Chloroform exposure in air and water in Swedish indoor swimming pools-urine as a biomarker of occupational exposure.

INTRODUCTION: Disinfection by-products are produced in water disinfected with chlorine-based products. One such group is trihalomethanes, and chloroform is the most abundant trihalomethane in swimming pool areas. Chloroform can be absorbed by inhalation, ingestion, and dermal absorption, and is classified as possibly carcinogenic. AIM: To investigate if chloroform concentrations in air and water affect the chloroform concentration in urine samples of exposed swimming pool workers. METHODS: Workers from 5 adventure indoor swimming pools carried personal chloroform air samplers and provided up to 4 urine samples during one workday. Chloroform concentrations were analyzed with a linear mixed model analysis to investigate a possible correlation between air and urine concentrations. RESULTS: The geometric mean chloroform concentration was 11 µg/m3 in air and 0.009 µg/g creatinine in urine among individuals with ≤2 h at work, 0.023 µg/g creatinine among those with >2-5 working hours, and 0.026 µg/g creatinine in the group with >5-10 working hours. A risk of higher levels of chloroform in urine was associated with longer hours at work (≤2 h versus >5-10 h, odds ratio [OR] 2.04, 95% confidence interval [CI] 1.25-3.34), personal chloroform concentrations in air (≤17.00 µg/m3 versus >28.00 µg/m3, OR 9.23, 95% CI 3.68-23.13) and working at least half the working day near the swimming pools (OR 3.16, 95% CI 1.33-7.55). Executing work tasks in the swimming pool water was not associated with higher chloroform concentrations in urine compared to only working on land (OR 0.82, 95% CI 0.27-2.45). CONCLUSION: There is an accumulation of chloroform concentrations in urine during a workday and a correlation between personal air and urine concentrations of chloroform among workers in Swedish indoor swimming pools.

Occupational Exposure to Trichloramine and Endotoxins: Adverse Health Effects Among Personnel in Adventure and Rehabilitation Swimming Pool Facilities.

OBJECTIVE: To study occupational exposure totrichloramine and endotoxins in air at adventure and rehabilitation swimming pool facilities from an adverse health effects perspective. METHODS: Air concentrations of trichloramine and endotoxins were measured in five adventure and 10 rehabilitation facilities. Respiratory and ocular symptoms were self-reported, and spirometry and fraction of exhaled nitric oxide (FEno) were measured. RESULTS: Compared to rehabilitation facilities, the mean trichloramine concentrations in the adventure facilities were higher, both personal (80 µg/m3 (n  = 41) vs 19 µg/m3 (n = 21)) and stationary (183 µg/m3 (n = 51) vs 23 µg/m3 (n = 32)), with higher frequency of ocular and respiratory symptoms. Low stationary endotoxin levels (<0.64 to 25 EU/m3) were found, compared to the reference value (90 EU/m3). CONCLUSIONS: Higher trichloramine concentrations in air and more ocular and respiratory symptoms in adventure facilities call for adequate occupational exposure limits.

Introducing a new design of digital tool to increase vibration risk assessments: challenges with education-based interventions.

Objectives. This study aimed to investigate whether introducing a digital risk assessment tool, the Swedish National Vibration Database, would increase the number of risk assessments on hand-arm and whole-body vibration. Employer and safety representatives from companies where vibration exposure is common were invited. Methods. Of the 2953 invited companies, 1916 were selected for educational intervention and the remaining 1037 companies served as a control group with no intervention. For the educational intervention, participating companies were further divided into two groups (group A, n = 26; group B, n = 47) that both received information regarding risk assessment, but group B was also informed about the digital tool. Both groups answered a questionnaire on risk assessment before the intervention and at the follow-up, 6 months later; the control group received the same questionnaire but no education (group C, n = 22). Results. Of the invited companies, only 2% chose to participate and 7% at follow-up. Seventy-eight percent of the participants had made some kind of risk assessment of vibration at follow-up. Conclusion. Due to the low participation rate among invited companies, this study is not able to draw any conclusions on whether the digital tool can be used to increase the number of risk assessments.

Occupational Exposure to Metalworking Fluid and the Effect on Health Symptoms-An Intervention Study.

Exposure to metalworking fluid has been shown to cause health problems among workers. The aim of this study was to compare health outcomes and levels of exposure among workers exposed to metalworking fluid before and after the implementation of preventive measures. The frequencies of self-reported symptoms, as well as the concentrations of measured substances were lower after the implementation of preventive measures. Logistic regression showed statistically significant differences in the report of irritations, a stuffy or runny nose, eye irritation, a hoarse or dry throat, and a cough with odds ratios of 0.31 (95% confidence interval [CI] 0.21 to 0.47), 0.12 (95% CI 0.05 to 0.29), 0.13 (95% CI 0.06 to 0.28), and 0.24 (95% CI 0.12 to 0.46), respectively. This confirms the efficiency of the performed intervention and highlights the importance of eliminating the recirculation of contaminated air.

Silica Dust Exposure Increases Risk for Rheumatoid Arthritis: A Swedish National Registry Case-Control Study.

OBJECTIVE: Rheumatoid arthritis (RA) is an inflammatory disease with unknown etiology. This study examines if silica dust exposure increases the risk for seropositive and seronegative RA. METHODS: A nationwide registry case-control study was conducted that included all cases of RA in Sweden between 2005 and 2016. In total, 31,139 cases with two matched controls were included. A JEM was used to estimate exposure. RESULTS: Silica dust exposure was associated with a statistically significant increase in odds ratio (OR) for seropositive (OR 1.22, 95% CI 1.05 to 1.40) and seronegative (OR 1.23, 95% CI 1.04 to 1.46) RA among men. CONCLUSION: This study found an increased OR for RA in silica-exposed men. The OR was equal for seropositive and seronegative RA. These findings further support the hypothesis that silica dust may be a trigger for RA.

Occupational Quartz Exposure in a Population of Male Individuals-Association With Risk of Developing Atrial Fibrillation.

OBJECTIVE: Occupational quartz exposure is a health risk, with increased risk of developing lung, autoimmune diseases, and elevated mortality in cardiovascular diseases. METHODS: The population was obtained from the period 2005 to 2016 and consisted of 5237 cases of patients with atrial fibrillation (AF). Quartz exposure information was obtained through a Swedish job exposure matrix. RESULTS: The risk of developing AF was increased for the quartz-exposed male population who were within a year of having commenced employment OR 1.54; (95% CI 1.06-2.24); this increased in the age group 20 to 55 (OR 2.05; CI 95% 1.02-4.10). CONCLUSION: Our main conclusion is that quartz dust exposure may be related to increased risk of AF in high exposed (above 0.05 mg/m mean quartz dust) in men aged 20 to 55 years.

Risks of developing ulcerative colitis and Crohn's disease in relation to silica dust exposure in Sweden: a case-control study.

OBJECTIVE: To determine whether occupational exposure to silica dust causes an increased risk of developing Crohn's disease (CD) and ulcerative colitis (UC). DESIGN: Case-control study of CD (K50) and UC (K51) from 2007 through 2016. Controls were matched to cases (2:1) based on age, sex and county at the time of diagnosis. A job exposure matrix was used to estimate the occupational silica exposure of all cases and controls. SETTING: Medical and occupational data from the National Outpatient Register were used to implement a case-control analysis, while the two controls used for each case were selected from the National Register of the Total Population. PARTICIPANTS: All men and women aged 20-65 years old who were diagnosed with CD (K50) and UC (K51) during the years of study were included and assigned two controls, resulting in 58 136 cases and 116 272 controls. MAIN OUTCOMES: Silica dust exposure correlates with an increased risk of developing UC in men and CD in women. RESULTS: The prevalence of UC was significantly higher in the group exposed to silica dust (OR 1.13, 95% CI 1.06 to 1.21) than in controls, particularly in individuals with over 5 years exposure. When stratified by sex, a significantly increased OR was detected for men (OR 1.33, 95% CI 1.05 to 1.22). This trend was also consistent with longer exposure times. The prevalence of UC was not increased in exposed women. The prevalence of CD was significantly increased among exposed women (OR 1.29, 95% CI 1.01 to 1.65), but not for exposed men. CONCLUSIONS: Silica dust exposure correlates with an increased risk of developing UC, especially in men, and the risk seems to increase with the duration and degree of exposure. Conversely, silica dust exposure correlates positively with the risk of developing CD in women.

Nerve Function Impairment After Acute Vibration Exposure.

OBJECTIVE: This study was to investigate the acute effects of hand-arm vibrations on the nerve functions of the hands, and the impact of the grip force applied to the vibrating tool during exposure. METHODS: Grip strength and perception of vibration, touch, and temperature were evaluated using quantitative sensory testing (QST) before and after vibration exposure in 21 occupationally unexposed individuals. The procedure was performed twice, with a higher grip force being applied during exposure on the second occasion. RESULTS: Vibration perception was significantly impaired after both exposures. Grip strength, perception of touch, and temperature were only significantly affected after the high grip force exposure. CONCLUSIONS: Exposure to hand-arm vibrations has acute effects on hand nerve function that are sensitive to the grip force applied during exposure.

Biomonitoring of Metal Exposure During Additive Manufacturing (3D Printing).

BACKGROUND: Additive manufacturing (AM) is a rapidly expanding new technology involving challenges to occupational health. Here, metal exposure in an AM facility with large-scale metallic component production was investigated during two consecutive years with preventive actions in between. METHODS: Gravimetric analyzes measured airborne particle concentrations, and filters were analyzed for metal content. In addition, concentrations of airborne particles <300 nm were investigated. Particles from recycled powder were characterized. Biomonitoring of urine and dermal contamination among AM operators, office personnel, and welders was performed. RESULTS: Total and inhalable dust levels were almost all below occupational exposure limits, but inductively coupled plasma mass spectrometry showed that AM operators had a significant increase in cobalt exposure compared with welders. Airborne particle concentrations (<300 nm) showed transient peaks in the AM facility but were lower than those of the welding facility. Particle characterization of recycled powder showed fragmentation and condensates enriched in volatile metals. Biomonitoring showed a nonsignificant increase in the level of metals in urine in AM operators. Dermal cobalt and a trend for increasing urine metals during Workweek Year 1, but not in Year 2, indicated reduced exposure after preventive actions. CONCLUSION: Gravimetric analyses showed low total and inhalable dust exposure in AM operators. However, transient emission of smaller particles constitutes exposure risks. Preventive actions implemented by the company reduced the workers' metal exposure despite unchanged emissions of particles, indicating a need for careful design and regulation of the AM environments. It also emphasizes the need for relevant exposure markers and biomonitoring of health risks.

Visualizing the Nonhomogeneous Structure of RAD51 Filaments Using Nanofluidic Channels.

RAD51 is the key component of the homologous recombination pathway in eukaryotic cells and performs its task by forming filaments on DNA. In this study we investigate the physical properties of RAD51 filaments formed on DNA using nanofluidic channels and fluorescence microscopy. Contrary to the bacterial ortholog RecA, RAD51 forms inhomogeneous filaments on long DNA in vitro, consisting of several protein patches. We demonstrate that a permanent "kink" in the filament is formed where two patches meet if the stretch of naked DNA between the patches is short. The kinks are readily seen in the present microscopy approach but would be hard to identify using conventional single DNA molecule techniques where the DNA is more stretched. We also demonstrate that protein patches separated by longer stretches of bare DNA roll up on each other and this is visualized as transiently overlapping filaments. RAD51 filaments can be formed at several different conditions, varying the cation (Mg(2+) or Ca(2+)), the DNA substrate (single-stranded or double-stranded), and the RAD51 concentration during filament nucleation, and we compare the properties of the different filaments formed. The results provide important information regarding the physical properties of RAD51 filaments but also demonstrate that nanofluidic channels are perfectly suited to study protein-DNA complexes.

The conquest of middle-earth: combining top-down and bottom-up nanofabrication for constructing nanoparticle based devices.

The development of top-down nanofabrication techniques has opened many possibilities for the design and realization of complex devices based on single molecule phenomena such as e.g. single molecule electronic devices. These impressive achievements have been complemented by the fundamental understanding of self-assembly phenomena, leading to bottom-up strategies to obtain hybrid nanomaterials that can be used as building blocks for more complex structures. In this feature article we highlight some relevant published work as well as present new experimental results, illustrating the versatility of self-assembly methods combined with top-down fabrication techniques for solving relevant challenges in modern nanotechnology. We present recent developments on the use of hierarchical self-assembly methods to bridge the gap between sub-nanometer and micrometer length scales. By the use of non-covalent self-assembly methods, we show that we are able to control the positioning of nanoparticles on surfaces, and to address the deterministic assembly of nano-devices with potential applications in plasmonic sensing and single-molecule electronics experiments.

Sensing conformational changes in DNA upon ligand binding using QCM-D. Polyamine condensation and Rad51 extension of DNA layers.

Biosensors, in which binding of ligands is detected through changes in the optical or electrochemical properties of a DNA layer confined to the sensor surface, are important tools for investigating DNA interactions. Here, we investigate if conformational changes induced in surface-attached DNA molecules upon ligand binding can be monitored by the quartz crystal microbalance with dissipation (QCM-D) technique. DNA duplexes containing 59-184 base pairs were formed on QCM-D crystals by stepwise assembly of synthetic oligonucleotides of designed base sequences. The DNA films were exposed to the cationic polyamines spermidine and spermine, known to condense DNA molecules in bulk experiments, or to the recombination protein Rad51, known to extend the DNA helix. The binding and dissociation of the ligands to the DNA films were monitored in real time by measurements of the shifts in resonance frequency (Δf) and in dissipation (ΔD). The QCM-D data were analyzed using a Voigt-based model for the viscoelastic properties of polymer films in order to evaluate how the ligands affect thickness and shear viscosity of the DNA layer. Binding of spermine shrinks all DNA layers and increases their viscosity in a reversible fashion, and so does spermidine, but to a smaller extent, in agreement with its lower positive charge. SPR was used to measure the amount of bound polyamines, and when combined with QCM-D, the data indicate that the layer condensation leads to a small release of water from the highly hydrated DNA films. The binding of Rad51 increases the effective layer thickness of a 59 bp film, more than expected from the know 50% DNA helix extension. The combined results provide guidelines for a QCM-D biosensor based on ligand-induced structural changes in DNA films. The QCM-D approach provides high discrimination between ligands affecting the thickness and the structural properties of the DNA layer differently. The reversibility of the film deformation allows comparative studies of two or more analytes using the same DNA layer as demonstrated here by spermine and spermidine.

UV transition moments of tyrosine.

To assist polarized-light spectroscopy for protein-structure analysis, the UV spectrum of p-cresol, the chromophore of tyrosine, was studied with respect to transition moment directions and perturbation by solvent environment. From linear dichroism (LD) spectra of p-cresol aligned in stretched matrices of poly(vinyl alcohol) and polyethylene, the lowest π-π* transition (Lb) is found to have pure polarization over its entire absorption (250-300 nm) with a transition moment perpendicular to the symmetry axis (C1-C4), both in polar and nonpolar environments. For the second transition (La), polarized parallel with the symmetry axis, a certain admixture of intensity with orthogonal polarization is noticed, depending on the environment. While the Lb spectrum in cyclohexane shows a pronounced vibrational structure, it is blurred in methanol, which can be modeled as due to many microscopic polar environments. With the use of quantum mechanical (QM) calculations, the transition moments and solvent effects were analyzed with the B3LYP and ωB97X-D functionals in cyclohexane, water, and methanol using a combination of implicit and explicit solvent models. The blurred Lb band is explained by solvent hydrogen bonds, where both accepting and donating a hydrogen causes energy shifts. The inhomogeneous solvent-shift sensitivity in combination with robust polarization can be exploited for analyzing tyrosine orientation distributions in protein complexes using LD spectroscopy.

Swi5-Sfr1 protein stimulates Rad51-mediated DNA strand exchange reaction through organization of DNA bases in the presynaptic filament.

The Swi5-Sfr1 heterodimer protein stimulates the Rad51-promoted DNA strand exchange reaction, a crucial step in homologous recombination. To clarify how this accessory protein acts on the strand exchange reaction, we have analyzed how the structure of the primary reaction intermediate, the Rad51/single-stranded DNA (ssDNA) complex filament formed in the presence of ATP, is affected by Swi5-Sfr1. Using flow linear dichroism spectroscopy, we observe that the nucleobases of the ssDNA are more perpendicularly aligned to the filament axis in the presence of Swi5-Sfr1, whereas the bases are more randomly oriented in the absence of Swi5-Sfr1. When using a modified version of the natural protein where the N-terminal part of Sfr1 is deleted, which has no affinity for DNA but maintained ability to stimulate the strand exchange reaction, we still observe the improved perpendicular DNA base orientation. This indicates that Swi5-Sfr1 exerts its activating effect through interaction with the Rad51 filament mainly and not with the DNA. We propose that the role of a coplanar alignment of nucleobases induced by Swi5-Sfr1 in the presynaptic Rad51/ssDNA complex is to facilitate the critical matching with an invading double-stranded DNA, hence stimulating the strand exchange reaction.

Minor-groove binding drugs: where is the second Hoechst 33258 molecule?

Hoechst 33258 binds with high affinity into the minor groove of AT-rich sequences of double-helical DNA. Despite extensive studies of this and analogous DNA binding molecules, there still remains uncertainty concerning the interactions when multiple ligand molecules are accommodated within close distance. Albeit not of direct concern for most biomedical applications, which are at low drug concentrations, interaction studies for higher drug binding are important as they can give fundamental insight into binding mechanisms and specificity, including drug self-stacking interactions that can provide base-sequence specificity. Using circular dichroism (CD), isothermal titration calorimetry (ITC), and proton nuclear magnetic resonance ((1)H NMR), we examine the binding of Hoechst 33258 to three oligonucleotide duplexes containing AT regions of different lengths: [d(CGCGAATTCGCG)]2 (A2T2), [d(CGCAAATTTGCG)]2 (A3T3), and [d(CGAAAATTTTCG)]2 (A4T4). We find similar binding geometries in the minor groove for all oligonucleotides when the ligand-to-duplex ratio is less than 1:1. At higher ratios, a second ligand can be accommodated in the minor groove of A4T4 but not A2T2 or A3T3. We conclude that the binding of the second Hoechst to A4T4 is not cooperative and that the molecules are sitting with a small separation apart, one after the other, and not in a sandwich structure as previously proposed.

Lifetime heterogeneity of DNA-bound dppz complexes originates from distinct intercalation geometries determined by complex-complex interactions.

Despite the extensive interest in structurally explaining the photophysics of DNA-bound [Ru(phen)(2)dppz](2+) and [Ru(bpy)(2)dppz](2+), the origin of the two distinct emission lifetimes of the pure enantiomers when intercalated into DNA has remained elusive. In this report, we have combined a photophysical characterization with a detailed isothermal titration calorimetry study to investigate the binding of the pure Δ and Λ enantiomers of both complexes with [poly(dAdT)](2). We find that a binding model with two different binding geometries, proposed to be symmetric and canted intercalation from the minor groove, as recently reported in high-resolution X-ray structures, is required to appropriately explain the data. By assigning the long emission lifetime to the canted binding geometry, we can simultaneously fit both calorimetric data and the binding-density-dependent changes in the relative abundance of the two emission lifetimes using the same binding model. We find that all complex-complex interactions are slightly unfavorable for Δ-[Ru(bpy)(2)dppz](2+), whereas interactions involving a complex canted away from a neighbor are favorable for the other three complexes. We also conclude that Δ-[Ru(bpy)(2)dppz](2+) preferably binds isolated, Δ-[Ru(phen)(2)dppz](2+) preferably binds as duplets of canted complexes, and that all complexes are reluctant to form longer consecutive sequences than triplets. We propose that this is due to an interplay of repulsive complex-complex and attractive complex-DNA interactions modulated by allosteric DNA conformation changes that are largely affected by the nature of the ancillary ligands.

Airway irritation among indoor swimming pool personnel: trichloramine exposure, exhaled NO and protein profiling of nasal lavage fluids.

PURPOSE: Occurrence of airway irritation among indoor swimming pool personnel was investigated. The aims of this study were to assess trichloramine exposure levels and exhaled nitric oxide in relation to the prevalence of airway symptoms in swimming pool facilities and to determine protein effects in the upper respiratory tract. METHODS: The presence of airway symptoms related to work was examined in 146 individuals working at 46 indoor swimming pool facilities. Levels of trichloramine, as well as exhaled nitric oxide, were measured in five facilities with high prevalence of airway irritation and four facilities with no airway irritation among the personnel. Nasal lavage fluid was collected, and protein profiles were determined by a proteomic approach. RESULTS: 17 % of the swimming pool personnel reported airway symptoms related to work. The levels of trichloramine in the swimming pool facilities ranged from 0.04 to 0.36 mg/m(3). There was no covariance between trichloramine levels, exhaled nitric oxide and prevalence of airway symptoms. Protein profiling of the nasal lavage fluid showed that the levels alpha-1-antitrypsin and lactoferrin were significantly higher, and S100-A8 was significantly lower in swimming pool personnel. CONCLUSIONS: This study confirms the occurrence of airway irritation among indoor swimming pool personnel. Our results indicate altered levels of innate immunity proteins in the upper airways that may pose as potential biomarkers. However, swimming pool facilities with high prevalence of airway irritation could not be explained by higher trichloramine exposure levels. Further studies are needed to clarify the environmental factors in indoor swimming pools that cause airway problems and affect the immune system.

Airway symptoms and biological markers in nasal lavage fluid in subjects exposed to metalworking fluids.

BACKGROUNDS: Occurrence of airway irritation among industrial metal workers was investigated. The aims were to study the association between exposures from water-based metal working fluids (MWF) and the health outcome among the personnel, to assess potential effects on the proteome in nasal mucous membranes, and evaluate preventive actions. METHODS: The prevalence of airway symptoms related to work were examined among 271 metalworkers exposed to MWF and 24 metal workers not exposed to MWF at the same factory. At the same time, air levels of potentially harmful substances (oil mist, morpholine, monoethanolamine, formaldehyde) generated from MWF was measured. Nasal lavage fluid was collected from 13 workers and 15 controls and protein profiles were determined by a proteomic approach. RESULTS: Airway symptoms were reported in 39% of the workers exposed to MWF although the measured levels of MWF substances in the work place air were low. Highest prevalence was found among workers handling the MWF machines but also those working in the same hall were affected. Improvement of the ventilation to reduce MWF exposure lowered the prevalence of airway problems. Protein profiling showed significantly higher levels of S100-A9 and lower levels of SPLUNC1, cystatin SN, Ig J and ß2-microglobulin among workers with airway symptoms. CONCLUSIONS: This study confirms that upper airway symptoms among metal workers are a common problem and despite low levels of MWF-generated substances, effects on airway immune proteins are found. Further studies to clarify the role of specific MWF components in connection to airway inflammation and the identified biological markers are warranted.

Cell surface binding and uptake of arginine- and lysine-rich penetratin peptides in absence and presence of proteoglycans.

Cell surface proteoglycans (PGs) appear to promote uptake of arginine-rich cell-penetrating peptides (CPPs), but their exact functions are unclear. To address if there is specificity in the interactions of arginines and PGs leading to improved internalization, we used flow cytometry to examine uptake in relation to cell surface binding for penetratin and two arginine/lysine substituted variants (PenArg and PenLys) in wildtype CHO-K1 and PG-deficient A745 cells. All peptides were more efficiently internalized into CHO-K1 than into A745, but their cell surface binding was independent of cell type. Thus, PGs promote internalization of cationic peptides, irrespective of the chemical nature of their positive charges. Uptake of each peptide was linearly dependent on its cell surface binding, and affinity is thus important for efficiency. However, the gradients of these linear dependencies varied significantly. Thus each peptide's ability to stimulate uptake once bound to the cell surface is reliant on formation of specific uptake-promoting interactions. Heparin affinity chromatography and clustering experiments showed that penetratin and PenArg binding to sulfated sugars is stabilized by hydrophobic interactions and result in clustering, whereas PenLys only interacts through electrostatic attraction. This may have implications for the molecular mechanisms behind arginine-specific uptake stimulation as penetratin and PenArg are more efficiently internalized than PenLys upon interaction with PGs. However, PenArg is also least affected by removal of PGs. This indicates that an increased arginine content not only improve PG-dependent uptake but also that PenArg is more adaptable as it can use several portals of entry into the cell.