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The purslane Portulaca oleraceae L. can tolerant high saline stress and has been developed as a seawater-irrigated vegetable. Currently, the underlying molecular mechanisms were still not well understood. In the present study, purslane seedlings were treated with 200 mM NaCl and then stems were subjected to transcriptome sequencing and metabolome analyses. Together with real-time quantitative PCR validation, the results showed that treatment with 200 mM NaCl depressed expression levels of genes for photosynthesis and aquaporins, suggesting that saline stress might inhibit photosynthesis and water uptake. However, expression levels of aminocyclopropane-1-carboxylic acid synthase, bifunctional L-3-cyanoalanine synthase/cysteine synthase and cyanoalanine synthase increased, suggesting elevation of ethylene production, which were further transduced by ethylene-responsive receptor and ethylene-insensitive 3 transcription factor. Metabolome analyses detected higher contents of pyrophosphate, D-galacturonic acid and elaidic acid in treatment with 200 mM NaCl than those in the control. These changes should positively regulate glycolysis, energy supply and integrity of cell membrane. Overall, changes of ethylene, pyrophosphate and elaidic acid might facilitate adaptation of purslane to saline stress.

A new lower tungsten divertor has been developed and installed in the EAST superconducting tokamak to replace the previous graphite divertor with power handling capability increasing from −2 to ∼10 MW m−2, aiming at achieving long-pulse H-mode operations in a full metal wall environment with the steady-state divertor heat flux of ∼10 MW m−2. A new divertor concept, ‘corner slot’ (CS) divertor, has been employed. By using the ‘corner effect’, a strongly dissipative divertor with the local buildup of high neutral pressure near the corner can be achieved, so that stable detachment can be maintained across the entire outer target plate with a relatively lower impurity seeding rate, at a separatrix density compatible with advanced steady-state core scenarios. These are essential for achieving efficient current drive with low-hybrid waves, a low core impurity concentration and thus a low loop voltage for fully non-inductive long-pulse operations. Compared with the highly closed small-angle-slot divertor in DIII-D, the new divertor in EAST exhibits the following merits: (1) a much simpler geometry with integral cassette body structure, combining vertical and horizontal target plates, which are more suitable for actively water-cooled W/Cu plasma facing components, facilitating installation precision control for minimizing surface misalignment, achieving high engineering reliability and lowering the capital cost as well; (2) it has much greater flexibility in magnetic configurations, allowing for the position of the outer strike point on either vertical or horizontal target plates to accommodate a relatively wide triangularity range, δ l = 0.4–0.6, thus enabling to explore various advanced scenarios. A water-cooled copper in-vessel coil has been installed under the dome. Five supersonic molecular beam injection systems have been mounted in the divertor to achieve faster and more precise feedback control of the gas injection rate. Furthermore, this new divertor allows for double null divertor operation and slowly sweeping the outer strike point across the horizontal and vertical target plates to spread the heat flux for long-pulse operations. Preliminary experimental results demonstrate the ‘corner effect’ and are in good agreement with simulations using SOLPS-ITER code including drifts. The EAST new divertor provides a test-bed for the closed divertor concept to achieve steady-state detachment operation at high power. Next step, a more closed divertor, ‘sharp-cornered slot’ divertor, building upon the current CS divertor concept, has been proposed as a candidate for the EAST upper divertor upgrade.

Divertor tungsten (W) erosion source during edge localized mode (ELM) bursts in EAST is investigated based on optical emission spectroscopy on atomic neutral tungsten line emission at 400.9 nm. Both temporal evolution and total source strength are analyzed and compared in three different ELM mitigation schemes including natural ELMs, resonant magnetic perturbations (RMP) and 2.45 GHz lower hybrid wave (LHW). The mitigation of W source caused by single ELM impact at the divertor target is mainly attributed to the reduction of the W source production in the ELM decay phase. However, with the increase of ELM frequency, the time-averaged intra-ELM W source during an ELM cycle rises and dominates the evolution of the total W source with increasing fraction, which exhibits strong correlation with the core W level in the RMP scheme. In the natural ELM scheme, the pedestal electron temperature is found not only to control the effective W sputtering yield at target during ELMs, thus influencing the intra-ELM W source, but also to be related to the delay time between the divertor ELM WI emission and the core extreme ultraviolet bolometer (XUV) signal which can scale with the parallel ion transit time. Furthermore, the delay time is found to have more consistent dependence on the pedestal plasma collisionality. The rise time of intra-ELM WI emission increases when RMP or LHW is applied, revealing a different ELM mitigation mechanism in comparison with the natural ELM scheme. The temporal profiles of the intra-ELM WI emission in different ELM mitigation schemes are compared and the potential mechanism is discussed. Besides, the ELM mitigation effects with RMP are found to be asymmetric at the outer and inner divertor targets, which is characterized by the different variations of the intra-ELM W source strength and the rise time of WI emission with the ramp-up of RMP coil current. Divertor partial detachment is achieved simultaneously with strong ELM mitigation under a suitable RMP phase difference without additional gas puff.

Slow freezing coupled with an AC electric field (ACEF) has been demonstrated to miniaturize the ice crystals of a 0.9% (w/v) NaCl solution in a prior study. The aim of this study was to assess the effect of ACEF on Sprague-Dawley (SD) rat liver in vitro during the slow cooling procedure. SD rat liver exposed to an oscillating electric field was frozen in a programmed freezer initially down to -30°C at a cooling rate of -1°C/min and continuing down to -80°C at a cooling rate of -5°C/min. The cryovials were finally transferred into liquid nitrogen for 7 days. The frequency range was 0-20 MHz, and peak field strength was 1,000 V/m. For the sham and electric-exposed groups, the freezing solution consisted of 0%, 2.5%, 5.0%, 7.5%, or 10% (v/v) dimethyl sulfoxide (DMSO) Dulbecco's modified Eagles' medium culture solution, and fresh tissue was selected as the control group. The changes in cell survival rate, adenosine triphosphate (ATP) content, and morphology of fresh and frozen-thawed liver tissue were examined. Compared with the sham group with 5.0% DMSO, the result showed that slow freezing coupled with 2.45 MHz or 5 MHz ACEF significantly increased the relative survival rate by 43.27% and 26.31% (P 0.001), respectively. However, ACEF exposure increased the ATP content compared with the sham group. Especially in 5% and 10% DMSO with 2.45 MHz ACEF exposure, the ATP content approximated the fresh group (7.3 ± 2.7 nmol/piece), corresponding to 94.52% and 80.82%. In addition, the cellular membrane and some organelles (e.g., mitochondria) in the electric-exposed group appeared to be more intact according to the transmission electron microscopy images. The underlying mechanism might be that the ACEF affects the formation and growth of the ice crystallization, and thus inhibits cryoinjury. These results show that ACEF would provide an efficient method for cryopreservation banking with a low concentration of CPA during the slow freezing process.

The lifetimes and decay channels of ultralong-range Rydberg molecules created in a dense BEC are examined by monitoring the time evolution of the Rydberg population using field ionization. Studies of molecules with values of principal quantum number, $n$, in the range $n=49$ to $n=72$ that contain tens to hundreds of ground state atoms within the Rydberg electron orbit show that their presence leads to marked changes in the field ionization characteristics. The Rydberg molecules have lifetimes of $\sim1-5\,\mu$s, their destruction being attributed to two main processes: formation of Sr$^+_2$ ions through associative ionization, and dissociation induced through $L$-changing collisions. The observed loss rates are consistent with a reaction model that emphasizes the interaction between the Rydberg core ion and its nearest neighbor ground-state atom. The measured lifetimes place strict limits on the time scales over which studies involving Rydberg species in cold, dense atomic gases can be undertaken and limit the coherence times for such measurements., Comment: 9 pages, 8 figures

Synopsis The photoexcitation rate for ultralong-range Rydberg molecules is sensitive to the initial spatial sepa-ration of the atoms from which they are formed. Here we explore the photoexcitation of different molecular vibrational states as a probe of spatial correlations in a cold atomic gas.

Synopsis The photoexcitation of ultralong-range Rydberg molecules is demonstrated as a probe of spatial cor-relations in a quantum gas on previously inaccessible length scales of 1400-3200 a 0. Measurements using cold (∼1 μK) (fermionic) 87Sr and (bosonic) 84Sr reveal the effects of quantum statistics on the pair correlation function g (2)(R).

The triple point of argon is a defined fixed point in ITS-90 for calibration of standard platinum resistance thermometers (SPRTs). A new triple point of argon system with multiple re-entrant wells ...

Aims: To screen and identify biosurfactant producers from petroleum-contaminated soil; to use response surface methodology (RSM) for medium optimization to enhance biosurfactant production; and to study the properties of the newly obtained biosurfactant towards pH, temperature and salinity. Methods and Results: We successfully isolated three biosurfactant producers from petroleum-contaminated soil and identified them through 16S rRNA sequence analysis, which exhibit the highest similarities to Acinetobacter beijerinckii (100%), Kocuria marina (99%) and Kineococcus marinus (99%), respectively. A quadratic response model was constructed through RSM designs, leading to a 57AE5% increase of the growth-associated biosurfactant production by Acinetobacter sp. YC-X 2 with an optimized medium: beef extract 3AE12 g l)1 ; peptone 20AE87 g l)1 ; NaCl 1AE04 g l)1 ; and n-hexadecane 1AE86 g l)1. Biosurfactant produced by Acinetobacter sp. YC-X 2 retained its properties during exposure to a wide range of pH values (5–11), high temperatures (up to 121C) and high salinities [up to 18% (w⁄ v) Na+ and Ca2+], which was more sensitive to Ca2+ than Na+ . Conclusions: Two novel biosurfactant producers were isolated from petroleumcontaminated soil. Biosurfactant from Acinetobacter sp. YC-X 2 has good properties to a wide range of pH, high temperature and high salinity, and its production was optimized successfully through RSM.

A triple-point-of-argon system was developed to realize the argon triple-point temperature defined in the International Temperature Scale of 1990 (ITS-90) and to calibrate long-stem standard platinum resistance thermometers (SPRTs). The system structure, the techniques for realization of the triple-point temperature, and the testing results are presented in this paper. In this system, there is a central argon cell that includes four re-entrant wells with an immersion depth of 160 mm which can be used to calibrate four long-stem SPRTs simultaneously. A high-accuracy temperature controller is used to control the realization process of the argon triple-point temperature plateau. The argon triple-point plateau can be easily realized with this system. The influence of different experimental parameters on the triple-point-of-argon plateau was investigated and is discussed. The testing results show that the duration of the plateau can be over 100 h with the temperature change less than 0.05 mK. The temperature homogeneity of the four re-entrant wells was tested. The immersion profile of the system was measured, and the measurement results were compared with the ITS-90 hydrostatic values. The uncertainty analysis shows that the uncertainty of the argon system is 0.25 mK (k = 2).

Many researchers had proved that both single amino acid composition and dipeptide composition can influence protein thermostability. We use ν-support vector machines approach to predict hyperthermophilic protein, thermophilic protein and mesophilic protein from single amino acid composition, dipeptide composition and the combination of the two factors. For the prediction accuracies, we conclude that, single amino acid composition is suitable for prediction of mesophilic protein; dipeptide composition is suitable for prediction of hyperthermophilic protein and thermophilic protein; when considering the combination of the two factors, the prediction accuracy of hyperthermophilic protein is 84.1%, thermophilic protein is 83.4%, mesophilic protein is 84.4%, average accuracy is 84.0%. It shows that the protein thermostability can be predicted properly based on the combination of single amino acid composition and dipeptide composition. Obviously, dipeptide composition is correlative significantly to protein thermostability based on the prediction accuracies.

To prevent short circuits, to improve stability, and to raise the upper temperature limit to the freezing point of copper (1084.62 °C), the high-temperature standard platinum resistance thermometer (HTSPRT) was redesigned. The most important change was an improvement in the structure of the sensor support. The strip support was replaced by a new specially designed cross support. The structure and design of the new HTSPRT are briefly described in this article. The test results of a group of thermometers are presented. The test included long-term drifts of the thermometers at the triple point of water and freezing point of silver during a period of a few hundred hours operation at 1085 °C, the short-term stability of R (tpw) and W (Ag) in a period of 5 days, and thermal cycles between 22 °C and 1085 °C. The test results show that the thermometer performance is improved, and the new HTSPRT can operate up to the freezing point of copper.

Europium (Eu)-doped zinc oxide (ZnO) nanocrystals are prepared by the sol–gel method. Compared with undoped ZnO nanocrystals, Eu-doped ZnO nanocrystals have enhanced ultraviolet (UV) lasing and reduced threshold at room temperature. The UV lasing properties of both the doped and the undoped samples were studied by the time-resolved and power-dependent photoluminescence (PL) experiments. The enhancement of UV lasing is due to the significantly reduced singly ionized oxygen vacancies with Eu doping, which is confirmed by both the PL spectra and the X-ray photoelectron spectra. It indicates that high-efficiency and low-threshold UV lasing sources can be fabricated from Eu-doped ZnO nanocrystals.

Segregation of phosphorus and molybdenum to grain boundaries in a commercial grade Ni-base (718 plus) superalloy subjected to different heat treatments has been examined using a field emission gun scanning transmission electron microscope with energy dispersive X-ray microanalysis. The results indicate that P and Mo concentrations at grain boundaries increase in linear manner with the square root of aging time.

Plasma etching is a critical tool in the fabrication of Si/SiGe heterostructure quantum devices, but it also presents challenges, including damage to etched feature sidewalls that affects device performance. Chemical and structural changes in device feature sidewalls associated with plasma-surface interactions are considered damage, as they affect band structure and electrical conduction in the active region of the device. Here the authors report the results of experiments designed to better understand the mechanisms of plasma-induced sidewall damage in modulation-doped Si/SiGe heterostructures containing a two-dimensional electron gas. Damage to straight wires fabricated in the heterostructure using plasma etching was characterized both by measuring the width of the nonconductive “sidewall depletion” region at the device sidewall and by measuring the noise level factor γH/N determined from spectra of the low frequency noise. Observed increases in sidewall depletion width with increasing etch depth are te...

The mechanisms for Fe(III) oxide reduction in Geobacter species are of interest because Fe(III) oxides are the most abundant form of Fe(III) in many soils and sediments and Geobacter species are prevalent Fe(III)-reducing microorganisms in many of these environments. Protein abundance in G. sulfurreducens grown on poorly crystalline Fe(III) oxide or on soluble Fe(III) citrate was compared with a global accurate mass and time tag proteomic approach in order to identify proteins that might be specifically associated with Fe(III) oxide reduction. A total of 2991 proteins were detected in G. sulfurreducens grown with acetate as the electron donor and either Fe(III) oxide or soluble Fe(III) citrate as the electron acceptor, resulting in 86% recovery of the genes predicted to encode proteins. Of the total expressed proteins 76% were less abundant in Fe(III) oxide cultures than in Fe(III) citrate cultures, which is consistent with the overall slower rate of metabolism during growth with an insoluble electron acceptor. A total of 269 proteins were more abundant in Fe(III) oxide-grown cells than in cells grown on Fe(III) citrate. Most of these proteins were in the energy metabolism category: primarily electron transport proteins, including 13 c-type cytochromes and PilA, the structural protein for electrically conductive pili. Several of the cytochromes that were more abundant in Fe(III) oxide-grown cells were previously shown with genetic approaches to be essential for optimal Fe(III) oxide reduction. Other proteins that were more abundant during growth on Fe(III) oxide included transport and binding proteins, proteins involved in regulation and signal transduction, cell envelope proteins, and enzymes for amino acid and protein biosynthesis, among others. There were also a substantial number of proteins of unknown function that were more abundant during growth on Fe(III) oxide. These results indicate that electron transport to Fe(III) oxide requires additional and/or different proteins than electron transfer to soluble, chelated Fe(III) and suggest proteins whose functions should be further investigated in order to better understand the mechanisms of electron transfer to Fe(III) oxide in G. sulfurreducens.

Segregation of phosphorus and molybdenum to grain boundaries in a commercial grade of 2·25Cr–1Mo steel subjected to different heat treatments has been examined using a field emission gun scanning transmission electron microscope with energy dispersive X-ray microanalysis. The results indicate that P and Mo concentrations at prior austenite grain boundaries increase with aging time. This follows the prediction of McLean's equilibrium segregation model, when modified to take account of the interaction energy between phosphorus and molybdenum.

In the interest of improving convenience and plateau duration, the use of melting points instead of freezing points for temperature fixed points in temperature calibration is considered. The questi...

Noninvasive diagnosis and prognostication of acute cellular rejection in the kidney allograft may help realize the full benefits of kidney transplantation. To investigate whether urine metabolites predict kidney allograft status, we determined levels of 749 metabolites in 1516 urine samples from 241 kidney graft recipients enrolled in the prospective multicenter Clinical Trials in Organ Transplantation-04 study. A metabolite signature of the ratio of 3-sialyllactose to xanthosine in biopsy specimen-matched urine supernatants best discriminated acute cellular rejection biopsy specimens from specimens without rejection. For clinical application, we developed a high-throughput mass spectrometry-based assay that enabled absolute and rapid quantification of the 3-sialyllactose-to-xanthosine ratio in urine samples. A composite signature of ratios of 3-sialyllactose to xanthosine and quinolinate to X-16397 and our previously reported urinary cell mRNA signature of 18S ribosomal RNA, CD3e mRNA, and interferon-inducible protein-10 mRNA outperformed the metabolite signatures and the mRNA signature. The area under the receiver operating characteristics curve for the composite metabolite-mRNA signature was 0.93, and the signature was diagnostic of acute cellular rejection with a specificity of 84% and a sensitivity of 90%. The composite signature, developed using solely biopsy specimen-matched urine samples, predicted future acute cellular rejection when applied to pristine samples taken days to weeks before biopsy. We conclude that metabolite profiling of urine offers a noninvasive means of diagnosing and prognosticating acute cellular rejection in the human kidney allograft, and that the combined metabolite and mRNA signature is diagnostic and prognostic of acute cellular rejection with very high accuracy.

Previous studies failed to detect c -type cytochromes in Pelobacter species despite the fact that other close relatives in the Geobacteraceae , such as Geobacter and Desulfuromonas species, have abundant c- type cytochromes. Analysis of the recently completed genome sequence of Pelobacter carbinolicus revealed 14 open reading frames that could encode c -type cytochromes. Transcripts for all but one of these open reading frames were detected in acetoin-fermenting and/or Fe(III)-reducing cells. Three putative c -type cytochrome genes were expressed specifically during Fe(III) reduction, suggesting that the encoded proteins may participate in electron transfer to Fe(III). One of these proteins was a periplasmic triheme cytochrome with a high level of similarity to PpcA, which has a role in Fe(III) reduction in Geobacter sulfurreducens . Genes for heme biosynthesis and system II cytochrome c biogenesis were identified in the genome and shown to be expressed. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels of protein extracted from acetoin-fermenting P. carbinolicus cells contained three heme-staining bands which were confirmed by mass spectrometry to be among the 14 predicted c -type cytochromes. The number of cytochrome genes, the predicted amount of heme c per protein, and the ratio of heme-stained protein to total protein were much smaller in P. carbinolicus than in G. sulfurreducens . Furthermore, many of the c -type cytochromes that genetic studies have indicated are required for optimal Fe(III) reduction in G. sulfurreducens were not present in the P. carbinolicus genome. These results suggest that further evaluation of the functions of c -type cytochromes in the Geobacteraceae is warranted.

In this manuscript, not only hexagonal CdS nanostructural materials could be prepared using facile solution chemistry, but cubic sphalerite CdS phase was found as well. In addition, both zero-dimensional nanospheres and one-dimensional CdS nanorods could be selectively grown through a use of amine. The present of organic amine plays a crucial role in the growth of CdS nanocrystals. Fascinatingly, a much low-temperature (60°C) synthesis of CdS nanorods were achieved via a sub-solvothemral route, this is the lowest temperature to synthesize well crystalline CdS nanorods from ethylenediamine (en) medium so far. Optical properties indicate the physical features are greatly dependent on the chemical synthesis procedures. With the use of en solvent, the green photoluminescence at about 550 nm is significantly enhanced and a blue shift was observed.

A numerical model for predicting microporosity formation in aluminium castings has been developed, in which the redistribution of hydrogen between solid and liquid phases, and Darcy flow in the mushy zone were taken into account. For simulating the nucleation and growth of hydrogen pores, the pore radius is assumed to be a function of the secondary dendrite arm spacing, whereas the pore growth assumes equilibrium hydrogen activity between pore, liquid and solid. One of the key features of the model is that it uses a two-stage approach to porosity prediction. In the first stage, the volume fraction of porosity is calculated based on the reduced pressure, whereas, in the second stage, at fractions solid greater than grain coalescence f SC, an increment in the fraction porosity is calculated based on the volume of liquid trapped within a continuous solid network, which is estimated using a correlation based on the Niyama criterion. The model has been implemented within the commercial software packag...

The objective of this study was to assess the protective effects of Ginkgo biloba leaf extracts (EGb) on trichloroethylene (TCE)-induced cytotoxicity and apoptosis in normal human epidermal keratinocytes (NHEK). Cytotoxicity was determined by neutral red uptake, and lipid peroxidation of the cells was assessed by malondialdehyde (MDA) and superoxide dismutase (SOD). Electron microscopy and flow cytometry were used to evaluate NHEK apoptosis. Treatment of NHEK with various concentrations of TCE caused a substantial decrease in cell viability. NR50 from the cytotoxicity assay was found to be 4.53 mM. TCE caused an increase in MDA, while an inhibition of SOD activity, in a concentration-dependent manner. Electron microscopic examination revealed typical morphologic changes of apoptosis in cells treated with TCE. Incubation of NHEK with TCE (0, 0.125, 0.5, 2.0 mM) for 4 h increased the proportion of apoptotic cells from control of 19.23% to nearly 44.35%. Pretreatment of EGb at 10–200 mg/l dose-dependently attenuated the cytotoxic effect of TCE, prevented TCE-induced elevation of lipid peroxidation and decline of antioxidant enzyme activities. Similar inhibition by EGb on TCE-mediated NHEK apoptosis was also observed. These results suggest that EGb can protect NHEK from TCE-induced cytotoxicity and apoptosis, which may be associated with the superoxide scavenging effect and enhancement of antioxidant enzyme activities.

The effect of human serum albumin (HSA), in its endogenous, free fatty acid free (FAF) and globulin free (GF) form, on the activity of CYP2C9 was studied in human liver microsomes using tolbutamide as the substrate. The widely used BSA was included to assess the differential effect of BSA and HSA. CYP2C9 activity was expressed as CLint (Vmax/Km). HSA(FAF) and BSA showed a concentration-dependent and biphasic (activation and inhibition) interaction with CYP2C9 activity. HSA(GF) and HSA exhibited an inhibitory effect, with an inhibition constant, Ki, of 19.9 microM (0.13% albumin) and 42.2 microM (0.35% albumin), respectively. Enzyme-kinetics revealed that the activation is accompanied by a decrease in Km values, while with inhibition Km values increased. A simplified method to calculate clearance, utilizing a single slope (V/S) determination based on V over the lowest linear range of [S] (designated as CLone) was assessed. Virtually identical values were obtained for CLint and CLone. The free-drug hypothesis was tested by comparing ratios of relative CLint/unbound fraction (FDH Test ratio). The FDH Test ratio for HSA was about 1, indicating that HSA binding of tolbutamide reduced the CYP2C9 activity in accord with the free-drug hypothesis. The FDH Test ratios for BSA and HSA(FAF) were 3.7 and 3.0, revealing a monophasic activation of CYP2C9. For 2%HSA(GF) the ratio of 0.3 confirmed inhibition. As revealed by their removal, free fatty acids and globulins, significantly alter the interaction of HSA with CYP2C9. In addition, HSA and BSA showed different effects on the oxidation of tolbutamide by CYP2C9.

A field experiment was carried out to study the effect of K nutrition and genotypic variation on the dry matter (DM) accumulation, and the K concentration, accumulation, uptake and utilization efficiency in barley (Hordeum vulgareL.). Successive increases in potassium nutrition had a significant effect on the dry matter and K accumulation either in the total or in various plant parts of barley at the tillering, stem elongation, heading and maturity growth stages. K nutrition also led to significantly higher grain yield with each unit K application than without K application. The yield increase due to K application was mainly due to the improvement in spike development from tillers. Dry matter and K accumulation in various plant parts varied significantly between genotypes at the main growth stages. Among the various plant parts, the stem contained the highest K concentration, had the highest K accumulation at maturity and changed considerably with the K level, while other plant parts remained relatively unchanged. Among the eleven genotypes, genotype 98-6 had the highest grain yield and the K use efficiency of this genotype was 10.4 kg grain per kg K applied. It could thus be used as a breeding line to breed barley varieties for higher productivity under rainfed conditions with low available soil potassium.

The effect of human serum albumin (HSA), alpha1-acid glycoprotein (alpha1-AGP), and alpha- and gamma-globulins on the in vitro metabolism of debrisoquine in human, hamster and rat liver microsomes was studied. Interaction of albumin with cytochrome P450 mediated phenytoin metabolism has been reported. Since plasma protein binding of phenytoin is high, in the present study a weakly protein bound drug, debrisoquine, was studied. Debrisoquine is a substrate of CYP2D6. The debrisoquine 4-hydroxylation was measured using a radio-TLC method. Among the four plasma proteins, alpha-globulins had the strongest inhibitory effect on the debrisoquine 4-hydroxylase activity. The inhibition with 2% alpha-globulins was 42+/-18% for human and higher for hamster and rat liver microsomes (65-71%). HSA had less effect than alpha-globulins. In the presence of HSA, the decrease in activity was between 18 and 35% for all liver microsomes studied. The debrisoquine 4-hydroxylase activity was not significantly changed by alpha1-AGP or gamma-globulins. Using an ultra-filtration method, the protein binding of debrisoquine to 4% HSA, 0.5% alpha1-AGP, 2% alpha-globulins and 2% gamma-globulins was found to be 22, 20, 22 and 5%, respectively. Since the observed inhibition is inconsistent with level of protein binding, it appears, particularly in the case of alpha-globulins, that the plasma proteins interact with CYP2D directly.

A pyrophosphate-dependent phosphofructokinase (PP i -PFK) and an ATP-dependent phosphofructokinase (ATP-PFK) from Thermotoga maritima have been cloned and characterized. The PP i -PFK is unique in that the K m and V max values indicate that polyphosphate is the preferred substrate over pyrophosphate; the enzyme in reality is a polyphosphate-dependent PFK. The ATP-PFK was not significantly affected by common allosteric effectors (e.g., phosphoenolpyruvate) but was strongly inhibited by PP i and polyphosphate. The results suggest that the control of the Embden-Meyerhof pathway in this organism is likely to be modulated by pyrophosphate and/or polyphosphate.

The distribution of phosphofructokinase phosphoryl donor subtypes (ATP-, ADP-, and pyrophosphate) in the deeply rooted phylogenetic lineages of thermophiles is of interest with regard to the evolution of phosphofructokinase activity and of the Embden–Meyerhof pathway. In this article we present the first biochemical description of a thermostable pyrophosphate-dependent phosphofructokinase from the hyperthermophilic bacterium Dictyoglomus thermophilum. The enzyme was not allosterically controlled by traditional modulators of phosphofructokinases and has significant activity with tripolyphosphate and polyphosphate. Kinetic parameters of the enzyme suggest it plays primarily a glycolytic role. The enzyme required Mg2+ for optimal activity, was partially activated by some monovalent and divalent cations, and was strongly inhibited by Cu2+. The sequence of the 21 N-terminal residues sug-gests that the enzyme is most similar to the pyrophosphate-dependent phosphofructokinases from Amycolatopsis methanolica and the hyperthermophilic crenarchaeon Thermoproteus tenax, enzymes which have been suggested to represent an ancient lineage of phosphofructoki-nases (Siebers et al. 1998). The unexpected finding of a pyrophosphate-dependent phosphofructokinase in Dictyoglomus thermophilum, which is phylogenetically related to Thermotoga maritima, previously shown to possess an ATP-dependent phosphofructokinase activity, is discussed.

Three steps in complex chemorheological studies are necessary for complete rheological characterization of filled rubbers during cross-linking reaction. They include: (i) kinetic studies of cross-linking reaction, (ii) rheological studies of green rubber compounds, and (iii) correlation between the rheological parameters and the degree of cure. Basic experiments in the steps (i)–(iii) and their modeling are presented in this paper on the example of a filled SBR compound with sulfur accelerated vulcanization. The approach provides a unique possibility to trace therheological properties of rubber compounds from the green to completely cured states.

Polybutadienes (PBs) of narrow molecular weight distributions with high 1,4-microstructure were synthesized by sec-butyllithium-initiated, living polymerization. The shoulder-like anomalous behavior of the storage modulus G′(ω) in the terminal zone for PBs was studied using size exclusion chromatography (SEC) and rheometric mechanical spectrometry (RMS). It was found that a high-molecular-weight fraction in the PBs was the main reason for the occurrence of the shoulders. The linear viscoelastic spectra were successfully obtained by the Pade–Laplace method from experimental dynamic data, using either the dynamic storage, G′(ω), or loss, G"(ω), modulus. Both the dynamic moduli and stress relaxation in the linear regime were then calculated with reasonable precision. The feasibility of time–temperature–molecular weight superposition for PBs using this method was demonstrated. The dependencies of relaxation spectra on temperature and molecular weight were obtained by a fitting procedure. Thus, with a known reference state, the relaxation spectrum for a given molecular weight PB at a given temperature can be predicted. The predicted dynamic moduli are in good agreement with experimental data. © 1999 John Wiley & Sons, Inc. J Appl Polym Sci 71: 1315–1324, 1999

The preparation of amylopection tris(phenylcarbamate) (ATP) chiral stationary phases(CSPs) coated on a small-particle (5μm, 120A) spherical silica support, according to a process that is reported, ...

Vulcanization kinetics for a natural rubber compound were studied by a kinetic approach using the cure meter and DSC methods. A simplified but realistic model reaction scheme was used to simulate induction, curing, and overcure periods continuously. Physically significant parameters of the model were extracted from isothermal experimental data using a cure meter. The calculation demonstrated a good correspondence with isothermal cure meter data over the temperature range studied. The length of induction time, variation of maximum modulus with temperature, and the reversion phenomena observed from cure curves can be predicted. DSC data were found to be incompatible with the cure meter test, because the complex vulcanization reaction system is multiexothermal and it is difficult to isolate the heat due to crosslinking. Hence, the cure meter technique is suggested for the study of crosslink formation. The kinetic approach provides a way to incorporate vulcanization kinetics into simulation of reactive processing operations. © 1996 John Wiley & Sons, Inc.

Vulcanization kinetics for a SBR compound was studied by using both curemeter and DSC methods by a kinetic approach. A simplified but realistic model reaction scheme was used to simulate both induction and curing periods simultaneously. Model parameters were extracted from isothermal curemeter experiments. The model prediction demonstrated a good agreement with isothermal curemeter data over a temperature range of 120°C to 180°C. The variation of equilibrium modulus with temperature, observed from cure curves, can also be predicted. However, DSC experiments showed a different reaction behavior in the curing period as compared to model calculations. This was explained by the assumption that the reaction heat observed in DSC is due to all possible exothermal reactions, and the formation of crosslinks is only a part of these reactions. Hence, the curemeter can provide a good indication of crosslink formation, while DSC displays the entire reaction heat released during the vulcanization process. The kinetic approach allows one to incorporate vulcanization kinetics into the practical simulation of reactive processing operations.

Background. The positive costimulatory proteins OX40 and OX40L and negative regulatory proteins programmed death (PD)-1, PD ligand 1, and PD ligand 2 have emerged as significant regulators of acute rejection in experimental transplantation models. Methods. We obtained 21 urine specimens from 21 renal allograft recipients with graft dysfunction and biopsy-confirmed acute rejection and 25 specimens from 25 recipients with stable graft function and normal biopsy results (stable). Urinary cell levels of mRNAs were measured using real-time quantitative polymerase chain reaction assays, and the levels were correlated with allograft status and outcomes. Results. Levels of OX40 mRNA (P

The simulations for linear devices using the 3D Monte-Carlo code ERO have a number of particularities of both physical and technical nature. Many physical effects introduced and tested for these simulations (elastic collisions, population of metastable states, etc.) can be relevant for tokamaks as well. The current status of these activities is summarized. As an example some simulations reproducing the experimental observations at PISCES-B and Pilot-PSI are presented. Main deviations and ideas for their origins are discussed. (c) 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

The effects of inducible expression of poly(ADP-ribose) polymerase (PADPRP) antisense RNA in HeLa cells were determined in order to gain further insight into the biological roles of the poly(ADP-ribosyl)ation modification of nuclear proteins. A recombinant expression plasmid was prepared with the mouse mammary tumor virus (MMTV) promoter upstream of the antisense-oriented PADPRP cDNA. Expression of the antisense RNA was under strict control, with negligible effects on cell growth being apparent in the absence of inducer. Consistent with the previously described stability of PADPRP (half-life of at least 2 days, in vivo), 48-72 h were required after induction of antisense RNA expression by dexamethasone for the abundant concentration of PADPRP, normally present in HeLa cells, to be reduced by greater than 80%. The depletion of endogenous PADPRP as mediated by induced antisense RNA expression was established by: (i) a progressive synthesis of antisense transcripts in cells as assessed by Northern analysis; (ii) an 80% decrease in activity of the enzyme; and (iii) a greater than 90% reduction in the cellular content of PADPRP protein, as demonstrated by both immunoblotting and immunohistochemical analysis in intact cells. Several biological parameters were monitored in cells depleted of PADPRP. The chromatin of PADPRP-depleted cells was shown to have an altered structure as assessed by deoxyribonuclease I susceptibility. Cell morphology was also altered, with multinucleated aggregates being evident 72 h after induction of antisense RNA expression. Cells depleted of PADPRP were not able to commence DNA strand break joining of damaged DNA. However, DNA repair capacity was re-established at later time periods, indicating that PADPRP may contribute to alterations in chromatin structure that occur initially in DNA strand break rejoining and that the concentration of the enzyme in nuclei exceeds the requirement for DNA repair/replication.

Amorphization of the (Fe, Co, Ni, Al, Si)-Yb multilayered alloy films under Xe+ and Ar+ irradiation at room temperature is studied. It is found that (Co, Ni, Al, Si)-Yb alloy films are amorphized or partially amorphized by ion beam mixing, while Fe-Yb alloy films are not amorphized at two selected compositions. The recrystallization temperatures of the (Co, Ni, Al, Si)-Yb amorphous alloys are also determined.

ACAD SINICA,INT CTR MAT PHYS,SHENYANG 110015,PEOPLES R CHINA. TSING HUA UNIV,DEPT MAT SCI & ENGN,BEIJING 100084,PEOPLES R CHINA.;LIU, BX (reprint author), CCAST,WORLD LAB,CTR CONDENSED MATTER & RADIAT PHYS,BEIJING 100080,PEOPLES R CHINA

AIM To investigate and compare the effect of ra-dio-frequency field exposure on stress response genes (Hsps) expression in three human glioma cell lines. METHODS Glioma cells were from MO54, A172 and T98 three human glioma cell lines. Exponentially growing cells were exposed to sham conditions or 1950 MHz continuous-wave for 1h. PC randomly determined which of the two waveguides was ex-posed. Specific absorption rates (SARs) were 1 and 10 W/kg. After treatment, the expression level of Hsp27 and p-Hs27 as well as Hsp70, was determined by Western blotting. RESULTS The Hsp27 expression level of A172, MO54 and T98 cells ex-posed to RF fields (SAR: 1 W/kg, 10 W/kg) for 1h showed no difference compared with sham-exposed cells. Phosphorylated Hsp27 level of A172, MO54 and T98 cells exposed to RF fields (SAR: 1 W/kg) for 1 h showed no differences compared with sham-exposed cells. However, exposure to an RF field at 10 W/kg for 1h slightly decreased the level of phosphorylated Hsp27 in MO54 cells other in A172 and T98 cells. For the expression of Hsp70 in A172, MO54 and T98 cells after expo-sure to RF fields (SAR: 1 W/kg and 10 W/kg) for 1 h, no obvi-ous changes in Hsp70 expression were observed in RF-exposed cells compared with sham-exposed cells. CONCLUSIONS Our results suggest 1950 MHz RF field has little or no apparent effect on HSP70 and HSP27 expression in MO54, A172 and T98 cells, it may only slightly inhibit the phosphorylation of Hsp27 at Serine 78 in MO54 cells.

This paper describes a technology which provides a method for the storage of natural gas, primarily for vehicular use, as a liquid solution with other hydrocarbons at ambient temperatures which achieves an energy storage density (on a volumetric basis) up to about 80% of gasoline. The solution is stored at pressures of about 2000 psig and provides approximately 3 times the storage density of compressed natural gas. Various operating scenarios may include differing hydrocarbon solvents, such as propane, LPG, or butanes, and different required storage temperatures, with lower temperatures allowing higher volume fractions of methane. With this technology, the low storage density of CNG and the very low storage temperatures of LNG are both overcome, thus dramatically improving the technical feasibility of large scale commercialization of natural gas powered vehicles.

The protein (AfpA, for archaeoflavoprotein) encoded by AF1518 in the genome of Archaeoglobus fulgidus was produced in Escherichia coli and characterized. AfpA was found to be a homodimer with a native molecular mass of 43 kDa and containing two noncovalently bound flavin mononucleotides (FMNs). The cell extract of A. fulgidus catalyzed the CO-dependent reduction of AfpA that was stimulated by the addition of ferredoxin. Ferredoxin was found to be a direct electron donor to purified AfpA, whereas rubredoxin was unable to substitute. Neither NADH nor NADPH was an electron donor. Ferricyanide, 2,6-dichlorophenolindophenol, several quinones, ferric citrate, bovine cytochrome c , and O 2 accepted electrons from reduced AfpA, whereas coenzyme F 420 did not. The rate of cytochrome c reduction was enhanced in the presence of O 2 suggesting that superoxide is a product of the interaction of reduced AfpA with O 2 . Although AF1518 was previously annotated as encoding a decarboxylase involved in coenzyme A biosynthesis, the results establish that AfpA is an electron carrier protein with ferredoxin as the physiological electron donor. The genomes of several diverse Archaea contained afpA homologs clustered with open reading frames annotated as homologs of genes encoding reductases involved in the oxidative stress response of anaerobes from the domain Bacteria . A potential role for AfpA in coupling electron flow from ferredoxin to the putative reductases is discussed. A search of the databases suggests that AfpA is the prototype of a previously unrecognized flavoprotein family unique to the domain Archaea for which the name archaeoflavoprotein is proposed.

Each of the genomic sequences of Methanosarcina acetivorans, Methanosarcina mazei, and Methanosarcina thermophila revealed two homologs of mtaA, three homologs of mtaB, and three homologs of mtaC encoding enzymes specific for methanogenesis from methanol. Two-dimensional gel electrophoretic analyses of polypeptides from M. thermophila established that methanol induces the expression of mtaA-1, mtaB-1, mtaB-2, mtaB-3, mtaC-1, mtaC-2, and mtaC-3 whereas mtaB-3 and mtaC-3 are constitutively expressed in acetate-grown cells. The gene product of one of three mttC homologs, encoding trimethylamine-specific methyltransferase I, was detected in methanol- but not acetate-grown M. thermophila. A postulated role for the multiple homologs is discussed.

The effects of human and bovine serum proteins on CYP3A activity, using testosterone as the probe substrate, were investigated in human liver microsomes. Serum albumin, alpha-globulins, and alpha(1)-acid glycoprotein (alpha(1)-AGP) of both species significantly inhibited testosterone 6beta-hydroxylation. When the inhibitory effects of serum proteins were compared with serum protein binding data, human alpha-globulins, with a ratio (relative metabolic activity/unbound fraction) of 0.3, showed higher, and bovine alpha(1)-AGP, with the ratio of 1.4, showed lower inhibitory effects than those expected from protein binding of testosterone. The effects of the other serum proteins were close to those expected from protein binding, according to the free drug hypothesis. The K(i) values obtained from the Dixon plots were 0.32% (w/v, 48 microM) for human serum albumin (HSA), 0.48% for human alpha-globulins, and 0.23% (52 microM) for human alpha(1)-AGP. K(i) values of bovine serum albumin, bovine alpha-globulins and bovine alpha(1)-AGP were 3-5 times higher than those of the respective human proteins. The results suggest a direct interaction of some of these serum proteins with the active site of the CYP3A isoform. Since the bovine serum proteins showed weaker inhibitory effects than human serum proteins, the wide use of BSA, which is viewed as interchangeable with HSA, needs to be cautioned.

WE REVIEW here the technical aspects and clinical application of a powerful and precise molecular tool: quantitative competitive-polymerase chain reaction (QC-PCR). Our design and development of a competitor DNA template for the quantification of granzyme B mRNA by QC-PCR are described to illustrate the principles and the practical utility of this test system for the molecular characterization of allograft rejection.

Instability of atherosclerotic plaques is due to macrophage inflammatory activity and can result in rupture of the plaque (Plutzky, 1999). We previously showed that long-chain n-3 fatty acids enter advanced atherosclerotic plaques and alter their morphology to one indicative of greater stability (Thies et al. 2003). Whether these fatty acids decrease inflammatory activity within advanced plaques is not known. The objectives of the present study were to confirm that long-chain n-3 fatty acids enter advanced atherosclerotic plaques and to examine the effect on plaque inflammation and on the expression of selected inflammatory genes. Patients (n 121) awaiting carotid endarterectomy were randomly assigned to n-3 fatty acid ethyl esters (Omacor; Pronova Biocare AS, Lysaker, Norway) or olive oil as placebo (both 2 g/d) until surgery (7–109 d; median 21 d). Carotid-plaque phospholipid-fatty acid composition was determined by GC. Plaques were subject to histological examination, which identified the extent of inflammation and instability. Levels of mRNA encoding several matrix metalloproteinases (MMP), the inflammatory cytokine IL-6 and intercellular adhesion molecule (ICAM)-1 were determined by real-time RT–PCR using 36B4 as the housekeeping gene. The n-3 fatty acid EPA was significantly higher in plaque phospholipids from patients in the Omacor group (0.83 (SE 0.05) v. 0.43 (SE 0.05); P