Residential indoor PM2.5 in wood stove homes: follow-up of the Libby changeout program.

UNLABELLED: In 2005 through 2008, a small rural mountain valley community engaged in a woodstove changeout program to address concerns of poor ambient air quality. During this program, we assessed changes to indoor air quality before and after the introduction of a new, lower emission woodstove. We previously reported a >70% reduction in indoor PM(2.5) concentrations in homes following the installation of a new Environmental Protection Agency's-certified stove within the home. We report here on follow-up of the experiences in these and other homes over three winters of sample collection. In 21 homes, we compared pre-changeout PM(2.5) concentrations [mean (s.d.) = 45.0 (33.0) µg/m(3)] to multiple post-changeout measures of PM(2.5) concentrations using a DustTrak. The mean reduction (and 95% confidence interval) from pre-changeout to post-changeout was -18.5 µg/m(3) (-31.9, -5.2), adjusting for ambient PM(2.5) , ambient temperature, and other factors. Findings across homes and across years were highly variable, and a subset of homes did not experience a reduction in PM(2.5) following changeout. Reductions were also observed for organic carbon, elemental carbon, and levoglucosan, but increases were observed for dehydroabietic acid and abietic acid. Despite overall improvements in indoor air quality, the varied response across homes may be due to factors other than the introduction of a new woodstove. PRACTICAL IMPLICATIONS: Biomass combustion is a common source of ambient PM(2.5) in many cold-climate communities. The replacement of older model woodstoves with newer technology woodstoves is a potential intervention strategy to improve air quality in these communities. In addition to ambient air, woodstove changeouts should improve residential indoor air quality. We present results from a multi-winter study to evaluate the efficacy of woodstove changeouts on improving indoor air quality. Reductions in indoor PM(2.5) were evident, but this observation was not consistent across all homes. These findings suggest that other factors beyond the introduction of an improved wood burning device are relevant to improving indoor air quality in wood burning homes.

Novel alkyl-modified anionic siloxanes as pseudostationary phases for electrokinetic chromatography: II. Selectivity studied by linear solvation energy relationships.

Anionic, water-soluble siloxane polymers modified with different lengths of alkyl chains have very different selectivity than sodium dodecyl sulfate (SDS) micelles when used as pseudostationary phases in electrokinetic chromatography. The siloxanes in this study are random copolymers with side chains bearing sulfonate groups and alkyl groups (C8, C12, or C18), with the proportion of alkyl groups between 10 and 25% of the total. The differences in selectivity have been studied by linear solvation energy relationships (LSERs). The siloxanes in general have been found to be more cohesive, less polar, more able to interact with solutes through n- and pi-electrons, and more able to accept hydrogen bonds than SDS micelles, while the ability to act as hydrogen bond donors is not significantly different than SDS micelles. In addition, the performance in a pH 7.0 Tris buffer has been investigated and the siloxanes were found to have higher methylene selectivities and more variable electrophoretic mobilities than in borate buffers.

Alkyl modified anionic siloxanes as pseudostationary phases for electrokinetic chromatography. I. Synthesis and characterization.

Anionic water soluble siloxane polymers have been synthesized and characterized for electrokinetic chromatography. Siloxane polymers are of interest in electrokinetic chromatography because of the wide variety of chemistries that can be developed based on these backbones, including much of the stationary phase chemistry developed in the last 30 years. The siloxanes in this study have a sulfonate functional group. The siloxanes have different length alkyl chains (C8, C12, C18) attached to the backbone in differing densities. The methylene selectivity generally increases with increasing alkyl chain length and with increasing alkyl chain density. The electrophoretic mobility appears to pass through a maximum as more alkyl chain is added to the siloxane backbone. The efficiency also would seem to pass through a maximum as more alkyl chain is added. The chemical selectivities of the siloxane polymers are very different from sodium dodecyl sulfate but are similar to each other.

Effect of pendant chain lengths and backbone functionalities on the chemical selectivity of sulfonated amphiphilic copolymers as pseudo-stationary phases in electrokinetic chromatography.

Amphiphilic copolymers of AMPS (2-acrylamido-2-methyl-1-propanesulfonic acid) and hydrophobic monomers with various chemical structures were synthesized, characterized and used as novel electrokinetic chromatography polymeric pseudo-stationary phases, showing significant chemical selectivity differences from that of the conventional monomeric pseudo-stationary phase, sodium lauryl sulphate. Copolymers of AMPS and methacrylates with different pendant chain lengths (C8, C12 and C18) were investigated and no significant difference in chemical selectivity was observed among them. However, the spacer bonding chemistry was shown to contribute to significant chemical selectivity difference, e.g. poly(AMPS-lauryl methacrylate) showed different chemical selectivity from poly(AMPS-lauryl methacrylamide). Linear solvation energy relationship analysis of 20 solutes by eight different polymeric pseudo-stationary phases was employed to investigate the solute molecule structural contributions to the retention. Hydrogen-bonding properties (described by system constants b and a) of poly(AMPS-alkyl methacrylamide) were found stronger than those of poly(AMPS-alkyl methacrylate).

A TRP homolog in Saccharomyces cerevisiae forms an intracellular Ca(2+)-permeable channel in the yeast vacuolar membrane.

The molecular identification of ion channels in internal membranes has made scant progress compared with the study of plasma membrane ion channels. We investigated a prominent voltage-dependent, cation-selective, and calcium-activated vacuolar ion conductance of 320 pS (yeast vacuolar conductance, YVC1) in Saccharomyces cerevisiae. Here we report on a gene, the deduced product of which possesses significant homology to the ion channel of the transient receptor potential (TRP) family. By using a combination of gene deletion and re-expression with direct patch clamping of the yeast vacuolar membrane, we show that this yeast TRP-like gene is necessary for the YVC1 conductance. In physiological conditions, tens of micromolar cytoplasmic Ca(2+) activates the YVC1 current carried by cations including Ca(2+) across the vacuolar membrane. Immunodetection of a tagged YVC1 gene product indicates that YVC1 is primarily localized in the vacuole and not other intracellular membranes. Thus we have identified the YVC1 vacuolar/lysosomal cation-channel gene. This report has implications for the function of TRP channels in other organisms and the possible molecular identification of vacuolar/lysosomal ion channels in other eukaryotes.

Synthesis and characterization of novel anionic siloxane polymers as pseudostationary phases for electrokinetic chromatography.

Four novel siloxane polymeric pseudostationary phases with three different ionic head groups have been synthesized and characterized for electrokinetic chromatography. Siloxane polymers are of interest in this application because of the wide range of chemistries that can be developed based on these backbones, including much of the chromatographic stationary phase chemistry developed in the last thirty years. All four of the siloxanes studied were synthesized by modification of a single methylhydrosiloxane polymer with highly acidic anionic functionalities. One of the siloxanes had both ionic groups and alkane chains attached to the siloxane backbone. The electrophoretic mobilities varied from being somewhat less than sodium dodecyl sulfate (SDS) to being much greater than SDS. The siloxanes substituted with ionic groups at all of the silicon sites showed significant nonequilibrium band broadening, severely limiting the efficiencies of these polymers. Substitution of 20% of the silicon sites with an alkyl group improved the efficiency of the separations and the peak symmetry. The chemical selectivities of the siloxane polymers are very different from SDS, but are similar to each other.

Sulfonated acrylamide copolymers as pseudo-stationary phases in electrokinetic chromatography.

Sulfonated copolymers were synthesized, characterized and used as separation media in electrokinetic chromatography. The polymers used were synthesized from AMPS (2-acrylamido-2-methyl-1-propanesulfonic acid) and LMAm (lauryl methacrylamide) in different mole ratios (from 100:0 to 60:40). Electrophoretic mobilities and methylene selectivities were calculated, which showed the expected correlation with the monomer ratios. The chemical selectivities for the separation of nine solutes by the copolymers were compared with that of sodium lauryl sulfate micelles, showing significant differences. No significant difference in chemical selectivities was observed for copolymers with different monomer ratios. No significant change of hydrophobic microdomain of copolymers was found in background buffers with different ionic strength values, based on the investigation of the retention factors, methylene selectivities and polymer effective mobilities. No change of hydrophobic microdomain of the copolymer solutions was found at copolymer concentrations from 0.17 to 3% (w/v), however, plots of k' versus polymer concentration suggested a different copolymer phase at lower concentrations (from 0 to 0.1%, w/v) from that at higher concentrations (from 0.17 to 3%, w/v). The copolymer with AMPS-LMAm (80:20) could be chosen as optimum copolymer as far as the methylene selectivity, peak symmetry and polymer mobility were concerned.

An anionic siloxane polymer as a pseudostationary phase for electrokinetic chromatography.

A novel polymeric pseudostationary phase for electrokinetic chromatography is introduced and characterized. Siloxane polymers are of interest for this application because of the range of chemistries that could be developed based on these backbones, and because successful development of siloxane polymers would make it possible to employ much of the stationary phase chemistry developed in the past thirty years. A commercially available water-soluble siloxane with a hydroxy-terminated alkyl group was converted to the sulfate derivative. This siloxane polymer is water-soluble, effectively eliminating this limitation associated with siloxane polymers. When employed as a pseudostationary phase, this compound provided rapid, efficient, and selective separations. The electrophoretic mobility of the polymer was less than sodium dodecyl sulfate (SDS) and poly(sodium 10-undecenylsulfate), providing a compressed migration time range, which is the main limiting factor for this polymer. The chemical selectivity of the siloxane sulfate was somewhat different than SDS micelles. The siloxane was employed in buffers modified with a large amount of acetonitrile to separate a number of polynuclear aromatic hydrocarbons. The addition of acetonitrile caused an apparent discontinuity in the electrophoretic mobility of the polymer, which may indicate a change in the structure with increasing organic solvent content.

Membrane-delimited coupling between sigma receptors and K+ channels in rat neurohypophysial terminals requires neither G-protein nor ATP.

Receptor-mediated modulation of ion channels generally involves G-proteins, phosphorylation, or both in combination. The sigma receptor, which modulates voltage-gated K+ channels, is a novel protein with no homology to other receptors known to modulate ion channels. In the present study patch clamp and photolabelling techniques were used to investigate the mechanism by which sigma receptors modulate K+ channels in peptidergic nerve terminals. The sigma receptor photoprobe iodoazidococaine labelled a protein with the same molecular mass (26 kDa) as the sigma receptor protein identified by cloning. The sigma receptor ligands pentazocine and SKF10047 modulated K+ channels, despite intra-terminal perfusion with GTP-free solutions, a G-protein inhibitor (GDPbetaS), a G-protein activator (GTPgammaS) or a non-hydrolysable ATP analogue (AMPPcP). Channels in excised outside-out patches were modulated by ligand, indicating that soluble cytoplasmic factors are not required. In contrast, channels within cell-attached patches were not modulated by ligand outside a patch, indicating that receptors and channels must be in close proximity for functional interactions. Channels expressed in oocytes without receptors were unresponsive to sigma receptor agonists, ruling out inhibition through a direct drug interaction with channels. These experiments indicate that sigma receptor-mediated signal transduction is membrane delimited, and requires neither G-protein activation nor protein phosphorylation. This novel transduction mechanism is mediated by membrane proteins in close proximity, possibly through direct interactions between the receptor and channel. This would allow for more rapid signal transduction than other ion channel modulation mechanisms, which in the present case of neurohypophysial nerve terminals would lead to the enhancement of neuropeptide release.

Polymeric and polymer-supported pseudostationary phases in micellar electrokinetic chromatography: performance and selectivity.

Several types of synthetic ionic polymers have been employed as pseudostationary phases in electrokinetic chromatography. The polymers have been shown to have some significant advantages and different chemical selectivity relative to conventional surfactant micelles. Polymeric phases are effective for the separation and analysis of hydrophobic and chiral compounds, and may be useful for the application of mass spectrometric detection. Additionally, the polymeric phases often demonstrate unique selectivity relative to micellar phases, and can be designed and synthesized to provide desired selectivity. This review covers efforts to develop and characterize the performance, characteristics, and selectivity of synthetic polymeric pseudostationary phases since their introduction in 1992. Some ideas for the future development of polymeric pseudostationary phases and the role they may play in electrokinetic separations are presented.

Evaluation of polymers based on a silicone backbone as pseudostationary phases for electrokinetic chromatography.

The feasibility of polymeric phases based on a silicone polymer backbone as pseudostationary phases for electrokinetic chromatography has been investigated. Silicone phases were studied because of the range of chemistries that could be developed based on these backbones, and because successful development of silicone phases would make it possible to employ much of the stationary phase chemistry developed in the past thirty years. Three silicone polymer structures have been investigated, but only one had sufficient aqueous solubility to permit application in electrokinetic chromatography. This phase was characterized by a variety of methods and was shown to be a mixture of partially hydrolyzed poly(bis-(3-cyanopropyl) siloxanes. When employed as a pseudostationary phase, this material provided selective and efficient separations. The electrophoretic mobility of the silicone polymer is greater than that of sodium dodecyl sulfate (SDS) micelles and poly(sodium 10-undecenylsulfate), providing an extended migration time range. A striking characteristic of the polymer is that the electrophoretic mobility is greater than typical electroosmotic mobilities. The chemical selectivity of the phase is significantly different from that of SDS micelles or poly(sodium 10-undecenylsulfate). The silicone phase is a more cohesive, basic and polar phase than SDS micelles. In buffers modified with a high concentration of organic solvents, the chromatographic properties of the silicone polymer are inferior to those of the poly(sodium 10-undecenylsulfate). The greatest limitation of silicone polymers for this application appears to be limited aqueous solubility, which will make it difficult to realize a family of such polymers with different chemical selectivities.

Polymers of sodium-N-undec-10-ene-1-oyl taurate and sodium-N-undec-10-ene-1-oyl aminoethyl-2-phosphonate as pseudostationary phases for electrokinetic chromatography.

The use of micelle polymers, a class of polysoaps with a polymerized hydrophobic interior and a charged hydrophillic exterior, as pseudostationary phases in electrokinetic chromatography has generated significant interest. Their stable structure has been shown to provide significant advantages over conventional micelles when used as pseudostationary phases. In previous studies, micelle polymers have had carboxylate and sulfate head groups. These chemistries have limitations: carboxylate micelle polymers precipitate out of solution at pH less than seven or eight and sulfate head groups are not stable to hydrolysis and are hydrolyzed during polymerization. Additionally, while the chemical selectivity of conventional micelles varies with head group chemistry, no significant differences in chemical selectivity were observed between analogous polymers with sulfate and carboxylate groups. To overcome the limitations of carboxylate and sulfate head groups, and to further investigate the chemical selectivity of micelle polymers, poly(sodium-N-undec-10-ene-1-oyl-taurate) and poly(sodium-N-undec-10-ene-1-oyl-ethyl-2-phosphonate) micellar polymers have been synthesized and characterized as pseudostationary phases. These polymers have amide functionality and stable, strongly acidic sulfonate and phosphonate head groups. These polymers did provide improved solubility at low pH, and are stable under the conditions studied. The chromatographic performance and chemical selectivity of the polymers has been studied by several methods, including linear solvation energy relationships. Poly(sodiumN-undec-10-ene-1-oyl-taurate) has greater electrophoretic mobility than other polymers of this type, and can be used for the separation of hydrophobic compounds. The polymers do exhibit unique selectivity, but the differences in selectivity are not significant for the majority of compounds studied.

Length polymorphism within the second variable region of the human immunodeficiency virus type 1 envelope glycoprotein affects accessibility of the receptor binding site.

Sequential mutations were introduced into the V2 region of human immunodeficiency virus (HIV) type 1 HXB2, affecting the length, charge, and number of potential glycosylation sites. The insertions had no effect on cytopathicity or on the ability of virus to replicate in peripheral blood mononuclear cells and established T-cell lines. However, deletion of amino acids 186 to 188, encoding a conserved glycosylation site, resulted in a nonviable virus, suggesting a minimal length requirement of 40 amino acids for a functional V2 loop. However, all amino acid insertions affected the sensitivity of the variants to neutralization by soluble CD4 and monoclonal antibodies specific for epitopes in the V3 and CD4 binding site regions. Furthermore, these mutant viruses showed resistance to neutralization by HIV-positive human sera. Soluble gp120 mutant glycoproteins showed increased affinities for soluble CD4 and monoclonal antibodies specific for a number of epitopes overlapping the CD4 binding site, confirming that length increases in V2 affect exposure of the CD4 binding site. In summary, these data demonstrate that differences in V2 length modulate immunoreactivity of the envelope glycoprotein and support an association between the V2 and CD4 binding site regions.

Multifocal demyelinating motor neuropathy: pathologic evidence of 'inflammatory demyelinating polyradiculoneuropathy'.

We report a case of multifocal demyelinating motor neuropathy in a patient with a 5-year history of progressive, asymmetric, predominantly motor weakness characterized by multifocal progression, multifocal conduction block, and lack of response to steroid therapy. Neuropathologic findings at autopsy showed an "inflammatory demyelinating polyradiculoneuropathy" in the motor cranial nerves and motor roots of peripheral nerves, an extensive deposition of IgG and focal accumulations of IgM in the peripheral nerve motor roots, and loss of motor neurons. These findings clearly document an inflammatory demyelinating polyradiculoneuropathy in multifocal demyelinating motor neuropathy, suggesting a close relation with chronic inflammatory demyelinating polyneuropathy.

Genetic modification of an entomopoxvirus: deletion of the spheroidin gene does not affect virus replication in vitro.

In the late stages of an entomopoxvirus infection, virions become embedded within a crystalline occlusion body or spheroid. Spheroids are composed primarily of a single polypeptide, spheroidin. We describe the construction of a genetically modified Amsacta moorei entomopoxvirus (AmEPV) in which the spheroidin gene coding sequences are deleted and replaced with those of a heterologous reporter gene encoding chloramphenicol acetyltransferase (CAT). A transfer vector, pAmCP1, was prepared containing a unique BamHI site in lieu of the spheroidin gene coding region, together with 1 kbp of upstream and downstream DNA sequence that flanks the spheroidin gene. The flanking sequences provide the transcriptional control signals and also guide homologous recombination so that the spheroidin gene coding region can be replaced with that of the foreign gene. The transfer vector was designed so that the translational start codon of the introduced foreign gene would be utilized. A recombinant virus, AmEPV.CAT, was produced by transfecting AmEPV-infected cells with the transfer vector encoding the CAT gene. The recombinant virus was isolated from wild-type virus by identifying plaques with a spheroidin-negative phenotype. Light microscopy and SDS-PAGE analysis demonstrated that no spheroids or spheroidin protein were produced in the recombinant virus-infected cells. The recombinant virus was able to replicate to high titres (10(7) p.f.u./ml) in insect cells indicating that the spheroidin gene is non-essential for AmEPV replication in vitro. Moderate levels of CAT were synthesized in recombinant virus-infected cells and temporal analyses indicated that CAT synthesis followed the pattern of spheroidin production suggesting that the spheroidin gene promoter was functioning under normal regulatory control in the genetically modified virus.

Replication and morphogenesis of Amsacta moorei entomopoxvirus in cultured cells of Estigmene acrea (salt marsh caterpillar).

A study of the sequence of morphogenic events occurring within Amsacta moorei entomopoxvirus-infected Estigmene acrea cells is presented. Stages in virion development, and the various cytopathic effects observed in these cells between 0 and 120 h post-infection (p.i.) are described. Events in the early stages of virion assembly (24 to 48 h p.i.), the formation of the outer viral membrane (48 to 72 h p.i.) and the development of occlusion bodies or spheroids (72 to 120 h p.i.) were identified. Cells grown in TC100 culture medium supported production of mature virus particles, the majority of which were either of the intracellular naked virion form, or mature virions incorporated into occlusion bodies. Only limited production of the extracellular enveloped form was observed in these cells.