Pilot study evaluating inhalation and dermal glyphosate exposure resulting from simulated heavy residential consumer application of Roundup®.

OBJECTIVES: The purpose of this study was to evaluate the individual contributions of inhalation and dermal exposures to urinary glyphosate levels following the heavy residential consumer application of a glyphosate-containing herbicide. METHODS: A pilot study was conducted in which each participant mixed and continuously spray-applied 16.3 gallons of a 0.96% glyphosate-containing solution for 100 min using a backpack sprayer. Twelve participants were divided evenly into two exposure groups, one equipped to assess dermal exposure and the other, inhalation exposure. Personal air samples (n = 12) and dermal patch samples (n = 24) were collected on the inhalation group participants and analyzed for glyphosate using HPLC-UV. Serial urine samples collected 30-min prior to application and 3-, 6-, 12-, 24-hr (inhalation and dermal groups) and 36-hr (dermal group only) post-application were analyzed for glyphosate and glyphosate's primary metabolite (AMPA) using HPLC-MS/MS. RESULTS: The mean airborne glyphosate concentration was 0.0047 mg/m3, and the mean concentrations of glyphosate for each applicator's four patch samples ranged from 0.04 µg/mm2 to 0.25 µg/mm2. In general, urinary glyphosate, AMPA, and total effective glyphosate levels were higher in the dermal exposure group than the inhalation exposure group, peaked within 6-hr following application, and were statistically indistinguishable from background at 24-hr post-application. CONCLUSIONS: This is the first study to characterize the absorption and biological fate of glyphosate in residential consumer applicators following heavy application. The results of this pilot study are consistent with previous studies that have shown that glyphosate is rapidly eliminated from the body, typically within 24 hr following application.

A meta-analysis of airborne asbestos fiber concentrations from work with or around asbestos-containing floor tile.

In this meta-analysis, exposures to airborne asbestos during work with or around floor tiles were characterized according to several variables: study, sample type, activity, and task. Personal breathing zone, bystander, and area sample exposure concentrations were differentiated and compared against current occupational exposure limits to asbestos. In total, 22 studies, including 804 personal, 57 bystander, and 295 area samples, were included in the analysis. The arithmetic mean airborne fiber concentrations were 0.05, 0.02, and 0.01 f/cm3 for personal, bystander, and area samples, respectively. Arithmetic mean time-weighted-average fiber concentrations over an 8-h working day were 0.02 and 0.01 f/cm3 for personal and bystander samples, respectively. Phase contrast microscopy (PCM) personal airborne fiber concentrations were highest for maintenance activities, followed by removal and installation. Tasks that involved buffing or burnishing, scoring or snapping, and scraping or lifting had the highest personal PCM concentrations, while stripping floor tile and removing it with chemical solvent had the lowest concentrations. Exposures associated with handling asbestos floor tiles, under working conditions normally encountered, do not generally produce airborne concentrations at levels that exceed the current OSHA PEL nor do they appear to approach the threshold cumulative asbestos dose concentrations that have been previously associated with an increased risk of asbestos-related disease.

PKCγ is required for ethanol-induced increases in GABA(A) receptor α4 subunit expression in cultured cerebral cortical neurons.

Ethanol exposure produces alterations in GABA(A) receptor function and expression associated with CNS hyperexcitability, but the mechanisms of these effects are unknown. Ethanol is known to increase both GABA(A) receptor α4 subunits and protein kinase C (PKC) isozymes in vivo and in vitro. Here, we investigated ethanol regulation of GABA(A) receptor α4 subunit expression in cultured cortical neurons to delineate the role of PKC. Cultured neurons were prepared from rat pups on postnatal day 0-1 and tested after 18 days. GABA(A) receptor α4 subunit surface expression was assessed using P2 fractionation and surface biotinylation following ethanol exposure for 4 h. Miniature inhibitory post-synaptic currents were measured using whole cell patch clamp recordings. Ethanol increased GABA(A) receptor α4 subunit expression in both the P2 and biotinylated fractions, while reducing the decay time constant in miniature inhibitory post-synaptic currents, with no effect on γ2 or δ subunits. PKC activation mimicked ethanol effects, while the PKC inhibitor calphostin C prevented ethanol-induced increases in GABA(A) receptor α4 subunit expression. PKCγ siRNA knockdown prevented ethanol-induced increases in GABA(A) receptor α4 subunit expression, but inhibition of the PKCß isoform with PKCß pseudosubstrate had no effect. We conclude that PKCγ regulates ethanol-induced alterations in α4-containing GABA(A) receptors.

Ethanol reduces GABAA alpha1 subunit receptor surface expression by a protein kinase Cgamma-dependent mechanism in cultured cerebral cortical neurons.

Prolonged ethanol exposure causes central nervous system hyperexcitability that involves a loss of GABAergic inhibition. We previously demonstrated that long-term ethanol exposure enhances the internalization of synaptic GABA(A) receptors composed of alpha1beta2/3gamma2 subunits. However, the mechanisms of ethanol-mediated internalization are unknown. This study explored the effect of ethanol on surface expression of GABA(A) alpha1 subunit-containing receptors in cultured cerebral cortical neurons and the role of protein kinase C (PKC) beta, gamma, and epsilon isoforms in their trafficking. Cultured neurons were prepared from rat pups on postnatal day 1 and maintained for 18 days. Cells were exposed to ethanol, and surface receptors were isolated by biotinylation and P2 fractionation, whereas functional analysis was conducted by whole-cell patch-clamp recording of GABA- and zolpidem-evoked responses. Ethanol exposure for 4 h decreased biotinylated surface expression of GABA(A) receptor alpha1 subunits and reduced zolpidem (100 nM) enhancement of GABA-evoked currents. The PKC activator phorbol-12,13-dibutyrate mimicked the effect of ethanol, and the selective PKC inhibitor calphostin C prevented ethanol-induced internalization of these receptors. Ethanol exposure for 4 h also increased the colocalization and coimmunoprecipitation of PKCgamma with alpha1 subunits, whereas PKCbeta/alpha1 association and PKCepsilon/alpha1 colocalization were not altered by ethanol exposure. Selective PKCgamma inhibition by transfection of selective PKCgamma small interfering RNAs blocked ethanol-induced internalization of GABA(A) receptor alpha1 subunits, whereas PKCbeta inhibition using pseudo-PKCbeta had no effect. These findings suggest that ethanol exposure selectively alters PKCgamma translocation to GABA(A) receptors and PKCgamma regulates GABA(A) alpha1 receptor trafficking after ethanol exposure.