Molecular cloning and characterization of a serine proteinase homolog prophenoloxidase-activating factor in the swimming crab Portunus trituberculatus

Serine proteinase homologues (SPHs), as one of prophenoloxiase-activating factors (PPAFs), play critical roles in innate immunity of crabs. Based on an EST from the eyestalk full length cDNA library, the complete cDNA (designated as PtSPH) and genomic DNA of SPH from the swimming crab Portunus trituberculatus were cloned in this study. The estimated molecular weight of mature PtSPH (354 amino acids) was 38.7 kDa and its isoelectric point was 5.08. Multiple sequence alignment revealed that PtSPH lacked a catalytic residue with a substitution of Ser in the active site triad to Gly. Phylogenetic analysis indicated PtSPH together with PPAFs of Callinectes sapidus (AAS60227), Eriocheir sinensis (ACU65942), Penaeus monodon (ABE03741, ACP19563) and Pacifastacus leniusculus (ACB41380), formed a distinct cluster which only included clip-SPHs. As the first analyzed genomic structure of PPAFs in crustaceans, two introns were found in the open reading frame region of this gene. The mRNA transcripts of PtSPH could be detected in all the examined tissues, and were higher expressed in the eyestalk than that in gill, hepatopancreas, haemocytes and muscle. Accompanied with the change in phenoloxidase (PO) activity and total haemocyte counts, the temporal expression of PtSPH gene in haemocytes after Vibrio alginolyticus challenge demonstrated a clear time-dependent expression pattern with two peaks within the experimental period of 32 h. These findings suggest that PtSPH is involved in the antibacterial defense mechanism of Portunus tritubercualtus crab. (C) 2010 Elsevier Ltd. All rights reserved.

Scophthalmus maximus cystatin B enhances head kidney macrophage-mediated bacterial killing

Cystatins form a large family of cysteine protease inhibitors found in a wide arrange of organisms. Studies have indicated that mammalian cystatins play important roles under both physiological and pathological conditions. However, much less is known about fish cystatins. In this report, we described the identification and analysis of a cystatin B homologue, SmCytB, from turbot Scophthalmus maximus. The open reading frame of SmCytB is 300 bp, which encodes a 99-residue protein that shares high levels of sequence identities with the cystatin B of a number of fish species and contains the conserved cysteine protease inhibitor motif of cystatin B. Constitutive expression of SmCytB is high in muscle, brain, heart and liver, and low in spleen. blood, gill and kidney. Bacterial infection upregulates SmCytB expression in kidney, spleen, liver and brain but not in muscle or heart. Functional analysis showed that recombinant SmCytB purified from Escherichia colt exhibits apparent cysteine protease inhibitor activity. Transient overexpression of SmCytB in head kidney macrophages enhances macrophage bactericidal activity probably through a nitric oxide-independent mechanism. These results indicate that SmCytB is involved in the immune defense of turbot against bacterial infection. (C) 2010 Elsevier Ltd. All rights reserved.

Comparison of effectiveness of different ashing auxiliaries for determination of phosphorus in natural waters, aquatic organisms and sediments by ignition method

An improved method for the determination of phosphorus in natural waters, aquatic organisms and sediments by ignition method is proposed. The recoveries of phosphorus (P) from selected inorganic and organic P-containing compound standards after ignition with different auxiliaries, such as MgSO4, Mg(NO3)(2), MgO2, Mg(Ac-2) and CaCl2, were compared. We found that the phosphorus from most compound standards could not be completely recovered when these compounds were ignited (450-500degreesC) with the MgSO4 as auxiliary and the baked residue was extracted with 0.2 mol l(-1) HCl for 30min at 80degreesC or at room temperature. P recoveries, for example, were poor, less than 85%, if pyrophosphate and metaphosphate were ignited with the addition Of MgSO4 prior to the extraction of the baked residue with 0.2 mol l(-1) HCl at 80degreesC for 30 min. In contrast, MgO2, Mg(Ac)(2) and CaCl2, as well as Mg(NO3)(2), could all yield complete P recoveries at routine ashing temperatures (450-500degreesC). The results demonstrate that MgC12 is a more effective auxiliary agent for the determination of phosphorus in natural waters, aquatic organisms and sediments by ignition method than MgSO4 which is commonly used. (C) 2003 Elsevier Science Ltd. All rights reserved.

Extraction of phosphorus in natural waters and sediments by ignition method

Phosphorus recoveries from selected inorganic and organic P-containing compounds after ignition with auxiliaries, such as MgSO4, Mg (NO3)(2), MgCl2, Mg (Ac)(2) and CaCl2 were studied. It was found that the phosphorus could not be completely recovered when most P-containing compounds were ignited with MgSO4 at temperature not higher than 500degreesC if the baked residue was extracted with 0.2 mol/L HCl for 30 min at 80degreesC or at room temperature. In contrast, MgCl2, Mg(Ac)(2) and CaCl2, as well as Mg(NO3)(2), could all yield complete P recoveries. We suggest that MgCl2 rather than MgSO4, which is usually used, should be utilized as ashing auxiliary agent in the P extraction by ignition method. Although Mg (NO3)(2) is a highly effective auxiliary agent, yet interference from MgSO4, danger of explosion, toxicity of nitrogen dioxide and more manipulation steps may limit its widespread utilization. It is suggested that if sediment is ignited with MgCl2, the extraction of residue with 0.2 mol/L HCl for 30 min at 80degreesC could give good result.

Residues of enrofloxacin, furazolidone and their metabolites in Nile tilapia (Oreochromis niloticus)

The residues of enrofloxacin and its metabolite in Nile tilapia (Oreochromis niloticus) were studied after oral dose of 50 mg/kg for 7 days. To find the differences between Nile tilapia and Chinese shrimp (Penaeus chinensis), the residues of enrofloxacin in P chinensis were also studied under the same conditions. The results showed that enrofloxacin metabolized into ciprofloxacin in both Nile tilapia and P chinensis, the maximal concentration of enrofloxacin in muscle, liver and plasma of Nile tilapia were 3.61 mu g/g, 5.96 mu g/g, 1.25 mu g/ml respectively, and ciprofloxacin in muscle was 0.22 mu g/g. The maximal concentration of enrofloxacin and ciprofloxacin in P chinensis were 1.68 mu g/g and 0.07 mu g/g respectively. The predicted withdrawal time for Nile tilapia was 22 days, and P. chinensis was 12 days under our experiment conditions. The residues of fitrazolidone [3-(5-nitrofurfurylidenamino)-2-oxazolidinone] and its main metabolite 3-amina-2-oxazolidinone (AOZ) in Nile tilapia were first determined by HPLC/MS. Results showed that after oral dose of 30 mg/kg for 7 days, the maximum concentration of farazolidone in Nile tilapia was 413 mu g/kg after 6 h, whereas AOZ residue reached its maximum (31 mu g/kg) right after stopping treatment. In contrast to the high metabolic rate of furazolidone, AOZ was very difficult to eliminate in vivo, thus the withdrawal time of furazolidone in Nile tilapia was 22 days at least. (c) 2005 Elsevier B.V. All rights reserved.

Corrosion behaviour of hot dip zinc and zinc-aluminium coatings on steel in seawater

A comparative investigation of hot dip Zn-25Al alloy, Zn-55Al-Si and Zn coatings on steel was performed with attention to their corrosion performance in seawater. The results of 2-year exposure testing of these at Zhoushan test site are reported here. In tidal and immersion environments, Zn-25Al alloy coating is several times more durable than zinc coating of double thickness. At long exposure times, corrosion rate for the Zn-25Al alloy coating remains indistinguishable from that for the Zn-55Al-Si coating of similar thickness in tidal zone, and is two to three times lower than the latter in immersion zone. The decrease in tensile strength suggested that galvanized and Zn-55Al-Si coated steel suffer intense pitting corrosion in immersion zone. The electrochemical tests showed that all these coatings provide cathodic protection to the substrate metal; the galvanic potentials are equal to - 1,050, - 1,025 and - 880 mV (SCE) for zinc, Zn-25Al alloy and Zn-55Al-Si coating, respectively, which are adequate to keep the steel inside the immunity region. It is believed that the superior performance of the Zn-25Al alloy coating is due to its optimal combination of the uniform corrosion resistance and pitting corrosion resistance. The inferior corrosion performance by comparison of the Zn coating mainly results from its larger dissolution rate, while the failure of the Zn-55Al-Si coating is probably related to its higher susceptibility to pitting corrosion in seawater.

Effect of sulfate-reducing bacteria on corrosion behavior of mild steel in sea mud

Microbiologically influenced corrosion (MIC) is very severe corrosion for constructions buried under sea mud environment. Therefore it is of great importance to carry out the investigation of the corrosion behavior of marine steel in sea mud. In this paper, the effect of sulfate-reducing bacteria (SRB) on corrosion behavior of mild steel in sea mud was studied by weight loss, dual-compartment cell, electronic probe microanalysis (EPMA), transmission electron microscopy (TEM).combined with energy dispersive X-ray analysis (EDX) and electrochemical impedance spectroscopy (EIS). The results showed that corrosion rate and galvanic current were influenced by the metabolic activity of SRB. In the environment of sea mud containing SRB, the original corrosion products, ferric (oxyhydr) oxide, transformed to iron sulfide. With the excess of the dissolved H2S, the composition of the protective layer formed of FeS transformed to FeS2 or other non-stoichiometric polysulphide, which changed the state of the former layer and accelerated the corrosion process.

Identification of phenolics and nucleoside derivatives in Gastrodia elata by HPLC-UV-MS

An HPLC-UV-MS method for simultaneous identification of predominant phenolics and minor nucleoside derivatives in Gastrodia elata was developed, which was based on their UV and MS characteristics summarized through a series of homemade reference standard experiments. Phenolics showed characteristic UV lambda(max) at 267 nm, [M + NH4](+) base peak in positive mode and [M - H](-) base peak in negative mode while nucleosides exhibited UV lambda(max) at 255 nm, [M + H](+), [M - H + 2H(2)O](-) or [M - H + CH3COOH](-). Phenolics conjugates mainly underwent the consecutive loss of gastrodin residue (- 268 U) and the combined loss of H2O and CO2 from the citric acid unit under negative MS/MS conditions whereas nucleosides simply lost the ribose (- 132 U) under positive MS/MS conditions. According to these characteristics, a special pattern under MS/MS conditions and reported compound data for G. elata in the literature, not only 15 phenolics were identified but also 6 nucleoside derivatives were identified. Among these compounds, seven phenolics and three nucleoside derivatives have not been reported yet from G. elata.

最近130ka来黄土物质化学风化强度的空间格局

Chemical weathering intensity of loess deposits is largely determined by three factors: chemical weathering in source regions, grain size and post-depositional weathering. The third factor is influenced by climatic conditions such as precipitation and temperature, and the dust sedimentation rate in the area of deposition. Previous studies have shown that the (CaO+MgO+Na2O)/TiO2 ratio of decarbonated residue from loess is independent of grain size changes and thus is a reliable proxy for chemical weathering. However, the validity of (CaO+MgO+Na2O)/TiO2 to describe changes in monsoon intensity requires further study. In this study, 48 sections over the last glacial-interglacial cycle on the Chinese Loess Plateau were sampled, and the major elemental concentrations of 248 decarbonated residue samples were measured to investigate the utility of the (CaO+MgO+Na2O)/TiO2 ratio as a proxy for changes in monsoon intensity. Results show that the (CaO+MgO+Na2O)/TiO2 ratio, is relatively more sensitive to climate change than other indexes independent of grain size, and is not affected substantially by sedimentation rate. Assuming the weathering regime is relatively stable in the loess source regions, the (CaO+MgO+Na2O)/TiO2 ratio is a reliable proxy for the intensity of summer monsoon. A decreasing (CaO+MgO+Na2O)/TiO2 ratio from northwest to southeast both in loess and paleosols indicates that the Chinese Loess Plateau is in the control of the East Asian summer monsoon during both interglacial and glacial times. In addition, the spatial distributions of (CaO+MgO+Na2O)/TiO2 ratios show a greater north-south gradient during interglacial periods than during glacial periods. This may suggest that the spatial precipitation gradient, controlled by the summer monsoon, is steeper during interglacials than in glacials.

扬子板块西部大陆地幔地球化学特征及壳幔演化－有关基性岩超基性岩的地球化学研究

The continental mantle geochemical characteristics and crust-mantle evolution in the west of Yangtze Plate was discussed through the study of some within-plate basic-ultrabasic rocks from Lower Proterozoic to Later Paleozoic in this paper. In the Lower Proterozoic, the plate subduction between the pre-Tethys Proterozoic Ocean Plate and paleo-Yangtze Plate induced some basic volcanic formed in the island arc-back arc surrounding, which were represented by Ailaoshan Group-Dibadu Formation-Dahongshan Group, and there existed EM I component in the mantle source. The Middle Proterozoic Caiziyuan peridotite was formed in the epicontinental basin at the ocean-land boundary or within-continent rift basin. Its mantle source could be metasomatized by the dehydration fluid of subducted plate, and much initial radioactive ~(143)Nd was added to the source. In the Later Proterozoic, some rifts at the epicontinent or within-continent was formed due to the pre-Tethys oceanic plate subduction, and within-plate hot-spot Dahongshan diabase came into being. The whole-rock isochronal age of diabase is 1066±110Ma, and its mantle source was enriched Nd isotope and trace element which was related to the primary volatile component from asthenosphere and mantle plume. Its mantle source was included "FOZO" component representing mantle plume. The layer ultramafic rocks located at the Panxi Rift in the Middle-Later Paleozoic were resulted from different period and source. The early ultramafic indicated the incipient action of Panxi Rift, which is residue of continental lithospheric partial melting. Its mantle source involved subducted material and had distinct EM II component. The Emeishan basalt in the Later Paleozoic was typical continental flood basalt and its source also contained EM II component. The subduction of paleo-Tethys Ocean Plate provided essential dynamic condition for the large-scale opening of Panxi Rift, while the mantle plume supplied much material for Emeishan basalt. However, the plume was contaminated by the metasomatized continental mantle lithosphere in its upwelling process, which resulted in the Sr isotopic and incompatible elemental enrichment, and the Nd isotope kept down the weak-depleted character of mantle plume. The magmatic history in the west of Yangtze Plate is the tectonic process between pre-Tethys, paleo-Tethys Oceanic Plate and Yangtze Plate in a long history. Due to the subduction of oceanic plate, the crustal source material took part in the crust-mantle evolution widely. the continental mantle lithosphere in the west of Yangtze Plate was metasomatized by the fluid released by the subducted plate and the primary volatile from deeper mantle, and the mantle source include obvious enriched component.

攀西地区红格、新街岩体的岩石地球化学特征

The mafic-ultramafic layered intrusions in the Panxi, China contain large V-Ti-magnetite deposits. These layered intrusions are related with the Emeishan continental flood basalts in space and time. Two layered intrusions, Hongge and Xinjie have clear PGE mineralization at the base of the intrusions. Thus the detailed investigations of these two intrusions not only have a geological but also have an economic significance. This thesis aims to characterize the elemental and Sr-Nd isotopic features of diverse rock zones within the intrusion on the basis of systematic studies of the major, trace element and isotope ratios, therefore to constrain the petrogenesis, mantle source and evolution of the Hongge and Xinjie intrusions. Generally, both Hongge and Xinjie intrusions show the same Fe-Ti-rich and Si-M-poor characteristics. They are also enriched in rare-earth elements (REE) and large-ion lithophile elements (LILE) as well as in Sr-Nd isotope ratios (Hongge: initial Sr = 0.7056-0.7076, ε_(Nd)(t) and (Nd/Sm)_N-ε_(Nd)(t) plots, the Hongge intrusion has a similar elemental and isotopic features to the Emeishan low-Ti (LT) basalts, whereas the Xinjie intrusion was close to the Emeishan high-Ti (HT) basalt. Therefore, the Hongge intrusion may be co-genetic with the LT basalt, formed by the partial melting of the spinel-garnet transition mantle that had a slight enriched isotope character. In contrast, the Xinjie intrusion and the HT basalts are probably derived from the garnet-phases mantle with a primitive isotope character. The involvement of the components of mantle wedge into the source is considered to be the major reason of the REE and LILE enrichment and Nd isotope depletion in the Xinjie intrusion. In contrast with the systematic variations in TiO_2 content, Mg#, transition elements (Ni, Cu, Co), REE concentrations, and La/Yb, La/Sm ratios from the lower zone to upper zone, the different rock zones of the Hongge intrusion have no clear Sr-Nd isotope variations. This suggests that the Hongge intrusions were formed by the crystal fractionation from the same magma source. The rhythm may be formed by slow injection of the co-genetic magma during the crystal fractionation. The increase in K_2O and Al_2O_3 contents, REE abundance, and the degree of the REE fractionation in the base of the intrusion, together with the relatively low ε_(Nd)(t) value, may imply that the base of the Hongge intrusion was contaminated with the local crust rocks. Xinjie intrusion shows the clearly elemental and isotopic differences in diverse cumulus cycles. The observation of the systematic variations in TiO_2 content, Mg# value, transition elements (Ni, Cu, Co), REE concentrations, and La/Yb, La/Sm ratios in first cycle was not occurred in second cumulus cycle. In addition, the ε_(Nd)(t) value in second cumulus cycle is apparently higher than that of the first one. Thus the abruptly elemental and isotopic changes at the base of second cycle demonstrate that there is considerable new and depleted magma addition to the residue magma after the crystallization of the first cycle. These features are very similar to those of the well-known PGE-rich Bushveld and Stillwater layered intrusions. The PGE mineralization in Xinjie intrusion is much better than in Hongge intrusion. Therefore, the layered intrusion similar to the Xinjie in Panxi area posses the better prospects for the PGE deposits.

高温高压下水热体系中黄铁矿-金原电池的腐蚀电化学原位测量研究

水流体-导电性矿物相互作用是自然界中水流体-固体相互作用的重要组成部分，是许多矿床形成和演化的核心过程。迄今为止，绝大多数地质和地球化学家们对导电性矿物在水流体中的溶解机制普遍存在模糊的认识，认为水流体中导电性矿物的溶解和定位是一种简单的化学溶解和沉淀或简单的氧化溶解和还原定位过程，而实质上导电性矿物（组合）在水流体中的溶解是一种由电极电位差驱使下的类似于金属腐蚀的复杂电化学过程。当具有不同电极电位的矿物在溶液中相互接触，就会形成短路原电池发生电化学腐蚀。其中，电极电位低的矿物作为原电池的阳极，其溶解会加剧，而电极电位高的矿物作为原电池的阴极，其溶解会受到抑制。鉴于前人对水流体-导电性矿物相互作用的腐蚀原电池反应机理的模糊认识，本论文工作基于腐蚀原电池观点对高温高压条件下NaCl水流体体系中黄铁矿与金之间的原电池反应进行了研究。 在本工作中，作者与所在的研究小组一道，首先自行研制了一套可用于高温高压水热体系中导电性矿物腐蚀电化学原位测量的高压釜反应装置。该装置主体部件选用在高温高压下具有高强度、抗腐蚀等优良性能的工业纯钛制成。在该装置中，通过将热电偶直接插入高压釜釜腔内，成功地实现了釜内流体温度的准确测量和精确控制。对高压釜不同部位的测温结果表明，沿高压釜的径向与轴向均存在显著的温度梯度，其中釜塞保护锥体处的温度比釜内流体的温度低约8℃，釜外壁中心处与釜内的温度差约10℃；对400℃保温条件下釜内流体温度的直接测量表明，釜内温度波动小于0.5℃。在该装置中，通过将脆性导电性矿物制作成锥形电极，采用耐高温无机绝缘材料制作密封部件，利用锥体自紧式密封技术，成功地解决了脆性电极和电极引线的高压密封与高温绝缘问题；通过引入另一根辅助性的矿物电极引线，成功地解决了原位监测实验过程中矿物电极与引线接触处是否进水的难题，确保了实验的可信度。可行性试验结果表明，该方法不仅可用于高温高压下水流体-导电性矿物，而且可广泛用于水流体-金属间相互作用的腐蚀电化学原位测量研究。 利用上述自行研制的腐蚀电化学实验装置，本工作对高温高压（250-400℃；10-35 MPa）NaCl水流体体系中黄铁矿-自然金原电池的热力学和动力学进行了实验研究。原位测量结果表明： （1）黄铁矿-金原电池的腐蚀电流变化与其开路电压的变化一致； （2）汽-液平衡条件下，黄铁矿-金原电池的开路电压和腐蚀电流在液相中比在汽相中要大； （3）在温度为400 C、压力远离临界点的过热蒸气和超临界区域，压力在实验研究的范围内（10-35 MPa）对黄铁矿-金原电池的开路电压和腐蚀电流均无显著的影响，随压力的增加两者仅略有增大，但当温度压力跨越临界点时，包括温压从汽-液平衡曲线同时进入超临界区以及温度恒定在400 C、压力跨越临界点时，黄铁矿-金原电池的开路电压和腐蚀电流在临界点附近均发生突变。 （4）在本工作中实验的温度、压力和水流体体系条件下，由黄铁矿与金构成的原电池在大多数情况下黄铁矿为阳极，因此在原电池短路时黄铁矿在水流体中发生氧化溶解，而金则为阴极，在原电池短路时金的溶解受到保护，仅在个别狭窄的温度压力范围内情况才相反。 上述原位测量结果与电极表面水流体的性质以及黄铁矿和金的能带结构密切相关，运用混合电位理论、Butler-Volmer方程以及半导体电化学的波动能级模型对实验结果能进行很好的解释。

Factors influencing the catalytic activity of SBA-15-supported copper nanoparticles in CO oxidation

Copper nanoparticles were deposited onto mesoporous SBA-15 support via two different routes: post-grafting method and incipient wet impregnation method. Both XRD and TEM reveal that the post-grafting can make Cu particles very small in size and highly dispersed into channels of SBA-15, while the impregnation method mainly forms large Cu particles on the external surface of SBA-15. TPR experiments show that CuO species formed by the post-grafting method is more reducible than that prepared by the impregnation method. The catalytic activity tests for CO oxidation manifests that the sample prepared by the post-grafting method has a much higher activity than that prepared by the impregnation method, with a lowering of 50 degrees C for T-50, showing a strong dependence of catalytic activity on the size and dispersion of Cu particles. Besides the preparation procedure, other factors including calcination temperature, reduction treatment, copper loading as well as the feed composition, have an important effect on the catalytic activity. The best performance was obtained when the catalyst was calcined at 500 degrees C and reduced at 550 degrees C. The calcination and reduction treatment at high temperature have been found to be necessary to completely remove the organic residue and to generate active metallic copper particles. (c) 2005 Elsevier B.V. All rights reserved.