Peak Fractions Research Articles

Exosite Interactions Impact Matrix Metalloproteinase Collagen Specificities

Members of the matrix metalloproteinase (MMP) family selectively cleave collagens in vivo. However, the substrate structural determinants that facilitate interaction with specific MMPs are not well defined. We hypothesized that type I-III collagen sequences located N- or C-terminal to the physiological cleavage site mediate substrate selectivity among MMP-1, MMP-2, MMP-8, MMP-13, and MMP-14/membrane-type 1 (MT1)-MMP. The enzyme kinetics for hydrolysis of three fluorogenic triple-helical peptides (fTHPs) was evaluated herein. The first fTHP contained consensus residues 769-783 from type I-III collagens, the second inserted α1(II) collagen residues 763-768 N-terminal to the consensus sequence, and the third inserted α1(II) collagen residues 784-792 C-terminal to the consensus sequence. Our analyses showed that insertion of the C-terminal residues significantly increased k(cat)/K(m) and k(cat) for MMP-1. MMP-13 showed the opposite behavior with a decreased k(cat)/K(m) and k(cat) and a greatly improved K(m) in response to the C-terminal residues. Insertion of the N-terminal residues enhanced k(cat)/K(m) and k(cat) for MMP-8 and MT1-MMP. For MMP-2, the C-terminal residues enhanced K(m) and dramatically decreased k(cat), resulting in a decrease in the overall activity. These changes in activities and kinetic parameters represented the collagen preferences of MMP-8, MMP-13, and MT1-MMP well. Thus, interactions with secondary binding sites (exosites) helped direct the specificity of these enzymes. However, MMP-1 collagen preferences were not recapitulated by the fTHP studies. The preference of MMP-1 for type III collagen appears to be primarily based on the flexibility of the hydrolysis site of type III collagen compared with types I and II. Further characterization of exosite determinants that govern interactions of MMPs with collagenous substrates should aid the development of pharmacotherapeutics that target individual MMPs.

Identification of Human Fumarylacetoacetate Hydrolase Domain-containing Protein 1 (FAHD1) as a Novel Mitochondrial Acylpyruvase

The human fumarylacetoacetate hydrolase (FAH) domain-containing protein 1 (FAHD1) is part of the FAH protein superfamily, but its enzymatic function is unknown. In the quest for a putative enzymatic function of FAHD1, we found that FAHD1 exhibits acylpyruvase activity, demonstrated by the hydrolysis of acetylpyruvate and fumarylpyruvate in vitro, whereas several structurally related compounds were not hydrolyzed as efficiently. Conserved amino acids Asp-102 and Arg-106 of FAHD1 were found important for its catalytic activity, and Mg(2+) was required for maximal enzyme activity. FAHD1 was found expressed in all tested murine tissues, with highest expression in liver and kidney. FAHD1 was also found in several human cell lines, where it localized to mitochondria. In summary, the current work identified mammalian FAHD1 as a novel mitochondrial enzyme with acylpyruvate hydrolase activity.

The tumultuous formation of the Hubble sequence at z > 1 examined with HST/Wide-Field Camera-3 observations of the Hubble Ultra Deep Field

We examine in this paper a stellar mass selected sample of galaxies at 1 < z < 3 within the Hubble Ultra Deep Field, utilising WFC3 imaging to study the rest-frame optical morphological distribution of galaxies at this epoch. We measure how apparent morphologies (disk, elliptical, peculiar) correlate with physical properties, such as quantitative structure and spectral-types. One primary result is that apparent morphology does not correlate strongly with stellar populations, nor with galaxy structure at this epoch, suggesting a chaotic formation history for Hubble types at z > 1. By using a locally defined definition of disk and elliptical galaxies based on structure and spectral-type, we find no true ellipticals at z > 2, and a fraction of 3.2+/-2.3% at 1.5 < z < 2. Local counterparts of disk galaxies are at a similar level of 7-10%, much lower than the 75% fraction at lower redshifts. We further compare WFC3 images with the rest-frame UV view of galaxies from ACS imaging, showing that galaxies imaged with ACS that appear peculiar often contain an `elliptical' like morphology in WFC3. We show through several simulations that this larger fraction of elliptical-like galaxies is partially due to the courser PSF of WFC3, and that the `elliptical' class very likely includes early-type disks. We also measure the merger history for our sample using CAS parameters, finding a redshift evolution increasing with redshift, and a peak merger fraction of ~30% at z~2 for the most massive galaxies with M_*> 10^{10} M_sol, consistent with previous results from ACS and NICMOS. We compare our results to semi-analytical model results and find a relatively good agreement between our morphological break-down and the predictions. Finally, we argue that the peculiars, ellipticals and peculiar ellipticals have similar properties, suggesting similar formation modes, likely driven by major mergers.

Crystal Structure of Human ISG15 Protein in Complex with Influenza B Virus NS1B

ISG15 (interferon-stimulated gene 15), the first ubiquitin-like protein (UBL) identified, has emerged as an important cellular antiviral factor. It consists of two UBL domains with a short linker between them. The covalent attachment of ISG15 to host and viral proteins to modify their functions, similar to ubiquitylation, is named ISGylation. Influenza B virus NS1B protein antagonizes human but not mouse ISGylation because NS1B exhibits species specificity; it only binds human and non-human primate ISG15. Previous studies have demonstrated that the N-terminal UBL domain and linker of ISG15 are required for the binding by NS1B and that the linker plays a large role in the species specificity, but the structural basis for them has not been elucidated. Here we report the crystal structure of human ISG15 in complex with NS1B at a resolution of 2.0 Å. A loop in the ISG15 N-terminal UBL domain inserts into a pocket in the NS1B dimer, forming a high affinity binding site. The nonspecific van der Waals contacts around the ISG15 linker form a low affinity site for NS1B binding. However, sequence alignment reveals that residues in the high affinity site are highly conserved in primate and non-primate ISG15. We propose that the low affinity binding around the ISG15 linker is important for the initial contact with NS1B and that the stable complex formation is largely contributed by the following high affinity interactions between ISG15 N-terminal UBL domain and NS1B. This provides a structural basis for the species-specific binding of ISG15 by the NS1B protein.

The effects of the hydrogen addition on the HCN profiles in fuel-rich-premixed, burner-stabilized C1–C3 alkane flames

The effects of hydrogen addition on HCN formation and consumption in fuel-rich, burner-stabilized methane, ethane and propane flames are reported. The HCN mole fraction was measured using quartz-microprobe sampling followed by direct absorption spectroscopy. Experiments were performed at equivalence ratio of 1.3, with 0 and 0.2 mole fraction of hydrogen in the fuel. The mass flux through the burner was varied for each mixture. The addition of hydrogen only modestly affects the HCN mole fractions. For the vast majority of the flames studied, the peak HCN fraction decreases by less than 20% upon hydrogen addition. The decrease in HCN fraction is smaller in ethane and propane flames compared to methane flames. Increasing the mass flux, independent of hydrogen fraction, results in a small increase in the HCN peak fraction, a shift of the HCN peak towards the burner and acceleration of the HCN consumption downstream. Burnout of HCN occurs faster in ethane and propane flames than in methane flames. Comparison of the experimental results with computations using two mechanisms for NCN oxidation shows that both mechanisms predict the HCN profiles reasonably well.

Crystal Structure of the E2 Domain of Amyloid Precursor Protein-like Protein 1 in Complex with Sucrose Octasulfate

Missense mutations in the amyloid precursor protein (APP) gene can cause familial Alzheimer disease. It is thought that APP and APP-like proteins (APLPs) may play a role in adhesion and signal transduction because their ectodomains interact with components of the extracellular matrix. Heparin binding induces dimerization of APP and APLPs. To help explain how these proteins interact with heparin, we have determined the crystal structure of the E2 domain of APLP1 in complex with sucrose octasulfate (SOS). A total of three SOS molecules are bound to the E2 dimer. Two SOSs are bound inside a narrow intersubdomain groove, and the third SOS is bound near the two-fold axis of the protein. Mutational analyses show that most residues interacting with SOS also contribute to heparin binding, although in varying degrees; a deep pocket, defined by His-376, Lys-422, and Arg-429, and an interfacial site between Lys-314 and its symmetry mate are most important in the binding of the negatively charged polysaccharide. Comparison with a lower resolution APP structure shows that all key heparin binding residues are conserved and identically positioned, suggesting that APLP1 and APP may bind heparin similarly. In transfected HEK-293 cells, mutating residues responsible for heparin binding causes little change in the proteolysis of APP by the secretases. However, mutating a pair of conserved basic residues (equivalent to Arg-414 and Arg-415 of APLP1) immediately adjacent to the heparin binding site affects both the maturation and the processing of APP.

Inhibition of RNase L and RNA-dependent Protein Kinase (PKR) by Sunitinib Impairs Antiviral Innate Immunity

RNase L and RNA-dependent protein kinase (PKR) are effectors of the interferon antiviral response that share homology in their pseudokinase and protein kinase domains, respectively. Sunitinib is an orally available, ATP-competitive inhibitor of VEGF and PDGF receptors used clinically to suppress angiogenesis and tumor growth. Sunitinib also impacts IRE1, an endoplasmic reticulum protein involved in the unfolded protein response that is closely related to RNase L. Here, we report that sunitinib is a potent inhibitor of both RNase L and PKR with IC(50) values of 1.4 and 0.3 μM, respectively. In addition, flavonol activators of IRE1 inhibited RNase L. Sunitinib treatment of wild type (WT) mouse embryonic fibroblasts resulted in about a 12-fold increase in encephalomyocarditis virus titers. However, sunitinib had no effect on encephalomyocarditis virus growth in cells lacking both PKR and RNase L. Furthermore, oral delivery of sunitinib in WT mice resulted in 10-fold higher viral titers in heart tissues while suppressing by about 2-fold the IFN-β levels. In contrast, sunitinib had no effect on viral titers in mice deficient in both RNase L and PKR. Also, sunitinib reduced mean survival times from 12 to 6 days in virus-infected WT mice while having no effect on survival of mice lacking both RNase L and PKR. Results indicate that sunitinib treatments prevent antiviral innate immune responses mediated by RNase L and PKR.

Characterization of voltage-dependent calcium channel blocking peptides from the venom of the tarantula Grammostola rosea

Voltage-dependent calcium channel blocking peptides were purified and sequenced from the venom of the tarantula, Grammostola rosea. cDNAs encoding the peptide sequences were cloned from the venom gland cDNA library. The electrophysiological effects of the peptides on several types of voltage-dependent calcium channels were evaluated using a Xenopus laevis oocyte expression system. A peptide contained in one of the HPLC peak fractions inhibited P/Q type voltage-dependent calcium channels (Cav2.1). The amino acid sequence of this peptide is identical to that of ω-grammotoxin SIA. A peptide from another discrete peak, which is identical to GsAFII except for one tryptophan residue in the C-terminus, inhibited L-type voltage-dependent calcium channels (Cav1.2). A novel peptide, named GTx1-15 (Accession number, AB201016), shows 76.5% sequence homology with the sodium channel blocker phrixotoxin 3, however, GTx1-15 preferentially inhibited T-type voltage-dependent calcium channels (Cav3.1). In silico secondary and tertiary structure prediction revealed that GTx1-15 and sodium channel blockers such as hainantoxin-IV, phrixotoxin 3, and ceratotoxin 2 show very similar β-strand composition, distribution of Optimal Docking Areas (continuous surface patches likely to be involved in protein–protein interactions), and surface electrostatic potential. These findings suggest that these peptide toxins evolved from common ancestors by gene duplication to maintain surface atmospheres appropriate for interaction with low-voltage-dependent ion channels.

Selective comprehensive multidimensional separation for resolution enhancement in high performance liquid chromatography. Part II: Applications

In this second paper of a two-part series, we demonstrate the utility of an approach to enhancing the resolution of select portions of conventional 1D-LC separations, which we refer to as selective comprehensive two-dimensional HPLC (sLC×LC), in three quite different example applications. In the first paper of the series we described the principles of this approach, which breaks the long-standing link in online multi-dimensional chromatography between the timescales of sampling the first dimension (1D) separation and the separation of fractions of 1D effluent in the second dimension. In the first example, the power of the sLC×LC approach to significantly reduce the analysis time and method development effort is demonstrated by selectively enhancing the resolution of critical pairs of peaks that are unresolved by a one-dimensional separation (1D-LC) alone. Transfer and subsequent 2D separations of multiple fractions of a particular 1D peak produces a two-dimensional chromatogram that reveals the coordinates of the peaks in the 2D separation space. The added time dimension of sLC×LC chromatograms also facilitates the application of sophisticated chemometric curve resolution algorithms to further resolve peaks that are otherwise chromatographically unresolved. This is demonstrated in this work by the targeted analysis of phenytoin in urban wastewater effluent using UV diode array detection. Quantitation by both standard addition and external calibration methods yielded results that were not statistically different from 2D-LC/MS/MS analysis of the same samples. Next, we demonstrate the utility of sLC×LC for reducing ion suppression due to matrix effects in electrospray ionization mass spectrometry through the analysis of cocaine in urban wastewater effluent. Finally, we explore the flexibility of the approach in its application to two select regions of a single 1D separation of triclosan and cocaine. The diversity of these applications demonstrates the power and versatility of the sLC×LC approach, which will benefit tremendously from further optimization and advances in valve technology that specifically address the needs of this new technique.

Selective comprehensive multi-dimensional separation for resolution enhancement in high performance liquid chromatography. Part I: Principles and instrumentation

An approach to enhancing the resolution of select portions of conventional one-dimensional high performance liquid chromatography (HPLC) separations was developed, which we refer to as selective comprehensive two-dimensional HPLC (sLC×LC). In this first of a series of two papers we describe the principles of this approach, which breaks the long-standing link in on-line multi-dimensional chromatography between the timescales of sampling the first dimension (1D) separation and the separation of fractions of 1D effluent in the second dimension. This allows rapid, high-efficiency separations to be used in the first dimension, while still adequately sampling 1D peaks. Transfer, transient storage, and subsequent second dimension (2D) separations of multiple fractions of a particular 1D peak produces a two-dimensional chromatogram that reveals the coordinates of the peak in both dimensions of the chromatographic space. Using existing valve technology we find that the approach is repeatable (%RSD of peak area <1.5%), even at very short first dimension sampling times – as low as 1s. We have also systematically studied the critical influence of the volume and composition of fractions transferred from the first to the second dimension of the sLC×LC system with reversed-phase columns in both dimensions, and the second dimension operated isocratically. We find that dilution of the transferred fraction, so that it contains 10–20% less organic solvent than the 2D eluent, generally mitigates the devastating effects of large transfer volumes on 2D performance in this type of system. Several example applications of the sLC×LC approach are described in the second part of this two-part series. We anticipate that future advances in the valve technology used here will significantly widen the scope of possible applications of the sLC×LC approach.

Experimental and kinetic modeling study of methyl butanoate and methyl butanoate/methanol flames at different equivalence ratios and C/O ratios

Premixed laminar methyl butanoate/oxygen/argon and methyl butanoate/methanol/oxygen/argon flames were studied with tunable synchrotron vacuum ultraviolet (VUV) photoionization and molecular-beam sampling mass spectrometry at 30 torr (4.0 kPa). Three flames were investigated in the experiment: MB (methyl butanoate) flame F1.54 ( ϕ = 1.54, C/O = 0.479), MB flame F1.67 ( ϕ = 1.67, C/O = 0.511) and MB/methanol flame F1.67M ( ϕ = 1.67, C/O = 0.479). By measuring the signal intensities at different distances from the burner surface, the mole fraction profiles of intermediates are derived. Experimental results show that the flame front shifts downstream and peak mole fractions of intermediates increase remarkably with the increase of equivalence ratio for pure MB fuel. When methanol is added, the peak mole fractions of most intermediates including those of soot precursors decrease remarkably at the same equivalence ratio, while peaks of soot precursors vary little (only slightly decreasing) at same C/O ratio. It is concluded that the formation of soot precursors is more sensitive to C/O ratio than to equivalence ratio. Besides, more CO 2 is produced near the burner surface in MB flame than that in MB/methanol flame, and this validates an early production of CO 2 in methyl ester oxidation. In addition, a modified MB detailed mechanism is used to model flame structure, and improved agreements between the experimental and predicted results are realized. Based on the simulation results, reaction flux and sensitivity are analyzed for CO 2 and C 3H 3, respectively.

Purification and anticancer activity investigation of pentacyclic triterpenoids from the leaves of Sinojackia sarcocarpa L.Q. Luo by high-speed counter-current chromatography

Sinojackia sarcocarpa L.Q. Luo has high aesthetic value, which is clinically used to treat diseases such as the variations of arthritis, thromboangiitis obliterans, angina pectoris as well as many other diseases. Pentacyclic triterpenoids are the main pharmacological effective compounds. High-speed counter-current chromatography method was performed on a Midi-DE centrifuge at 25°C. The solvent system was n-hexane–ethylacetate–methanol–water (10 : 5 : 3 : 1, v/v). Peak fractions were collected according to the elution profile for subsequent high-performance liquid chromatography analysis. The structure identification was performed by ultraviolet, infrared, mass spectrometry, 1H-NMR and 13C-NMR. Compound 2α,3α,19β,23β-tetrahydroxyurs-12-en-28-oic acid and 2α,3α,23β-trihydroxyurs-12-en-28-oic acid were purified from the plant of Styracaceaen genius for the first time, whose purities were 96.57% and 97.33%, respectively. Compared with the same dose of oral 5-fluorouracil with 57.6% inhibition rate, the S180 tumour inhibition rates of 20 mg kg−1 d−1 two compounds were 59.5% and 48.9%, respectively.

Structure of the Membrane-tethering GRASP Domain Reveals a Unique PDZ Ligand Interaction That Mediates Golgi Biogenesis

Biogenesis of the ribbon-like membrane network of the mammalian Golgi requires membrane tethering by the conserved GRASP domain in GRASP65 and GRASP55, yet the tethering mechanism is not fully understood. Here, we report the crystal structure of the GRASP55 GRASP domain, which revealed an unusual arrangement of two tandem PDZ folds that more closely resemble prokaryotic PDZ domains. Biochemical and functional data indicated that the interaction between the ligand-binding pocket of PDZ1 and an internal ligand on PDZ2 mediates the GRASP self-interaction, and structural analyses suggest that this occurs via a unique mode of internal PDZ ligand recognition. Our data uncover the structural basis for ligand specificity and provide insight into the mechanism of GRASP-dependent membrane tethering of analogous Golgi cisternae.

The Structural Basis of Endocannabinoid Oxygenation by Cyclooxygenase-2

The cyclooxygenases (COX-1 and COX-2) oxygenate arachidonic acid (AA) in the committed step of prostaglandin biogenesis. Substitutions of I434V, H513R, and I523V constitute the only differences in residues lining the cyclooxygenase channel between COX-1 and COX-2. These changes create a hydrophobic pocket in COX-2, with Arg-513 located at the base of the pocket, which has been exploited in the design of COX-2-selective inhibitors. Previous studies have shown that COX-2, but not COX-1, can oxygenate endocannabinoid substrates, including 2-arachidonoyl glycerol (2-AG). To investigate the isoform-specific structural basis of endocannabinoid binding to COX-2, we determined the crystal structure of the 2-AG isomer 1-arachidonoyl glycerol (1-AG) in complex with wild type and R513H murine (mu) COX-2 to 2.2 and 2.35 Å, respectively, and R513H muCOX-2 in complex with AA to 2.45 Å resolution. The 2,3-dihydroxypropyl moiety of 1-AG binds near the opening of the cyclooxygenase channel in the space vacated by the movement of the Leu-531 side chain, validating our previous hypothesis implicating the flexibility of the Leu-531 side chain as a determinant for the ability of COX-2 to oxygenate endocannabinoid substrates. Functional analyses carried out to compliment our structural findings indicated that Y355F and R513H muCOX-2 constructs had no effect on the oxygenation of 1-AG and 2-AG, whereas substitutions that resulted in a shortened side chain for Leu-531 had only modest effects. Both AA and 1-AG bind to R513H muCOX-2 in conformations similar to those observed in the co-crystal structures of these substrates with wild type enzyme.

Mapping the IκB Kinase β (IKKβ)-binding Interface of the B14 Protein, a Vaccinia Virus Inhibitor of IKKβ-mediated Activation of Nuclear Factor κB

The IκB kinase (IKK) complex regulates activation of NF-κB, a critical transcription factor in mediating inflammatory and immune responses. Not surprisingly, therefore, many viruses seek to inhibit NF-κB activation. The vaccinia virus B14 protein contributes to virus virulence by binding to the IKKβ subunit of the IKK complex and preventing NF-κB activation in response to pro-inflammatory stimuli. Previous crystallographic studies showed that the B14 protein has a Bcl-2-like fold and forms homodimers in the crystal. However, multi-angle light scattering indicated that B14 is in monomer-dimer equilibrium in solution. This transient self-association suggested that the hydrophobic dimerization interface of B14 might also mediate its interaction with IKKβ, and this was investigated by introducing amino acid substitutions on the dimer interface. One mutant (Y35E) was entirely monomeric but still co-immunoprecipitated with IKKβ and blocked both NF-κB nuclear translocation and NF-κB-dependent gene expression. Therefore, B14 homodimerization is nonessential for binding and inhibition of IKKβ. In contrast, a second monomeric mutant (F130K) neither bound IKKβ nor inhibited NF-κB-dependent gene expression, demonstrating that this residue is required for the B14-IKKβ interaction. Thus, the dimerization and IKKβ-binding interfaces overlap and lie on a surface used for protein-protein interactions in many viral and cellular Bcl-2-like proteins.

Origin Single-stranded DNA Releases Sld3 Protein from the Mcm2–7 Complex, Allowing the GINS Tetramer to Bind the Mcm2–7 Complex

The replication fork helicase in eukaryotic cells is comprised of Cdc45, Mcm2-7, and GINS (CMG complex). In budding yeast, Sld3, Sld2, and Dpb11 are required for the initiation of DNA replication, but Sld3 and Dpb11 do not travel with the replication fork. Sld3 and Cdc45 bind to early replication origins during the G(1) phase of the cell cycle, whereas Sld2, GINS, polymerase ε, and Dpb11 form a transient preloading complex that associates with origins during S phase. We show here that Sld3 binds tightly to origin single-stranded DNA (ssDNA). CDK-phosphorylated Sld3 binds to origin ssDNA with similar high affinity. Origin ssDNA does not disrupt the interaction between Sld3 and Dpb11, and origin ssDNA does not disrupt the interaction between Sld3 and Cdc45. However, origin ssDNA substantially disrupts the interaction between Sld3 and Mcm2-7. GINS and Sld3 compete with one another for binding to Mcm2-7. However, in a mixture of Sld3, GINS, and Mcm2-7, origin ssDNA inhibits the interaction between Sld3 and Mcm2-7, whereas origin ssDNA promotes the association between GINS and Mcm2-7. We also show that origin single-stranded DNA promotes the formation of the CMG complex. We conclude that origin single-stranded DNA releases Sld3 from Mcm2-7, allowing GINS to bind Mcm2-7.

DNA Binding Restricts the Intrinsic Conformational Flexibility of Methyl CpG Binding Protein 2 (MeCP2)

Mass spectrometry-based hydrogen/deuterium exchange (H/DX) has been used to define the polypeptide backbone dynamics of full-length methyl CpG binding protein 2 (MeCP2) when free in solution and when bound to unmethylated and methylated DNA. Essentially the entire MeCP2 polypeptide chain underwent H/DX at rates faster than could be measured (i.e. complete exchange in ≤10 s), with the exception of the methyl DNA binding domain (MBD). Even the H/DX of the MBD was rapid compared with that of a typical globular protein. Thus, there is no single tertiary structure of MeCP2. Rather, the full-length protein rapidly samples many different conformations when free in solution. When MeCP2 binds to unmethylated DNA, H/DX is slowed several orders of magnitude throughout the MBD. Binding of MeCP2 to methylated DNA led to additional minor H/DX protection, and only locally within the N-terminal portion of the MBD. H/DX also was used to examine the structural dynamics of the isolated MBD carrying three frequent mutations associated with Rett syndrome. The effects of the mutations ranged from very little (R106W) to a substantial increase in conformational sampling (F155S). Our H/DX results have yielded fine resolution mapping of the structure of full-length MeCP2 in the absence and presence of DNA, provided a biochemical basis for understanding MeCP2 function in normal cells, and predicted potential approaches for the treatment of a subset of RTT cases caused by point mutations that destabilize the MBD.

A numerical study on the effects of pressure and gravity in laminar ethylene diffusion flames

The effects of pressure and gravity on sooting characteristics and flame structure were studied numerically in coflow ethylene–air laminar diffusion flames between 0.5 and 5 atm. Computations were performed by solving the unmodified and fully-coupled equations governing reactive, compressible, gaseous mixtures which include complex chemistry, detailed radiation heat transfer, and soot formation/oxidation. Soot formation/oxidation was modeled using an acetylene-based, semi-empirical model which has been verified with previously published experimental data to correctly capture many of the observed trends at normal-gravity. Calculations for each pressure considered were performed for both normal- and zero-gravity conditions to help separate the effects of pressure and buoyancy on soot formation. Based on the numerical predictions, pressure and gravity were observed to significantly influence the flames through their effects on buoyancy and reaction rates. The zero-gravity flames have higher soot concentrations, lower temperatures and broader soot-containing zones than normal-gravity flames at the same pressure. The zero-gravity flames were also found to be longer and wider. Differences were observed between the two levels of gravity when pressure was increased. The zero-gravity flames displayed a stronger dependence of the maximum soot yield on pressure from 0.5 to 2 atm and a weaker dependence from 2 to 5 atm as compared to the normal-gravity flames. In addition, flame diameter decreased with increasing pressure under normal-gravity while it increased with pressure in the zero-gravity cases. Changing the prescribed wall boundary condition from fixed-temperature to adiabatic significantly altered the numerical predictions at 5 atm. When the walls were assumed to be adiabatic, peak soot volume fractions and temperatures increased in both the zero- and normal-gravity flames, emphasizing the importance of heat conduction to the burner rim on flame structure.

Erratum to: Investigation on Stability of Transporter Protein, Glucuronide Transporter from Escherichia coli

Original Article has an error in the caption of Figure 2. In the last sentense of the caption, ‘‘heptameric’’ should be read as ‘‘pentameric’’. Corrected caption for figure 2 is given below. Fig. 2 Size-exclusion HPLC elution profile of GusB with moleculer size marker standards. The GusB–DDM complex was found to have a Stokes radius of 57 A. Six soluble proteins of known Stokes radius were used for calibration: ferritin (63 A), catalase (52 A), aIdorase (46 A), bovine serum albumin (35 A), ovalbumin (28 A) and chymotrypsinogen (22 A). Inset is a nagatively stained transmission electron microscopic image of the peak fraction. Scale bar, 500 nm. Based on the elution profile and TEM image, GusB in the presence of DDM most probably stays as a pentameric oligomer.

Utilization of carbon dioxide for polymer electrolytes [I]: Effect of supercritical treatment conditions on ionic conduction in amorphous polyether/salt mixtures

Supercritical carbon dioxide (scCO 2) as a treatment medium has a possibility to realize excellent room temperature conductivity more than 10 −4 S/cm for polymer electrolytes in the dry state. In this study, a typical high ion-conductive polyether-based electrolyte which consists of poly-[ethylene oxide- co-2-(2-methoxyethoxy)ethyl glycidyl ether] (P(EO/EM)) and lithium perchlorate (LiClO 4) was used as a model sample for the scCO 2 treatment. We found the suitable scCO 2 treatment conditions (pressure, temperature and time) for high conductivity. The conductivity of sample treated at 7.5 MPa and 40 °C for 40 min was more than 100-times higher than that of original without the treatment, and the value decreased only 20% after 30 days. DSC measurement revealed that the decrease in glass transition temperature ( T g) is caused by the scCO 2-treatment. The change of ionic association in the scCO 2-treated samples was confirmed using FT-IR measurement. The scCO 2 treatment gave rise to increase in peak fraction of free ClO 4 − anions (620–625 cm −1) and peak shift of ν(C–O–C) mode to lower frequency region (1060–1070 cm −1) depending on ether–Li + interactions.