Reflections on the OECD guidelines for in vitro skin absorption studies.

At the 8th conference of Occupational and Environmental Exposure of the Skin to Chemicals (OEESC) (16-18 September 2019) in Dublin, Ireland, several researchers performing skin permeation assays convened to discuss in vitro skin permeability experiments. We, along with other colleagues, all of us hands-on skin permeation researchers, present here the results from our discussions on the available OECD guidelines. The discussions were especially focused on three OECD skin absorption documents, including a recent revision of one: i) OECD Guidance Document 28 (GD28) for the conduct of skin absorption studies (OECD, 2004), ii) Test Guideline 428 (TGD428) for measuring skin absorption of chemical in vitro (OECD, 2004), and iii) OECD Guidance Notes 156 (GN156) on dermal absorption issued in 2011 (OECD, 2011). GN156 (OECD, 2019) is currently under review but not finalized. A mutual concern was that these guidance documents do not comprehensively address methodological issues or the performance of the test, which might be partially due to the years needed to finalize and update OECD documents with new skin research evidence. Here, we summarize the numerous factors that can influence skin permeation and its measurement, and where guidance on several of these are omitted and often not discussed in published articles. We propose several improvements of these guidelines, which would contribute in harmonizing future in vitro skin permeation experiments.

Chloroanisoles may explain mold odor and represent a major indoor environment problem in Sweden.

Indoor mold odor is associated with adverse health effects, but the microbial volatiles underlying mold odor are poorly described. Here, chloroanisoles were studied as potential key players, being formed by microbial metabolism of chlorophenols in wood preservatives. Using a three-stage approach, we (i) investigated the occurrence of chloroanisoles in buildings with indoor air quality problems, (ii) estimated their frequency in Sweden, and (iii) evaluated the toxicological risk of observed chloroanisole concentrations. Analyses of 499 building materials revealed several chloroanisole congeners in various types of buildings from the 1950s to 1970s. Evaluation of Swedish records from this time period revealed three coinciding factors, namely an unprecedented nationwide building boom, national regulations promoting wood preservatives instead of moisture prevention, and use of chlorophenols in these preservatives. Chlorophenols were banned in 1978, yet analysis of 457 indoor air samples revealed several chloroanisole congeners, but at median air levels generally below 15 ng/m(3) . Our toxicological evaluation suggests that these concentrations are not detrimental to human health per se, but sufficiently high to cause malodor. Thereby, one may speculate that chloroanisoles in buildings contribute to adverse health effects by evoking odor which, enhanced by belief of the exposure being hazardous, induces stress-related and inflammatory symptoms.

The Global Landscape of Occupational Exposure Limits--Implementation of Harmonization Principles to Guide Limit Selection.

Occupational exposure limits (OELs) serve as health-based benchmarks against which measured or estimated workplace exposures can be compared. In the years since the introduction of OELs to public health practice, both developed and developing countries have established processes for deriving, setting, and using OELs to protect workers exposed to hazardous chemicals. These processes vary widely, however, and have thus resulted in a confusing international landscape for identifying and applying such limits in workplaces. The occupational hygienist will encounter significant overlap in coverage among organizations for many chemicals, while other important chemicals have OELs developed by few, if any, organizations. Where multiple organizations have published an OEL, the derived value often varies considerably-reflecting differences in both risk policy and risk assessment methodology as well as access to available pertinent data. This article explores the underlying reasons for variability in OELs, and recommends the harmonization of risk-based methods used by OEL-deriving organizations. A framework is also proposed for the identification and systematic evaluation of OEL resources, which occupational hygienists can use to support risk characterization and risk management decisions in situations where multiple potentially relevant OELs exist.

Neurobehavioral performance in volunteers after inhalation of white spirits with high and low aromatic content.

The content of aromatic hydrocarbons in solvent mixtures, such as white spirits (WS), has been assumed a major contributor to the neurotoxic effects of these compounds. Hence, dearomatized WS have been introduced to the market rapidly in the last decade. Studies investigating other aromatic hydrocarbons (toluene) and animal models have supported the aforementioned assumption, but the current study is the first one to compare acute neurobehavioral effects of exposure to aromatic and dearomatized WS (aWS, daWS) content in human volunteers at current occupational exposure limit values. In a pseudo-randomized crossover design, six female and six male healthy volunteers were exposed to aWS and daWS at two concentrations (100 and 300 mg/m(3)) and to clean air for 4 h at rest. During each of the five exposure conditions, volunteers performed five neurobehavioral tasks that were selected following a multidisciplinary approach that accounted for findings from the cognitive neurosciences and mechanisms of solvent toxicity. Two of the tasks indicated performance changes during aromatic WS exposure, the working memory (WM) and the response shifting task, but both effects are difficult to interpret due to low mean accuracy in the WM task and due to a lack of dose-response relationship in the response shifting task. Healthy human volunteers showed weak and inconsistent neurobehavioral impairment after 4-h exposures to 100 and 300 mg/m(3) aromatic or dearomatized WS. Our multidisciplinary approach of selecting neurobehavioral test methods may guide the test selection strategies in future studies.

Acute effects of exposure to vapors of 3-methyl-1-butanol in humans.

UNLABELLED: The secondary alcohol 3-methyl-1-butanol (3MB, isoamyl alcohol) is used, for example, as a solvent in a variety of applications and as a fragrance ingredient. It is also one of the microbial volatile organic compounds (MVOCs) found in indoor air. There are little data on acute effects. The aim of the study was to assess the acute effects of 3MB in humans. Thirty healthy volunteers (16 men and 14 women) were exposed in random order to 1 mg/m(3) 3MB or clean air for 2 h at controlled conditions. Ratings with visual analogue scales revealed slightly increased perceptions of eye irritation (P = 0.048, Wilcoxon) and smell (P < 0.0001) compared with control exposure. The other ratings were not significantly affected (irritation in nose and throat, dyspnea, headache, fatigue, dizziness, nausea, and intoxication). No significant exposure-related effects were found in blinking frequency, tear film break-up time, vital staining of the eye, nasal lavage biomarkers, lung function, and nasal swelling. In conclusion, this study suggests that 3MB is not a causative factor for health effects in damp and moldy buildings. PRACTICAL IMPLICATIONS: 3-Methyl-1-butanol (3MB) is one of the most commonly reported MVOCs in damp and moldy buildings and in occupational settings related to agriculture and composting. Our study revealed no irritation effects at 1 mg/m3, a concentration higher than typically found in damp and moldy buildings. Our study thus suggests that 3MB is not a causative factor for health effects in damp and moldy buildings.

Acute effects of exposure to 1 mg/m(3) of vaporized 2-ethyl-1-hexanol in humans.

The objective was to assess acute effects from controlled exposure of volunteers to 2-ethyl-1-hexanol, a volatile organic compound that is often found in indoor air. Sixteen males and fourteen females were in random order exposed to 1 mg/m(3) of vapors of 2-ethyl-1-hexanol or to clean air (control exposure) in an exposure chamber during 2 h at rest. The subjects performed symptom ratings on Visual Analog Scales. During exposure to 2-ethyl-1-hexanol subjective ratings of smell and eye discomfort were minimally but significantly increased. Ratings of nasal irritation, throat irritation, headache, dyspnoea, fatigue, dizziness, nausea, and intoxication were not significantly affected. No exposure-related effects on measurement of blinking frequency by electromyography, measurement of the eye break-up time, vital staining of the eye, nasal lavage biomarkers, transfer tests, spirometric and rhinometric measures were seen. No differences in response were seen between sexes or between atopics and non-atopics. Practical Implications It is important to assess acute effects in volatile organic compounds like 2-ethyl-1-hexanol. 2-ethyl-1-hexanol is often found in indoor air generated by degradation of plastic building materials or in new buildings. There are associations between 2-ethyl-1-hexanol in indoor air and respiratory effects, eye irritation, headache, and blurred vision. A controlled chamber exposure study in acute effects was performed. In conclusion, this study showed weak subjective symptom of irritation in the eyes.

The 2008 ICOH Workshop on Skin Notation.

BACKGROUND: On 29 March 2008 the International Commission on Occupational Health (ICOH) Scientific Committee on Occupational and Environmental Dermatoses organized a Skin Notation Workshop hosted by the 11th International Percutaneous Penetration Perspectives Conference (La Grande Motte, France). Skin notation (S) was chosen as a topic for discussion because this is the only example of existing regulation in the field of dermal risk assessment. The issue was discussed in a previous workshop held in Siena, Italy in 2006 with the objective of focussing on the problems related to S, the different assignment criteria and the attempts to improve the S system made by various international and governmental agencies. A position paper was subsequently published. OBJECTIVES: The workshop in France was a continuation of this activity with the aim of evaluating how the different strategies can improve S. METHODS AND DISCUSSION: The Workshop was divided into two sessions. The first was dedicated to lectures focused on different aspects of S. In the second session participants discussed key issues with the aim of exploring the actions needed to improve international S. systems.

Washout kinetics of ethanol from the airways following inhalation of ethanol vapors and use of mouthwash.

Introduction: Breath analyzers are commonly used to test for alcohol intoxication, i.e., elevated systemic levels of ethanol, at workplaces and among vehicle drivers. However, local low-dose exposure to ethanol in the mouth or airways may temporarily increase the breath-alcohol concentration (BrAC) without the systemic ethanol level being affected, leading to false positive test results. The aim of this study was to assess the impact of local ethanol exposure on the BrAC.Methods: Eleven healthy adults (six women) were exposed to on average 856 mg/m3 ethanol vapor for 15 min, followed by repeat collection of exhaled breath in Tedlar bags. One hour later, the subjects washed their mouth for 30 s with a typical mouthwash containing 22% ethanol and post-exposure breaths were again collected repeatedly. Negligible systemic uptake of ethanol was confirmed by analysis of blood sampled before, between and after the exposures. Ethanol in breath and blood was analyzed by gas chromatography.Results: No or very low levels (less than 0.002 mg/g) of ethanol were detected in blood at any time point, indicating negligible systemic uptake. The decline in breath was mono-exponential after both exposures with average half times of 0.4 (range 0.3-0.8) min after inhalation exposure and 1.9 (1.1-3.0) min after mouthwash. BrAC levels in the first sample, collected a few seconds after exposure, were 0.14 (0.07-0.13) mg/L after inhalation and 4.4 (2.7-6.0) mg/L after mouth wash. On average, it took 0.5 (0.06-0.7) min and 11 (6-15) min, respectively, for the BrAC to fall below the Swedish statutory limit of 0.1 mg/L air.Conclusion: In practice, use of breath analysis should not be a problem even if the subject inhaled ethanol vapors before the test. In contrast, use of ethanol-containing mouthwash results in a false positive test if sampling is done within 15 min.

Acute effects of exposure to vapours of dioxane in humans.

Information on the acute effects associated with the handling of 1,4-dioxane is sparse. Our aim was to evaluate the acute effects of 1,4-dioxane vapours. In a screening study, six healthy volunteers rated symptoms on a visual analogue scale (VAS), while exposed to stepwise increasing levels of 1,4-dioxane, from 1 to 20 ppm. The initial study indicated no increased ratings at any of the exposure levels; we decided to use 20 ppm (72 mg/m3) as a tentative no observed adverse effect level (NOAEL). In the main study, six female and six male healthy volunteers were exposed to 0 (control exposure) and 20 ppm 1,4-dioxane vapour, for 2 hours at rest. The volunteers rated 10 symptoms on VAS before, during, and after the exposure. Blink frequency was monitored during exposure. Pulmonary function, and nasal swelling, was measured before, and at 0 and 3 hours after exposure. Inflammatory markers in plasma (C-reactive protein, and interleukin-6) were measured before and at 3 hours after exposure. In conclusion, exposure to 20 ppm 1,4-dioxane for 2 hours did not significantly affect symptom ratings, blink frequency, pulmonary function, nasal swelling, or inflammatory markers in the plasma of the 12 volunteers in our study.

Glutathione transferase T1 phenotype affects the toxicokinetics of inhaled methyl chloride in human volunteers.

The aim of the present study was to investigate how the genetic polymorphism in glutathione transferase T1 (GSTT1) affects the metabolism and disposition of methyl chloride in humans in vivo. The 24 volunteers (13 males and 11 females) who participated in the study were recruited from a group of 208 individuals previously phenotyped for GSTT1 by measuring the glutathione transferase activity with methyl chloride in lysed erythrocytes ex vivo. Eight individuals with high (+/+), eight with medium (+/0) and eight with no (0/0) GSTT1 activity were exposed to methyl chloride gas (10 p.p.m.) in an exposure chamber for 2 h. Uptake and disposition was studied by measuring the concentration of methyl chloride in inhaled air, exhaled air and blood. A two-compartment model with two elimination pathways corresponding to exhalation and metabolism was fitted to experimental data. The average net respiratory uptake of methyl chloride was 243, 158, and 44 micromol in individuals with high, intermediate and no GSTT1 activity, respectively. Metabolic clearance was high (4.6 l/min) in the +/+ group, intermediate (2.4 l/min) in the +/0 group, and close to zero in 0/0 individuals, while the exhalation clearance was similar in the three groups. No exposure related increase in urinary S-methyl cysteine was detected. However, gender and the GSTTl phenotype seemed to affect the background levels. In conclusion, GSTT1 appears to be the sole determinant of methyl chloride metabolism in humans. Thus, individuals with nonfunctional GSTT1 entirely lack the capacity to metabolize methyl chloride.

Demonstration of bilateral projection of the central retina of the monkey with horseradish peroxidase neuronography.

In the primate, ganglion cells of the temporal retina project ipsilaterally and those of the nasal retina, contralaterally into the optic tract. The vertical meridian passing through the fovea defines the border between these two populations of ganglion cells and has been demonstrated in four Macaque monkeys after unilateral injection of horseradish peroxidase into the dorsal lateral geniculate nucleus and examination of the pattern of retrograde labeling of those ganglion cells projecting to the injected side. A median 1 degree vertical strip in which ipsi- and contralaterally projecting ganglion cells intermingle was found, confirming the report by Stone et al. ('73). In addition, occasional extrafoveal labeled ganglion cells were found as far as 2 degrees from the vertical midline in the otherwise unlabeled hemiretinae. These ganglion cells were not numerous and had somata of all sizes, suggesting that they do not constitute a separate class of ganglion cells as found in the temporal retina of the cat. In contrast to the description by Stone et al. ('73), the strip of vertical overlap did not show a constant width through the fovea, since mixing of labeled and unlabeled ganglion cells was found in a band approximately 1/2 degree wide along both the nasal and temporal rims of the foveal pit which is 500 mum (2 degrees) in diameter. Beyond these 1/2 degree arcs, the appropriate hemiretina was either completely unlabeled, or contained virtually every ganglion cell labeled on the side projecting to the injected dorsal lateral geniculate nucleus. The scattered labeled ganglion cells rimming an otherwise unlabeled hemifovea represent a possible anatomical basis for the phenomenon of "macular" or "foveal sparing" in which unilateral damage to the occipital cortex produces homonymous hemianopsia with sparing of a small island of centralmost vision extending about 1 degree from the foveal center. From this study, it is not possible to define the receptive fields or specific photoreceptor connections of the ganglion cells labeled with horseradish peroxidase, so that at the present time quantitative correlations cannot be made between the numbers of ganglion cells remaining on the affected side of the fovea and the extent of preservation of visual function in the spared zone. The presence of labeled ganglion cells rimming the fovea in its entirety is compatible with the sequence of foveal development in late prenatal life. After lateral displacement both nasally and temporally of ganglion cells which initially lay in the median vertical overlap strip of 1 degree, in the adult retina a strip approximately 1/2 degree wide around the perimeter of the foveola should contain a mixture of ipsi- and contralaterally projecting ganglion cells. The total population of ganglion cells beyond this 1/2 degree band should be completely ipsi- or contralateral in their projection patterns, as is observed...

Orthograde and retrograde axoplasmic transport during acute ocular hypertension in the monkey.

Orthograde and retrograde axoplasmic transport have been studied in the optic nerve heads of 37 Macaca fascicularis eyes with normal or elevated intraocular pressure (IOP) produced by cannulation of the anterior chamber. Orthograde transport was labeled by 3H-amino acids injected intravitreally and incorporated into retinal ganglion cell proteins. Retrograde transport was studied in the same eyes by injecting horseradish peroxidase (HRP) into one or both optic tracts and dorsal lateral geniculate nuclei (dLGN). Both tracers accumulated in the lamina scleralis (LS) of eyes maintained at pressures of 25 to 150 mm. Hg for 12 to 28 hours (pressure in normal controls = 10 to 14 mm. Hg) but the HRP technique was markedly more sensitive. The degree of retrograde transport obstruction in the LS appeared to be directly proportional to both the height and the duration of elevated IOP. In one experiment, the blockades of orthograde and retrograde transport induced at 50 mm. Hg were demonstrated to be reversible. Serial reconstructions of radioautographs and peroxidase-reacted sections of the optic nerve heads demonstrated that the orthograde and retrograde transport obstructions were coincidental anatomically by light microscopy in the LS and occurred most prominently in the temporal quadrants of the nerve head. These transport obstructions occurred at moderate elevations of IOP (25 TO 50 mm. Hg) despite (1) elevated arterial PO2 levels during inhalation of 100 percent oxygen and (2) intact nerve head capillary circulation, as demonstrated by perfusion with nucleated avian erythrocytes.

Physiologically based toxicokinetic modeling of inhaled ethyl tertiary-butyl ether in humans.

A physiologically based toxicokinetic (PBTK) model was developed for evaluation of inhalation exposure in humans to the gasoline additive, ethyl tertiary-butyl ether (ETBE). PBTK models are useful tools to relate external exposure to internal doses and biological markers of exposure in humans. To describe the kinetics of ETBE, the following compartments were used: lungs (including arterial blood), liver, fat, rapidly perfused tissues, resting muscles, and working muscles. The same set of compartments and, in addition, a urinary excretion compartment were used for the metabolite tertiary-butyl alcohol (TBA). First order metabolism was assumed in the model, since linear kinetics has been shown experimentally in humans after inhalation exposure up to 50 ppm ETBE. Organ volumes and blood flows were calculated from individual body composition based on published equations, and tissue/blood partition coefficients were calculated from liquid/air partition coefficients and tissue composition. Estimates of individual metabolite parameters of 8 subjects were obtained by fitting the PBTK model to experimental data from humans (5, 25, 50 ppm ETBE, 2-h exposure; Nihlén et al., Toxicol. Sci., 1998; 46, 1-10). The PBTK model was then used to predict levels of the biomarkers ETBE and TBA in blood, urine, and exhaled air after various scenarios, such as prolonged exposure, fluctuating exposure, and exposure during physical activity. In addition, the interindividual variability in biomarker levels was predicted, in the eight experimentally exposed subjects after a working week. According to the model, raising the work load from rest to heavy exercise increases all biomarker levels by approximately 2-fold at the end of the work shift, and by 3-fold the next morning. A small accumulation of all biomarkers was seen during one week of simulated exposure. Further predictions suggested that the interindividual variability in biomarker levels would be higher the next morning than at the end of the work shift, and higher for TBA than for ETBE. Monte Carlo simulations were used to describe fluctuating exposure scenarios. These simulations suggest that ETBE levels in blood and exhaled air at the end of the working day are highly sensitive to exposure fluctuations, whereas ETBE levels the next morning and TBA in urine and blood are less sensitive. Considering these simulations, data from the previous toxicokinetic study and practical issues, we suggest that TBA in urine is a suitable biomarker for exposure to ETBE and gasoline vapor.

Controlled ethyl tert-butyl ether (ETBE) exposure of male volunteers. I. Toxicokinetics.

Ethyl tert-butyl ether (ETBE) might replace methyl tert-butyl ether (MTBE), a widely used additive in unleaded gasoline. The aim of this study was to evaluate uptake and disposition of ETBE, and eight healthy male volunteers were exposed to ETBE vapor (0, 5, 25, and 50 ppm) during 2 h of light physical exercise. ETBE and the proposed metabolites tert-butyl alcohol (TBA) and acetone were analyzed in exhaled air, blood, and urine. Compared to a previous MTBE study (A. Nihlen et al., 1998b, Toxicol. Appl. Pharmacol. 148, 274-280) lower respiratory uptake of ETBE (32-34%) was seen as well as a slightly higher respiratory exhalation (45-50% of absorbed ETBE). The kinetic profile of ETBE could be described by four phases in blood (average half-times of 2 min, 18 min, 1.7 h, and 28 h) and two phases in urine (8 min and 8.6 h). Postexposure half-times of TBA in blood and urine were on average 12 and 8 h, respectively. The 48-h pulmonary excretion of TBA accounted for 1.4-3.8% of the absorbed ETBE, on an equimolar basis. Urinary excretion of ETBE and TBA was low, below 1% of the ETBE uptake, indicating further metabolism of TBA or other routes of metabolism and elimination. The kinetics of ETBE and TBA were linear up to 50 ppm. Based upon blood profile, levels in blood and urine, and kinetic profile we suggest that TBA is a more appropriate biomarker for ETBE than the parent ether itself. The acetone level in blood was higher after ETBE exposures compared to control exposure, and acetone is probably partly formed from ETBE.

Controlled ethyl tert-butyl ether (ETBE) exposure of male volunteers. II. Acute effects.

The aim of this study was to evaluate acute effects of ethyl tert-butyl ether (ETBE) in man after short-term exposure. ETBE may in the future replace methyl tert-butyl ether, a widely used oxygenate in unleaded gasoline. Eight healthy male volunteers were exposed to ETBE vapor for 2 h at four levels (0, 5, 25, and 50 ppm) during light physical exercise. The subjects rated irritative symptoms, discomfort, and central nervous system effects in a questionnaire. Ocular (eye redness, tear film break-up time, conjunctival epithelial damage, and blinking frequency), nasal (acoustic rhinometry and analysis of inflammatory markers and cells in nasal lavage fluid), and pulmonary (peak expiratory flow, forced expiratory volume in 1 s, forced vital capacity, vital capacity, and transfer factor) measurements were performed. Significantly increased ratings of solvent smell (p = 0.001, repeated-measures ANOVA) were seen during exposures and correlated to exposure levels. Furthermore, significantly elevated ratings of discomfort in throat and airways were seen during and after 50 ppm compared to the control exposure (p = 0.02). Increased nasal swelling (p = 0.001) and blinking frequency (p = 0.01) were noted at all exposure levels, but their magnitudes were not related to exposure levels. A slightly impaired pulmonary function was seen at 25 and 50 ppm, since forced vital capacity (p = 0.02) and vital capacity (p = 0.04) differed significantly from the clean air exposure. Although the impairments seemed to fall within normal inter- and intraindividual variation and have no clinical relevance as such, it cannot be excluded that other individuals may react more severely than eight healthy male volunteers in this study.

Physiologically based pharmacokinetic modeling of inhalation exposure of humans to dichloromethane during moderate to heavy exercise.

Dichloromethane (methylene chloride, DCM) is metabolized via two pathways in humans: mixed-function oxidases (MFO) and glutathione-S:-transferase (GST). Most likely, the carcinogenicity for DCM is related to metabolic activation of DCM via the GST pathway. However, as the two pathways are competing, the metabolic capacity for the MFO pathway in vivo is also of interest in risk assessment for DCM. Past estimates of MFO metabolism are based on the in vitro activity of tissue samples. The aim of the present study was to develop a population model for DCM in order to gain more knowledge on the variability of DCM inhalation toxicokinetics in humans, with main emphasis on the MFO metabolic pathway. This was done by merging published in vitro data on DCM metabolism and partitioning with inhalation toxicokinetic data (Astrand et al., 1975, Scand. J. Work.Environ. Health 1, 78-94) from five human volunteers, using the MCMC technique within a population PBPK model. Our results indicate that the metabolic capacity for the MFO pathway in humans is slightly larger than previously estimated from four human liver samples. Furthermore, the interindividual variability of the MFO pathway in vivo is smaller among our five subjects than indicated by the in vitro samples. We also derive a Bayesian estimate of the population distribution of the MFO metabolism (median maximum metabolic rate 28, 95% confidence interval 12-66 micromol/min) that is a compromise between the information from the in vitro data and the toxicokinetic information present in the experimental data.

Toxicokinetic interactions between orally ingested chlorzoxazone and inhaled acetone or toluene in male volunteers.

The aim of this study was to examine if the drug chlorzoxazone has any influence on the toxicokinetics of acetone and toluene. Chlorzoxazone is mainly metabolized by the same enzyme (Cytochrome P450 2E1) as ethanol and many other organic solvents. Ten male volunteers were exposed to solvent vapor (2 h, 50 watt) in an exposure chamber. Each subject was exposed to acetone only (250 ppm), acetone + chlorzoxazone, toluene (50 ppm) only, toluene + chlorzoxazone, and chlorzoxazone only. Chlorzoxazone (500 mg) was taken as two tablets 1 h prior to solvent exposure. Samples of blood, urine and exhaled air were collected before, during and until 20 h post exposure. The samples were analyzed by head-space gas chromatography (acetone and toluene) and high-performance liquid chromatography (chlorzoxazone, 6-hydroxychlorzoxazone and hippuric acid). The time-concentration curves of acetone and toluene in blood were fitted to one- and four-compartment toxicokinetic models, respectively. Intake of chlorzoxazone was associated with slight but significant increases in the area under the blood concentration-time curve (AUC) and steady state concentration of acetone in blood, along with non significant tendencies to an increased half time in blood and an increased AUC in urine. Except for a delayed excretion of hippuric acid in urine, no effects on the toluene toxicokinetics were seen after chlorzoxazone treatment. Small increases in chlorzoxazone plasma levels were seen after exposure compared to chlorzoxazone alone. These interactions, although statistically significant, seem to be small compared to the interindividual variability on metabolism and toxicokinetics.

Pulmonary reactions after exposure to 3-methylfuran vapour, a fungal metabolite.

A case of obstructive pulmonary reaction with flu-like symptoms after exposure to 3-methylfuran is described. This compound is produced by fungi, and can be found in buildings with mould growth. Previous studies have shown that exposure to the substance might increase the prevalence of respiratory symptoms, and is pneumotoxic to animals at high concentrations.

Bayesian estimation of variability in adipose tissue blood flow in man by physiologically based pharmacokinetic modeling of inhalation exposure to toluene.

Due to the lipophilicity of many xenobiotics, the perfusion of fat tissue is of special interest in physiologically based pharmacokinetic (PBPK) modeling. In order to estimate inter- and intra-individual variability in fat tissue blood flow with exercise, a population PBPK model for toluene was fitted to experimental data from subjects exposed to toluene vapors (Carlsson, A., 1982. Exposure to toluene: uptake, distribution and elimination in man. Scand. J. Work Environ. Health 8, 43-55). Six male volunteers were exposed to 80 ppm toluene for two hours during rest and moderate to heavy exercise (50-150 W). Extensive data collection was made, including sampling of arterial blood, exhaled breath and subcutaneous fat tissue. The model was simultaneously fitted to the time courses of toluene in arterial blood, exhaled breath, and subcutaneous fat in the six individuals by Markov chain Monte Carlo (MCMC) simulation. In order to describe the experimental observations in subcutaneous fat accurately, the fat compartment was split in two. According to the analysis, the increased perfusion of perirenal fat associated with physical workload was best described if it was set to the same, elevated, level during all exercise levels, rather than scaled directly to the increase in oxygen uptake. No increase in subcutaneous fat perfusion could be detected at these exposure conditions.

PBPK model for butadiene metabolism to epoxides: quantitative species differences in metabolism.

We have developed a physiologically based pharmacokinetic (PBPK) model for 1,3-butadiene (BD) and its first reactive metabolite 1,2-epoxybutene-3 (EB). This model contrasts with other published ones, in that it incorporates three important features: (I) reduced alveolar ventilation, based on experimental observations on a number of vapors and gases; (II) intrahepatic first-pass hydrolysis of EB, based on experimental observations with BD-EB, ethylene-ethylene oxide, and styrene-styrene oxide; (III) a two-substrate Michealis-Menten kinetic description of EB conjugation with GSH. We believe these features are essential for a correct toxicokinetic description of BD. The model was validated against a number of published experimental observations on BD, EB, and liver glutathione (GSH), kinetics made in vivo with rats and mice, including EB exhalation upon BD exposure and liver GSH depletion at high exposure levels of BD. According to our model, the relative internal doses of EB (expressed as the relation between steady-state concentrations or AUCs in mixed venous blood) are: mouse 1.6, rat 1.0, man 0.3. In the mouse, GSH depletion occurs after 6-9 h exposure at high concentrations resulting in a shift of the relative internal dose from 1.6 to between 2 and 3. The clear but relatively small mouse-rat difference in internal EB doses can only partly explain the marked species difference in cancer response between mice and rats exposed to BD.