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Pulsations generated by personal sampling pumps modulate the airflow through the sampling trains, thereby varying sampling efficiencies, and possibly invalidating collection or monitoring. The purpose of this study was to characterize pulsations generated by personal sampling pumps relative to a nominal flow rate at the inlet of different respirable cyclones. Experiments were conducted using a factorial combination of 13 widely used sampling pumps (11 medium and 2 high volumetric flow rate pumps having a diaphragm mechanism) and 7 cyclones [10-mm nylon also known as Dorr-Oliver (DO), Higgins-Dewell (HD), GS-1, GS-3, Aluminum, GK2.69, and FSP-10]. A hot- wire anemometer probe cemented to the inlet of each cyclone type was used to obtain pulsation readings. The three medium flow rate pump models showing the highest, a midrange, and the lowest pulsations and two high flow rate pump models for each cyclone type were tested with dust-loaded filters (0.05, 0.21, and 1.25 mg) to determine the effects of filter loading on pulsations. The effects of different tubing materials and lengths on pulsations were also investigated. The fundamental frequency range was 22–110 Hz and the magnitude of pulsation as a proportion of the mean flow rate ranged from 4.4 to 73.1%. Most pump/cyclone combinations generated pulse magnitudes >10% (48 out of 59 combinations), while pulse shapes varied considerably. Pulsation magnitudes were not considerably different for the clean and dust-loaded filters for the DO, HD, and Aluminum cyclones, but no consistent pattern was observed for the other cyclone types. Tubing material had less effect on pulsations than tubing length; when the tubing length was 183 cm, pronounced damping was observed for a pump with high pulsation (>60%) for all tested tubing materials except for the Tygon Inert tubing. The findings in this study prompted a further study to determine the possibility of shifts in cyclone sampling efficiency due to sampling pump pulsations, and those results are reported subsequently.

This second, and concluding, part of this study evaluated changes in sampling efficiency of respirable size-selective samplers due to air pulsations generated by the selected personal sampling pumps characterized in Part I (Lee E, Lee L, Möhlmann C et al. Evaluation of pump pulsation in respirable size-selective sampling: Part I. Pulsation measurements. Ann Occup Hyg 2013). Nine particle sizes of monodisperse ammonium fluorescein (from 1 to 9 μm mass median aerodynamic diameter) were generated individually by a vibrating orifice aerosol generator from dilute solutions of fluorescein in aqueous ammonia and then injected into an environmental chamber. To collect these particles, 10-mm nylon cyclones, also known as Dorr-Oliver (DO) cyclones, were used with five medium volumetric flow rate pumps. Those were the Apex IS, HFS513, GilAir5, Elite5, and Basic5 pumps, which were found in Part I to generate pulsations of 5% (the lowest), 25%, 30%, 56%, and 70% (the highest), respectively. GK2.69 cyclones were used with the Legacy [pump pulsation (PP) = 15%] and Elite12 (PP = 41%) pumps for collection at high flows. The DO cyclone was also used to evaluate changes in sampling efficiency due to pulse shape. The HFS513 pump, which generates a more complex pulse shape, was compared to a single sine wave fluctuation generated by a piston. The luminescent intensity of the fluorescein extracted from each sample was measured with a luminescence spectrometer. Sampling efficiencies were obtained by dividing the intensity of the fluorescein extracted from the filter placed in a cyclone with the intensity obtained from the filter used with a sharp-edged reference sampler. Then, sampling efficiency curves were generated using a sigmoid function with three parameters and each sampling efficiency curve was compared to that of the reference cyclone by constructing bias maps. In general, no change in sampling efficiency (bias under ±10%) was observed until pulsations exceeded 25% for the DO cyclone. However, for three models of pumps producing 30%, 56%, and 70% pulsations, substantial changes were confirmed. The GK2.69 cyclone showed a similar pattern to that of the DO cyclone, i.e. no change in sampling efficiency for the Legacy producing 15% pulsation and a substantial change for the Elite12 producing 41% pulsation. Pulse shape did not cause any change in sampling efficiency when compared to the single sine wave. The findings suggest that 25% pulsation at the inlet of the cyclone as measured by this test can be acceptable for the respirable particle collection. If this test is used in place of that currently in European standards (EN 1232–1997 and EN 12919-1999) or is used in any International Organization for Standardization standard, then a 25% pulsation criterion could be adopted. This work suggests that a 10% criterion as currently specified in the European standards for testing may be overly restrictive and not able to be met by many pumps on the market. Further work is recommended to determine which criterion would be applicable to this test if it is to be retained in its current form.

This study introduces two semi-quantitative methods, Structured Subjective Assessment (SSA) and Control of Substances Hazardous to Health (COSHH) Essentials, in conjunction with two-dimensional Monte Carlo simulations for determining prior probabilities. Prior distribution using expert judgment was included for comparison. Practical applications of the proposed methods were demonstrated using personal exposure measurements of isoamyl acetate in an electronics manufacturing facility and of isopropanol in a printing shop. Applicability of these methods in real workplaces was discussed based on the advantages and disadvantages of each method. Although these methods could not be completely independent of expert judgments, this study demonstrated a methodological improvement in the estimation of the prior distribution for the Bayesian decision analysis tool. The proposed methods provide a logical basis for the decision process by considering determinants of worker exposure. [Supplementary materials are availab...

Foamy viruses are retroviruses whose Pol protein is synthesized without Gag from a spliced mRNA. Unlike orthoretroviruses, reverse transcription occurs during viral assembly, leading to DNA-containing virions. When prototype foamy virus Pol is expressed as an orthoretroviral-like Gag-Pol fusion protein, reverse transcription also occurs late in viral replication, as measured by the timing of reverse transcriptase sensitivity to the inhibitor 3′-azido-3′deoxythymidine (AZT). Thus, timing of reverse transcription is intrinsic to Pol itself.

The US Occupational Safety and Health Administration (OSHA) and Mine Safety and Health Administration do not regulate cleavage fragments of amphibole and serpentine minerals as asbestos, even when particles meet the dimensional criteria for counting under standard phase-contrast microscopy methods. The OSHA ID-160 method cautions that discriminatory counting is difficult and should not be attempted unless necessary and no procedure is provided for differentiation. A standard published by the American Society for Testing and Materials (ASTM International D7200-06) includes an attempt to codify a procedure but recognizes that the procedure should be validated in an inter-laboratory study. The US National Institute for Occupational Safety and Health has carried out such a study with multiple laboratories using slides made from riebeckite and crocidolite, grunerite and amosite, tremolite and tremolite asbestos, and actinolite and actinolite asbestos using two different measurement aids (graticules). The asbestos fibers had dimensions consistent with those reported for air samples from actual amphibole asbestos operations, and the cleavage fragments were also dimensionally consistent with those found in non-asbestos mining and milling operations. The procedure for discriminating asbestos fibers from other mineral particles in the ASTM Standard calls for the recognition of characteristics supposedly common to asbestos. For the asbestos fibers created in this study, these characteristics were found not to be common and generally a function of length. More importantly, different laboratories did not recognize these features consistently. Laboratories were much more consistent in measuring dimensions, but excessive overlap in the lengths of asbestos fibers and cleavage fragments rendered length a poor criterion for discrimination. The ASTM discrimination procedure as written could not be supported on the basis of this study. Width was a much more consistent parameter for distinguishing the asbestos and non-asbestos fibers in this study and inclusion of aspect ratio, while considered important by some researchers, did not refine the discrimination further. The ability of the majority of microscopists in this study to discriminate fibers and cleavage fragments through measurement of particle widths was determined and found to be within limits of uncertainty typical for air sampling measurements. A width criterion might be a very simple and useful aid where discrimination between asbestos and non-asbestos fibers in fiber counting by phase-contrast microscopy is required for further investigation. Recognition of asbestos features can also be retained as excessive recognition by some laboratories will lead to a conservative decision for additional investigation.

A study was conducted to examine whether there are significant differences between organic vapor concentrations measured using charcoal tubes with three different configurations: uncovered sample holder (open tube), SKC, and Buck brand covered sample holders. A fractional factorial experimental design was used with the following factors and levels: vapor (n-hexane vs. m-xylene), pump type (pulsating vs. continuous), exposure profile (variable vs. constant), flow rate (30 mL/min vs. 200 mL/min), duration (30 min vs. 80 min), and sample placement (mannequin vs. free hanging). Two of each sampler configuration (six total) were placed in an exposure chamber, and a dynamic test-atmosphere generation system was used to prepare atmospheres containing approximately 12–15 ppm n-hexane or m-xylene with exposure profiles and sampling conducted according to a run sheet generated for the experimental design. A total of 24 runs were completed with six samplers per run, yielding 144 samples that were analyzed by gas chromatography/flame ionization detector. Concentration results for each pair of SKC and Buck covered sample holders were averaged and normalized by dividing by the average result for the open tube sampler from the same run to eliminate the effect of daily variation in chamber concentrations. The resulting ratio of covered sample tube holder and open tube concentrations was used as the response variable. Results of analysis of variance using the general linear model (MINITAB 16) identified statistically significant main effects and/or interactions for pump type, exposure profile, flow rate, and sample holder. However, the magnitude of the effects was generally less than 10%, and overall mean concentration ratios were 0.989 and 1.02 for the Buck and SKC sample holders, respectively. These results show good agreement between covered sample holder results and open tube measurements and demonstrate that exposure assessment errors resulting from the use of covered sorbent tube sample holders for organic vapor monitoring are relatively small (

Individual membrane protein spanning sequences can promote protein export. To help define the sequence features necessary for this action, we identified mutations disrupting export mediated by the first spanning sequence (TM1) of the Escherichia coli serine chemoreceptor. Mutant spanning sequences were generated and characterized using beta-galactosidase and alkaline phosphatase gene fusions. The protein export function of TM1 was remarkably tolerant of single charged residues, and the introduction of pairs of charged amino acids was necessary to eliminate export. The results are accommodated by a model in which export requires a stretch of uncharged residues whose summed hydrophobicity exceeds a particular threshold value. This threshold approximates the minimum hydrophobicity required for cleavable signal sequence function. In addition, the threshold was near the minimum hydrophobicity observed for wild-type spanning sequences in a collection of topologically characterized membrane proteins.

A laboratory study was performed to measure quartz in coal dust using high-flow rate samplers (CIP10-R, GK2.69 cyclone, and FSP10 cyclone) and low-flow rate samplers [10-mm nylon and Higgins-Dewell type (BGI4L) cyclones] and to determine whether an increased mass collection from high-flow rate samplers would affect the subsequent quartz measurement by Fourier transform infrared (FTIR) and X-ray diffraction (XRD) analytical procedures. Two different sizes of coal dusts, mass median aerodynamic diameter 4.48 μm (Coal Dust A) and 2.33 μm (Coal Dust B), were aerosolized in a calm air chamber. The mass of coal dust collected by the samplers was measured gravimetrically, while the mass of quartz collected by the samplers was determined by FTIR (NIOSH Manual of Analytical Method 7603) and XRD (NIOSH Manual of Analytical Method 7500) after one of two different indirect preparations. Comparisons between high-flow rate samplers and low-flow rate samplers were made by calculating mass concentration ratios of coal dusts, net mass ratios of coal dusts, and quartz net mass. Mass concentrations of coal dust from the FSP10 cyclone were significantly higher than those from other samplers and mass concentrations of coal dust from 10-mm nylon cyclone were significantly lower than those from other samplers, while the CIP10-R, GK2.69, and BGI4L samplers did not show significant difference in the comparison of mass concentration of coal dusts. The BGI4L cyclone showed larger mass concentration of ∼9% compared to the 10-mm nylon cyclone. All cyclones provided dust mass concentrations that can be used in complying with the International Standard Organization standard for the determination of respirable dust concentration. The amount of coal dust collected from the high-flow rate samplers was found to be higher with a factor of 2-8 compared to the low-flow rate samplers but not in direct proportion of increased flow rates. The high-flow rate samplers collected more quartz compared to low-flow rate samplers in the range of 2-10. There was no significant difference between the per cent (%) quartz in coal dust between the FTIR and XRD analyses. The findings of this study indicated that the increased mass of quartz collected with high-flow rate samplers would provide precise analytical results (i.e. significantly above the limit of detection and/or limit of quantification) compared to the mass collected with low-flow rate samplers, especially in environments with low concentrations of quartz or where short sampling times are desired.

The objectives of this study were to evaluate mounting methods for fiber examination of air sample filters by phase contrast microscopy (PCM) and to evaluate differences in fiber counts that might be due to fiber movement. Acetone/triacetin (AT) with various amounts of triacetin and acetone/Euparal (AE) where the mounting medium was placed between the cleared filter wedge and the coverslip were tested as a function of time. Field sample slides collected from a taconite iron-ore processing mill, a tremolitic talc-ore processing mill, and from around a crusher in a meta-basalt stone quarry were prepared with relocatable coverslips to revisit the same field areas on the slides. For each slide, three or four field areas were randomly selected and pictures were taken every 2 weeks to determine any sign of fiber movement over time. For 11 AT slides (named as AT-3.5) prepared with 3.5 μl of the mounting medium according to the NIOSH 7400 method, no fiber movements were detected over 59 weeks. On the other hand, AT slides prepared with larger quantities (10, 15, and 20 μl) of the mounting medium (named as AT-10) and AE slides prepared with ∼10 μl mounting medium showed fiber movement from the eighth day at the earliest. Fiber movement began earlier for the slides mounted with excess triacetin than for those mounted with Euparal. The sample slide storage method, either vertically or horizontally, did not seem to accelerate fiber movement. Additionally, two other modified methods, dimethylformamide solution/Euparal (mDE) and dimethylformamide solution/triacetin (mDT), were also prepared where the mounting medium was placed between the cleared filter wedge and the glass slide. The findings of fiber movements were similar; when 3.5 μl of triacetin was used for the mDT slides, fiber movements were not detected, while fibers on slides prepared with 10 μl triacetin (mDT-10) moved around. No fiber movements were observed for the mDE slides at any time during 59 weeks. Once fiber movement started, fibers moved over distances measured from 4 μm and up to1000 μm within 22 weeks. However, since then, no further fiber movements have been observed in any field sample slides. Additional sample slides, two Amosite and two chrysotile, were prepared from Union for International Cancer Control (UICC) samples using the AT method with 5 μl triacetin mounting medium. Fiber movements were also observed in these samples; chrysotile fibers began to migrate in 3 weeks, while Amosite fiber movement started after 3 months. Although fiber movement was observed for the AT-10, AE, and mDT-10 sample slides, fiber counts were not significantly different from AT-3.5 and mDE samples that exhibited no fiber movement. Although fiber counts would not be significantly changed by fiber movement, the type and amount of mounting medium for sample slide preparation remains critical for issues such as quality assurance and training of analysts by revisiting the same fibers.

The Control of Substances Hazardous to Health (COSHH) Essentials model was evaluated using full-shift exposure measurements of five chemical components in a mixture [acetone, ethylbenzene, methyl ethyl ketone, toluene, and xylenes] at a medium-sized plant producing paint materials. Two tasks, batch-making and bucket-washing, were examined. Varying levels of control were already established in both tasks and the average exposures of individual chemicals were considerably lower than the regulatory and advisory 8-h standards. The average exposure fractions using the additive mixture formula were also less than unity (batch-making: 0.25, bucket-washing: 0.56) indicating the mixture of chemicals did not exceed the combined occupational exposure limit (OEL). The paper version of the COSHH Essentials model was used to calculate a predicted exposure range (PER) for each chemical according to different levels of control. The estimated PERs of the tested chemicals for both tasks did not show consistent agreement with exposure measurements when the comparison was made for each control method and this is believed to be because of the considerably different volatilities of the chemicals. Given the combination of health hazard and exposure potential components, the COSHH Essentials model recommended a control approach ‘special advice’ for both tasks, based on the potential reproductive hazard ascribed to toluene. This would not have been the same conclusion if some other chemical had been substituted (for example styrene, which has the same threshold limit value as toluene). Nevertheless, it was special advice, which had led to the combination of hygienic procedures in place at this plant. The probability of the combined exposure fractions exceeding unity was 0.0002 for the batch-making task indicating that the employees performing this task were most likely well protected below the OELs. Although the employees involved in the bucket-washing task had greater potential to exceed the threshold limit value of the mixture (P > 1 = 0.2375), the expected personal exposure after adjusting for the assigned protection factor for the respirators in use would be considerably lower (P > 1 = 0.0161). Thus, our findings suggested that the COSHH essentials model worked reasonably well for the volatile organic chemicals at the plant. However, it was difficult to override the reproductive hazard even though it was meant to be possible in principle. Further, it became apparent that an input of existing controls, which is not possible in the web-based model, may have allowed the model be more widely applicable. The experience of using the web-based COSHH Essentials model generated some suggestions to provide a more user-friendly tool to the model users who do not have expertise in occupational hygiene.

The performance of two thoracic samplers, the GK2.69 cyclone and the CATHIA-T sampler, and the GK3.51 cyclone was investigated in the field against the standard cowled sampler (current NIOSH 7400 method) to determine the effect of thoracic sampling. The CATHIA-T sampler and the GK2.69 cyclone were operated at 7 and 1.6 l min(-1), respectively. The GK3.51 sampler is related to the GK2.69 cyclone, but designed to give a thoracic cut at a flow rate of 3.2 l min(-1). A total of 136 area samples were obtained from a tremolitic talc processing mill and 148 area samples were obtained within a quarry in which metamorphosed volcanic rocks were being crushed for construction stone. Sample slides were prepared using the dimethyl formamide/Euparal technique and relocatable cover slips. NIOSH 7400 'A' counting rules were used to examine fibers. Additionally, counters were asked to record the number of fibers where a fiber meets the 'A' rules and is wider than 3-mum physical diameter in order to estimate the proportion of extra-thoracic fibers. A few slides from each sampler type were randomly selected and fiber widths for those fibers satisfying the counting rules were measured to determine median width ratios of each thoracic sampler to the cowled sampler. Overall, the combined results of this study and the previous study by the same authors (Lee et al., 2008) showed lower fiber concentrations for the CATHIA-T sampler and higher concentrations for the GK2.69 cyclone and the GK3.51 cyclone compared to the standard cowled sampler. The proportion of extra-thoracic fibers (3-mum physical diameter) on the filters collected with each type of thoracic samplers was comparable to the proportion of such fibers collected with the cowled sampler. The most consistent result over this study and our previous study is that both cyclones gave higher fiber concentrations than the CATHIA-T sampler. However, the estimated width ratios of each cyclone type to the cowled sampler were similar to or equal to 1 indicating no separation of fiber bundles due to cyclone effects. An inverse relationship between the fiber concentration ratios of each thoracic sampler type to the cowled sampler and the relative sample loading rate compared to the cowled sampler was observed. The difference in fiber concentration ratios might be a function of sample loading rather than an effect of thoracic size selection. Therefore, it is not recommended that any of these samplers be used to replace the cowled sampler in measurements intended for comparison with limit values arising from risk assessments developed from cowled sampler measurements without considering this possible effect.

The merits of using physiologic sampling pumps (PSPs) instead of using constant-flow sampling pumps, i.e., "traditional sampling pumps" (TSPs), are discussed. A novel PSP that overcomes shortcomings of previous PSP designs is presented. Calibrated valves are used to obviate pump inertia that has limited the system response and accuracy of prior work. Technologies that provide minute ventilation (V[combining dot above](E)) of subjects in real time may therefore be used to the limit of their own accuracies to sample inhalation exposures. Analysis of the design and data from a prototype are presented to show how air sampling can be modulated to follow breathing.

A study was conducted to evaluate a portable respiratory inductive plethysmograph (RIP) as a means to estimate minute ventilation (V(E)) for use in controlling the flow rate of a physiologic sampling pump (PSP). Specific aims were to: (1) evaluate the ability of the portable RIP system to measure V(E) using a direct (individual) fixed-volume calibration method (Direct RIP model), (2) develop and evaluate the performance of indirect (group) regression models for V(E) prediction using output data from the portable RIP and subject demographic characteristics (Indirect RIP model), and (3) compare V(E) estimates from indirect and direct portable RIP calibration with indirect estimation models published previously. Nine subjects (19-44 years) were divided into calibration (n = 6) and test (n = 3) datasets and performed step-tests on three different days while wearing the portable RIP and breathing through a pneumotachometer (reference). Minute ventilation and portable RIP output including heart rate, breathing rate, and a motion index were recorded simultaneously during the 80 minute sessions. Calibration data were used to develop a regression model for V(E) prediction that was subsequently applied to the test dataset. Direct calibration of the portable RIP system produced highly variable estimates of V(E) (R2 = 0.62, average % error = 15 +/- 50) while Indirect RIP model results were highly correlated with the reference (R2 = 0.80-0.88) and estimates of total volume were within 10% of reference values on average. Although developed from a limited dataset, the Indirect RIP model provided an alternative approach to estimation of V(E) and total volume with accuracy comparable to previously published models.

Mining or processing asbestos minerals can liberate isolated fibers or fiber bundles regulated as airborne asbestos fibers. Coarsely crystalline amphibole minerals are more common than asbestos in many geologic environments, and disturbance can result in the release of prismatic or acicular single crystals or cleavage fragments resembling asbestos fibers or fiber bundles but that are not currently regulated as asbestos. Bulk samples of six coarsely crystalline amphiboles and their five asbestos analogs were processed to maximize the number of particles meeting the criterion for counting under the current U.S. National Institute for Occupational Safety and Health Method 7400 "A" counting rules (> 5 microm long with an aspect ratio >or= 3:1) and also within the respirable width range, i.e. < 3 microm width. The length distributions of the particles produced showed substantial overlap between cleavage fragments and asbestos fibers. Available data sets generally confirmed the relevance of the size distributions of particles generated from reference materials to airborne particles. The length criterion in the current ASTM International standard D7200-06 causes a large proportion (e.g., 40% grunerite and 39% tremolite) of the non-asbestiform particles to be considered potential asbestos. An alternative procedure may be to use a distinction based on width alone as some, but not the majority of, cleavage fragments were thinner than 1 microm (e.g., 9% of actinolite and 20% of grunerite particles), and not many amphibole asbestos particles were wider (e.g., 5% of crocidolite and 18% of amosite particles). This proposal would need further testing. This research should not be considered as addressing any controversy with regard to the toxicity of non-asbestiform amphibole particles of similar dimensions to asbestos particles.

Foamy viruses (FV) differ from orthoretroviruses in many aspects of their replication cycle. A major difference is in the mode of Pol expression, regulation, and encapsidation into virions. Orthoretroviruses synthesize Pol as a Gag-Pol fusion protein so that Pol is encapsidated into virus particles through Gag assembly domains. However, as FV express Pol independently of Gag from a spliced mRNA, packaging occurs through a distinct mechanism. FV genomic RNA contains cis -acting sequences that are required for Pol packaging, suggesting that Pol binds to RNA for its encapsidation. However, it is not known whether Gag is directly involved in Pol packaging. Previously our laboratory showed that sequences flanking the three glycine-arginine-rich (GR) boxes at the C terminus of FV Gag contain domains important for RNA packaging and Pol expression, cleavage, and packaging. We have now shown that both deletion and substitution mutations in the first GR box (GR1) prevented neither the assembly of particles with wild-type density nor packaging of RNA genomes but led to a defect in Pol packaging. Site-directed mutagenesis of GR1 indicated that the clustered positively charged amino acids in GR1 play important roles in Pol packaging. Our results suggest that GR1 contains a Pol interaction domain and that a Gag-Pol complex is formed and binds to RNA for incorporation into virions.

Foamy viruses (FVs) assemble using pathways distinct from those of orthoretroviruses. FV capsid assembly takes place near the host microtubule-organizing center (MTOC). Assembled capsids then migrate by an unknown mechanism to the trans-Golgi network to colocalize with the FV glycoprotein, Env. Interaction with Env is required for FV capsid egress from cells; the amino terminus of FV Gag contains a cytoplasmic targeting/retention signal that is responsible for targeting assembly to the MTOC. A mutant Gag was constructed by addition of a myristylation (M) signal in an attempt to target assembly to the plasma membrane and potentially overcome the dependence upon Env for budding (S. W. Eastman and M. L. Linial, J. Virol. 75:6857-6864, 2001). Using this and additional mutants, we now show that assembly is not redirected to the plasma membrane. Addition of an M signal leads to gross morphological defects. The aberrant particles still assemble near the MTOC but do not produce infectious virus. Although extracellular Gag can be detected in a pelletable form in the absence of Env, the mutant particles contain very little genomic RNA and are less dense. Our analyses indicate that the amino terminus of Gag contains an Env interaction domain that is critical for bona fide egress of assembled capsids.

Several recommendations have been made to the effect that the most appropriate health-based size-selective criterion for fibers is the thoracic convention of the International Organization for Standardization (ISO). The performance of two thoracic samplers, the CATHIA-T (37-mm filter) and the GK2.69 cyclone (37-mm filter), was investigated against the standard 25-mm cowled sampler (current NIOSH 7400 standard method) to determine the effect of thoracic sampling on field results. A total of 270 samples: 80 field and 10 field blank samples for each sampler type, were taken from seven stations in the processing mill of an iron ore mine whose ore contains amphibole minerals. Slides were prepared using the dimethyl formamide/Euparal technique and relocatable cover slips. Two counters examined the slides according to NIOSH 7400 counting A rules with phase contrast microscopes. Prior to counting the sample slides, four reference slides were randomly selected and counted three times on different days to compare the coefficient of variation (CV) between and within counters. Also, seven reference slides were chosen to explore variability between the two microscopes. The average CV between counters (0.148) showed slightly higher than the average CVs within counters (0.072 for Counter 1 and 0.119 for Counter 2). The average CV between the two microscopes was 0.147. Compared to the standard cowled sampler, the overall fiber concentration was lower for the CATHIA-T sampler (CATHIA-T/Cowled = 0.63) and higher for the GK2.69 cyclone (GK2.69/Cowled = 1.66). The result for the CATHIA-T sampler is as expected from laboratory trials, but the result for the GK2.69 cyclone is not as expected. In conclusion, the CATHIA-T sampler has a potential advantage as a high-flow static sampler for screening coarse particles. However, these findings resulted from one field sampling site that contains amphibole minerals, not all of which are asbestiform. Thus, additional field samples from other environments might be helpful to confirm the performance of these samplers.

Foamy viruses (FV) comprise a subfamily of retroviruses. Orthoretroviruses, such as human immunodeficiency virus type 1, synthesize Gag and Pol from unspliced genomic RNA. However, FV Pol is expressed from a spliced mRNA independently of Gag. FV pol splicing uses a 3′ splice site located at the 3′ end of gag , resulting in a shared exon between gag and pol . Previously, our laboratory showed that C-terminal Gag premature termination codon (PTC) mutations in the 3′ shared exon led to greatly decreased levels of Pol protein (C. R. Stenbak and M. L. Linial, J. Virol. 78 :9423-9430, 2004). To further characterize these mutants, we quantitated the levels of unspliced gag and spliced pol mRNAs using a real-time PCR assay. In some of the PTC mutants, the levels of spliced pol mRNA were reduced as much as 30-fold, whereas levels of unspliced gag RNA were not affected. Substitutions of a missense codon in place of a PTC restored normal levels of spliced pol mRNA. Disrupting Upf proteins involved in nonsense-mediated mRNA decay (NMD) did not affect Pol protein expression. Introduction of an exonic splicing enhancer downstream of the PTC mutation restored pol splicing to the wild-type level. Taken together, our results show that the PTC mutation itself is responsible for decreased levels of pol mRNA but that mechanisms other than NMD might be involved in downregulating Pol expression. The results also suggest that normal pol splicing utilizes a suboptimal splice site seen for other spliced mRNAs in most retroviruses, in that introduced exonic enhancer elements can increase splicing efficiency.

Asbestos is a form of certain serpentine or amphibole minerals that has crystallized in a particular habit known as asbestiform.( 1 ) Amphibole minerals are often encountered in metamorphic geologi...

We previously showed that the yeast three-hybrid system provides a genetic assay of both RNA and protein components for avian retroviral RNA encapsidation. In the current study, we used this assay to precisely define cis -acting determinants involved in avian leukosis sarcoma virus packaging RNA binding to Gag protein. In vivo screening of Rous sarcoma virus mutants was performed with randomly mutated minimal packaging sequences (MΨ) made using PCR amplification after cotransformation with GagΔPR protein into yeast cells. Colonies with low β-galactosidase activity were analyzed to locate mutations in MΨ sequences affecting binding to Gag proteins. This genetic assay delineated secondary structural elements that are important for efficient RNA binding, including a single-stranded small bulge containing the initiation codon for uORF3, as well as adjacent stem structures. This implies a possible tertiary structure favoring the high-affinity binding sites for Gag. In most cases, results from the three-hybrid assay were well correlated with those from the viral RNA packaging assays. The results from random mutagenesis using the rapid three-hybrid binding assay are consistent with those from site-directed mutagenesis using in vivo packaging assays.

Recent laboratory research indicates physiologic sampling of gas and vapor may provide more representative estimates of personal exposures than traditional methods. Modifications to the physiologic sampling pump (PSP) used in that research are described which extend its usefulness to size-selective sampling of particulates. PSPs used in previous research varied motor speed to keep sampling proportional to the subject's inhalation. This caused airflow and particle velocities through the collection device to continually change making those pumps unsuitable for sampling particulates. The modified implementation of the PSP pulls a constant airflow into and through a cyclone, then uses valves to either direct the airflow through, or divert the airflow around, the sampling filter. By using physiologic inputs to regulate the fraction of each second that air flows through the sampling filter, samples may be collected in proportion to inhalation rate. To evaluate the performance of a functional prototype 5 different sizes of monodisperse aerosols of ammonium fluorescein were generated by a vibrating orifice aerosol generator and introduced into a calm air chamber. To simulate different inhalation rates the valves of the PSP were energized using 9 different duty cycles. Efficiency curves are presented and compared to a standard respirable convention by bias mapping. The performance of the modified cyclone used in the PSP sampling head compared favorably with a commercially available cyclone of the same model, operating at a constant airflow (± 10% over almost all the size distributions of concern). The new method makes physiologic sampling of the respirable fraction of particulates feasible.

Recently, physiologic sampling pumps (PSPs), which can adjust their sampling rates in proportion to wearers' minute ventilation (V[combining dot above](E)), have been proposed to better estimate exposure to airborne contaminants in the workplace. A laboratory evaluation was conducted to compare the performance of a new PSP with a traditional sampling pump (TSP) in an exposure chamber. Fifteen subjects (aged 19-36 years) performed two replicate sessions of step-tests for correlated and uncorrelated exposure scenarios on four separate days. When exposed to a scenario in which subject V[combining dot above](E) is highly correlated with m-xylene concentration over the sampling period (r = 0.93), the PSP-measured time-weighted average (TWA) concentrations are higher than TSP-measured concentrations (average ratio of PSP to TSP = 1.18). The ratio of PSP- and TSP-measured TWA concentrations for the uncorrelated scenario (r = 0.02) is closer to one, as expected, with an average value of 0.94. The test results of the linear mixed model further indicate that the performance of the PSP is unaffected by the anthropometric and physiological characteristics of the wearer. Potential differences in exposure estimates resulting from the use of the two instruments were examined in light of various schemes which can potentially occur in the field. With the capability of estimating the total volume of air inhaled over the sampling period with improved accuracy, PSPs show promise in reducing the inherent uncertainty in current risk assessment approaches that entail constant-flow (TSP) sampling approaches.

Methods for estimating airborne contaminant concentrations at specific locations within enclosed spaces, such as mathematical models and computational fluid dynamics (CFD), often are validated against directly measured concentrations. However, concentration variation with time introduces uncertainty into the measured concentration. Failure to determine monitoring time requirements can lead to errors in quantifying representative concentrations, which are likely to be attributed to errors in the method being validated. In the current study, to obtain the representative concentrations at multiple locations with a direct reading instrument, we used the standard deviation ratio (SDR) method to determine the required minimum monitoring time within a specified precision limit. To demonstrate the use of the SDR approach in constructing precision confidence intervals, tracer gas concentrations at nine sampling locations in an experimental room were measured to obtain population parameters. Three flow rates of 0.9, 3.3 and 5.5 m(3) min(-1) were employed and contaminant concentrations were measured using a photoionization analyser. Monitoring time requirements varied substantially with location within the room and were strongly dependent upon the flow rate of air through the room. The proposed method would be very useful for industrial hygienists and indoor air researchers who sometimes need to obtain several hundred measured concentrations for validation purposes or to perform tests under repeatable conditions in enclosed spaces. This study also showed that the proposed method can be used to devise efficient indoor monitoring strategies.