Evaluation of different culture conditions for high-level soluble expression of human cyclin A(2) with pET vector in BL21 (DE3) and spectroscopic characterization of its inclusion body structure

In this paper, we evaluated various parameters of culture condition affecting high-level soluble expression of human cyclin A, in Escherichia coli BL21(DE3), and demonstrated that the highest protein yield was obtained using TB(no glycerol) + 0.5% glucose medium at 25 degrees C. By single immobilized metal ion affinity chromatography, we got highly purified human cyclin A(2) with a yield ranged from 20 to 30 mg/L. By amyloid-diagnostic dye ThT binding and Fourier transform infrared spectroscopy, we observed a significant decrease in alpha-helix content and an increase in beta-sheet structure in cyclin A(2) inclusion body in comparison to its native protein, and confirmed the resemblance of the internal organization of cyclin A(2) inclusion body and amyloid fibrils.

Effect of pretreatment of black carbon on electrocatalytic activity of Pt/C catalyst for methanol oxidation

Direct methanol fuel cell (DMFC) has attracted wide attention due to its many advantages. However, its practical application is limited by the low electrocatalytic activity of the anodic Pt/C catalyst usually used for the methanol oxidation. In this paper, in order to increase the electrocatalytic performance of the Pt/C catalyst for the methanol oxidation, the black carbon, usually used as the supporter, was pretreated with CO2, air, HNO3 or H2O2. The cyclic voltarnmetric results indicated that the current densities of the anodic peak of methanol oxidation at the Pt/C catalysts with the black carbon pretreated with CO2,air, HN03, H202 and untreated black carbon were 39, 33, 32, 20 and 18 mA center dot cm(-2), respectively, illustrating that among the above five kinds of the Pt/C catalysts, the Pt/C catalyst with the black carbon pretreated with CO2 shows the best electrocatalytic activity and stability for the methanol oxidation. Its main reason is that the CO2 pretreatment could reduce the content of the oxygen-containing groups on the surface of the black carbon and increase the content of graphite in the black carbon, leading to the low resistance of the black carbon and the increase in the dispersion extent of the Pt particles in the Pt/C catalyst.

LDPE/carbon black conductive composites: Influence of radiation crosslinking on PTC and NTC properties

The electrical resistivity of low-density polyethylene/carbon black composites irradiated by Co-60 gamma-rays was investigated as a function of temperature. The experimental results obtained by scanning electron microscopy, solvent extraction techniques, and pressure-specific volume-temperature analysis techniques showed that the positive temperature coefficient (PTC) and negative temperature coefficient (NTC) effects of the composites were influenced by the irradiation dose, network forming (gel), and soluble fractions (Sol). The NTC effect was effectively eliminated when the radiation dose reached 400 kGy. The results showed that the elimination of the NTC effect was related to the difference in the thermal expansion of the gel and Sol regions. The thermal expansion of the sol played an important role in both increasing the PTC intensity and decreasing the NTC intensity at 400 kGy.

Studies on the PTC/NTC effect of carbon black filled low density polyethylene composites

The positive temperature coefficient (PTC) and negative temperature coefficient (NTC) effect of carbon black (CB) filled low density polyethylene (LDPE) composites was studied using electrical resistivity spectra, DSC, tensile mechanical analysis (TMA) and small-angle X-ray scattering (SAXS) techniques. The three LDPEs used have a similar crystallinity and different melting index (MI). The experimental results indicate that the CB has no significant effect on the crystallinity and the long spacing of crystalline domains of LDPE. Based upon the TMA and dynamic elastic modulus spectra, it can be concluded that the PTC effect is related to the thermal expansion of the polymer matrix, and the NTC effect is caused by a decrease of the elastic modulus of the polymer at high temperatures. The NTC effect can be reduced by enhancing either the elastic modulus or the interaction between carbon black and matrix. (C) 1997 Elsevier Science Ltd.

Electrical and dynamic mechanical behavior of carbon black filled polymer composites

The effect of processing conditions on the electrical and dynamic behavior of carbon black (CB) filled ethylene/ethylacrylate copolymer (EEA) composites was investigated. The compounds were prepared by two methods, solution blending and mechanical mixing. Compared with the solution counterpart, the mechanical composites have a strong positive temperature coefficient (PTC) effect and a high dynamic elastic modulus, which results from the good dispersion state of carbon black in EEA, i.e. the strong interaction between carbon black and EEA. It can be concluded that the strong interaction between polymer and carbon black is essential for composites to have a high PTC intensity, good electrical reproducibility and high dynamic elastic modulus. Copyright (C) 1996 Published by Elsevier Science Ltd.

Studies on the electrical conductivity of carbon black filled polymers

The conductivity mechanism for a carbon black (CB) filled high-density polyethylene (HDPE) compound was investigated in this work. From the experimental results obtained, it can be seen that the relation between electrical current density (J) and applied voltage across the sample (V) coincides with Simmons's equation (i.e., the electrical resistivity of the compound decreases with the applied voltage, especially at the critical voltage). The minimum electrical resistivity occurs near the glass transition temperature (T-g) of HDPE (198 K). It can be concluded that electron tunneling is an important mechanism and a dominant transport process in the HDPE/CB composite. A new model of carbon black dispersion in the matrix was established, and the resistivity was calculated by using percolation and quantum mechanical theories. (C) 1996 John Wiley & Sons, Inc.

CDNA cloning and mRNA expression of the lipopolysaccharide- and beta-1,3-glucan-binding protein gene from scallop Chlamys farreri

Lipopolysaccharide and beta-1,3-glucan-binding protein (LGBP) play a crucial role in the innate immune response of invertebrates as a pattern recognition protein (PRP). The scallop LGBP gene was obtained from Chlamys farreri challenged by Vibrio anguillarum by randomly sequencing cDNA clones from a whole body cDNA library, and by fully sequencing a clone with homology to known LGBP genes. The scallop LGBP consisted of 1876 nucleotides with a canonical polyadenylation signal sequence AATAAA and a poly(A) tail, encoding a polypeptide of 440 amino acids with the estimated molecular mass of 47.16 kDa and a predicted isoelectric point of 5.095. The deduced amino acid sequence showed a high similarity to that of invertebrate recognition proteins from blue shrimp, black tiger shrimp, mosquito, freshwater crayfish, earthworms, and sea urchins, with conserved features including a potential polysaccharide-binding motif, a glucanase motif, and N-glycosylation sites. The temporal expression of LGBP genes in healthy and V. anguillarum-challenged C farreri scallop, measured by real-time semiquantitative reverse transcription polymerase chain reaction (PCR), showed that expression was up-regulated initially, followed by recovery as the stimulation cleared. Results indicated that scallop LGBP was a constitutive and inducible acute-phase protein that could play a critical role in scallop-pathogen interaction. (C) 2004 Elsevier B.V. All rights reserved.

Triploid and tetraploid Zhikong scallop, Chlamys farreri Jones et Preston, produced by inhibiting polar body I

Triploid scallops are valuable for aquaculture because of their enlarged adductor muscle, and tetraploids are important for the commercial production of triploids. We tested tetraploid induction in the zhikong scallop by inhibiting polar body I in newly fertilized eggs. The ploidy of resultant embryos was determined by chromosome counting at 2- to 4-cell stage and by flow cytometry thereafter. Embryos from the control groups were mostly diploids (79%), along with some aneuploids. Embryos from the treated groups were 13% diploids, 18% triploids, 26% tetraploids, 13% pentaploids, and 36% aneuploids. Tetraploids, pentaploids, and most aneuploids suffered heavy mortality during the first week and became undetectable among the larvae at day 14. Five tetraploids (2%) were found among a sample of 267 spat from one of the replicates, and none was detected at day 450. The adductor muscle of triploid scallops was 44% heavier (P < .01) than that of diploids, confirming the value of the triploid technology in this species.

A key gene of the RNA interference pathway in the black tiger shrimp, Penaeus monodon: Identification and functional characterisation of Dicer-1

RNA interference (RNAi) is an evolutionarily conserved mechanism by which double-stranded RNA (dsRNA) initiates post-transcriptional silencing of homologous genes. Here we report the amplification and characterisation of a full length cDNA from black tiger shrimp (Penaeus monodon) that encodes the bidentate RNAase III Dicer, a key component of the RNAi pathway. The full length of the shrimp Dicer (Pm Dcr1) cDNA is 7629 bp in length, including a 51 untranslated region (UTR) of 130 bp, a 3' UTR of 77 bp, and an open reading frame of 7422 bp encoding a polypeptide of 2473 amino acids with an estimated molecular mass of 277.895 kDa and a predicted isoelectric point of 4.86. Analysis of the deduced amino acid sequence indicated that the mature peptide contains all the seven recognised functional domains and is most similar to the mosquito (Aedes aegypti) Dicer-1 sequence with a similarity of 34.6%. Quantitative RT-PCR analysis showed that Pm Dcr1 mRNA is most highly expressed in haemolymph and lymphoid organ tissues (P 0.05). However, there was no correlation between Pm Dcr1 mRNA levels in lymphoid organ and the viral genetic loads in shrimp naturally infected with gill-associated virus (GAV) and Mourilyan virus (P > 0.05). Treatment with synthetic dsRNA corresponding to Pm Dcr1 sequence resulted in knock-down of Pm Dcr1 mRNA expression in both uninfected shrimp and shrimp infected experimentally with GAV. Knock-down of Pm Dcr1 expression resulted in more rapid mortalities and higher viral loads. These data demonstrated that Dicer is involved in antiviral defence in shrimp. (c) 2007 Elsevier Ltd. All rights reserved.

The complete mitochondrial genomes of the whistling duck (Dendrocygna javanica) and black swan (Cygnus atratus): dating evolutionary divergence in Galloanserae

Galloanserae is an ancient and diverse avian group, for which comprehensive molecular evidence relevant to phylogenetic analysis in the context of molecular chronology is lacking. In this study, we present two additional mitochondrial genome sequences of Galloanserae (the whistling duck, Dendrocygna javanica, and the black swan, Cygnus atratus) to broaden the scope of molecular phylogenetic reconstruction. The lengths of the whistling duck's and black swan's mitochondrial genomes are 16,753 and 16,748 bases, respectively. Phylogenetic analyses suggest that Dendrocygna is more likely to be in a basal position of the branch consisting of Anatinae and Anserinae, an affiliation that does not conform to its traditional classification. Bayesian approaches were employed to provide a rough timescale for Galloanserae evolution. In general, a narrow range of 95% confidence intervals gave younger estimates than those based on limited genes and estimated that at least two lineages originated before the Coniacian epoch around 90 MYA, well before the Cretaceous-Tertiary boundary. In addition, these results, which were compatible with estimates from fossil evidence, also imply that the origin of numerous genera in Anseriformes took place in the late Oligocene to early Miocene. Taken together, the results presented here provide a working framework for future research on Galloanserae evolution, and they underline the utility of whole mitochondrial genome sequences for the resolution of deep divergence.

Black carbon and polycyclic aromatic hydrocarbons (PAHs) in surface sediments of China's marginal seas

This study investigates the distribution of black carbon (BC) and its correlation with total polycyclic aromatic hydrocarbons (I PAH) pound in the surface sediments of China's marginal seas. BC content ranges from < 0.10 to 2.45 mg/g dw (grams dry weight) in the sediments studied, and varied among the different coastal regions. The Bohai Bay sediments had the highest BC contents (average 2.18 mg/g dw), which comprises a significant fraction (27%-41%) of the total organic carbon (TOC) preserved in the sediments. In comparison, BC in the surface sediments of the North Yellow Sea, Jiaozhou Bay, East China Sea and the South China Sea is less abundant and accounted for an average of 6%, 8%, 14% and 5%, respectively, of the sedimentary organic carbon pool. The concentration of I PAH pound in the surface sediments ranges from 41 to 3 667 ng/g dw and showed large spatial variations among the sampling sites of different costal regions. The Bohai Bay has the highest I PAH pound values, ranging from 79 to 3 667 ng/g dw. This reflects the high anthropogenically contaminated nature of the sediments in the bay. BC is positively correlated to TOC but a strong correlation is not found between BC and I PAH pound in the surface sediments studied, suggesting that BC and PAHs preserved in the sediments are derived from different sources and controlled by different biogeochemical processes. Our study suggests that the abundance of BC preserved in the sediments could represent a significant sink pool of carbon cycling in China's marginal seas.