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The eukaryotic genome is organized into nucleosomes, the fundamental units of chromatin. The positions of nucleosomes on DNA regulate protein-DNA interactions and in turn influence DNA-templated events. Despite the increasing number of genome-wide maps of nucleosome position, how global changes in gene expression relate to changes in nucleosome position is poorly understood. We show that in nucleosome occupancy mapping experiments in maize (Zea mays), particular genomic regions are highly susceptible to variation introduced by differences in the extent to which chromatin is digested with micrococcal nuclease (MNase). We exploited this digestion-linked variation to identify protein footprints that are hypersensitive to MNase digestion, an approach we term differential nuclease sensitivity profiling (DNS-chip). Hypersensitive footprints were enriched at the 5' and 3' ends of genes, associated with gene expression levels, and significantly overlapped with conserved noncoding sequences and the binding sites of the transcription factor KNOTTED1. We also found that the tissue-specific regulation of gene expression was linked to tissue-specific hypersensitive footprints. These results reveal biochemical features of nucleosome organization that correlate with gene expression levels and colocalize with functional DNA elements. This approach to chromatin profiling should be broadly applicable to other species and should shed light on the relationships among chromatin organization, protein-DNA interactions, and genome regulation., (© 2014 American Society of Plant Biologists. All rights reserved.)

Genome-scale mapping of pre-replication complex proteins has not been reported in mammalian cells. Poor enrichment of these proteins at specific sites may be due to dispersed binding, poor epitope availability or cell cycle stage-specific binding. Here, we have mapped sites of biotin-tagged ORC and MCM protein binding in G1-synchronized populations of Chinese hamster cells harboring amplified copies of the dihydrofolate reductase (DHFR) locus, using avidin-affinity purification of biotinylated chromatin followed by high-density microarray analysis across the DHFR locus. We have identified several sites of significant enrichment for both complexes distributed throughout the previously identified initiation zone. Analysis of the frequency of initiations across stretched DNA fibers from the DHFR locus confirmed a broad zone of de-localized initiation activity surrounding the sites of ORC and MCM enrichment. Mapping positions of mononucleosomal DNA empirically and computing nucleosome-positioning information in silico revealed that ORC and MCM map to regions of low measured and predicted nucleosome occupancy. Our results demonstrate that specific sites of ORC and MCM enrichment can be detected within a mammalian initiation zone, and suggest that initiation zones may be regions of generally low nucleosome occupancy where flexible nucleosome positioning permits flexible pre-RC assembly sites.

Previous work has demonstrated that the shield gas composition in gas metal arc welding can have a considerable effect on hexavalent chromium [Cr(VI)] concentration in the fume and on ozone concentrations near the arc. Normally a single shield gas is used. This paper describes a double shroud torch that allows used of concentric shield gases of different compositions. A solid stainless steel wire was used for welding. The double shroud torch used secondary shield gases containing small amounts of the reducing agents NO and C2H4. The Cr(VI) concentration in the fume and ozone concentration at a fixed point relative to the arc were measured and compared with results when using a single shield gas. Use of the reducing agents in secondary shielding using the double shroud torch was found to offer advantages for ozone concentration reduction compared with use in a conventional torch, but this was not found to be an advantage for reducing Cr(VI) concentrations.

Hexavalent chromium [Cr(VI)] and ozone are produced in many arc-welding processes. Cr(VI) is formed when welding with chromium-containing alloys and is a suspected carcinogen. Ozone is formed by the action of ultraviolet light from the arc on oxygen and can cause severe irritation to the eyes and mucous membranes. Previous work has demonstrated that reduction of sodium and potassium in manual metal arc-welding electrodes leads to substantial reductions in Cr(VI) concentrations in the fume as well as a reduction in the fume formation rate. In this paper replacement of potassium by lithium in a tubular wire welding electrode (self-shielding flux-cored) is shown to give reductions in Cr(VI) concentrations and fume formation rates. Previous work has also demonstrated that use of a tubular wire (metal cored) containing 1% zinc can, under certain conditions, result in a reduction in Cr(VI) formation rate and in ozone concentration near the arc but with a rise in the total fume formation rate. The effects of different shield gases and different levels of zinc are examined. An experimental chromium-containing tubular wire with 1% zinc was used with the following shield gases: argon, Argoshield 5, Argoshield 20, Helishield 101, Ar + 2% CO2, Ar + 5% CO2, Ar + 1% O2 and Ar + 2% O2. The wire gave > 98% reduction in Cr(VI) formation rate compared to the control wire provided the shield gas contained no oxygen. When the shield gas did contain oxygen, 1% zinc enhanced Cr(VI) formation rate, resulting in more than double the rates measured when welding with the control wire. Experiments with zinc concentrations, from 0.018 to 0.9% using Helishield 101, gave results indicating that there is an optimum zinc concentration from the point of view of Cr(VI) reduction. Implications of the use of lithium or zinc on the overall exposure risk are discussed.

Objective: To determine the prevalence of hearing loss associated with occupational noise exposure and other risk factors., Design: A cross-sectional study involving 269 exposed and 99 non-exposed subjects (non-industrial noise exposed subjects) randomly selected. Current noise exposure was estimated using both sound level meter and noise-dosimeter. Past noise exposure was estimated by interview questionnaire. Otoscopic examination and conventional frequency (0.25-8 kHz) audiometry were used to assess the hearing loss in each subject., Results: 75% (202 subjects) from the exposed group were exposed to a daily Leq above the permissible level of 85 dB(A) and most (61%) of these did not and had never used any form of hearing protection. Hearing loss was found to be bilateral and symmetrical in both groups. Bivariate analysis showed a significant hearing loss in the exposed vs non-exposed subjects with a characteristic dip at 4 kHz. Thirty eight percent of exposed subjects had hearing impairment, which was an 8-fold higher rate than that found for non-exposed subjects. Multivariate analysis indicated exposure to noise was the primary, and age the secondary predictor of hearing loss. Odds of hearing impairment were lower for a small sub-group of exposed workers using hearing protection (N=19) in which logistic regression analysis showed the probability of workers adopting hearing protective devices increased with noise exposure, education, and awareness of noise control. Hearing loss was also greater amongst those who used headphones to listen to recorded cassettes., Conclusion: Gross occupational exposure to noise has been demonstrated to cause hearing loss and the authors believe that occupational hearing loss in Saudi Arabia is a widespread problem. Strategies of noise assessment and control are introduced which may help improve the work environment.

The objective was to investigate the reliability and effects of age and noise on high-frequency hearing thresholds. A cross-sectional study was used involving 187 exposed and 52 non-industrial noise-exposed subjects selected randomly from noise-exposed and non-industrial noise-exposed subjects, respectively. Each subject was tested with both conventional-frequency (0.25-8 kHz) and high-frequency (10-18 kHz) audiometry. Test-retest results showed that high-frequency audiometry (HFA) was as reliable as the conventional procedure. Although the inter-subject variation was large, the intra-subject variation was small, indicating that HFA can be used more reliably than the conventional procedure to monitor individual cases over time. Both the hearing threshold at high frequencies and the upper frequency limit deteriorated as a function of age and frequency. The exposed subjects had significantly higher hearing thresholds than the non-exposed subjects at all the high frequencies tested, the difference between the two groups being greatest at 14 kHz. Multivariate analysis indicated that age was the primary predictor and noise exposure the secondary predictor of hearing thresholds in a high frequency range (10-18 kHz). In contrast, multivariate analysis indicated the reverse order-noise exposure as the primary predictor, then age-for a conventional frequency range (0.25-8 kHz). The results of this study suggest that HFA might be used as an early indicator for noise-induced hearing loss and acoustic trauma rather than audiometry at a conventional frequency (4 kHz), particularly for younger groups.

Prediction of fume formation rate during metal arc welding and the composition of the fume are of interest to occupational hygienists concerned with risk assessment and to manufacturers of welding consumables. A model for GMAW (DC electrode positive) is described based on the welder determined process parameters (current, wire feed rate and wire composition), on the surface area of molten metal in the arc and on the partial vapour pressures of the component metals of the alloy wire. The model is applicable to globular and spray welding transfer modes but not to dip mode. Metal evaporation from a droplet is evaluated for short time increments and total evaporation obtained by summation over the life of the droplet. The contribution of fume derived from the weld pool and spatter (particles of metal ejected from the arc) is discussed, as are limitations of the model. Calculated droplet temperatures are similar to values determined by other workers. A degree of relationship between predicted and measured fume formation rates is demonstrated but the model does not at this stage provide a reliable predictive tool.

Inhalation provocation tests were used to assess whether the volatile products of an activated resin had caused occupational asthma in a non-random sample of six asthmatic coal miners. The resin system uses the polymerization of polyester and styrene under the influence of the cross-linking agent dibenzoyl peroxide to secure roof, wall and floor bolts in mine tunnels. The tests were conducted sequentially in a double-blind fashion over a 'dose' range which extended just beyond the maximum likely to have been experienced occupationally during a single day's work. The tests were monitored by symptoms, changes in the forced expiratory volume in 1 s (FEV1) and changes in airway responsiveness. All subjects completed the series of tests without any significant decrements in FEV1 or significant increases in airway responsiveness. We conclude that the use of this resin system is not likely to have been the cause of the asthma in the test subjects, nor in the larger group of miners of which they were a sample, but neither possibility is fully excluded and the participants may not have been adequately representative of other asthmatic coal miners.

This paper describes the relationships between ultra-violet emission, ozone generation and CrVI production in MIG welding which were measured as a function of shield gas flow rate, welding voltage, electrode stick-out and shield gas composition using an automatic welding rig that permitted MIG welding under reproducible conditions. The experimental results are interpreted in terms of the physico-chemical processes occurring in the micro- and macro-environments of the arc as part of research into process modification to reduce occupational exposure to ozone and CrVI production rates in MIG welding. We believe the techniques described here, and in particular the use of what we have termed u.v.-ozone measurements, will prove useful in further study of ozone generation and CrVI formation and may be applied in the investigation of engineering control of occupational exposure in MIG and other welding process such as Manual Metal Arc (MMA) and Tungsten Inert Gas (TIG).

A 46-year-old endoscopy nurse developed symptoms suggestive of occupational asthma after seven years of exposure to glutaraldehyde. An initial inhalation challenge test at the endoscopy suite caused a very dramatic immediate fall in FEV1 from 3.6 to 1.5 litres. To further evaluate the case and establish a threshold for the response, a technique was devised to allow a series of double-blind inhalation challenges with activated glutaraldehyde vapour at exposure levels of 0.01-0.32 ppm. The results suggested marked dual asthmatic reactions following challenges with 0.032 ppm glutaraldehyde; however, similar 'reactions' were observed on control days. These were associated with an increase in airway responsiveness to methacholine, with the PD20FEV1 falling from > 6400 micrograms to 135 micrograms. The interpretation of these results was potentially confounded by an intercurrent respiratory tract infection and by technically poor FEV1 recordings, so the challenge series was repeated three weeks later. The second series of double-blind inhalation challenges with carefully controlled exposures to glutaraldehyde (up to 0.32 ppm for 10 min) gave rise to no obvious asthmatic reactions, in marked contrast to the results of the unblinded workplace challenge. There was a slight increase in airway responsiveness, with the PD20FEV1 falling from > 6400 micrograms to 1850 micrograms. These results illustrate the potential for misdiagnosis of occupational asthma when unblinded challenge tests are used and show that, even with sophisticated investigatory techniques, a clear-cut diagnostic result may be elusive.