On the oscillatory phenomenon in an oil/water interface

A simple theoretical model is presented for simulating the self-sustained oscillations of electric potential and pH at an oil/water interface appearing in a two-phase system composed of 2-nitropropane solution containing picrate acid and an aqueous solution of cetyltrimethylammonium bromide. In the present model, a well-known condition necessary for the occurrence of self-sustained oscillations, i.e., the presence of a positive feedback process far from equilibrium, is taken into account in a set of kinetic equations to describe simplified characters of the following two processes: (i) a cooperative formation of ion pair complexes at the interface, and (ii) supply of picrate anions and cetyltrimethylammonium cations to the interface accompanied by release of ion pair complexes to the organic phase. The numerical solutions of the present equations are shown to reproduce fairly well the characteristic properties of the oscillation of electric potential and pH such as wave forms and frequencies.     

Solid-Phase Artificial Chaperone-Assisted Refolding using Insoluble β-Cyclodextrin–Acrylamide Copolymer Beads

Solid-phase refolding methods are advantageous since they facilitate both separation of solid additives from the refolded protein and recycling of the additives. -Cyclodextrin–acrylamide copolymer hydrogel beads were used as a matrix for detergents in solid-phase artificial chaperone-assisted refolding and improved the yield of lysozyme (up to 65%) and carbonic anhydrase B (up to 80%), compared with conventional solid host matrices.Revisions received 29 September 2004     

Vascular Smooth Muscle Cells Stimulate Platelets and Facilitate Thrombus Formation through Platelet CLEC-2: Implications in Atherothrombosis

The platelet receptor CLEC-2 is involved in thrombosis/hemostasis, but its ligand, podoplanin, is expressed only in advanced atherosclerotic lesions. We investigated CLEC-2 ligands in vessel walls. Recombinant CLEC-2 bound to early atherosclerotic lesions and normal arterial walls, co-localizing with vascular smooth muscle cells (VSMCs). Flow cytometry and immunocytochemistry showed that recombinant CLEC-2, but not an anti-podoplanin antibody, bound to VSMCs, suggesting that CLEC-2 ligands other than podoplanin are present in VSMCs. VSMCs stimulated platelet granule release and supported thrombus formation under flow, dependent on CLEC-2. The time to occlusion in a FeCl₃-induced animal thrombosis model was significantly prolonged in the absence of CLEC-2. Because the internal elastic lamina was lacerated in our FeCl₃-induced model, we assume that the interaction between CLEC-2 and its ligands in VSMCs induces thrombus formation. Protein arrays and Biacore analysis were used to identify S100A13 as a CLEC-2 ligand in VSMCs. However, S100A13 is not responsible for the above-described VSMC-induced platelet activation, because S100A13 is not expressed on the surface of normal VSMCs. S100A13 was released upon oxidative stress and expressed in the luminal area of atherosclerotic lesions. Suspended S100A13 did not activate platelets, but immobilized S100A13 significantly increased thrombus formation on collagen-coated surfaces. Taken together, we proposed that VSMCs stimulate platelets through CLEC-2, possibly leading to thrombus formation after plaque erosion and stent implantation, where VSMCs are exposed to blood flow. Furthermore, we identified S100A13 as one of the ligands on VSMCs.     

Comparison of WTC dust size on macrophage inflammatory cytokine release in vivo and in vitro

Background

The WTC collapse exposed over 300,000 people to high concentrations of WTC-PM; particulates up to ∼50 mm were recovered from rescue workers’ lungs. Elevated MDC and GM-CSF independently predicted subsequent lung injury in WTC-PM-exposed workers. Our hypotheses are that components of WTC dust strongly induce GM-CSF and MDC in AM; and that these two risk factors are in separate inflammatory pathways.

Methodology/Principal Findings

Normal adherent AM from 15 subjects without WTC-exposure were incubated in media alone, LPS 40 ng/mL, or suspensions of WTC-PM_10–53 or WTC-PM_2.5 at concentrations of 10, 50 or 100 µg/mL for 24 hours; supernatants assayed for 39 chemokines/cytokines. In addition, sera from WTC-exposed subjects who developed lung injury were assayed for the same cytokines. In the in vitro studies, cytokines formed two clusters with GM-CSF and MDC as a result of PM_10–53 and PM_2.5. GM-CSF clustered with IL-6 and IL-12(p70) at baseline, after exposure to WTC-PM_10–53 and in sera of WTC dust-exposed subjects (n = 70) with WTC lung injury. Similarly, MDC clustered with GRO and MCP-1. WTC-PM_10–53 consistently induced more cytokine release than WTC-PM_2.5 at 100 µg/mL. Individual baseline expression correlated with WTC-PM-induced GM-CSF and MDC.

Conclusions

WTC-PM_10–53 induced a stronger inflammatory response by human AM than WTC-PM_2.5. This large particle exposure may have contributed to the high incidence of lung injury in those exposed to particles at the WTC site. GM-CSF and MDC consistently cluster separately, suggesting a role for differential cytokine release in WTC-PM injury. Subject-specific response to WTC-PM may underlie individual susceptibility to lung injury after irritant dust exposure.     

Soil organic matter dynamics in density and particle-size fractions following destruction of tropical rainforest and the subsequent establishment of Imperata grassland in Indonesian Borneo using stable carbon isotopes

Background and aims

Large portions of the deforested areas in Southeast Asia have been ultimately replaced by the invasive grass Imperata cylindrica, but the dynamics of soil organic matter (SOM) during such land transitions are poorly understood. This study presents SOM dynamics in density and particle-size fractions following rainforest destruction and the subsequent establishment and persistence of Imperata grassland.

Methods

We examined soil C stock and natural ¹³C abundance in these fractions to depths of 100 cm. We predicted future soil C storage and evaluated C turnover rates in these fractions using a simple exponential model. Because soil texture strongly affects soil C storage, two chronosequences of soils differing in soil texture were compared (n?=?1 in each chronosequence).

Results

The clay-associated SOM increased in all soil layers (0–100 cm) along the forest-to-grassland chronosequence, whereas light-fraction SOM in the surface soil layer (0–5 cm) decreased.

Conclusions

In the surface layer, all SOM fractions exhibited rapid replacement of forest-derived C to grassland-derived C, indicating fast turnover. Meanwhile, δ¹³C values of the light fraction in the surface layer indicated that forest-derived charcoal and/or occluded low-density organic matter constituted unexpectedly large proportions of the light fraction. Mathematical modelling (0–50 cm) showed that grassland-derived C in the clay and silt fractions in all soil layers increased almost linearly for at least 50 years after grassland establishment. In the meantime, the forest-derived C stock in the clay fraction constituted 82 % of the total stable C pool at 0–50-cm depths even under steady-state conditions (t = ∞), indicating that residue of forest-derived SOM associated with clay largely contributed to preserving the soil C pool. Comparing soils with different soil textures, clay and silt particles in coarse-textured soil exhibited a substantially higher degree of organo-mineral interactions per unit volume of clay or silt compared to fine-textured soils.     

Site-specific conjugation of oligonucleotides to the C-terminus of recombinant protein by expressed protein ligation

We developed a convenient method for synthesizing homogeneous DNA-protein conjugates. The method is based on expressed protein ligation of intein-fusion proteins and oligonucleotides derivatized with a cysteine. A range of cysteinyl oligonucleotides were synthesized by using a new reagent 1 and were successfully applied to expressed protein ligation to attach the oligonucleotides specifically at the C-terminus of a recombinant protein.     

Successful control of aggregation and folding rates during refolding of denatured lysozyme by adding N-methylimidazolium cations with various N'-substituents

The present study aimed to obtain more effective refolding agents and to understand the influence of their chemical structures on their function as refolding agents. To achieve these aims, we investigated the effects of a large variety of N'-substituted N-methylimidazolium chlorides on the oxidative refolding of lysozyme in a high throughput manner. Among the molecules examined, N-methylimidazolium cations with a short N'-alkyl chain, such as an N'-ethyl or N'-butyl chain, significantly enhanced the refolding yield compared to conventional refolding additives such as arginine hydrochloride and Triton X-100. Detailed kinetic analyses revealed that the effective cations selectively decreased the aggregation rate constant (kA) without any large decreases in the folding rate constant (kN). However, when the hydrophobicity of the N'-substituent of the cations was increased, the desirable properties of the short N'-alkyl chain-type cations for protein refolding were diminished. Furthermore, increases in the N'-alkyl chain length to an N'-octyl or N'-dodecyl chain drastically decreased the kA values, thereby increasing the ratio of kN to kA, despite the very small kN values and resulting in enhanced refolding yields. Thus, by tuning the chemical structure of the N'-substituents of N-methylimidazolium chloride, five effective refolding agents (N'-ethyl-, N'-propyl-, N'-butyl-, N'-pentyl- and N'-isobutyl-N-methylimidazolium chlorides) were successfully obtained, and the kinetic parameters of folding and aggregation during the refolding process could be controlled using three different modes.