145 research outputs found
Live-cell topology assessment of URG7, MRP6âââ and SP-C using glycosylatable green fluorescent protein in mammalian cells.
Experimental tools to determine membrane topology of a protein are rather limited in higher eukaryotic organisms. Here, we report the use of glycosylatable GFP (gGFP) as a sensitive and versatile membranetopology reporter in mammalian cells. gGFP selectively loses its fluorescence upon N-linked glycosylationin the ER lumen. Thus, positive fluorescence signal assigns location of gGFP to the cytosol whereas nofluorescence signal and a glycosylated status of gGFP map the location of gGFP to the ER lumen. By usingmammalian gGFP, the membrane topology of disease-associated membrane proteins, URG7, MRP6102,SP-C(Val) and SP-C(Leu) was confirmed. URG7 is partially targeted to the ER, and inserted in Cinform.MRP6102and SP-C(Leu/Val) are inserted into the membrane in Coutform. A minor population of untarget-ed SP-C is removed by proteasome dependent quality control system
Production of Secondary Organic Aerosol During Aging of Biomass Burning Smoke From Fresh Fuels and Its Relationship to VOC Precursors
After smoke from burning biomass is emitted into the atmosphere, chemical and physical processes change the composition and amount of organic aerosol present in the aged, diluted plume. During the fourth Fire Lab at Missoula Experiment, we performed smog-chamber experiments to investigate formation of secondary organic aerosol (SOA) and multiphase oxidation of primary organic aerosol (POA). We simulated atmospheric aging of diluted smoke from a variety of biomass fuels while measuring particle composition using high-resolution aerosol mass spectrometry. We quantified SOA formation using a tracer ion for low-volatility POA as a reference standard (akin to a naturally occurring internal standard). These smoke aging experiments revealed variable organic aerosol (OA) enhancements, even for smoke from similar fuels and aging mechanisms. This variable OA enhancement correlated well with measured differences in the amounts of emitted volatile organic compounds (VOCs) that could subsequently be oxidized to form SOA. For some aging experiments, we were able to predict the SOA production to within a factor of 2 using a fuel-specific VOC emission inventory that was scaled by burn-specific toluene measurements. For fires of coniferous fuels that were dominated by needle burning, volatile biogenic compounds were the dominant precursor class. For wiregrass fires, furans were the dominant SOA precursors. We used a POA tracer ion to calculate the amount of mass lost due to gas-phase oxidation and subsequent volatilization of semivolatile POA. Less than 5% of the POA mass was lost via multiphase oxidation-driven evaporation during up to 2 hr of equivalent atmospheric oxidation
Reactive intermediates revealed in secondary organic aerosol formation from isoprene
Isoprene is a significant source of atmospheric organic aerosol; however, the oxidation pathways that lead to secondary organic aerosol (SOA) have remained elusive. Here, we identify the role of two key reactive intermediates, epoxydiols of isoprene (IEPOX = ÎČ-IEPOX + ÎŽ-IEPOX) and methacryloylperoxynitrate (MPAN), which are formed during isoprene oxidation under low- and high-NO_x conditions, respectively. Isoprene low-NO_x SOA is enhanced in the presence of acidified sulfate seed aerosol (mass yield 28.6%) over that in the presence of neutral aerosol (mass yield 1.3%). Increased uptake of IEPOX by acid-catalyzed particle-phase reactions is shown to explain this enhancement. Under high-NO_x conditions, isoprene SOA formation occurs through oxidation of its second-generation product, MPAN. The similarity of the composition of SOA formed from the photooxidation of MPAN to that formed from isoprene and methacrolein demonstrates the role of MPAN in the formation of isoprene high-NO_x SOA. Reactions of IEPOX and MPAN in the presence of anthropogenic pollutants (i.e., acidic aerosol produced from the oxidation of SO_2 and NO_2, respectively) could be a substantial source of âmissing urban SOAâ not included in current atmospheric models
Secondary aerosol formation from photochemical aging of aircraft exhaust in a smog chamber
Field experiments were performed to investigate the effects of photo-oxidation on fine particle emissions from an in-use CFM56-2B gas turbine engine mounted on a KC-135 Stratotanker airframe. Emissions were sampled into a portable smog chamber from a rake inlet installed one-meter downstream of the engine exit plane of a parked and chocked aircraft. The chamber was then exposed to sunlight and/or UV lights to initiate photo-oxidation. Separate tests were performed at different engine loads (4, 7, 30, 85 %). Photo-oxidation created substantial secondary particulate matter (PM), greatly exceeding the direct PM emissions at each engine load after an hour or less of aging at typical summertime conditions. After several hours of photo-oxidation, the ratio of secondary-to-primary PM mass was on average 35 ± 4.1, 17 ± 2.5, 60 ± 2.2, and 2.7 ± 1.1 for the 4, 7, 30, and 85 % load experiments, respectively. The composition of secondary PM formed strongly depended on load. At 4 % load, secondary PM was dominated by secondary organic aerosol (SOA). At higher loads, the secondary PM was mainly secondary sulfate. A traditional SOA model that accounts for SOA formation from single-ring aromatics and other volatile organic compounds underpredicts the measured SOA formation by ~60 % at 4 % load and ~40 % at 85 % load. Large amounts of lower-volatiliy organic vapors were measured in the exhaust; they represent a significant pool of SOA precursors that are not included in traditional SOA models. These results underscore the importance of accounting for atmospheric processing when assessing the influence of aircraft emissions on ambient PM levels. Models that do not account for this processing will likely underpredict the contribution of aircraft emissions to local and regional air pollution
High-spatial-resolution mapping and source apportionment of aerosol composition in Oakland, California, using mobile aerosol mass spectrometry
We investigated spatial and temporal patterns in the concentration and
composition of submicron particulate matter (PM1) in Oakland,
California, in the summer of 2017 using an aerosol mass spectrometer mounted
in a mobile laboratory. We performed ââŒâ160 h of mobile sampling in the
city over a 20-day period. Measurements are compared for three adjacent
neighborhoods with distinct land uses: a central business district
(downtown), a residential district (West Oakland), and a major
shipping port (port). The average organic aerosol (OA) concentration is
5.3 ”g mâ3 and contributes ââŒâ50 % of the PM1
mass. OA concentrations in downtown are, on average,
1.5 ”g mâ3 higher than in West Oakland and port. We
decomposed OA into three factors using positive matrix factorization:
hydrocarbon-like OA (HOA; 20 % average contribution), cooking OA (COA;
25 %), and less-oxidized oxygenated OA (LO-OOA; 55 %). The collective
45 % contribution from primary OA (HOA + COA) emphasizes the
importance of primary emissions in Oakland. The dominant source of primary OA
shifts from HOA-rich in the morning to COA-rich after lunchtime. COA in
downtown is consistently higher than West Oakland and port due to a large
number of restaurants. HOA exhibits variability in space and time. The
morning-time HOA concentration in downtown is twice that in port, but port
HOA increases more than two-fold during midday, likely because trucking
activity at the port peaks at that time. While it is challenging to
mathematically apportion traffic-emitted OA between drayage trucks and cars,
combining measurements of OA with black carbon and CO suggests that while
trucks have an important effect on OA and BC at the port, gasoline-engine
cars are the dominant source of traffic emissions in the rest of Oakland.
Despite the expectation of being spatially uniform, LO-OOA also exhibits
spatial differences. Morning-time LO-OOA in downtown is roughly 25 %
(ââŒâ0.6 ”g mâ3) higher than the rest of Oakland. Even as
the entire domain approaches a more uniform photochemical state in the
afternoon, downtown LO-OOA remains statistically higher than West Oakland and
port, suggesting that downtown is a microenvironment with higher
photochemical activity. Higher concentrations of particulate sulfate (also of
secondary origin) with no direct sources in Oakland further reflect higher
photochemical activity in downtown. A combination of several factors (poor
ventilation of air masses in street canyons, higher concentrations of
precursor gases, higher concentrations of the hydroxyl radical) likely
results in the proposed high photochemical activity in downtown. Lastly,
through Van Krevelen analysis of the elemental ratios (H â C, O â C)
of the OA, we show that OA in Oakland is more chemically reduced than several
other urban areas. This underscores the importance of primary emissions in
Oakland. We also show that mixing of oceanic air masses with these primary
emissions in Oakland is an important processing mechanism that governs the
overall OA composition in Oakland.</p
A dual-chamber method for quantifying the effects of atmospheric perturbations on secondary organic aerosol formation from biomass burning emissions
Biomass burning (BB) is a major source of atmospheric pollutants. Field and laboratory studies indicate that secondary organic aerosol (SOA) formation from BB emissions is highly variable. We investigated sources of this variability using a novel dual-smog-chamber method that directly compares the SOA formation from the same BB emissions under two different atmospheric conditions. During each experiment, we filled two identical Teflon smog chambers simultaneously with BB emissions from the same fire. We then perturbed the smoke with UV lights, UV lights plus nitrous acid (HONO), or dark ozone in one or both chambers. These perturbations caused SOA formation in nearly every experiment with an average organic aerosol (OA) mass enhancement ratio of 1.78 ± 0.91 (mean ± 1Ï). However, the effects of the perturbations were highly variable ranging with OA mass enhancement ratios ranging from 0.7 (30% loss of OA mass) to 4.4 across the set of perturbation experiments. There was no apparent relationship between OA enhancement and perturbation type, fuel type, and modified combustion efficiency. To better isolate the effects of different perturbations, we report dual-chamber enhancement (DUCE), which is the quantity of the effects of a perturbation relative to a reference condition. DUCE values were also highly variable, even for the same perturbation and fuel type. Gas measurements indicate substantial burn-to-burn variability in the magnitude and composition of SOA precursor emissions, even in repeated burns of the same fuel under nominally identical conditions. Therefore, the effects of different atmospheric perturbations on SOA formation from BB emissions appear to be less important than burn-to-burn variability
Production of Secondary Organic Aerosol During Aging of Biomass Burning Smoke From Fresh Fuels and Its Relationship to VOC Precursors
After smoke from burning biomass is emitted into the atmosphere, chemical and physical processes change the composition and amount of organic aerosol present in the aged, diluted plume. During the fourth Fire Lab at Missoula Experiment, we performed smog-chamber experiments to investigate formation of secondary organic aerosol (SOA) and multiphase oxidation of primary organic aerosol (POA). We simulated atmospheric aging of diluted smoke from a variety of biomass fuels while measuring particle composition using high-resolution aerosol mass spectrometry. We quantified SOA formation using a tracer ion for low-volatility POA as a reference standard (akin to a naturally occurring internal standard). These smoke aging experiments revealed variable organic aerosol (OA) enhancements, even for smoke from similar fuels and aging mechanisms. This variable OA enhancement correlated well with measured differences in the amounts of emitted volatile organic compounds (VOCs) that could subsequently be oxidized to form SOA. For some aging experiments, we were able to predict the SOA production to within a factor of 2 using a fuel-specific VOC emission inventory that was scaled by burn-specific toluene measurements. For fires of coniferous fuels that were dominated by needle burning, volatile biogenic compounds were the dominant precursor class. For wiregrass fires, furans were the dominant SOA precursors. We used a POA tracer ion to calculate the amount of mass lost due to gas-phase oxidation and subsequent volatilization of semivolatile POA. Less than 5% of the POA mass was lost via multiphase oxidation-driven evaporation during up to 2 hr of equivalent atmospheric oxidation
Practical and clinical utility of non-invasive vagus nerve stimulation (nVNS) for the acute treatment of migraine. A post hoc analysis of the randomized, sham-controlled, double-blind PRESTO trial
Background: The PRESTO study of non-invasive vagus nerve stimulation (nVNS; gammaCoreÂź) featured key primary and secondary end points recommended by the International Headache Society to provide Class I evidence that for patients with an episodic migraine, nVNS significantly increases the probability of having mild pain or being pain-free 2 h post stimulation. Here, we examined additional data from PRESTO to provide further insights into the practical utility of nVNS by evaluating its ability to consistently deliver clinically meaningful improvements in pain intensity while reducing the need for rescue medication. Methods: Patients recorded pain intensity for treated migraine attacks on a 4-point scale. Data were examined to compare nVNS and sham with regard to the percentage of patients who benefited by at least 1 point in pain intensity. We also assessed the percentage of attacks that required rescue medication and pain-free rates stratified by pain intensity at treatment initiation. Results: A significantly higher percentage of patients who used acute nVNS treatment (n = 120) vs sham (n = 123) reported a â„ 1-point decrease in pain intensity at 30 min (nVNS, 32.2%; sham, 18.5%; P = 0.020), 60 min (nVNS, 38.8%; sham, 24.0%; P = 0.017), and 120 min (nVNS, 46.8%; sham, 26.2%; P = 0.002) after the first attack. Similar significant results were seen when assessing the benefit in all attacks. The proportion of patients who did not require rescue medication was significantly higher with nVNS than with sham for the first attack (nVNS, 59.3%; sham, 41.9%; P = 0.013) and all attacks (nVNS, 52.3%; sham, 37.3%; P = 0.008). When initial pain intensity was mild, the percentage of patients with no pain after treatment was significantly higher with nVNS than with sham at 60 min (all attacks: nVNS, 37.0%; sham, 21.2%; P = 0.025) and 120 min (first attack: nVNS, 50.0%; sham, 25.0%; P = 0.018; all attacks: nVNS, 46.7%; sham, 30.1%; P = 0.037). Conclusions: This post hoc analysis demonstrated that acute nVNS treatment quickly and consistently reduced pain intensity while decreasing rescue medication use. These clinical benefits provide guidance in the optimal use of nVNS in everyday practice, which can potentially reduce use of acute pharmacologic medications and their associated adverse events. Trial registration: ClinicalTrials.gov identifier: NCT02686034
VaporâWall Deposition in Chambers: Theoretical Considerations
In order to constrain the effects of vaporâwall deposition on measured secondary organic aerosol (SOA) yields in laboratory chambers, researchers recently varied the seed aerosol surface area in toluene oxidation and observed a clear increase in the SOA yield with increasing seed surface area (Zhang, X.; et al. Proc. Natl. Acad. Sci. U.S.A. 2014, 111, 5802). Using a coupled vaporâparticle dynamics model, we examine the extent to which this increase is the result of vaporâwall deposition versus kinetic limitations arising from imperfect accommodation of organic species into the particle phase. We show that a seed surface area dependence of the SOA yield is present only when condensation of vapors onto particles is kinetically limited. The existence of kinetic limitation can be predicted by comparing the characteristic time scales of gas-phase reaction, vaporâwall deposition, and gasâparticle equilibration. The gasâparticle equilibration time scale depends on the gasâparticle accommodation coefficient α_p. Regardless of the extent of kinetic limitation, vaporâwall deposition depresses the SOA yield from that in its absence since vapor molecules that might otherwise condense on particles deposit on the walls. To accurately extrapolate chamber-derived yields to atmospheric conditions, both vaporâwall deposition and kinetic limitations must be taken into account
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