Detection of anti-human serum albumin antibody using surface plasmon resonance biosensor

The biosensor based on surface plasmon resonance(SPR) technology is a very useful tool to study the interaction between biomolecles. The main advantages of this technique is to "visualize" macromolecular interactions directly in real time, and in a label-free mode rather than indirect methods like enzyme-linked immunosorbent assays (ELISAs). We immobilize human serum albumin (HSA) to the carboxymethyldextran-modified sensor chip surface covalently to detect the activity of anti-HSA in serum, and regenerate the surface with .1 mol/L phosphoric acid. The results show that SPR biosensor can detect the activity of anti-HSA in real-time quickly and the sensor chip can be used over 100 cycles.

In situ external reflection FTIR spectroelectrochemical investigation of poly(o-phenylenediamine) film coated on a platinum electrode

The electrochemically deposited poly(o-phenylenediamine) film on a Pt electrode has been investigated utilizing in situ external reflection FTIR spectroelectrochemistry technique. The prepared ladder polymer film is found to be partially ring-opened. The dopant ClO4- is evidenced to orient in such a way that more than one oxygen atom attach to the charge sites of the polymer. This suggests that positive charges of oxidized polymer are partially delocalized over the whole chains. The proton movement observed during the oxidation reaction is associated with the solvated MeCN molecule. It is proposed that the proton diffusion, dissolvation and protonation of the film may be essential to the electrochemical reduction reaction of the film. Copyright (C) 1996 Elsevier Science Ltd.

ELECTROCHEMISTRY AND IN-SITU IR MICROSPECTROELECTROCHEMISTRY IN A VERSATILE THIN-LAYER REFLECTION FOURIER-TRANSFORM IR MICROSPECTROELECTROCHEMICAL CELL

A simple, convenient and versatile thin layer reflection Fourier transform IR microspectroelectrochemical (FTIRMSEC) cell has been described and characterized. Electrochemistry and in situ FTIR microspectroscopy were studied by using the hexacyanoferrate redox couple in aqueous sulphate solution, indicating that this type of cell is characteristic of both micro- or ultramicroelectrode and thin layer spectroelectrochemistry. Furthermore, the application of this FTIRMSEC cell to IR for characterization of the products of electrochemical reactions was carried out for the oxidation of (mesotetraphenylporphinato)manganese(III) perchlorate in dichloromethane + tetrabutylammonium perchlorate solution. Finally, the advantages and problems of this type of cell compared with a conventional optically transparent thin layer FTIR spectroelectrochemical cell were discussed.

Tectonic geomorphology of the Ryukyu Trench-Arc-Backarc System: geological-geophysical exploration and mapping

OKINAWA TROUGH; BASIN

A potent anti-oxidant property: fluorescent recombinant alpha-phycocyanin of Spirulina

To express and product a fluorescent antioxidant holo-alpha-phycocyanin (PC) of Spirulina platensis (Sp) with His-tag (rHHPC; recombinant holo-alpha-phycocyaninof Spirulina platensis with His-tag) in 5-l bench scale. A vector harbouring two cassettes was constructed: cpcA along with cpcE-cpcF in one cassette; ho1-pcyA in the other cassette. Lyases CpcE/F of Synechocystis sp. PCC6803 (S6) could catalyse the 82 site Cys in apo-alpha-PC of Sp linking with bilin chromophores, and rHHPC was biosynthesized in Escherichia coli BL21. The constant feeding mode was adopted, and transformant reached the biomass of rHHPC up to 0.55 g l(-1) broth in 5-litre bench scale. rHHPC was purified by Ni2+ affinity column conveniently. The absorbance and the fluorescence emission spectra of rHHPC had lambda(max) at 621 and 650 nm, respectively. The IC50 values of rHHPC were 277.5 +/- 25.8 mu g ml(-1) against hydroxyl radicals and 20.8 +/- 2.2 mu g ml(-1) against peroxyl radicals. Combinational biosynthesis of rHHPC was feasible, and the constant feeding mode was adopted to produce good yields of rHHPC. Fluorescent rHHPC with several unique qualitative and quantitative features was effective on scavenging hydroxyl and peroxyl radicals. A potent antioxidant rHHPC was co-expressed, produced and characterized for nutritional and pharmacological values, which would help to develop phycobiliproteins' applications in their fluorescent and biological activities.

A novel anti-tumor protein extracted from Meretrix meretrix Linnaeus induces cell death by increasing cell permeability and inhibiting tubulin polymerization

Discovery and development of new pharmaceuticals from marine organisms are attracting increasing interest. Several agents derived from marine organisms are under preclinical and clinical evaluation as potential anticancer drugs. We extracted and purified a novel anti-tumor protein from the coelomic fluid of Meretrix meretrix Linnaeus by ammonium sulphate fractionation, ion exchange and hydrophobic interaction chromatography. The molecular weight of the highly purified protein, designated MML, was 40 kDa as determined by SDS-PAGE analysis. MML exhibited significant cytotoxicity to several cancer cell types, including human hepatoma BEL-7402, human breast cancer MCF-7 and human colon cancer HCT116 cells. However, no inhibitory effect was found when treating murine normal fibroblasts NIH3T3 and benign human breast MCF-10A cells with MML. The cell death induced by MML was characterized by cell morphological changes. The induction of apoptosis of BEL-7402 cells by MML was weak by DNA ladder assay. The possible mechanisms of its anti-tumor effect might be the changes in cell membrane permeability and inhibition of tubulin polymerization. MML may be developed as a novel, highly selective and effective anti-cancer drug.

Molecular cloning, genomic organization and functional analysis of an anti-lipopolysaccharide factor from Chinese mitten crab Eriocheir sinensis

Anti-lipopolysaccharide factor (ALF) represents one kind of basic proteins, which binds and neutralizes LPS and exhibits strong antibacterial activity against Gram-negative R-type bacteria. The ALF gene of Chinese mitten crab Eriocheir sinensis (Milne Edwards, 1853) (denoted as EsALF) was identified from haemocytes by expressed sequence tag (EST) and PCR approaches. The full-length cDNA of EsALF consisted of 700 nucleotides with a canonical polyadenylation signal-sequence AATAAA, a polyA tail, and an open-reading frame of 363 bp encoding 120 amino acids. The high similarity of EsALF-deduced amino acid sequence shared with the ALFs from other species indicated that EsALF should be a member of ALF family. The mRNA expression of EsALF in the tissues of heart, gonad, gill, haemocytes, eyestalk and muscle was examined by Northern blot analysis and mRNA transcripts of EsALF were mainly detected in haemocytes, heart and gonad. The temporal expression of EsALF in haemocytes after Vibrio anguillarum challenge was recorded by quantitative real-time RT-PCR. The relative expression level of EsALF was up-regulated rapidly at 2 h post-injection and reached 3-fold to that in blank group. After a drastic decrease to the original level from 4 to 8h, the expression level increased again and reached 4-fold to that in the blank group at 12 h post-injection. The genomic DNA sequence of EsALF gene consists of 1174bp containing three exons and two introns. The coding sequence of the EsALF mature peptide was cloned and expressed in Escherichia coli BL21(DE3)-pLysS to further elucidate its biological functions. The purified recombinant product showed bactericidal activity against both Gram-positive (G(+)) and Gram-negative (G(-)) bacteria, which demonstrated that the rEsALF was a broad-spectrum antibacterial peptide. All these results indicated that EsALF was an acute-phase protein involved in the immune responses of Chinese mitten crab, and provided a potential therapeutic agent for disease control in aquaculture. (c) 2007 Elsevier Ltd. All rights reserved.

A novel polypeptide from shark cartilage with potent anti-angiogenic activity

Using guanidine-HCl extraction, acetone precipitation, ultra-filtration and chromatography, a novel polypeptide with potent anti-angiogenic activity was purified from cartilage of the shark, Prionace glauca. N-terminal amino acid sequence analysis and SDS-PAGE revealed that the substance is a novel polypeptide with MW 15500 (PG155). The anti-angiogenic effects of PG155 were evaluated using zebrafish embryos model in vivo. Treatment of the embryos with 20 mu g/ml PG155 resulted in a significant reduction in the growth of subintestinal vessels (SIVs). A higher dose resulted in almost complete inhibition of SIV growth, as observed by endogenous alkaline phosphatase (EAP) staining assay. An in vitro transwell experiment revealed that the polypeptide inhibited vascular endothelial growth factor (VEGF) induced migration and tubulogenesis of human umbilical vein endothelial cells (HUVECs). Exposure of HUVECs in 20 mu g/ml PG155 significantly decreased the density of migrated cells. Almost complete inhibition of cell migration was found when HUVECs were treated with 40-80 mu g/ml PG155. PG155 (20 mu g/ml) markedly inhibited the tube formation of HUVECs and a dose-dependent effect was also found when treatment of HUVECs with PG155 at the concentration from 20 to 160 mu g/ml.

The second anti-lipopolysaccharide factor (EsALF-2) with antimicrobial activity from Eriocheir sinensis

The anti-lipopolysaccharide factor CALF) is a small basic protein that can bind and neutralize lipopolysaccharide (LPS), mediating degranulation and activation of an intracellular coagulation cascade. In the present study, cDNA of the second Eriocheir sinensis ALF (designated as EsALF-2) was cloned and the full-length cDNA of EsALF-2 was of 724 bp, consisting of an open reading frame (ORF) of 363 bp encoding a polypeptide of 120 amino acids. The deduced amino acid of EsALF-2 shared 82% similarity with EsALF-1 from E. sinensis and about 53-65% similarity with ALFs from other crustaceans. The potential tertiary structures of EsALF-1 and EsALF-2 contained two highly conserved-cysteine residues to define the LPS binding site, but the N-terminal of EsALF-1 formed a single additional alpha-helix compared to EsALF-2, implying that EsALF-1 and EsALF-2 might represent different biological functions in E. sinensis. The mRNA transcript of EsALF-2 was detected in all examined tissues of healthy crabs, including haemocytes, hepatopancreas, gill, muscle, heart and gonad, which suggested that EsALF-2 could be a multifunctional molecule for the host immune defense responses and thereby provided systemic protection against pathogens. The mRNA expression of EsALF-2 was up-regulated after Listonelln anguillarum and Pichia pastoris challenge and the recombinant protein of EsALF-2 showed antimicrobial activity against L. anguillarum and P. pastoris. indicating that EsALF-2 was involved in the immune defense responses in Chinese mitten crab against L. anguillarum and P. pastoris. These results together indicated that there were abundant and diverse ALFs in E. sinensis with various biological functions and these ALFs would provide candidate promising therapeutic or prophylactic agents in health management and diseases control of crab aquaculture. (C) 2010 Elsevier Ltd. All rights reserved.

Propagators and ridge jumps in a back-arc basin, the West Philippine Basin

We explore the tectono-magmatic processes in the western West Philippine Basin, Philippine Sea Plate, using bathymetric data acquired in 2003 and 2004. The northwestern part of the basin formed through a series of northwestward propagating rifts. We identify at least five sequences of propagating rifts, probably triggered by mantle flow away from the mantle thermal anomaly that is responsible for the origin of the Benham and Urdenata plateaus. Gravitational forces caused by along-axis topographic gradient and a similar to 30 degrees ridge reorientation appear to also be driving the rift propagations. The along-axis mantle flow appears to be reduced and deflected along the Luzon-Okinawa fracture zone, because the spreading system remained stable west of this major fault zone. North-east of the Benham plateau, a left-lateral fracture zone has turned into a NE-SW-trending spreading axis. As a result, a microplate developed at the triple junction.

Sedimentary processes and development of the Zenisu deep-sea channel, Philippine Sea

Zenisu deep-sea channel originated from a volcanic arc region, Izu-Ogasawara Island Arc, and vanished in the Shikoku Basin of the Philippine Sea. According to the swath bathymetry, the deep-sea channel can be divided into three,segments. They are Zenisu canyon, E-W fan channel and trough-axis channel. A lot of volcanic detritus were deposited in the Zenisu Trough via the deep-sea channel because it originated from volcanic arc settings. On the basis of the swath bathymetry, submersible and seismic reflection data, the deposits are characterized by turbidite and debrite deposits as those in the other major deep-sea channels. Erosion or few sediments were observed in the Zenisu canyon, whereas a lot of turbidites and debrites occurred in the E-W channel and trough axis channel. Cold seep communities, active fault and fluid flow were discovered along the lower slope of the Zenisu Ridge. Vertical sedimentary sequences in the Zenisu Trough consist of the four post-rift sequence units of the Shikoku Basin, among which Units A and B are two turbidite units. The development of Zenisu canyon is controlled by the N-S shear fault, the E-W fan channel is related to the E-W shear fault, and the trough-axis channel is related to the subsidence of central basin.

Metasomatism of the peridotites from southern Mariana fore-arc: Trace element characteristics of clinopyroxene and amphibole

Modal composition and mineral composition of harzburgites from the southern Mariana fore-arc show that they are highly refractory. There are a few modals of clinopyroxene (0.7 vol %) in harzburgites. Two types of amphibole are found in these harzburgites: magnesiohornblende accompanied by clinopyroxene with higher Al2O3 content (> 7%) and lower Mg-#; tremolite around orthopyroxene with lower Al2O3 content (< 2%) and higher Mg-#. Trace element of clinopyroxene and two types of amphibole are analyzed. Primitive mantle-normalised REE patterns for clinopyroxene and magnesio hornblende are very similar and both show HREE enrichment relative to LREE, while magnesiohornblende has higher content of trace element than clinopyroxene. The contents of trace element of tremolite are much lower than those of magnesiohornblende. Clinopyroxene shows enrichment of most of the trace element except HREE and Ti relative to clinopyroxene in abyssal peridotites. Petrology and trace element characteristic of clinopyroxene and two types of amphibole indicate that southern Mariana fore-arc harzburgites underwent two stages of metasomatism. The percolation of a hydrous melt led to mobility of Al, Ca, Fe, Mg, Na, and large amounts of trace element. LILE and LREE can be more active in hydrous melt than HREE and Ti, and the activities of most of the trace element except some of LILE are influenced by temperature and pressure.

The characteristics of BSRs and their derived heat flow on the profile 973 in the northeastern South China Sea

Processing of a recently acquired seismic line in the northeastern South China Sea by Project 973 has been conducted to study the character and the distribution of gas hydrate Bottom-Simulating Reflectors (BSRs) in the Hengchun ridge. Analysis of different-type seismic profiles shows that the distribution of BSRs can be revealed to some extents by single-channel profile in this area, but seismic data processing plays an important role to resolve the full distribution of BSRs in this area. BSR' s in the northeastern South China Sea have the typical characteristics of BSRs on worldwide continental margins: they cross sediment bed reflections, they are generally parallel to the seafloor and the associated reflections have strong amplitude and a negative polarity. The characteristics of BSRs in this area are obvious and the BSRs indicate the occurrence of gas hydrate-bearing sediments in the northeastern South China Sea. The depth of the base of the gas-hydrate stability zone was calculated using the phase stability boundary curve of methane hydrate and gas hydrate with mixture gas composition and compared with the observed BSR depth. If a single gradient geothermal curve is used for the calculation, the base of the stability zone for methane hydrate or gas hydrate with a gas mixture composition does not correspond to the depth of the BSRs observed along the whole seismic profile. The geothermal gradient therefore changes significantly along the profile. The geothermal gradient and heat flow were estimated from the BSR data and the calculations show that the geothermal gradient and heat flow decrease from west to east, with the increase of the distance from the trench and the decrease of the distance to the island arc. The calculated 2 heat flow changes from 28 to 64 mW/m(2), which is basically consistent with the measured heat flow in southwestern offshore Taiwan.