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The apportionment of the contribution of wood smoke emitted particles to the total concentration of particulate matter in a region has been greatly aided by the development of new analytical methods. These analytical methods quantitatively determine organic marker compounds unique to wood combustion such as levoglucosan and dehydroabietic acid. These markers have generally been determined in 24-hour averaged samples. We have developed an instrument based on the collection of particles on an inert filter, desorption of the organic material in an inert atmosphere with subsequent GC separation and MS detection of the desorbed compounds. The GC-MS Organic Aerosol Monitor (OAM) instrument has been used in three field studies. An unexpected finding from these studies was the quantification of the contribution of secondary organic aerosols from gases present in wood smoke in addition to primary wood smoke emitted particles. The identification of this secondary material was made possible by the collection of hourly averaged data that allowed for the time patterns of black carbon, organic material, and wood smoke marker compounds to be included and compared in a Positive Matrix Factorization (PMF) analysis. Most of the organic markers associated with wood smoke (levoglucosan, stearic acid and dehydroabietic acid) are associated with primary wood smoke emissions, but a fraction of the levoglucosan and stearic acid are also associated with secondary organic material formed from gaseous precursors in wood smoke. Additionally, this secondary material was shown to be present in each in of the three urban area where wood smoke burning occurs. There is a need for additional studies to better understand the contribution of secondary particulate formation from both urban and wildfires.iImplications/i: This paper presents results from three field studies which show that in addition to the formation of primary particulate matter from the combustion of wood smoke and secondary particulate matter is also formed from the gaseous compounds emitted with the wood smoke. This material is identified in the studies of wood combustion reported here by the identification and quantification of specific organic marker compounds related to wood combustion and is shown to and represents a contributor nearly as large as the primary emitted material and better quantifying the impact of wood combustion on airborne fine particulate matter.

During August and September 2012, a study was conducted to determine the sources of PM2.5 adjacent to the I-710 Long Beach Freeway. The site is directly affected by the emissions from heavy diesel traffic flowing from major container ports about 10 km south of the sampling site. Hourly average data were obtained for particulate species including PM2.5, black carbon and UV absorbing carbon, EC, fine particulate nonvolatile and semi-volatile organic material (NVOM and SVOM), sulfate, nitrate, chloride, ammonium ion, and Na ion, and for related factors including O3, CO, NOX, SO2, and total traffic flow on the I-710. A total of 520 hourly averaged data sets with 15 measured variables were analyzed by EPA-PMF v5.0. The data were best described by a 10-factor solution. Based on the composition and diurnal patterns of the factors, they were assigned to three diesel-related factors (two of which appeared to represent traffic from the ports and one general freeway diesel factor), a light-duty, spark-ignition vehicle-related factor, three secondary factors (one of which was associated with O3 formation processes), and three factors dominated by sulfate, SO2, and chloride, respectively. The diurnal patterns for these last three factors are strongly correlated. Meteorological and refinery upset data indicate that they are associated with emissions from a nearby refinery. The results of the PMF analysis were combined with nephelometer light scattering, corrected for coarse particle scattering and estimated aerosol water content in a multilinear regression analysis to identify visibility degradation sources. Major contributors were the aerosol water content, and the secondary PMF factors associated with either Nitrate and NVOM or NVOM and SVOM. The use of hourly average data made possible the identification of factors associated with gasoline vehicle emissions and both port and non-port diesel emissions.Implications: Hourly averaged data were obtained for PM2.5, its components and factors related to primary emissions and the formation of secondary material at a near freeway sampling location adjacent to the I-710 freeway just south of the Long Beach Boulevard entrance and 10 km north of the Ports of Long Beach and Los Angeles. The major objective of the study was to determine the impact of traffic from the ports at the monitoring site. This manuscript reports on the PMF analysis of the data set. Factors related to both diesel traffic originating from the ports and diesel traffic from non-port origins were identified. The diesel traffic originating from the ports was responsible for 9% of the total traffic and 95% of the BC measured at the sampling site. The non-port diesel traffic was responsible for 15% of the total traffic and 5% of the BC. While the Port 1 diesel traffic coming from the ports contributed a large fraction of the BC, this source contributed only 2% of the CO and 5% of the NOX at the sampling site. The impact of these traffic sources on light scattering was also small. Analysis of sources of sulfate and SO2 at the sampling site indicated that these species did not come from port activities of ships at or approaching the port, but rather from upset flare events at a nearby oil refinery.

A study was conducted on the Brigham Young University campus during January and February 2015 to identify winter-time sources of fine particulate material in Utah Valley, Utah. Fine particulate mass and components and related gas-phase species were all measured on an hourly averaged basis. Light scattering was also measured during the study. Included in the sampling was the first-time source apportionment application of a new monitoring instrument for the measurement of fine particulate organic marker compounds on an hourly averaged basis. Organic marker compounds measured included levoglucosan, dehydroabietic acid, stearic acid, pyrene, and anthracene. A total of 248 hourly averaged data sets were available for a positive matrix factorization (PMF) analysis of sources of both primary and secondary fine particulate material. A total of nine factors were identified. The presence of wood smoke emissions was associated with levoglucosan, dehydroabietic acid, and pyrene markers. Fine particulate secondary nitrate, secondary organic material, and wood smoke accounted for 90% of the fine particulate material. Fine particle light scattering was dominated by sources associated with wood smoke and secondary ammonium nitrate with associated modeled fine particulate water.The identification of sources and secondary formation pathways leading to observed levels of PM

Particulate matter (PM) is among the most harmful air pollutants to human health, but due to its complex chemical composition is poorly characterized. A large fraction of PM is composed of organic compounds, but these compounds are not regularly monitored due to limitations in current sampling and analysis techniques. The Organic Aerosol Monitor (GC-MS OAM) combines a collection device with thermal desorption, gas chromatography and mass spectrometry to quantitatively measure the carbonaceous components of PM on an hourly averaged basis. The GC-MS OAM is fully automated and has been successfully deployed in the field. It uses a chemically deactivated filter for collection followed by thermal desorption and GC-MS analysis. Laboratory tests show that detection limits range from 0.2 to 3 ng for 16 atmospherically relevant compounds, with the possibility for hundreds more. The GC-MS OAM was deployed in the field for semi-continuous measurement of the organic markers, levoglucosan, dehydroabietic acid, and polycyclic aromatic hydrocarbons (PAHs) from January to March 2015. Results illustrate the significance of this monitoring technique to characterize the organic components of PM and identify sources of pollution.

During August and September 2012, a study was conducted to determine the sources of PM2.5 adjacent to the I-710 Long Beach Freeway. The site is directly affected by the emissions from heavy diesel traffic flowing from major container ports about 10 km south of the sampling site. Hourly average data were obtained for particulate species including PM2.5, black carbon and UV absorbing carbon, EC, fine particulate nonvolatile and semi-volatile organic material (NVOM and SVOM), sulfate, nitrate, chloride, ammonium ion, and Na ion, and for related factors including O3, CO, NOX, SO2, and total traffic flow on the I-710. A total of 520 hourly averaged data sets with 15 measured variables were analyzed by EPA-PMF v5.0. The data were best described by a 10-factor solution. Based on the composition and diurnal patterns of the factors, they were assigned to three diesel-related factors (two of which appeared to represent traffic from the ports and one general freeway diesel factor), a light-duty, spark-ignition vehicle-related factor, three secondary factors (one of which was associated with O3 formation processes), and three factors dominated by sulfate, SO2, and chloride, respectively. The diurnal patterns for these last three factors are strongly correlated. Meteorological and refinery upset data indicate that they are associated with emissions from a nearby refinery. The results of the PMF analysis were combined with nephelometer light scattering, corrected for coarse particle scattering and estimated aerosol water content in a multilinear regression analysis to identify visibility degradation sources. Major contributors were the aerosol water content, and the secondary PMF factors associated with either Nitrate and NVOM or NVOM and SVOM. The use of hourly average data made possible the identification of factors associated with gasoline vehicle emissions and both port and non-port diesel emissions.

This study examined the effect of neurodegeneration, and its interaction with Alzheimer's disease (AD) cerebrospinal fluid biomarkers, on longitudinal verbal learning and memory performance in cognitively unimpaired (CU) late middle-aged adults. Three hundred and forty-two CU adults (cognitive baseline mean age = 58.4), with cerebrospinal fluid and structural MRI, completed 2–10 (median = 5) cognitive assessments. Learning and memory were assessed using the Rey Auditory Verbal Learning Test (RAVLT). We used sequential comparison of nested linear mixed effects models to analyze the data. Model selection preserved a significant ptau181/Aβ42 × global atrophy × age interaction; individuals with less global atrophy and lower ptau181/Aβ42 levels had less learning and delayed recall decline than individuals with more global atrophy and/or higher levels of ptau181/Aβ42. The hippocampal volume × age × ptau181/Aβ42 interaction was not significant. Findings suggest that in a sample of CU late middle-aged adults, individuals with AD biomarkers, global atrophy, or both evidence greater verbal learning and memory decline than individuals without either risk factor.

Knowledge of black carbon (BC) emission factors from ships is important from human health and environmental perspectives. A study of instruments measuring BC and fuels typically used in marine operation was carried out on a small marine engine. Six analytical methods measured the BC emissions in the exhaust of the marine engine operated at two load points (25% and 75%) while burning one of three fuels: a distillate marine (DMA), a low sulfur, residual marine (RMB-30) and a high-sulfur residual marine (RMG-380). The average emission factors with all instruments increased from 0.08 to 1.88 gBC/kg fuel in going from 25 to 75% load. An analysis of variance (ANOVA) tested BC emissions against instrument, load, and combined fuel properties and showed that both engine load and fuels had a statistically significant impact on BC emission factors. While BC emissions were impacted by the fuels used, none of the fuel properties investigated (sulfur content, viscosity, carbon residue and CCAI) was a primary driver for BC emissions. Of the two residual fuels, RMB-30 with the lower sulfur content, lower viscosity and lower residual carbon, had the highest BC emission factors. BC emission factors determined with the different instruments showed a good correlation with the PAS values with correlation coefficients R2 >0.95. A key finding of this research is the relative BC measured values were mostly independent of load and fuel, except for some instruments in certain fuel and load combinations.

A novel approach for measurement of stable carbon isotopic ratio of atmospheric aerosols was developed by tandem operation of two instruments: a Sunset Organic Carbon-Elemental Carbon (OC-EC) analyzer and an online Carbon Dioxide Stable Isotope Analyzer (LGR, CCIA-36d). Sensitivity, accuracy and measurement uncertainty of the CCIA was comprehensively investigated using the standard reference CO_2 gas with known concentration and isotopic ratio. Drift in CCIA measurement due to varying CO_2 and water vapor concentration was evaluated and a humidity stabilizer was designed and developed to control the water vapor concentration of exhaust gas flow from OC-EC prior to entering the CCIA. A Keeling approach was applied to separate the ratio in the samples from the mixture of PM sample-produced CO_2 and reference gas and we developed a protocol to derive the isotopic composition of the particle samples. A lithium carbonate standard (in powder form) from National Institute of Standards and Technology (NIST) was used to validate measurement of δ^(13)C ratios by CCIA. Offline measurement on ambient aerosol and diesel exhaust aerosols produced comparable results of isotopic ratio with literature values. This study demonstrates the utility of this tandem operation for carbon isotopic measurement of atmospheric particles with better than 1.0 per mille precision as a cost-effective alternative of conventional Isotopic Ratio Mass Spectrometer (IR-MS).

During the summer of 2012 and 2013, we measured carbon monoxide (CO), carbon dioxide (CO2), ozone (O3), nitrogen oxides (NOx), reactive nitrogen (NOy), peroxyacetyl nitrate (PAN), aerosol scattering (σsp) and absorption, elemental and organic carbon (EC and OC), and aerosol chemistry at the Mount Bachelor Observatory (2.8 km above sea level, Oregon, US). Here we analyze 23 of the individual plumes from regional wildfires to better understand production and loss of aerosols and gaseous species. We also developed a new method to calculate enhancement ratios and Modified Combustion Efficiency (MCE), which takes into account possible changes in background concentrations during transport. We compared this new method to existing methods for calculating enhancement ratios. The MCE values ranged from 0.79–0.98, ΔO3/ΔCO ranged from 0.01–0.07 ppbv ppbv–1, Δσsp/ΔCO ranged from 0.23–1.32 Mm–1 (at STP) ppbv–1, ΔNOy/ΔCO ranged from 2.89–12.82 pptv ppbv–1, and ΔPAN/ΔCO ranged from 1.46–6.25 pptv ppbv–1. A comparison of three different methods to calculate enhancement ratios (ER) showed that the methods generally resulted in similar Δσsp/ΔCO, ΔNOy/ΔCO, and ΔPAN/ΔCO; however, there was a significant bias between the methods when calculating ΔO3/ΔCO due to the small absolute enhancement of O3 in the plumes. The ΔO3/ΔCO ERs calculated using two common methods were biased low (~20–30%) when compared to the new proposed method. Two pieces of evidence suggest moderate secondary particulate formation in many of the plumes studied: 1) mean observed ΔOC/ΔCO2 was 0.028 g particulate-C gC–1 (as CO2)—27% higher than the midpoint of the biomass burning emission ratio range reported by a recent review—and 2) single scattering albedo (ω) was relatively constant at all MCE values, in contrast with results for fresh plumes. The observed NOx, PAN, and aerosol nitrate represented 6–48%, 25–57%, and 20–69% of the observed NOy in the aged plumes, respectively, and other species represented on average 11% of the observed NOy.

One potential obstacle limiting our ability to clarify ADHD etiology is the heterogeneity within the disorder, as well as in typical samples. In this study, we utilized a community detection approach on 106 children with and without ADHD (aged 7–12 years), in order to identify potential subgroups of participants based on the connectivity of the reward system. Children with ADHD were compared to typically developing children within each identified community, aiming to find the community-specific ADHD characteristics. Furthermore, to assess how the organization in subgroups relates to behavior, we evaluated delay-discounting gradient and impulsivity-related temperament traits within each community. We found that discrete subgroups were identified that characterized distinct connectivity profiles in the reward system. Importantly, which connections were atypical in ADHD relative to the control children were specific to the community membership. Our findings showed that children with ADHD and typically developing children could be classified into distinct subgroups according to brain functional connectivity. Results also suggested that the differentiation in “functional” subgroups is related to specific behavioral characteristics, in this case impulsivity. Thus, combining neuroimaging data and community detection might be a valuable approach to elucidate heterogeneity in ADHD etiology and examine ADHD neurobiology.

In this study concentrations of organic (OC) and elemental carbon (EC) from free tropospheric (FT) fine particulate matter (PM) were measured from March to September, 2012 with a Semi-Continuous OC/EC carbon aerosol analyzer at the top of Mt. Bachelor (2.8 km a.s.l) in Central Oregon, U.S. The average concentrations of OC and EC in the FT were low (OC: 1.87 ± 6.10, EC: 0.07 ± 0.26 μg m −3 ; average ± SD) but much higher during specific pollution episodes. During springtime the highest OC and EC concentrations were measured for dry free tropospheric air masses, whereas during summertime the highest OC and EC concentrations were typically measured for more humid air masses that were uplifted from the boundary layer (BL). The highest OC and EC concentrations were measured during biomass burning episodes (3 h average OC: up to 146.0 μg m −3 , EC up to 5.5 μg m −3 ). Elevated OC and EC concentrations were also measured during Asian Long Range Transport (LRT) episodes (OC: up to 3.6 μg m −3 , EC up to 1.1 μg m −3 ). In addition, between episodes, an increase in OC was seen in the afternoon, possibly due to SOA formation. This SOA can then be exported to the FT via diurnal ventilation of the BL. For Asian LRT episodes the OC/EC ratios varied between 8 and 34, with an average of 17.9. For local biomass burning emissions OC/EC ratios were between 25 and 30. Higher OC/EC ratios (30–40) were observed for the biomass burning plumes originating from longer distances, possibly due to SOA formation.

A significant need exists to better characterize air pollution and its sources. This especially pertains to fine particulate matter (PM2.5). PM2.5 is chemically complex and its sources of emission and secondary production are highly variable. PM2.5 complexity is largely due to the organic fraction, which ranges from 10 to 90% of its total mass. However, the organic compounds in PM2.5 have generally not been monitored in the field due to limitations in available sampling techniques. A source apportionment study was conducted at the Neil Armstrong Academy campus, 5194 Highbury Pkwy, West Valley City, UT 84120, during January and February 2016. Among the objectives of the study was to identify winter-time sources of fine particulate material in West Valley City, Utah. Fine particulate mass and components, particulate organic marker and related gas phase species were all measured on an hourly average basis. The following hourly averaged data were used in the Positive Matrix Factorization (PMF) analysis, fine particulate mass, cations and anions, carbonaceous material, organic marker compounds, gas phase species including NOX, NO2 and CO and mass spectrometric measurements of gas phase methanol and C8 aromatics. A total of 557 hourly averaged data sets with 23 components were available for an EPA Positive Matrix Factorization (v5.0) analysis of the data set. The data were best described by a solution with 7 factors. Two of the factors were associated with emissions from mobile sources, diesel and automotive. One factor was associated with wood smoke emissions. Three factors were associated with the formation of secondary fine particulate material. The larger of the three was associated with the production of secondary nitrate and accounted for 70% of the PM2.5, while the other two were associated with secondary formation of particulate organic material from wood smoke associated gas phase organic compounds but accounted for only 3% of the total PM2.5 mass. The final factor was associated with fine particulate sulfate and SO2. Meteorological back-trajectory data clearly indicted the source was associated with emissions from a copper smelter to the west of the sampling site.

Current research in connectomics highlights that self-organized functional networks or "communities" of cortical areas can be detected in the adult brain. This perspective may provide clues to mechanisms of treatment response in psychiatric conditions. Here we examine functional brain community topology based on resting-state fMRI in adult Attention-Deficit/Hyperactivity Disorder (ADHD; n = 22) and controls (n = 31). We sought to evaluate ADHD patterns in adulthood and their modification by short term stimulants administration. Participants with ADHD were scanned one or two weeks apart, once with medication and once without; comparison participants were scanned at one time-point. Functional connectivity was estimated from these scans and community detection applied to determine cortical network topology. Measures of change in connectivity profile were calculated via a graph measure, termed the Node Dissociation Index (NDI). Compared to controls, several cortical networks had atypical connectivity in adults with ADHD when withholding stimulants, as measured by NDI. In most networks stimulants significantly reduced, but did not eliminate, differences in the distribution of connections between key brain systems relative to the control sample. These findings provide an enriched model of connectivity in ADHD and demonstrate how stimulants may exert functional effects by altering connectivity profiles in the brain.

Wall shear stresses (WSS) exerted by blood flow on cardiac tissues modulate growth and development of the heart. To study the role of hemodynamic conditions on cardiac morphogenesis, here, we present a methodology that combines imaging and finite element modeling to quantify the in vivo blood flow dynamics and WSS in the cardiac outflow tract (OFT) of early chicken embryos (day 3 out of 21-day incubation period). We found a distinct blood flow field and heterogeneous distribution of WSS in the chicken embryonic heart OFT, which have physiological implications for OFT morphogenesis.

Background: Several neurodegeneration (N) metrics using structural MRI are used for the purpose of Alzheimer's disease (AD)-related staging, including hippocampal volume, global atrophy, and an “AD signature” composite consisting of thickness or volumetric estimates derived from regions impacted early in AD. This study sought to determine if less user-intensive estimates of global atrophy and hippocampal volume were equivalent to a thickness-based AD signature from FreeSurfer for defining N across the AD continuum (i.e., individuals who are amyloid-positive (A+)). Methods: Cognitively unimpaired (CU) late middle-aged and older adults, as well as A+ mild cognitive impairment (MCI) and A+ AD dementia individuals, with available CSF and structural MRI scan

In order to study particulate matter transport and transformation in the Megacity environment, fine particulate carbon was measured simultaneously at two supersites, suburban T1 and rural T2, downwind of Mexico City during the MILAGRO field campaign in March 2006. Organic carbon (OC), element carbon (EC), and total carbon (TC=OC+EC) were determined in near real-time using a Sunset semi-continuous OCEC field analyzer. The semi-empirical EC tracer method was used to derive primary organic carbon (POC) and secondary organic carbon (SOC). Diurnal variations of primary and secondary carbon were observed at T1 and T2, which resulted from boundary layer inversion and impacted by local traffic patterns. The majority of organic carbon particles at T1 and T2 were secondary. The SOCTC% (SOC%=SOC/TC×100%) at T1 ranged from 0.5–93.8% with an average of 63.5±17.2%. The SOCTC% at T2 ranged from 9.3–98.1% with an average of 67.4±12.4%. The average EC to PM2.5 percentage (ECPM%=EC/PM2.5×100%) and OCPM% were 6.0% and 20.0% over the whole sampling time at T1. The POC to PM percentage (POCPM%) and SOCPM% were 3.7% and 16.3%, respectively at the same site. The maximum ECPM% was 21.2%, and the maximum OCPM% was 57.2% at T1. The maximum POCPM% was 12.9%, and the maximum SOCPM% was 49.7% at T1. Comparison of SOC and POC at T1 and T2 showed similar characteristics under favorable meteorological conditions, which indicated that transport from T1 towards T2 took place. Strong correlations between EC and carbon monoxide (CO) and odd nitrogen species (NO and NOx) were observed at T1. This indicated that EC had nearby sources, such as local traffic emissions. The EC/CO ratio derived by linear regression analysis, with units of μg C/m3 and μg/m3, respectively, was 0.004 at T1. Correlations were also seen between OC and SOC vs. the sum of oxidants, such as O3 and NO2, suggesting the secondary nature of carbons observed at T1.

As part of a major atmospheric chemistry and aerosol field program carried out in March 2006, a study was conducted in the area to the north and northeast of Mexico City to investigate the evolution of aerosols and their associated optical properties in the first few hours after their emission. The focus of the T1-T2 aerosol study was to investigate changes in the specific absorption αABS (absorption per unit mass, with unit of m2 g−1) of black carbon as it aged and became coated with compounds such as sulfate and organic carbon, evolving from an external to an internal mixture. Such evolution has been reported in previous studies. The T1 site was located just to the north of the Mexico City metropolitan area; the T2 site was situated approximately 35 km farther to the northeast. Nephelometers, particle soot absorption photometers, photoacoustic absorption spectrometers, and organic and elemental carbon analyzers were used to measure the optical properties of the aerosols and the carbon concentrations at each of the sites. Radar wind profilers and radiosonde systems helped to characterize the meteorology and to identify periods when transport from Mexico City over T1 and T2 occurred. Organic and elemental carbon concentrations at T1 showed diurnal cycles reflecting the nocturnal and early morning buildup from nearby sources, while concentrations at T2 appeared to be more affected by transport from Mexico City. Specific absorption during transport periods was lower than during other times, consistent with the likelihood of fresher emissions being found when the winds blew from Mexico City over T1 and T2. The specific absorption at T2 was larger than at T1, which is also consistent with the expectation of more aged particles with encapsulated black carbon being found at the more distant location. In situ measurements of single scattering albedo with an aircraft and a ground station showed general agreement with column-averaged values derived from rotating shadowband radiometer data, although some differences were found that may be related to boundary-layer evolution.

Reference Material (RM) 8785 Air Particulate Matter on Filter Media is intended primarily for use in the evaluation of analytical methods used to characterize the carbon composition of atmospheric fine particulate matter (PM) for national air quality monitoring programs. This RM consists of a fine fraction (nominally < 2.5 μ m aerodynamic diameter) of standard reference material (SRM) 1649a Urban Dust resuspended in air and filtered onto quartz-fiber filters. RM 8785 will also provide the atmospheric-chemistry and ocean-sciences communities with a means to intercompare methods and laboratories for the measurement of elemental (black) carbon. RM 8785 has values assigned for total carbon, elemental carbon, and organic carbon mass fractions measured according to two thermal–optical methods: the Interagency Monitoring of Protected Visual Environments (IMPROVE) and Speciation Trends Network–National Institute of Occupational Safety and Health (STN–NIOSH) protocols. Each filter is uniquely identified by its air...

A commercial semi-continuous elemental and organic carbon (ECOC) analyzer has been developed by Sunset Laboratory Inc. for the analysis of atmospheric particulate matter. The field deployable instrument is an alternative to off-line laboratory-based analysis of samples collected in the field. Although the Sunset Laboratory analyzer provides time-resolved particulate ECOC measurements using a methodology comparable to the laboratory-based NIOSH 5040 ECOC measurement, the performance of the semi-continuous ECOC analyzer has not been fully validated in the past. As part of the St. Louis-Midwest Supersite, two semi-continuous Sunset Labs ECOC field analyzers were operated to measure fine particle organic and elemental carbon for the entire year of 2002. The dual instrument operation provided a means to have a true continuous ECOC measurement, which would not be possible with only one instrument since the instrument cannot continue sample collection during the sample analysis period. In parallel with the semi-continuous ECOC analyzers, daily 24-h integrated PM2.5 samples were collected for ECOC analysis in the laboratory. Both the 24-h integrated filter samplers and the semi-continuous ECOC instruments were operated downstream of an organics denuder to remove positive adsorption artifacts associated with semi-volatile organic compounds. Comparison of 24-h averaged semi-continuous fine particle total carbon (TC), organic carbon (OC) and elemental carbon (EC) showed good agreement with the daily integrated off-line measurements. The linear regression of the averaged semi-continuous TC on the integrated off-line TC had a coefficient of regression (R2) of 0.89 and a regression slope of 0.97±0.02 when at least 20 h of the day were measured. The OC measurements had a R2 equal to 0.90 and a regression slope of 0.93±0.02. The EC measurements had a poorer R2 due to the very low levels of EC at the sampling site but a regression slope of 0.95±0.05. The accuracy of using a single semi-continuous ECOC instrument, configured to measure ECOC every other hour to represent the daily 24-h average, was also evaluated.

Because of increased interest in the marine and atmospheric sciences in elemental carbon (EC), or black carbon (BC) or soot carbon (SC), and because of the difficulties in analyzing or even defining this pervasive component of particulate carbon, it has become quite important to have appropriate reference materials for intercomparison and quality control. The NIST "urban dust" Standard Reference Material(®) SRM 1649a is useful in this respect, in part because it comprises a considerable array of inorganic and organic species, and because it exhibits a large degree of ((14)C) isotopic heterogeneity, with biomass carbon source contributions ranging from about 2 % (essentially fossil aliphatic fraction) to about 32 % (polar fraction). A primary purpose of this report is to provide documentation for the new isotopic and chemical particulate carbon data for the most recent (31 Jan. 2001) SRM 1649a Certificate of Analysis. Supporting this is a critical review of underlying international intercomparison data and methodologies, provided by 18 teams of analytical experts from 11 institutions. Key results of the intercomparison are: (1) a new, Certified Value for total carbon (TC) in SRM 1649a; (2) (14)C Reference Values for total carbon and a number of organic species, including for the first time 8 individual PAHs; and (3) elemental carbon (EC) Information Values derived from 13 analytical methods applied to this component. Results for elemental carbon, which comprised a special focus of the intercomparison, were quite diverse, reflecting the confounding of methodological-matrix artifacts, and methods that tended to probe more or less refractory regions of this universal, but ill-defined product of incomplete combustion. Availability of both chemical and (14)C speciation data for SRM 1649a holds great promise for improved analytical insight through comparative analysis (e.g., fossil/biomass partition in EC compared to PAH), and through application of the principle of isotopic mass balance.

Several advantages and disadvantages have been cited for image collection with a slow-scan CCD camera. Here we explore its use for cryo-EM single particle reconstruction and present two practical examples. The icosahedral adenovirus (Ad) type 2 (∼150 MDa) was reconstructed from 396 particle images. The Fourier shell correlation (FSC) 0.5 threshold and the Fourier shell phase residual (FSPR) 45°criterion yielded 17 AA resolution for the ordered viral capsid. Visual comparison with the filtered Ad2 crystallographic hexon confirmed a resolution range of 15–17 A. The asymmetric DNA-PKcs protein (470 kDa) was reconstructed from 9,473 particle images, using a previously published reconstruction based on class-sum images as an orientational search model [Chiu et al. (1998) J. Mol. Biol. 284:1075–1081]. FSC and FSPR methods yielded 17 A resolution for the new DNA-PKcs reconstruction, indicating a small but noticeable improvement over that of the class-sum based reconstruction. Despite the lack of symmetry for DNA-PKcs and its lower image contrast compared to Ad2 (0.8% vs. 2.5%), the same resolution was obtained for both particles by averaging significantly more DNA-PKcs images. Use of the CCD camera enables the microscopist to adjust the electron beam strength interactively and thereby maximize the image contrast for beam sensitive samples. On-line Fourier transformation also allows routine monitoring of drift and astigmatism during image collection, resulting in a high percentage of micrographs suitable for image processing. In conclusion, our results show that digital image collection with the YAG-scintillator slow-scan CCD camera is a viable approach for 3D reconstruction of both symmetric and asymmetric particles. Microsc. Res. Tech. 49:224–232, 2000. © 2000 Wiley-Liss, Inc.

The DNA-dependent protein kinase (DNA-PK) plays an important role in mammalian DNA double-strand break repair and immunoglobulin gene rearrangement. The DNA-PK holoenzyme is activated by assembly at DNA ends and is comprised of DNA-PKcs, a 460 kDa protein kinase catalytic subunit, and Ku, a 70 kDa/80 kDa heterodimeric DNA-targeting component. We have solved the three-dimensional structure of DNA-PKcs to approximately 21 A resolution by analytically combining images of nearly 9500 individual particles extracted from cryo-electron micrographs. The DNA-PKcs protein has an open, pseudo 2-fold symmetric structure with a gap separating a crown-shaped top from a rounded base. Columns of density are observed to protrude into the gap from both the crown and the base. Measurements of the enclosed volume indicate that the interior of the protein is largely hollow. The structure of DNA-PKcs suggests that its association with DNA may involve the internalization of double-stranded ends.

In mature striated muscle, intermediate filaments (IFs) are associated with the periphery of Z-discs and sites of myofibril-membrane attachment. Previously T. Schultheiss, Z. X. Lin, H. Ishikawa, I. Zamir, C. J. Stoeckert and H. Holtzer (1991) J. Cell Biol. 114, 953)reported that the disruption of IF organization in cultured chick myotubes had no detectable effect on muscle cell structure. Cultured muscle is not, however, under the mechanical loads characteristic of muscle in situ. The dorsal myotomal muscle (DMM) of the Xenopus tadpole provides an accessible model system in which to study the effects of mutant IF proteins on an intact, functional muscle. DNAs encoding truncated forms of Xenopus vimentin or desmin were injected into fertilized Xenopus eggs. Embryos were allowed to develop to the tadpole stage and then examined by confocal or electron microscopy. DMM cells containing the truncated IF polypeptides displayed disorganized IF systems. While the alignment of Z-lines appeared unaffected, cells accumulating mutant IF polypeptides displayed abnormal organization at the intersomite junction. Myocyte termini are normally characterized by deep invaginations of the sarcolemma. In myocytes expressing mutated IF polypeptides, these membrane invaginations were reduced or completely absent. Furthermore, the attachment of myofibrils to the junctional membrane was often aberrant or completely disrupted. These results suggest that in active muscle IFs play an important role in the organization and/or stabilization of myofibril-membrane attachment sites.

In most myogenic systems, synthesis of the intermediate filament (IF) protein vimentin precedes the synthesis of the muscle-specific IF protein desmin. In the dorsal myotome of the Xenopus embryo, however, there is no preexisting vimentin filament system and desmin's initial organization is quite different from that seen in vimentin-containing myocytes (Cary and Klymkowsky, 1994. Differentiation. In press.). To determine whether the organization of IFs in the Xenopus myotome reflects features unique to Xenopus or is due to specific properties of desmin, we used the injection of plasmid DNA to drive the synthesis of vimentin or desmin in myotomal cells. At low levels of accumulation, exogenous vimentin and desmin both enter into the endogenous desmin system of the myotomal cell. At higher levels exogenous vimentin forms longitudinal IF systems similar to those seen in vimentin-expressing myogenic systems and massive IF bundles. Exogenous desmin, on the other hand, formed a reticular IF meshwork and non-filamentous aggregates. In embryonic epithelial cells, both vimentin and desmin formed extended IF networks. Vimentin and desmin differ most dramatically in their NH2-terminal "head" regions. To determine whether the head region was responsible for the differences in the behavior of these two proteins, we constructed plasmids encoding chimeric proteins in which the head of one was attached to the body of the other. In muscle, the vimentin head-desmin body (VDD) polypeptide formed longitudinal IFs and massive IF bundles like vimentin. The desmin head-vimentin body (DVV) polypeptide, on the other hand, formed IF meshworks and non-filamentous structures like desmin. In embryonic epithelial cells DVV formed a discrete filament network while VDD did not. Based on the behavior of these chimeric proteins, we conclude that the head domains of vimentin and desmin are structurally distinct and not interchangeable, and that the head domain of desmin is largely responsible for desmin's muscle-specific behaviors.

To study vimentin filament organization in vivo we injected Xenopus oocytes, which have no significant vimentin system of their own, with in vitro-synthesized RNAs encoding Xenopus vimentins. Exogenous vimentins were localized primarily to the cytoplasmic surface of the nucleus and to the subplasma membrane "cortex." In the cortex of the animal hemisphere, wild-type vimentin forms punctate structures and short filaments. In contrast, long anastomosing vimentin filaments are formed in the vegetal hemisphere cortex. This asymmetry in the organization of exogenous vimentin is similar to that of the endogenous keratin system (Klymkowsky, M. W., L. A. Maynell, and A. G. Polson. 1987. Development (Camb.). 100:543-557), which suggests that the same cellular factors are responsible for both. Before germinal vesicle breakdown, in the initial stage of oocyte maturation, large vimentin and keratin filament bundles appear in the animal hemisphere. As maturation proceeds, keratin filaments fragment into soluble oligomers (Klymkowsky, M. W., L. A. Maynell, and C. Nislow. 1991. J. Cell Biol. 114:787-797), while vimentin filaments remain intact and vimentin is hyperphosphorylated. To examine the role of MPF kinase in the M-phase reorganization of vimentin we deleted the conserved proline of vimentin's single MPF-kinase site; this mutation had no apparent effect on the prophase or M-phase behavior of vimentin. In contrast, deletion of amino acids 19-68 or 18-61 of the NH2-terminal "head" domain produced proteins that formed extended filaments in the animal hemisphere of the prophase oocyte. We suggest that the animal hemisphere cortex of the prophase oocyte contains a factor that actively suppresses the formation of extended vimentin filaments through a direct interaction with vimentin's head domain. During maturation this "suppressor of extended filaments" appears to be inactivated, leading to the formation of an extended vimentin filament system.

Semi-volatile organic carbonaceous material (SVOC) in fine particles is not reliably measured with conventional semicontinuous carbon monitors because semi-volatile carbonaceous material is lost from the collection media during sample collection. Two modifications of a Sunset Laboratory carbon aerosol monitor allowing for the determination of semi-volatile fine particulate organic material are described. Collocated conventional and modified instruments were operated simultaneously using a common inlet. Comparisons were made with integrated PC-BOSS data for quartz filter retained nonvolatile organic carbon (NVOC) and elemental carbon (EC), SVOC, and total carbon (TC = SVOC + NVOC + EC) and good agreement was observed between TC concentrations during studies conducted in Rubidoux, CA. Precision of the comparison was sigma = +/-1.5 microg-C/m3 (+/-8%). On the basis of experiments performed with the modified Sunset monitor, a dual-oven Sunset monitor was developed and extensively tested in Lindon, UT; Riverside, CA; and in environmental exposure chambers. The precision for the measurement of TC with the dual-oven instrument was sigma = +/-1.4 microg-C/m3 (+/-13%).

The field-deployable Sunset Semi-Continuous Organic Carbon/Elemental Carbon (Sunset OCEC) aerosol analyzer utilizes the modified National Institute for Occupational Safety and Health thermal-optical method to determine total carbon (TC), organic carbon (OC), and elemental carbon (EC) at near real-time. Two sets of OC and EC are available: thermal OC and EC, and optical OC and EC. The former is obtained by the thermal-optical approach, and the latter is obtained by directly determining EC optically and deriving optical OC from TC. However, the performance of the Sunset OCEC is not yet fully characterized. Two collocated Sunset OCEC analyzers, Unit A and Unit B, were used to determine the pooled relative standard deviation (RSD) and limit of detection (LOD) between September 18 and November 6, 2007 in Richland, WA. The LOD of Unit A was approximately 0.2 microgC/m3 (0.1 microgC/cm2) for TC, optical OC, and thermal OC, and 0.01 microgC/m3 (0.01 microgC/cm2) for optical EC. Similarly, Unit B had an LOD of approximately 0.3 microgC/m3 (0.2 microgC/cm2) for TC, optical OC, and thermal OC, and 0.02 microgC/m3 (0.01 microgC/cm2) for optical EC. The LOD for thermal EC is estimated to be 0.2 microgC/m3 (0.1 microgC/cm2) for both units. The pooled RSDs were 4.9% for TC (carbon mass loadings 0.6-6.0 microgC/cm2), 5.6% for optical OC (carbon mass loadings 0.6-5.4 microgC/cm2), 5.3% for thermal OC (carbon mass loadings 0.6-5.3 microgC/ cm2), and 9.6% for optical EC (carbon mass loadings 0-1.4 microgC/cm2), which indicates good precision between the instruments. The RSD for thermal EC is higher at 24.3% (carbon mass loadings 0-1.2 microgC/cm2). Low EC mass loadings in Richland contributed to the poor RSD of EC. The authors found that excessive noise from the nondispersive infrared (NDIR) laser in the Sunset OCEC analyzer could result in a worsened determination of OC and EC. It is recommended that a "quieter" NDIR laser and detector be used in the Sunset OCEC analyzer to improve quantification. Future work should re-evaluate the precision of the EC parameters in an environment favorable for EC collection. Investigation among quantification differences using various thermal-optical protocols to determine OC and EC is also in need.

This chapter summarizes our studies on the three toll-like receptor pathways, namely TLR4, TLR2, and TLR3, induced by lipopolysaccharides (LPS), peptidoglycan (PGN), and double-stranded RNA (dsRNA) in antigen presenting cells (APC). The particular emphasis is on the activation of human innate immune responses via cytokine and chemokine production. Three different measurements have been performed on monocytic and dendritic cells as model APCs: (i) the expression of various cytokine and chemokine genes by real-time PCR, (ii) the release of the cytokines and chemokines by ELISA, and (iii) gene expression analysis by cytokine and chemokine pathway-specific and whole genome microarrays. Real-time PCR and ELISA enable us to identify cytokines and chemokines that are produced specifically upon LPS, PGN, or dsRNA stimulation. Subsequently, microarray studies and appropriate validation experiments help us to identify genes involved in the upstream pathways that cause the induction of cytokines and chemokines. It is evident that TLR4-LPS, TLR2-PGN, and TLR3-dsRNA pathways are distinguished by the specific set of cytokines and chemokines they induce as well as by the upstream signaling events.

This chapter summarizes our studies on the three toll-like receptor pathways, namely TLR4, TLR2, and TLR3, induced by lipopolysaccharides (LPS), peptidoglycan (PGN), and double-stranded RNA (dsRNA) in antigen presenting cells (APC). The particular emphasis is on the activation of human innate immune responses via cytokine and chemokine production. Three different measurements have been performed on monocytic and dendritic cells as model APCs: (i) the expression of various cytokine and chemokine genes by real-time PCR, (ii) the release of the cytokines and chemokines by ELISA, and (iii) gene expression analysis by cytokine and chemokine pathway-specific and whole genome microarrays. Real-time PCR and ELISA enable us to identify cytokines and chemokines that are produced specifically upon LPS, PGN, or dsRNA stimulation. Subsequently, microarray studies and appropriate validation experiments help us to identify genes involved in the upstream pathways that cause the induction of cytokines and chemokines. It is evident that TLR4-LPS, TLR2-PGN, and TLR3-dsRNA pathways are distinguished by the specific set of cytokines and chemokines they induce as well as by the upstream signaling events.

Exposure to beryllium (Be) induces a delayed-type hypersensitivity immune reaction in the lungs of susceptible individuals, which leads to the onset of Be sensitivity and Chronic Beryllium Disease (CBD). Although some mechanistic aspects of CBD have begun to be characterized, very little is known about the molecular mechanisms by which Be activates the host immune response. To gain insight into the cellular response to Be exposure, we have performed global microarray analysis using a mixture of peripheral blood mononuclear and dendritic cells (PBMC/DCs) from a non-CBD source to identify genes that are specifically upregulated in response to BeSO(4) stimulation, compared to a control metal salt, Al(2)(SO(4))(3). We identified a number of upregulated immunomodulatory genes, including several chemokines in the MIP-1 and GRO families. Using PBMC/DCs from three different donors, we demonstrate that BeSO(4) stimulation generally exhibits an increased rate of both chemokine mRNA transcription and release compared to Al(2)(SO(4))(3) exposure, although variations among the individual donors do exist. We show that MIP-1 alpha and MIP-1 beta neutralizing antibodies can partially inhibit the ability of BeSO(4) to stimulate cell migration of PBMC/DCs in vitro. Finally, incubation of PBMC/DCs with BeSO(4) altered the binding of the transcription factor RUNX to the MIP-1 alpha promoter consensus sequence, indicating that Be can regulate chemokine gene activation. Taken together, these results suggest a model in which Be stimulation of PBMC/DCs can modulate the expression and release of different chemokines, leading to the migration of lymphocytes to the lung and the formation of a localized environment for development of Be disease in susceptible individuals.

Rapid, accurate, and sensitive detection of biothreat agents requires a broad-spectrum assay capable of discriminating between closely related microbial or viral pathogens. Moreover, in cases where a biological agent release has been identified, forensic analysis demands detailed genetic signature data for accurate strain identification and attribution. To date, nucleic acid sequences have provided the most robust and phylogentically illuminating signature information. Nucleic acid signature sequences are not often linked to genomic or extrachromosomal determinants of virulence, a link that would further facilitate discrimination between pathogens and closely related species. Inextricably coupling genetic determinants of virulence with highly informative nucleic acid signatures would provide a robust means of identifying human, livestock, and agricultural pathogens. By means of example, we present here an overview of two general applications of microarray-based methods for: (1) the identification of candidate virulence factors; and (2) the analysis of genetic polymorphisms that are coupled to Bacillus anthracis virulence factors using an accurate, low cost solid-phase mini-sequencing assay. We show that microarray-based analysis of gene expression can identify potential virulence associated genes for use as candidate signature targets, and, further, that microarray-based single nucleotide polymorphism assays provide a robust platform for the detection and identification of signature sequences in a manner independent of the genetic background in which the signature is embedded. We discuss the strategy as a general approach or pipeline for the discovery of virulence-linked nucleic acid signatures for biothreat agents.

The ataxia-telangiectasia mutated (ATM) gene product plays a role in responding to double stand DNA breaks. Some biochemical studies of ATM function have been hampered by lack of an efficient expression system and abundant purified ATM protein. We report the construction of a vaccinia virus expressing ATM, vWR-ATM, which was used to produce large amounts of functional FLAG-tagged ATM protein (FLAG-ATM) in HeLa cells. Kinase activity of the purified FLAG-ATM was dependent on manganese and inhibited with wortmannin. Using the FLAG-ATM recombinant protein, GST-p53 serine 15 phosphorylation increased in the presence of damaged DNA. PHAS-1 phosphorylation was found to be DNA independent. Purified FLAG-ATM was recovered in the autophosphorylated form, as demonstrated by phosphorylation of ATM serine 1981. As shown by atomic force microscopy, FLAG-ATM bound to linear DNA both at broken ends and in mid-strands. Vaccinia virus is the most efficient ATM expression system described to date.

Several findings have revealed a likely role for DNA ligase IV, and interacting protein XRCC4, in the final steps of mammalian DNA double-strand break repair. Recent evidence suggests that the human DNA ligase IV protein plays a critical role in the maintenance of genomic stability. To identify protein–protein interactions that may shed further light on the molecular mechanisms of DSB repair and the biological roles of human DNA ligase IV, we have used the yeast two-hybrid system in conjunction with traditional biochemical methods. These efforts have resulted in the identification of a physical association between the DNA ligase IV polypeptide and the human condensin subunit known as hCAP-E. The hCAP-E polypeptide, a member of the Structural Maintenance of Chromosomes (SMC) super-family of proteins, coimmunoprecipitates from cell extracts with DNA ligase IV. Immunofluorescence studies reveal colocalization of DNA ligase IV and hCAP-E in the interphase nucleus, whereas mitotic cells display colocalization of both polypeptides on mitotic chromosomes. Strikingly, the XRCC4 protein is excluded from the area of mitotic chromosomes, suggesting the formation of specialized DNA ligase IV complexes subject to cell cycle regulation. We discuss our findings in light of known and hypothesized roles for ligase IV and the condensin complex.

The human neurodegenerative and cancer predisposition condition ataxia-telangiectasia is characterized at the cellular level by radiosensitivity, chromosomal instability, and impaired induction of ionizing radiation-induced cell cycle checkpoint controls. Recent work has revealed that the gene defective in ataxia-telangiectasia, termed ATM , encodes an ≈350-kDa polypeptide, ATM, that is a member of the phosphatidylinositol 3-kinase family. We show that ATM binds DNA and exploit this to purify ATM to near homogeneity. Atomic force microscopy reveals that ATM exists in two populations, with sizes consistent with monomeric and tetrameric states. Atomic force microscopy analyses also show that ATM binds preferentially to DNA ends. This property is similar to that displayed by the DNA-dependent protein kinase catalytic subunit, a phosphatidylinositol 3-kinase family member that functions in DNA damage detection in conjunction with the DNA end-binding protein Ku. Furthermore, purified ATM contains a kinase activity that phosphorylates serine-15 of p53 in a DNA-stimulated manner. These results provide a biochemical assay system for ATM, support genetic data indicating distinct roles for DNA-dependent protein kinase and ATM, and suggest how ATM may signal the presence of DNA damage to p53 and other downstream effectors.

Ku, the DNA binding component of DNA-dependent protein kinase (DNA-PK), is a heterodimer composed of 70 and 86 kDa subunits, known as Ku70 and Ku80 respectively . Defects in DNA-PK subunits have been shown to result in a reduced capacity to repair DNA double-strand breaks. Assembly of the Ku heterodimer is required to obtain DNA end binding activity and association of the DNA-PK catalytic subunit. The regions of the Ku subunits responsible for heterodimerization have not been clearly defined in vivo . A previous study has suggested that the C-terminus of Ku80 is required for interaction with Ku70. Here we examine Ku subunit interaction using N- and C-terminal Ku80 deletions in a GAL4-based two-hybrid system and an independent mammalian in vivo system. Our two-hybrid study suggests that the central region of Ku80, not its C-terminus, is capable of mediating interaction with Ku70. To determine if this region mediates interaction with Ku70 in mammalian cells we transfected xrs-6 cells, which lack endogenous Ku80, with epitope-tagged Ku80 deletions carrying a nuclear localization signal. Immunoprecipitation from transfected cell extracts revealed that the central domain identified by the GAL4 two-hybrid studies stabilizes and co-immunoprecipitates with endogenous xrs-6 Ku70. The central interaction domain maps to the internally deleted regions of Ku80 in the mutant cell lines XR-V9B and XR-V15B. These findings indicate that the internally deleted Ku80 mutations carried in these cell lines are incapable of heterodimerization with Ku70.

The DNA-dependent protein kinase (DNA-PK) is required for DNA double-strand break (DSB) repair and immunoglobulin gene rearrangement and may play a role in the regulation of transcription. The DNA-PK holoenzyme is composed of three polypeptide subunits: the DNA binding Ku70/86 heterodimer and an ≈460-kDa catalytic subunit (DNA-PKcs). DNA-PK has been hypothesized to assemble at DNA DSBs and play structural as well as signal transduction roles in DSB repair. Recent advances in atomic force microscopy (AFM) have resulted in a technology capable of producing high resolution images of native protein and protein–nucleic acid complexes without staining or metal coating. The AFM provides a rapid and direct means of probing the protein–nucleic acid interactions responsible for DNA repair and genetic regulation. Here we have employed AFM as well as electron microscopy to visualize Ku and DNA-PK in association with DNA. A significant number of DNA molecules formed loops in the presence of Ku. DNA looping appeared to be sequence-independent and unaffected by the presence of DNA-PKcs. Gel filtration of Ku in the absence and the presence of DNA indicates that Ku does not form nonspecific aggregates. We conclude that, when bound to DNA, Ku is capable of self-association. These findings suggest that Ku binding at DNA DSBs will result in Ku self-association and a physical tethering of the broken DNA strands.

The tail domain of the intermediate filament (IF) protein vimentin is unnecessary for IF assembly in vitro. To study the role of vimentin's tail in vivo, we constructed a plasmid that directs the synthesis of a ‘myc-tagged’ version of the Xenopus vimentin-1 tail domain in bacteria. This polypeptide, mycVimTail, was purified to near homogeneity and injected into cultured Xenopus A6 cells. In these cells the tail polypeptide co-localized with actin even in the presence of cytochalasin. Two myc-tagged control polypeptides argue for the specificity of this interaction. First, a similarly myc-tagged lamin tail domain localizes to the nucleus, indicating that the presence of the myc tag did not itself confer the ability to co-localize with actin (Hennekes and Nigg (1994) J. Cell Sci. 107, 1019–1029). Second, a myc-tagged polypeptide with a molecular mass and net charge at physiological pH (i.e. -4) similar to that of the mycVimTail polypeptide, failed to show any tendency to associate with actin-containing structures, indicating that the interaction between mycVimTail and actin-containing structures was not due to a simple ionic association. Franke (1987; Cell Biol. Int. Rep. 11, 831) noted a similarity in the primary sequence between the tail of the type I keratin DG81A and vimentin. To test whether the DG81A tail interacted with actin-containing structures, we constructed and purified myc-tagged DG81A tail polypeptides. Unexpectedly, these keratin tail polypeptides were largely insoluble under physiological conditions and formed aggregates at the site of injection. While this insolubility made it difficult to determine if they associated with actin-containing structures, it does provide direct evidence that the tails of vimentin and DG81A differ dramatically in their physical properties. Our data suggest that vimentin's tail domain has a highly extended structure, binds to actin-containing structures and may mediate the interaction between vimentin filaments and microfilaments involved in the control of vimentin filament organization (Hollenbeck et al. (1989) J. Cell Sci. 92, 621; Tint et al. (1991) J. Cell Sci. 98, 375).

Microarray analysis of the DNA-dependent protein kinase catalytic subunit knock-out mouse: global effects on gene regulation and the cellular response to ionizing radiation

Diesel exhaust has been classified a probable human carcinogen, and the National Institute for Occupational Safety and Health (NIOSH) has recommended that employers reduce workers' exposures. Because diesel exhaust is a chemically complex mixture containing thousands of compounds, some measure of exposure must be selected. Previously used methods involving gravimetry or analysis of the soluble organic fraction of diesel soot lack adequate sensitivity and selectivity for low-level determination of particulate diesel exhaust; a new analytical approach was therefore needed. In this paper, results of investigation of a thermal-optical technique for the analysis of the carbonaceous fraction of particulate diesel exhaust are discussed. With this technique, speciation of organic and elemental carbon is accomplished through temperature and atmosphere control and by an optical feature that corrects for pyrolytically generated carbon, or "char,' which is formed during the analysis of some materials. The thermal-optical method was selected because the instrument has desirable design features not present in other carbon analysers. Although various carbon types are determined by the method, elemental carbon is the superior marker of diesel particulate matter because elemental carbon constitutes a large fraction of the particulate mass, it can be quantified at low levels and its only significant source in most workplaces is the diesel engine. Exposure-related issues and sampling methods for particulate diesel exhaust also are discussed.

The rate of neutralization of sulfuric acid aerosol by ammonia gas has been measured in a laboratory flow reactor for particle diameters between 0.3 and 1.4 micrometers and for relative humidities between 8 and 80%. The rates were between 21 and 70% of the rates expected if diffusion of NH/sub 3/ to the aerosol droplets was the rate-determining factor, and the reaction coefficient, which is the fraction of gas-particle collisions resulting in a chemical reaction, increased with increasing particle size. A simple model of H/sub 2/SO/sub 4/ aerosol generation and neutralization in the atmosphere showed that sulfate aerosol more acidic than NH/sub 4/HSO/sub 4/ should only be present during periods of rapid oxidation of the precursor SO/sub 2/ and high ratios of (SO/sub 2/) to (NH/sub 3/) or when the equilibrium vapor pressure of NH/sub 3/ over the partially neutralized H/sub 2/SO/sub 4/ droplet exceeds the ambient NH/sub 3/ partial pressure.