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.

Evaluation of 28 marine algae from the Qingdao coast for antioxidative capacity and determination of antioxidant efficiency and total phenolic content of fractions and subfractions derived from Symphyocladia latiuscula (Rhodomelaceae)

The extracts obtained from 28 species of marine algae were evaluated for their antioxidant activity (AA) versus the positive controls butylated hydroxytoluene (BHT), gallic acid (GA), and ascorbic acid (AscA). Most of the tested samples displayed antioxidant activity to various degrees. Among them, the extract of Symphyocladia latiuscula exhibited the strongest AA, which was comparable to BHT, GA, and AscA in radical scavenging activity, as shown in the DPPH (alpha,alpha-diphenyl-beta-picrylhydrazyl) assay, and higher than those of the positive controls in beta-carotene-linoleate assay system. In addition, the ethyl acetate-soluble fraction isolated from the crude extract of S. latiuscula exhibited the highest antioxidant activity in both assay systems. This fraction was further fractionated into seven subfractions (F1-F7) by vacuum liquid chromatography (VLC). F1 and F4 were found to be the most effective subfractions in scavenging DPPH radical assay and in the beta-carotene-linoleate assay, respectively. The total phenolic content (TPC) and reducing power (RP) for all of the extracts, fractions, and subfractions (F1-F7) were also determined. The TPC of the 28 extracts ranged from 0.10 to 8.00 gallic acid equivalents (mg/g seaweed dry weight) while the RP ranged from 0.07 to 11.60 ascorbic acid equivalents (mg center dot g(-1) seaweed dry weight). Highly positive relationships between AA and TPC as well as between AA and RP were found for the extracts and fractions, while for the subfractions F1-F7 only weak or no such relations were found. The results obtained from this study indicate that further analysis is needed of those marine algal species that contain the most antioxidant activity in order to identify the active principles.

Evaluation of antioxidant property of extract and fractions obtained from a red alga, Polysiphonia urceolata

Antioxidant activity (AA), total phenolic content, and reducing power of the crude extract, fractions, and subfractions derived from a red alga, Polysiphonia urceolata, were evaluated and determined. The antioxidative activity was measured using the alpha,alpha-diphenyl-beta-pierylhydrazyl (DPPH) radical scavenging assay and the P-carotene-linoleate assay systems, and compared with that of butylated hydroxytoluene (BHT), gallic acid (GA), and ascorbic acid (AscA). The results showed that the crude extract and the ethyl acetate-soluble fraction exhibited higher AA than BHT in the DPPH assay model, at all of four concentration levels tested (from 0.4 to 50 mu g/ml), while, in the beta-carotene-linoleate assay system, the crude extract and the ethyl acetate-soluble fraction exhibited similar or, in most cases, higher AA than GA and AscA at the same concentrations (from 10 to 200 mu g/ml). The ethyl acetate-soluble fraction was further fractionated into seven subfractions F1-F7 by silica gel vacuum liquid chromatography. F1 was found to be the most effective subfraction in both assay systems. The total phenolic content and reducing power were determined using the Folin-Ciocalteu and the potassium ferricyanide reduction methods, respectively. Statistical analysis indicated a significant association between the antioxidant potency and total phenolic content as well as between the antioxidant potency and reducing power. (c) 2005 Elsevier Ltd. All rights reserved.

Experimental study on the fractionation of yttrium from holmium during the coprecipitation with calcium carbonates in seawater solutions

The partitioning of Y and Ho between CaCO3 (calcite and aragonite respectively) and seawater was experimentally investigated at 25 degrees C and I atm. Both Y and Ho were observed to be strongly partitioned into the overgrowths of calcite or aragonite. Their partition coefficients, D-Y and D-Ho, were determined to be similar to 520-1400 and similar to 700-1900 in calcite, similar to 1200-2400 and similar to 2400-4300 in aragonite, respectively. Y fractionates from Ho during the coprecipitation with either calcite or aragonite. Within our experimental conditions, the fractionation factor, k = D-Y/D-Ho, was determined to be similar to 0.62-0.77 in calcite and similar to 0.50-0.57 in aragonite, respectively. The aqueous complexation of Y and Ho, which is a function of solution chemistry, probably plays an important role in both the partitioning and the fractionation. Further analyses suggest that the difference in covalency between Y and Ho associated with changes in their coordination environments is the determinant factor to the Y-Ho fractionation in the H2CO3-CaCO3 System.

Phytoplankton biomass size structure and its regulation in the Southern Yellow Sea (China): Seasonal variability

Phytoplankton size structure plays a significant role in controlling the carbon flux of marine pelagic ecosystems. The mesoscale distribution and seasonal variation of total and size-fractionated phytoplankton biomass in surface waters. as measured by chlorophyll a (Chl a), was studied in the Southern Yellow Sea using data from four cruises during 2006-2007. The distribution of Chl a showed a high degree of spatial and temporal variation in the study area. Chl a concentrations were relatively high in the summer and autumn, with a mean of 142 and 1.27 mg m(-3), respectively. Conversely, in the winter and spring. the average Chl a levels were only 098 and 0.99 mg m(-3) Total Chl a showed a clear decreasing gradient from coastal areas to the open sea in the summer, autumn and winter cruises. Patches of high Chl a were observed in the central part of the Southern Yellow Sea in the spring due to the onset of the phytoplankton bloom. The eutrophic coastal waters contributed at least 68% of the total phytoplankton biomass in the surface layer. Picophytoplankton showed a consistent and absolute dominance in the central region of the Southern Yellow Sea (>40%) in all of the cruises, while the proportion of microphytoplankton was the highest in coastal waters The relative proportions of pico- and nanophytoplankton decreased with total biomass, whereas the proportion of the micro-fraction increased with total biomass. Relationships between phytoplankton biomass and environmental factors were also analysed. The results showed that the onset of the spring bloom was highly dependent on water column stability. Phytoplankton growth was limited by nutrient availability in the summer due to the strong thermocline. The combined effects of P-limitation and vertical mixing in the autumn restrained the further increase of phytoplankton biomass in the Surface layer. The low phytoplankton biomass in winter was caused by vertical dispersion due to intense mixing. Compared with the availability of nutrients. temperature did not seem to cause direct effects on phytoplankton biomass and its size structure. Although interactions of many different environmental factors affected phytoplankton distributions. hydrodynamic conditions seemed to be the dominant factor. Phytoplankton size structure was determined mainly by the size-differential capacity in acquiring resource. Short time scale events, such as the spring bloom and the extension of Yangtze River plume, can have substantial influences, both on the total Chl a concentration and on the size structure of the phytoplankton. (C) 2009 Elsevier Ltd. All rights reserved.

Fast microzooplankton grazing on fast-growing, low-biomass phytoplankton: a case study in spring in Chesapeake Bay, Delaware Inland Bays and Delaware Bay

Dilution experiments were performed to examine the growth and grazing mortality rates of picophytoplankton (< 2 mu m), nanophytoplankton (2-20 mu m), and microphytoplankton (> 20 mu m) at stations in the Chesapeake Bay (CB), the Delaware Inland Bays (DIB) and the Delaware Bay (DB), in early spring 2005. At station CB microphytoplankton, including chain-forming diatoms were dominant, and the microzooplankton assemblage was mainly composed of the tintinnid Tintinnopsis beroidea. At station DIB, the dominant species were microphytoplanktonic dinoflagellates, while the microzooplankton community was mainly composed of copepod nauplii and the oligotrich ciliate Strombidium sp. At station DB, nanophytoplankton were dominant components, and Strombidium and Tintinnopsis beroidea were the co-dominant microzooplankton. The growth rate and grazing mortality rate were 0.13-3.43 and 0.09-1.92 d(-1) for the different size fractionated phytoplankton. The microzooplankton ingested 73, 171, and 49% of standing stocks, and 95, 70, and 48% of potential primary productivity for total phytoplankton at station CB, DIB, and DB respectively. The carbon flux for total phytoplankton consumed by microzooplankton was 1224.11, 100.76, and 85.85 mu g C 1(-1) d(-1) at station CB, DIB, and DB, respectively. According to the grazing mortality rate, carbon consumption rate and carbon flux turn over rates, microzooplankton in study area mostly preferred to graze on picophytoplankton, which was faster growing but was lowest biomass component of the phytoplankton. The faster grazing on Fast-Growing-Low-Biomass (FGLB) phenomenon in coastal regions is explained as a resource partitioning strategy. This quite likely argues that although microzooplankton grazes strongly on phytoplankton in these regions, these microzooplankton grazers are passive.

Study on extraction of agaropectin from Gelidium amansii and its anticoagulant activity

Gelidium amansii agar was fractionated on DEAE-cellulose and four fractions were obtained sequentially. The yields of 1.0 mol/L NaCl fraction and 2.5 mol/L NaCl fraction were 2.80% and 2.03%. They are highly sulfated agar, and named as agaropectin with sulfate content being 22.8% and 32.5%, respectively. The anticoagulant experiment results show that agaropectin could effectively prolong the coagulation time in a dose-dependent manner in vitro. Agaropection could be absorbed and effectively prolong the plasma coagulation time in vivo. After intragastric administration at the doses of 100, 200, and 400 mg/kg.d in rats for 15 days, TT (thrombin time), CT (coagulation time), PT (prothrombin time), and APTT (activated partial thromboplastin time) could be effectively prolonged and the plasma Fib level could be significantly lowered.

SIZE STRUCTURES OF MICROPLANKTON BIOMASS AND PRODUCTION IN JIAOZHOU BAY, CHINA

Seasonal investigations of size-fractionated biomass and production were carried out from February 1992 to May 1993 in Jiaozhou Bay, China. Microplankton assemblages were separated into three fractions: pico-(0.7-2 mu m), nano- (2-20 mu m) and netplankton (20-200 mu m). The biomass was measured as chlorophyll a (Chl a), particulate organic carbon (POC) and particulate organic nitrogen (PON). The production was determined by C-14 and N-15 tracer techniques. The seasonal patterns in biomass, though variable, were characterized by higher values in spring and lower values in autumn and summer (for Chl a only). The seasonal patterns in production, on the other hand, were more clear with higher values occurring in summer and spring, and lower values occurring in autumn and winter. Averaged over the whole study period, the respective proportions of total biomass accounted for by net-, nano- and picoplankton were 26, 45 and 29% for Chl a, 32, 33 and 35% for POC, and 26, 32 and 42% for PON. The contributions to total primary production by net-, nano- and picoplankton were 31, 35 and 34%, respectively. The respective proportions of total NH4+-N uptake accounted for by net-, nano- and picoplankton were 28, 33 and 39% in the daytime, and 10, 29 and 61% at night. The respective contributions to total NO3--N uptake by net-, nano- and picoplankton were 37, 40 and 23% in the daytime, and 13, 23 and 64% at night. Some comprehensive ratios, including C/N biomass ratio, Chl a/C ratio, C uptake/Chl a ratio, C:N uptake ratio and the f-ratio, were also calculated size separately, and their biological and ecological meanings are discussed.

北极Gakkel超慢速扩张脊大洋橄榄岩的地球化学特征及其意义

Gakkel Ridge in Arctic Ocean is the ulstraslow spreading ridge in the world with a full spreading rate decreasing from 14 mm/yr in the western end to 7mm/yr in the eastern end. To study the histories of partial melting and melt referilization occurred in the oceanic mantle beneath Gakkel Ridge, both extremely fresh and altered abyssal peridotites from two dredge hauls (PS66-238 and HLY0102-D70) have been selected in this research. Major and trace element data of the residual minerals suggest that all samples have been refertilized by late enriched melts after low to moderate degrees (3-12%) of partial melting in the stability field of spinel, whereas some samples also inherited signatures of partial melting in stability field of garnet. Os isotopic compositions of Gakkel samples have not been significantly affected by late processes, e.g., seawater alteration and melt refertilzaiton. Samples from both dredge hauls have similar range of 187Os/188Os, from strongly unradiogenic (~0.114) in the harzburgites to approximating the inferred values of PUM (primitive upper mantle) in some lherzolites (~0.129). Inherited ancient depletion events in the harzburgites with Re-depletion age up to 2 billion years are unrelevant to the recent genesis of MORB (mid-ocean ridge basalts) beneath Gakkel Ridge. Comparisons of highly siderophile elements (HSEs) between the fresh and altered samples suggested both Pd and Re were affected and thus are mobile during seawater alteration, whereas the other HSEs (i.e., Os, Ir, Ru an Pt) are stable. The fractionated HSEs patterns in the harzburgites suggest both PPGEs (Pt and Pd) and Re can be fractionated from IPGEs (Os, Ir and Ru) at low degree of partial melting, which might be due to physical dredging of sulfide melts by silicate melts rather than equilibrium partitioning between residues and silicate melts. Inferred HSEs budget of the PUM confirm the previous study that both Ru/Ir and Pd/Ir are suprachondritic in the PUM. Some modifications of late-veneer hypothesis are required in light of the unique PUM composition. HSEs and Os isotopic compositions of Gakkel abyssal peridotites indicate the oceanic mantle is highly heterogeneous within a scale of one dredge haul (<5 km). Both depleted and fertile mantle domains are likely to be mechanically juxtaposed in the asthenosphere in a state of ‘plum pudding’. Widely distribution of ancient depleted components in the asthenosphere suggests that DMM (depleted MORB mantle) should not be synonymous with the MORB source. The later is just the fertile part of the former, i.e., the depleted components in the DMM do not or contribute little to the genesis of MORB.

荣成基底片麻岩原岩岩石成因及其构造意义

Extensive high to ultrahigh pressure metamorphic rocks are outcropped in the the Dabie-Sulu UHP orogenic belt. Disputes still exist about for protolith nature of metamorphic rocks, petrogenesis, tectonic setting, and influence on upper mantle during the Triassic deep subduction. In this study, a combined study of petrology, geochemistry, isotope geochemistry and zircon chronology was accomplished for high-grade gneisses in the basement of the ultrahigh-pressure metamorphic Rongcheng terrane to reveal protolith nature and petrogenesis of the gneisses and to disucss the magmatic succession along the northern margin of the Yangtze block in Neoproterozoic. Gneisses in the Rongcheng terrane are characterized by negative Nb, Ta, P and Ti anomalies, relatively low Sr/Y ratios and relatively high Ba/La, Ba/Nb and Ba/Zr ratios, mostly displaying geochemical affinity to Phanerozoic volcanic arc. Neoproterozoic protolith ages (0.7 ~ 0.8 Ga) and Paleoproterozoic average crustal residence time (1.92 ~ 2.21 Ga) favour a Yangtze affinity. The gneisses mostly display characteristics of enrichment of LREE, flat heavy rare earth elements (REE) patterns, moderately fractionation between LREE and HREE and slight negative or positive Eu anomalies, probably reflecting that melting took place in the middle to low crust (26 ~ 33 km), where amphibole fractionated from the melts and/or inherited from source material as major mineral phases in the source area. Sr-Nd isotopic composition of the gneisses supports this conclusion. According to εNd(t) and εHf(t) values, the gneisses can be divided into three groups. Gneisses of group I have the highest εNd(t) and εHf(t) values, corresponding to the range of -6 ~ -3 and -2.9 ~ 13.4, respectively. This suggests obvious influx of depleted mantle or juvenile crust in the formation of protoliths. Gneisses of group II have medium εNd(t) (-9 ~ -7) and εHf(t) values (-15.8 ~ -1.4), corresponding to relatively high TDM2(Nd) (1.99 ~ 2.31 Ga) and TDM2(Hf) (1.76 ~ 2.67 Ga) , respectively. This suggests these gneisses were formed by partial melting of Paleoproterozoic crust. Gneisses of group III have the lowest εNd(t) (-15 ~ -10) and εHf(t) values (-15.8 ~ -1.4), corresponding to the largest TDM2(Nd) (1.99 ~ 2.31 Ga) and TDM2(Hf) ( 1.76 ~ 2.67 Ga), respectively. This indicates that gneisses of group III were formed by remelting of Archean crustal material and further demonstrates existence of an Archean basement probably of the Yangtze affinity beneath the Rongcheng terrane. Gneisses of three groups have also certain different geochemical characteristics. Contents of REEs and trace elements reduce gradually from group I to group III. Zirconium saturation temperatures also show similar tendency. Compared to gneisses of group II and group III, gneisses of group I display geochemical feature similar to extensional tectonic setting, having relatively little influence by the source area. Therefore, geochemical characteristics for gneisses of group I can indictate that the protoliths of the Rongcheng gneisses formed in an extensional rifting tectonic setting. This conclusion is supported by the results of eclogites and gabbros previously reported in the Dabie-Sulu orogenic belt. Statistical results of the protolith ages of the Rongcheng gneisses show two age peaks around ~728 Ma and ~783 Ma with an about 50 Ma gap. Extensive magatism in abou 750 Ma along the northern margin of the Yangtze block can hardly be observed in the Rongcheng terrane. This phenomenon likely suggests discontinuous Neoproterozoic magmatism along the northern margin of the Yangtze block.

Purification and characterization of recombinant truncated human interleukin-11 expressed as fusion protein in Escherichia coli

Mature human interleukin-11 (HuIL-11) is a cytokine consisting of 178 amino acid residues that results from scission of the N-terminal signal peptide, consisting of 21 amino acid residaues, from the corresponding nascent polypeptide. A DNA fragment encoding a truncated HuIL-11 (trHuIL-11), with an additional 5 amino acid residues removed from the N-terminus, was cloned into vector pGEX-2T between the BamHI site and the EcoRI site. Upon transformation with Escherichia coli BL21, the construct over-produced a glutathione S-transferase (GST)-fused protein in a soluble form after IPTG induction. The fusion protein was initially fractionated with butyl-Sepharose 4 fast flow column and by affinity chromatography using a GSH-Sepharose 4B column. On-site enzymatic release with thrombin gave the target protein at 96% purity as judged by SDS-PAGE and HPLC. Expression of the interleukin as a GST-fused protein thus greatly improved downstream processing. Subsequent biological activity assay suggested that trHuIL-11 had similar activity profile to the naturally produced sample and may be a promising candidate for further development as biopharmaceutical.