Lack of detectable sex differences in the mitochondrial function of Caenorhabditis elegans.

BACKGROUND: Sex differences in mitochondrial function have been reported in multiple tissue and cell types. Additionally, sex-variable responses to stressors including environmental pollutants and drugs that cause mitochondrial toxicity have been observed. The mechanisms that establish these differences are thought to include hormonal modulation, epigenetic regulation, double dosing of X-linked genes, and the maternal inheritance of mtDNA. Understanding the drivers of sex differences in mitochondrial function and being able to model them in vitro is important for identifying toxic compounds with sex-variable effects. Additionally, understanding how sex differences in mitochondrial function compare across species may permit insight into the drivers of these differences, which is important for basic biology research. This study explored whether Caenorhabditis elegans, a model organism commonly used to study stress biology and toxicology, exhibits sex differences in mitochondrial function and toxicant susceptibility. To assess sex differences in mitochondrial function, we utilized four male enriched populations (N2 wild-type male enriched, fog-2(q71), him-5(e1490), and him-8(e1498)). We performed whole worm respirometry and determined whole worm ATP levels and mtDNA copy number. To probe whether sex differences manifest only after stress and inform the growing use of C. elegans as a mitochondrial health and toxicologic model, we also assessed susceptibility to a classic mitochondrial toxicant, rotenone. RESULTS: We detected few to no large differences in mitochondrial function between C. elegans sexes. Though we saw no sex differences in vulnerability to rotenone, we did observe sex differences in the uptake of this lipophilic compound, which may be of interest to those utilizing C. elegans as a model organism for toxicologic studies. Additionally, we observed altered non-mitochondrial respiration in two him strains, which may be of interest to other researchers utilizing these strains. CONCLUSIONS: Basal mitochondrial parameters in male and hermaphrodite C. elegans are similar, at least at the whole-organism level, as is toxicity associated with a mitochondrial Complex I inhibitor, rotenone. Our data highlights the limitation of using C. elegans as a model to study sex-variable mitochondrial function and toxicological responses.

Quantitation of Total PFAS Including Trifluoroacetic Acid with Fluorine Nuclear Magnetic Resonance Spectroscopy.

Fluorine nuclear magnetic resonance (19F-NMR) spectroscopy has been shown to be a powerful tool capable of quantifying the total per- and polyfluoroalkyl substances (PFAS) in a complex sample. The technique relies on the characteristic terminal -CF3 shift (-82.4 ppm) in the alkyl chain for quantification and does not introduce bias due to sample preparation or matrix effects. Traditional quantitative analytical techniques for PFAS, such as liquid chromatography-mass spectrometry (LC-MS) and combustion ion chromatography (CIC), contain inherent limitations that make total fluorine analysis challenging. Here, we report a sensitive 19F-NMR method for the analysis of total PFAS, with a limit of detection of 99.97 nM, or 50 µg/L perfluorosulfonic acid. To demonstrate the capabilities of 19F-NMR, the technique was compared to two commonly used methods for PFAS analysis: total oxidizable precursor (TOP) assay and LC-high resolution MS analysis for targeted quantification and suspect screening. In both cases, the 19F-NMR analyses detected higher total PFAS quantities than either the TOP assay (63%) or LC-MS analyses (65%), suggesting that LC-MS and TOP assays can lead to underreporting of PFAS. Importantly, the 19F-NMR detected trifluoroacetic acid at a concentration more than five times the total PFAS concentration quantified using LC-MS in the wastewater sample. Therefore, the use of 19F-NMR to quantify the total PFAS in highly complex samples can be used to complement classic TOP or LC-MS approaches for more accurate reporting of PFAS contamination in the environment.

Comparing Equilibrium Concentrations of Polychlorinated Biphenyls Based on Passive Sampling and Bioaccumulation in Water Column Deployments.

Biomonitoring at contaminated sites undergoing cleanup, including Superfund sites, often uses bioaccumulation of anthropogenic contaminants by field-deployed organisms as a metric of remedial effectiveness. Bioaccumulation studies are unable to assess the equilibrium status of the organisms relative to the contaminants to which they are exposed. Establishing equilibrium provides a reproducible benchmark on which scientific and management decisions can be based (e.g., comparison with human dietary consumption criteria). Unlike bioaccumulating organisms, passive samplers can be assessed for their equilibrium status. In our study, over a 3-year period, we compared the bioaccumulation of selected polychlorinated biphenyls (PCBs) by mussels in water column deployments at the New Bedford Harbor Superfund site (New Bedford, MA, USA) to codeployed passive samplers. Based on comparisons to the calculated passive sampler equilibrium concentrations, the mussels were not at equilibrium, and the subsequent analysis focused on evaluating approaches for estimating equilibrium bioaccumulation. In addition, a limited evaluation of metal bioaccumulation by the exposed mussels and a metal passive sampler was performed. In general, mussel and passive sampler accumulation of PCBs was significantly correlated; however, surprisingly, agreement on the magnitude of accumulation was optimal when bioaccumulation and passive sampler uptake were not corrected for nonequilibrium conditions. A subsequent comparison of four approaches for estimating equilibrium mussel bioaccumulation using octanol-water partition coefficients (KOW ), triolein-water partition coefficients (KTW ), and two types of polymer-lipid partition coefficients demonstrated that field-deployed mussels were not at equilibrium with many PCBs. A range of estimated equilibrium mussel bioaccumulation concentrations were calculated, with the magnitude of the KOW -based values being the smallest and the polymer-lipid partition coefficient-based values being the largest. These analyses are intended to assist environmental scientists and managers to interpret field deployment data when transitioning from biomonitoring to passive sampling. Environ Toxicol Chem 2023;42:317-332. Published 2022. This article is a U.S. Government work and is in the public domain in the USA.

Mild pentachlorophenol-mediated uncoupling of mitochondria depletes ATP but does not cause an oxidized redox state or dopaminergic neurodegeneration in Caenorhabditis elegans.

Aims: Mitochondrial dysfunction is implicated in several diseases, including neurological disorders such as Parkinson's disease. However, there is uncertainty about which of the many mechanisms by which mitochondrial function can be disrupted may lead to neurodegeneration. Pentachlorophenol (PCP) is an organic pollutant reported to cause mitochondrial dysfunction including oxidative stress and mitochondrial uncoupling. We investigated the effects of PCP exposure in Caenorhabditis elegans, including effects on mitochondria and dopaminergic neurons. We hypothesized that mild mitochondrial uncoupling by PCP would impair bioenergetics while decreasing oxidative stress, and therefore would not cause dopaminergic neurodegeneration. Results: A 48-hour developmental exposure to PCP causing mild growth delay (∼10 % decrease in growth during 48 h, covering all larval stages) reduced whole-organism ATP content > 50 %, and spare respiratory capacity âˆ¼ 30 %. Proton leak was also markedly increased. These findings suggest a main toxic mechanism of mitochondrial uncoupling rather than oxidative stress, which was further supported by a concomitant shift toward a more reduced cellular redox state measured at the whole organism level. However, exposure to PCP did not cause dopaminergic neurodegeneration, nor did it sensitize animals to a neurotoxic challenge with 6-hydroxydopamine. Whole-organism uptake and PCP metabolism measurements revealed low overall uptake of PCP in our experimental conditions (50 µM PCP in the liquid exposure medium resulted in organismal concentrations of < 0.25 µM), and no measurable production of the oxidative metabolites tetra-1,4-benzoquinone and tetrachloro-p-hydroquinone. Innovation: This study provides new insights into the mechanistic interplay between mitochondrial uncoupling, oxidative stress, and neurodegeneration in C. elegans. These findings support the premise of mild uncoupling-mediated neuroprotection, but are inconsistent with proposed broad "mitochondrial dysfunction"-mediated neurodegeneration models, and highlight the utility of the C. elegans model for studying mitochondrial and neurotoxicity. Conclusions: Developmental exposure to pentachlorophenol causes gross toxicological effects (growth delay and arrest) at high levels. At a lower level of exposure, still causing mild growth delay, we observed mitochondrial dysfunction including uncoupling and decreased ATP levels. However, this was associated with a more-reduced cellular redox tone and did not exacerbate dopaminergic neurotoxicity of 6-hydroxydopamine, instead trending toward protection. These findings may be informative of efforts to define nuanced mitochondrial dysfunction-related adverse outcome pathways that will differ depending on the form of initial mitochondrial toxicity.

The variability of volatile organic compounds in the indoor air of clinical environments.

The development of clinical breath-analysis is confounded by the variability of background volatile organic compounds (VOCs). Reliable interpretation of clinical breath-analysis at individual, and cohort levels requires characterisation of clinical-VOC levels and exposures. Active-sampling with thermal-desorption/gas chromatography-mass spectrometry recorded and evaluated VOC concentrations in 245 samples of indoor air from three sites in a large National Health Service (NHS) provider trust in the UK over 27 months. Data deconvolution, alignment and clustering isolated 7344 features attributable to VOC and described the variability (composition and concentration) of respirable clinical VOC. 328 VOC were observed in more than 5% of the samples and 68 VOC appeared in more than 30% of samples. Common VOC were associated with exogenous and endogenous sources and 17 VOC were identified as seasonal differentiators. The presence of metabolites from the anaesthetic sevoflurane, and putative-disease biomarkers in room air, indicated that exhaled VOC were a source of background-pollution in clinical breath-testing activity. With the exception of solvents, and waxes associated with personal protective equipment (PPE), exhaled VOC concentrations above 3µg m-3are unlikely to arise from room air contamination, and in the absence of extensive survey-data, this level could be applied as a threshold for inclusion in studies, removing a potential environmental confounding-factor in developing breath-based diagnostics.

In Situ Investigation of Performance Reference Compound-Based Estimates of PCB Equilibrated Passive Sampler Concentrations and Cfree in the Marine Water Column.

Low-density polyethylene sheets are used as passive samplers for aquatic environmental monitoring to measure the freely dissolved concentration (Cfree ) of hydrophobic organic contaminants (HOCs). Freely dissolved HOCs in water will partition into the polyethylene until a thermodynamic equilibrium is achieved; that is, the HOC's activity in the passive sampler is the same as its activity in the surrounding environment. One way to evaluate the equilibrium status or estimate the uptake kinetics is by using performance reference compounds (PRCs). A fractional equilibrium (feq ) can be determined for target HOCs, under the assumption that PRC desorption from the passive sampler occurs at the same rate as for the unlabeled target HOCs. However, few investigations have evaluated how effectively and accurately PRCs estimate target contaminant Cfree under in situ conditions. In the present study, polyethylene passive samplers were preloaded with 6 13 C-labeled polychlorinated biphenyls (PCBs) as PRCs; deployed in New Bedford Harbor, Massachusetts, USA; and collected after 30-, 56-, 99-, and 129-d deployments. Using this unique temporal sampling design, PRC results from each deployment were fit to a diffusion model to estimate the Cfree of 27 PCB congeners and compare the results between the different deployment times. Smaller PCBs had variable concentrations over the 4 deployments, whereas mid-molecular weight PCBs had consistent Cfree measurements for all deployments (relative standard deviation <20%). High-molecular weight PCBs had the largest Cfree estimates after 30 d; these estimates and their standard deviations decreased with longer deployment times. These findings suggest that when targeting PCBs with more than 6 chlorines or contaminants with a log octanol-water partition coefficient ≥6.5, a deployment time longer than 30 d may be prudent. Environ Toxicol Chem 2020;39:1165-1173. © 2020 SETAC.

Using performance reference compounds to compare mass transfer calibration methodologies in passive samplers deployed in the water column.

Performance reference compounds (PRCs) are often added to passive samplers prior to field deployments to provide information about mass transfer kinetics between the sampled environment and the passive sampler. Their popularity has resulted in different methods of varying complexity to estimate mass transfer and better estimate freely dissolved concentrations (Cfree ) of targeted compounds. Three methods for describing a mass transfer model are commonly used: a first-order kinetic method, a nonlinear least squares fitting of sampling rate, and a diffusion method. Low-density polyethylene strips loaded with PRCs and of 4 different thicknesses were used as passive samplers to create an array of PRC results to assess the comparability and reproducibility of each of the methods. Samplers were deployed in the water column at 3 stations in New Bedford Harbor (MA, USA). Collected data allowed Cfree comparisons to be performed in 2 ways: 1) comparison of Cfree derived from one thickness using different methods, and 2) comparison of Cfree derived by the same method using different thicknesses of polyethylene. Overall, the nonlinear least squares and diffusion methods demonstrated the most precise results for all the PCBs measured and generated Cfree values that were often statistically indistinguishable. Relative standard deviations (RSDs) for total PCB measurements using the same thickness and varying model types ranged from 0.04 to 12% and increased with sampler thickness, and RSDs for estimates using the same method and varying thickness ranged from 8 to 18%. Environmental scientists and managers are encouraged to use these methods when estimating Cfree from passive sampling and PRC data. Environ Toxicol Chem 2018;37:2089-2097. Published 2018 Wiley Periodicals Inc. on behalf of SETAC. This article is a US government work and, as such, is in the public domain in the United States of America.

Evaluating the Relationship between Equilibrium Passive Sampler Uptake and Aquatic Organism Bioaccumulation.

This Critcal Review evaluates passive sampler uptake of hydrophobic organic contaminants (HOCs) in water column and interstitial water exposures as a surrogate for organism bioaccumulation. Fifty-seven studies were found where both passive sampler uptake and organism bioaccumulation were measured and 19 of these investigations provided direct comparisons relating passive sampler uptake and organism bioaccumulation. Polymers compared included low-density polyethylene (LDPE), polyoxymethylene (POM), and polydimethylsiloxane (PDMS), and organisms ranged from polychaetes and oligochaetes to bivalves, aquatic insects, and gastropods. Regression equations correlating bioaccumulation (CL) and passive sampler uptake (CPS) were used to assess the strength of observed relationships. Passive sampling based concentrations resulted in log-log predictive relationships, most of which were within one to 2 orders of magnitude of measured bioaccumulation. Mean coefficients of determination (r2) for LDPE, PDMS, and POM were 0.68, 0.76, and 0.58, respectively. For the available raw, untransformed data, the mean ratio of CL and CPS was 10.8 ± 18.4 (n = 609). Using passive sampling as a surrogate for organism bioaccumulation is viable when biomonitoring organisms are not available. Passive sampling based estimates of bioaccumulation provide useful information for making informed decisions about the bioavailability of HOCs.

Using performance reference compound-corrected polyethylene passive samplers and caged bivalves to measure hydrophobic contaminants of concern in urban coastal seawaters.

Low-density polyethylene (PE) passive samplers containing performance reference compounds (PRCs) were deployed at multiple depths in two urban coastal marine locations to estimate dissolved concentrations of hydrophobic organic contaminants (HOCs), including dichlorodiphenyltrichloroethane (DDT) and its metabolites, polychlorinated biphenyl (PCB) congeners, and polybrominated flame retardants. PE samplers pre-loaded with PRCs were deployed at the surface, mid-column, and near bottom at sites representing the nearshore continental shelf off southern California (Santa Monica Bay, USA) and a mega commercial port (Los Angeles Harbor). After correcting for fractional equilibration using PRCs, concentrations ranged up to 100 pg L(-1) for PCBs and polybrominated diphenyl ethers (PBDEs), 500 pg L(-1) for DDMU and 300 pg L(-1) for DDNU, and to 1000 pg L(-1) for p,p'-DDE. Seawater concentrations of DDTs and PCBs increased with depth, suggesting that bed sediments serve as the source of water column HOCs in Santa Monica Bay. In contrast, no discernable pattern between surface and near-bottom concentrations in Los Angeles Harbor was observed, which were also several-fold lower (DDTs: 45-300 pg L(-1), PCBs: 5-50 pg L(-1)) than those in Santa Monica Bay (DDTs: 2-1100 pg L(-1), PCBs: 2-250 pg L(-1)). Accumulation by mussels co-deployed with the PE samplers at select sites was strongly correlated with PE-estimated seawater concentrations, providing further evidence that these samplers are a viable alternative for monitoring of HOC exposure. Fractional equilibration observed with the PRCs increased with decreasing PRC molar volume indicating the importance of target compound physicochemical properties when estimating water column concentrations using passive samplers in situ.

Preparation of N-alkylbis(3-aminopropyl)amines by the catalytic hydrogenation of N-alkylbis(cyanoethyl)amines.

An improved process for the preparation of N-alkylbis(3-aminopropyl)amines is described. These triamines are of interest as monomers for the condensation polymerization with esterified carbohydrate diacids (aldaric acids) to generate the corresponding poly(4-alkyl-4-azaheptamethylene aldaramides). The triamine synthesis is comprised of two efficient steps and requires no chromatographic purification. Bisconjugate addition of alkylamines to acrylonitrile followed by catalytic hydrogenation of the N-alkylbis(cyanoethyl)amines over Raney nickel yields the target N-alkylbis(3-aminopropyl)amines. Much less solvent was used in the bisconjugate addition step then previously reported, and in the second step, a relatively low-pressure catalytic hydrogenation (50 psi of hydrogen) was employed using Raney nickel as the catalyst in a 7 N methanolic ammonia solvent system to afford the N-alkylbis(3-aminopropyl)amines of high purity in nearly quantitative yield.