cl remained in The LiCl product. It is interested that the chlorination reaction can be applied to the determinations of phase diagram by thermal analysis method and the preparation of Al-Li alloy by electrolysis in molten salt.

Microprojectile bombardment of Laminaria japonica gametophytes and rapid propagation of transgenic lines within a bubble-column bioreactor

A human acidic fibroblast growth factor gene, hafgf, was successfully transferred into Laminaria japonica (kelp) gametophytes via microprojectile bombardment using the biolistic PDS-1000/He gene gun. Following phosphinothricin screening, PCR detection and Southern blot analysis, transgenic L. japonica gametophytes were cultivated in an illuminated bubble-column bioreactor to optimize growth conditions. A maximal final dry cell density of 1,695 mg l(-1) was obtained in a batch culture having an initial dry cell density of 129.75 mg l(-1). This was achieved using an aeration rate of 1.08 l air min(-1) l(-1) culture in a medium containing 1.5 mM inorganic nitrate and 0.15 mM phosphate. In addition, the relationship between different nitrogen sources and growth of transgenic gametophytes indicated that both urea and sodium nitrate were effective nitrogen sources for cell growth, while ammonium ions inhibited growth of these gametophytes.

Purification and partial characterization of Mn superoxide dismutase from muscle tissue of the shrimp Macrobrachium nipponense

Superoxide dismutase (SOD; EC 1.15.1.1) is an enzyme that protects against oxidative stress from superoxide radicals in living cells. This enzyme had been isolated, purified and partially characterized from muscle tissue of the shrimp Macrobrachium nipponense. The purification was achieved by heat treatment, ammonium sulfate fractionated precipitation and column chromatograph on DEAE-cellulose 32. Some physiological and biochemical characterization of it was tested. The molecular weight of it was about 21.7 kDa, as judged by SDS-polyacrylamide gel electrophoresis. The purified enzyme had an absorption peak of 278 nm in ultraviolet region, and the enzyme remained stable at 25-45 degreesC within 90 min. However, it was rapidly inactivated at higher temperature. Treatment of the enzyme with 1 mM ZnCl2, SDS and 1 mM or 10 mM mercaptoethanol showed some increasing activity. However, the enzyme activity was obviously inhibited by 10 mM CaCl2, CuSO4, ZnCl2 and 1 mM CaCl2 and 10 mM K2Cr2O7. SOD activity did not show significantly variation after incubated with 1 mM CaCl2, EDTA and 10 MM SDS. The enzyme was insensitive to cyanide and contained 1.03 +/- 0.14 atoms of manganese per subunit shown in atomic absorption spectroscopy, which revealed that purified SOD was Mn superoxide dismutase. (C) 2004 Elsevier B.V. All rights reserved.

Integrated cultivation of the red alga Kappaphycus alvarezii and the pearl oyster Pinctada martensi

The effect of simultaneously cultivating the pearl oyster Pinctada martensi and the red alga Kappaphycus alvarezii on growth rates of both species was investigated in laboratory and field studies conducted from December 1993 to June 1995. The two study sites were in subtidal areas 100 km apart off the east coast of Hainan Island, China. Pearl oysters were cultivated in the center of an algal farm and red alga was cultivated in the center of the pearl oyster farm. These field experiments showed higher growth rates of both P. martensi and K. alvarezii in a co-culture system than in a monospecies culture system. Laboratory studies showed that the algae removed nitrogenous wastes released by pearl oysters. Algae treated with pearl oyster wastes grew much faster than those without oyster wastes. Algae treated with the seawater to which NH4Cl, NaNO3 and NaNO2 were added grew at the same rate as those treated with natural seawater containing oyster nitrogenous wastes, suggesting that enhanced growth of algae in the co-culture system was largely due to nitrogenous metabolites of the pearl oysters. In the co-culture, growth of pearl oysters was positively influenced by the presence of rapidly growing algae but when seawater temperature decreased below 20 degrees C, the algae grew slowly and there was no measurable benefit of mixed culture to either algae or pearl oyster.

Purification and characterization of agarases from a marine bacterium Vibrio sp. F-6

Marine bacterium Vibrio sp. F-6, utilizing agarose as a carbon source to produce agarases, was isolated from seawater samples taken from Qingdao, China. Two agarases (AG-a and AG-b) were purified to a homogeneity from the cultural supernatant of Vibrio sp. F-6 through ammonium sulfate precipitation, Q-Sepharose FF chromatography, and Sephacryl S-100 gel filtration. Molecular weights of agarases were estimated to be 54.0 kDa (AG-a) and 34.5 kDa (AG-b) by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The optimum pH values for AG-a and AG-b were about 7.0 and 9.0, respectively. AG-a was stable in the pH range of 4.0-9.0 and AG-b was stable in the pH range of 4.0-10.0. The optimum temperatures of AG-a and AG-b were 40 and 55 degrees C, respectively. AG-a was stable at temperature below 50 degrees C. AG-b was stable at temperature below 60 degrees C. Zn2+, Mg2+ or Ca2+ increased AG-a activity, while Mn2+, Cu2+ or Ca2+ increased AG-b activity. However, Ag+, Hg2+, Fe3+, EDTA and SDS inhibited AG-a and AG-b activities. The main hydrolysates of agarose by AG-a were neoagarotetraose and neoagarohexaose. The main hydrolysates of agarose by AG-b were neoagarooctaose and neoagarohexaose. When the mixture of AG-a and AG-b were used, agarose was mainly degraded into neoagarobiose.

Physiological and immune responses of zhikong scallop Chlamys farreri to the acute viral necrobiotic virus infection

The Zhikong Scallop, Chlamys farreri, is one of the most Important bivalve mollusks cultured in northern China However, mass mortality of the cultured C farreri has posed a serious threat to the maricultural Industry in recent years. Acute Viral Necrobiotic Virus (AVNV) is believed as an important etiological agent causing the scallop mass mortalities To understand the mechanism behind the AVNV associated scallop disease and mortality, we assessed the physiological and immune responses of C farreri to the virus infection using oxygen consumption rate, ammonium-nitrogen excretion rate, hemocyte copper, zinc superoxide dismutase gene expression, and plasma superoxide dismutase activity and alkaline phosphatase activity as indicators Scallops challenged by AVNV at 25 C developed typical disease signs 2 days after virus injection Before the disease manifested, scallop oxygen consumption and NH4+-N excretion rates rose and then fell back. Real-time PCR revealed that the hemocyte cytosol Cu, Zn SOD gene expression was upregulated followed by recovery The plasma SOD activity, however, augmented consistently following virus injection Moreover, plasma AKP activity first lowered and then elevated gradually to the highest level at 24 h post virus injection Scallops challenged by AVNV at 17 degrees C neither developed notable disease nor showed obvious responses that could be associated with the virus infection. While the results suggested a correlation between the elevated seawater temperature and the AVNV infection associated C farreri mortalities, they also indicated that the viral infection provoked multiple physiological and immune responses in the host scallops (C) 2010 Elsevier Ltd All rights reserved

Seasonal and interannual variability of primary and export production in the South China Sea: a three-dimensional physical-biogeochemical model study

To investigate the seasonal and interannual variations in biological productivity in the South China Sea (SCS), a Pacific basin-wide physical - biogeochemical model has been developed and used to estimate the biological productivity and export flux in the SCS. The Pacific circulation model, based on the Regional Ocean Model Systems (ROMS), is forced with daily air-sea fluxes derived from the NCEP (National Centers for Environmental Prediction) reanalysis between 1990 and 2004. The biogeochemical processes are simulated with a carbon, Si(OH)(4), and nitrogen ecosystem (CoSiNE) model consisting of silicate, nitrate, ammonium, two phytoplankton groups (small phytoplankton and large phytoplankton), two zooplankton grazers (small micrograzers and large mesozooplankton), and two detritus pools. The ROMS-CoSiNE model favourably reproduces many of the observed features, such as ChI a, nutrients, and primary production (PP) in the SCS. The modelled depth-integrated PP over the euphotic zone (0-125 m) varies seasonally, with the highest value of 386 mg C m (-2) d (-1) during winter and the lowest value of 156 mg C m (-2) d (-1) during early summer. The annual mean value is 196 mg C m (-2) d (-1). The model-integrated annual mean new production (uptake of nitrate), in carbon units, is 64.4 mg C m (-2) d (-1) which yields an f-ratio of 0.33 for the entire SCS. The modelled export ratio (e-ratio: the ratio of export to PP) is 0.24 for the basin-wide SCS. The year-to-year variation of biological productivity in the SCS is weaker than the seasonal variation. The large phytoplankton group tends to dominate over the smaller phytoplankton group, and likely plays an important role in determining the interannual variability of primary and new production.

Nitrogen isotopic fractionation during a simulated diatom spring bloom: importance of N-starvation in controlling fractionation

N isotope fractionation (epsilon) was first determined during ambient NO3- depletion in a simulated diatom spring bloom. After 48 h of N-starvation, NH4+ was resupplied to the diatoms in small pulses to simulate grazer-produced N and then epsilon was determined. Large variations in epsilon values were observed: from 2.0-3.6 to 14-0 parts per thousand during NO3- and NH4+ uptake, respectively. This is the first study reporting an epsilon value as low as 0 to 2 parts per thousand for NH4+ uptake and we suggest that greater N demand after N-starvation may have drastically reduced NH3 efflux out of the cells. Thus the N status of the phytoplankton and not the ambient NH4+ concentration may be the important factor controlling epsilon, because, when N-starvation increased, epsilon values for NH4+ uptake decreased within 30 h. This study may thus have important implications for interpreting the delta(15)N of particulate N in nutrient-depleted regimes in temperate coastal oceans.

The status and characteristics of eutrophication in the Yangtze River (Changjiang) estuary and the adjacent East China Sea, China

Eutrophication has become increasingly serious and noxious algal blooms have been of more frequent occurrence in the Yangtze River Estuary and in the adjacent East China Sea. In 2003 and 2004, four cruises were undertaken in three zones in the estuary and in the adjacent sea to investigate nitrate (NO3-N), ammonium (NH4-N), nitrite (NO2-N), soluble reactive phosphorus (SRP), dissolved reactive silica (DRSi), dissolved oxygen (DO), phytoplankton chlorophyll a (Chl a) and suspended particulate matter (SPM). The highest concentrations of DIN (NO3-N+NH4-N+NO2-N), SRP and DRSi were 131.6, 1.2 and 155.6 mu M, respectively. The maximum Chl a concentration was 19.5 mg m(-3) in spring. An analysis of historical and recent data revealed that in the last 40 years, nitrate and SRP concentrations increased from 11 to 97 mu M and from 0.4 to 0.95 mu M, respectively. From 1963 to 2004, N:P ratios also increased from 30-40 up to 150. In parallel with the N and P enrichment, a significant increase of Chl a was detected, Chl a maximum being 20 mg m(-3), nearly four times higher than in the 1980s. In 2004, the mean DO concentration in bottom waters was 4.35 mg l(-1), much lower than in the 1980s. In comparison with other estuaries, the Yangtze River Estuary was characterized by high DIN and DRSi concentrations, with low SRP concentrations. Despite the higher nutrient concentrations, Chl a concentrations were lower in the inner estuary (Zones 1 and 2) than in the adjacent sea (Zone 3). Based on nutrient availability, SPM and hydrodynamics, we assumed that in Zones 1 and 2 phytoplankton growth was suppressed by high turbidity, large tidal amplitude and short residence time. Furthermore, in Zone 3 water stratification was also an important factor that resulted in a greater phytoplankton biomass and lower DO concentrations. Due to hydrodynamics and turbidity, the open sea was unexpectedly more sensitive to nutrient enrichment and related eutrophication processes.

Seasonal variation in metabolism of cultured Pacific oyster, Crassostrea gigas, in Sanggou Bay, China

Rates of respiration and excretion of the Pacific oyster, Crassostrea gigas, were measured seasonally from June 2002 to July 2003 under ambient conditions of food, water temperature, pH, and salinity in Sanggou Bay, an important mariculture coast in north China. The aim of this study is to obtain fundamental data for further establishing an energy budget model and assessing the carrying capacity for cultivation of C. gigas in north China. Oysters were collected monthly or bimonthly from the integrated culture areas of bivalve and kelp in the bay. Oxygen consumption and ammonium and phosphorus excretion rates were measured, and ratios of O/N and NIP were calculated. One-way ANOVA was applied to determine differences among these parameters that act as a function of seasonal variation. All the physiological parameters yielded highly significant variations with season (P<0.01) The rate of respiration varied seasonally, with the highest oxygen consumption rate in July and the lowest rate in January, ranging from 0.07 to 2.13 mg O-2 h(-1) g(-1) dry tissue weight (DW). Maximum and minimum ammonium excretion rates were recorded in August and January, respectively, ranging from 0.51 to 5.40 mu mol NH4-N h(-1) g(-1) DW. Rates of phosphorus excretion varied from 0.11 (in January) to 0.64 (in July) mu mol PO4-P h(-1) g(-1) DW. The O/N and N/P ratios changed from 9.2 (in January) to 59.8 (in July) and from 4.6 (in January) to 10.9 (in August), respectively. For each season, the allometric relationship between the physiological response (e.g., rate of oxygen consumption, ammonium and phosphorus excretion) and DW of the animal was estimated using the formula: Y=a x DWb. (C) 2005 Elsevier B.V. All rights reserved.

Effects of nitrogen source and concentration on growth rate and fatty acid composition of Ellipsoidion sp (Eustigmatophyta)

In order to study the effects of different nitrogen source and concentration on the growth rate and fatty acid composition, a marine microalga Ellipsoidion sp. with a high content of eicosapentaenoic acid (EPA) was cultured in media with different nitrogen sources and concentrations. During the pre-logarithmic phase, the alga grew faster with ammonium as N source than with nitrate, but the reverse applied during the post-logarithmic phase. The alga grew poorly in N-free medium or medium with urea as the sole N source. In the same growth phase, ammonium medium resulted in higher yield of total lipid, but the EPA yield did not differ significantly different from that using nitrate medium. The maximum growth rate occurred in medium containing 1.28 mmol L-1 sodium nitrate, while maximum EPA and total lipid contents were reached at 1.92 mmol L-1, when EPA accounted for 27.9% total fatty acids. The growth rate kept stable when NH4Cl ranged from 0.64 to 2.56 mmol L-1, and the maximum content of total lipid and EPA occurred in the medium with 2.56 mmol L-1 NH4Cl. The EPA content was higher in the pre- than post-logarithmic phase, though the total lipid content was lower. The highest EPA content expressed as percent total fatty acid was 27.9% in nitrate medium and and 39.0% in ammonium medium.