Construction of shuttle, expression vector of human tumor necrosis factor alpha (hTNF-alpha) gene and its expression in a cyanobacterium, Anabaena sp. PCC 7120

The construction of the shuttle, expression vector of human tumor necrosis factor alpha (hTNF-alpha) gene and its expression in a cyanobacterium Anabaena sp. PCC 7120 was reported. The 700-bp hTNF cDNA fragments have been recovered from plasmid pRL-rhTNF, then inserted downstream of the promoter PpsbA in the plasmid pRL439. The resultant intermediary plasmid pRL-TC has further been combined with the shuttle vector pDC-8 to get the shuttle, expression vector pDC-TNF. The expression of the rhTNF gene in Escherichia coil has been analyzed by SDS-PAGE and thin-layer scanning, and the results show that the expressed TNF protein with these two vectors is 16.9 percent (pRL-TC) and 15.0 percent (pDC-TNF) of the total proteins in the cells, respectively, while the expression level of TNF gene in plasmid pRL-rhTNF is only 11.8 percent. Combined with the participation of the conjugal and helper plasmids, pDC-TNF has been introduced into Anabaena sg PCC 7120 by triparental conjugative transfer, and the stable transgenic strains have been obtained. The existence of the introduced plasmid pDC-TNF in recombinant cyanobacterial cells has been demonstrated by the results of the agarose electrophoresis with the extracted plasmid samples and Southern blotting with alpha-(32)p labeled hTNF cDNA probes, while the expression of the hTNF gene in Anabaena sp. PCC 7120 has been confirmed by the results of Western blotting with extracted protein samples and human TNF-alpha monoclonal antibodies. The cytotoxicity assays using the mouse cancer cell line L929 proved the cytotoxicity of the TNF in the crude extracts from the transgenic cyanobacterium Anabaena sp. PCC 7120.

Molecular cloning, expression and characterization of a chitosanase from Microbacterium sp.

A gene encoding a chitosanase (mschito) was cloned from Microbacterium, sp. OU01. The ORF consists of 801 bp which encoded a polypeptide of 266 amino acid residues. The deduced amino acid sequence shows 98% identity to that of the chitosanase reported in Pseudomonas sp. A-01. In addition, the fusion protein containing MSCHITO was expressed in E. coli and purified using Ni-NTA affinity chromatography. The purified rMSCHITO protein degraded the chitosan (the degree of deacetylation of 99%) and produced a mixture of chitooligosaccharides. The MSCHITO is thus an endo-chitosanase.

Thermodynamics of the displacive mechanism of alpha(1) transformation in a beta

Thermodynamics of the displacive mechanism of plate-shaped phase alpha(1) was analyzed in beta'Cu-Zn alloys. It was proposed that the displacive transformation of the alpha(1) plate took place in the solute-depleted region formed in the parent phase during the incubation period. The thermodynamic analysis indicated that the driving force of alpha(1) transformation, Delta G, increased with the reduction of x(d), the solute concentration of the depleted region. And, Delta G could overcome-the transformation barrier with solute depletion to a certain degree. In addition, x(d) was higher than the equilibrium concentration in the phase diagram. Therefore, the shear formation of alpha(1) plate in the solute-depleted region was thermodynamically supported.

Spatial and Temporal Variation of LURR and Its Implication for the Tendency of Earthquake Occurrence in Southern California

Based on the theory of LURR and its recent development, spatial and temporal variation of Y/Y-c (value of LURR/critical value of LURR) in the Southern California region during the period from 1980 through March, 2001 was studied. According to the previous study on the fault system and stress field in Southern California, we zoned the Southern California region into 11 parts in each of which the stress field is almost uniform. With the time window of one year, time moving step of three months, space window of a circle region with a radius of 100 km and space moving step of 0.25 degree in latitude and longitude direction, the evolution of Y/Y-c were snapshot. The scanning results show that obvious Y/Y-c anomalies occurred before 5/6 of strong earthquakes considered with a magnitude of 6.5 or greater. The critical regions of Y/Y-c are near the epicenters of the strong earthquakes and the Y/Y-c anomalies occur months to years prior to the earthquakes. The tendency of earthquake occurrence in the California region is briefly discussed on the basis of the examination of Y/Y-c.

Lateral motion and departure of vapor bubbles in nucleate pool boiling on thin wires in microgravity

A space experiment on bubble behavior and heat transfer in subcooled pool boiling phenomenon has been performed utilizing the temperature-controlled pool boiling (TCPB) device both in normal gravity in the laboratory and in microgravity aboard the 22(nd) Chinese recoverable satellite. The fluid is R113 at 0.1 MPa and subcooled by 26 degrees C nominally. A thin platinum wire of 60 mu m in diameter and 30mm in length is simultaneously used as heater and thermometer. Only the lateral motion and the departure of discrete vapor bubbles in nucleate pool boiling are reported and analyzed in the present paper. A scale analysis on the Marangoni convection surrounding a bubble in the process of subcooled nucleate pool boiling leads to formulas of the characteristic velocity of the lateral motion and its observability. The predictions consist with the experimental observations. Considering the Marangoni effect, a new qualitative model is proposed to reveal the mechanism underlying the bubble departure processes and a quantitative agreement can also be acquired.

Dynamics of discrete bubble in nucleate pool boiling on thin wires in microgravity

A space experiment on bubble behavior and heat transfer in subcooled pool boiling phenomenon has been performed utilizing the temperature-controlled pool boiling (TCPB) device both in normal gravity in the laboratory and in microgravity aboard the 22(nd) Chinese recoverable satellite. The fluid is degassed R113 at 0.1 MPa and subcooled by 26 degrees C nominally. A thin platinum wire of 60 mu m in diameter and 30 mm in length is simultaneously used as heater and thermometer. Only the dynamics of the vapor bubbles, particularly the lateral motion and the departure of discrete vapor bubbles in nucleate pool boiling are reported and analyzed in the present paper. It's found that these distinct behaviors can be explained by the Marangoni convection in the liquid surrounding vapor bubbles. The origin of the Marangoni effect is also discussed.

Study on the Capacity Degradation of Bucket Foundation in Liquefied Sand Layer under Cyclic Loads

The capacity degradation of bucket foundation in liquefied sand layer under cyclic loads such as equivalent dynamic ice-induced loads is studied. A simplified numerical model of liquefied sand layer has been presented based on the dynamic centrifuge experiment results. The ice-induced dynamic loads are modeled as equivalent sine cyclic loads, the liquefaction degree in different position of sand layer and effects of main factors are investigated. Subsequently, the sand resistance is represented by uncoupled, non-linear sand springs which describe the sub-failure behavior of the local sand resistance as well as the peak capacity of bucket foundation under some failure criterion. The capacity of bucket foundation is determined in liquefied sand layer and the rule of capacity degradation is analyzed. The capacity degradation in liquefied sand layer is analyzed comparing with that in non-liquefied sand layer. The results show that the liquefaction degree is 0.9 at the top and is only 0.06 at the bottom of liquefied sand layer. The numerical results are agreement well with the centrifugal experimental results. The value of the degradation of bucket capacity is 12% in numerical simulating whereas it is 17% in centrifugal experiments.

Photoionization of H atoms in few-cycle laser pulses

Photoionization of hydrogen atoms in few-cycle laser pulses is studied numerically. The total ionization probability, the. instantaneous ionization probability; and the partial ionization probabilities in a pair of opposite directions are obtained. The partial ionization probabilities are not always equal to each other which is termed as inversion asymmetry. The variation of asymmetry degree with the CE phase, the pulse duration and the pulse intensity is studied. It is found that the pulse intensity affects the asymmetry degree in many aspects. Firstly, the asymmetry is more distinct at higher intensities than that at lower intensities when the pulse duration exceeds 4 cycles; secondly, the maximal asymmetry in lower intensities varies with the CE phase visibly while at higher intensities riot; thirdly, the partial ionization probabilities equal to each other for some special CE phases. For lower pulse intensities, the corresponding value of CE phase is always 0.5 pi and 1.5 pi, while for higher pulse intensities, the corresponding value varies with the pulse intensity. Similar phenomena were observed in a recent experiment using few-cycle radio-frequency (RF) pulses.

Methylation and gene mutation in eukaryotic DNA

5-methylcytosine (m(5)C) as a rare base exists in eukaryotic genomes, which is a normal constitution in many eukaryotic DNA and the existence of m(5)C is a feature of eukaryotic DNA. Under regular physiological conditions, cytosine of eukaryotic DNA is usually methylated. Up to the present, many people consider that the m(5)C may be mutation hotspots by the deamination leading to gene mutation. Our study indicated that the spontaneous mutation caused by the transition of G.C --> A.T, in eukaryotic DNA, may result from the tautomer changing of base pairs and may also be cause by other factor actions, however it could not be caused by the deamination of m(5)C.

Response of growth and fatty acid compositions of Nannochloropsis sp to environmental factors under elevated CO2 concentration

Nannochloropsis sp. was grown with different levels of nitrate, phosphate, salinity and temperature with CO2 at 2,800 mu l l(-1). Increased levels of NaNO3 and KH2PO4 raised protein and polyunsaturated fatty acids (PUFAs) contents but decreased carbohydrate, total lipid and total fatty acids (TFA) contents. Nannochloropsis sp. grew well at salinities from 22 to 49 g l(-1), and lowering salinity enhanced TFA and PUFAs contents. TFA contents increased with the increasing temperature but PUFAs contents decreased. The highest eicosapentaenoic acid (EPA, 20:5 omega 3) content based on the dry mass was above 3% under low N (150 mu M NaNO3) or high N (3000 mu M NaNO3) condition. Excessive nitrate, low salinity and temperature are thus favorable factors for improving EPA yields in Nannochloropsis sp.

Optimization of growth and fatty acid composition of a unicellular marine picoplankton, Nannochloropsis sp., with enriched carbon sources

A unicellular marine picoplankton, Nannochloropsis sp., was grown under CO2-enriched photoautotrophic or/and acetate-added mixotrophic conditions. Photoautotrophic conditions with enriched CO2 of 2800 mul CO2 l(-1) and aeration gave the highest biomass yield (634 mg dry wt l(-1)), the highest total lipid content (9% of dry wt), total fatty acids (64 mg g(-1) dry wt), polyunsaturated fatty acids (35% total fatty acids) and eicosapentaenoic acid (EPA, 20:5omega3) (16 mg g(-1) dry wt or 25% of total fatty acids). Mixotrophic cultures gave a greater protein content but less carbohydrates. Adding sodium acetate (2 mM) decreased the amounts of the total fatty acids and EPA. Elevation of CO2 in photoautotrophic culture thus enhances growth and raises the production of EPA in Nannochloropsis sp.

Photon localization in amorphous photonic crystal

The photon localization in disordered two-dimensional photonic crystal is studied theoretically. It is found that the mean transmission coefficient in the photonic band decreases exponentially as the disorder degree increases, reflecting the occurrence of Anderson localization. The strength of photon localization can be controlled by tuning the disorder degree in the photonic crystal. We think the variation regular of the transmission coefficient in our disordered system is equivalent to that of the scaling theory of localization.

Expression of syncytin in leukemia and lymphoma cells

Syncytin is a placenta-specific protein and generally believed to play a pivotal role in syncytiotrophoblast morphogenesis. In this study, transcripts of this gene were quantified by real-time RT-PCR and the translated products were measured by an indirect immunofluorescence assay. Results showed that syncytin was found to be expressed in all nine leukemia and lymphoma cell lines studied albeit at different levels and in 43 peripheral blood samples of 57 leukemia or lymphoma patients. Neither the transcripts nor the translated syncytin was detected in blood samples of normal individuals. In conclusion, peripheral blood syncytin may serve as a marker for leukemia and lymphoma. ©

LITHIUM PASSIVATION OF ZN AND CD ACCEPTORS IN P-TYPE GAAS

We report lithium passivation of the shallow acceptors Zn and Cd in p-type GaAs which we attribute to the formation of neutral Li-Zn and Li-Cd complexes. Similar to hydrogen, another group-I element, lithium strongly reduces the concentration of free holes when introduced into p-type GaAs. The passivation is inferred from an increase of both the hole mobility and the resisitivity throughout the bulk of the sample. It is observed most clearly for Li concentrations comparable to the shallow-acceptor concentration. In addition, compensation of shallow acceptors by randomly distributed donors is present in varying degree in the Li-diffused samples. Unlike hydrogenation of n-type GaAs, Li doping shows no evidence of neutralizing shallow donors in GaAs.

Extinction of lean near-limit methane/air flames at elevated pressures under normal- and reduced-gravity

perimentally at evaluated pressures and under normal- and micro-gravity conditions utilizing the 3.5 s drop tower of the National Microgravity Laboratory of China. The results showed that under micro-gravity conditions the natural convection is minimized and the flames become more planar and symmetric compared to normal gravity. In both normal- and micro-gravity experiments and for a given strain rate and fuel concentration, the flame luminosity was found to enhance as the pressure increases. On the other hand, at a given pressure, the flame luminosity was determined to weaken as the strain rate decreases. At a given strain rate, the fuel concentration at extinction was found to vary non-monotonically with pressure, namely it first increases and subsequently decreases with pressure. The limit fuel concentration peaks around 3 and 4 atm under normal- and micro-gravity, respectively. The extinction limits measured at micro-gravity were in good agreement with predictions obtained through detailed numerical simulations but they are notably lower compared to the data obtained under normal gravity. The simulations confirmed the non-monotonic variation of flammability limits with pressure, in agreement with previous studies. Sensitivity analysis showed that for pressures between one and 5 atm, the near-limit flame response is dominated by the competition between the main branching, H + O2 ? OH + O, and the pressure sensitive termination, H+O2+M? HO2 + M, reaction. However, for pressures greater than 5 atm it was determined that the HO2 kinetics result in further chain branching in a way that is analogous to the third explosion limit of H2/O2 mixtures. 2010 The Combustion Institute. Published by Elsevier Inc. All rights reserved.