Erica D. Bruce

PBDE developmental effects on embryonic zebrafish.

Polybrominated diphenyl ethers (PBDEs) have become ubiquitous environmental contaminants with potential for bioaccumulation and maternal-fetal transfer that has led to regulatory bans and/or phasing out of several technical mixtures of PBDEs. In the present study, six PBDE congeners (BDE 28, BDE 47, BDE 99, BDE 100, BDE 153, BDE 183) were evaluated for developmental effects on embryonic zebrafish. These congeners were chosen because they are environmentally relevant and cover a wide range of physical-chemical properties. Alterations in behavior, physical malformations, and mortality were scored daily until 168 h postfertilization (hpf). A concentration-dependent increase in spontaneous movement indicated an early onset of behavioral responses to PBDE exposures. Spontaneous movement was affected the most by BDE 47 and BDE 28, whereas BDE 183 did not alter behavior at any concentration tested. Swimming rates were significantly increased by BDE 28 at 96 and 120 hpf, but decreased swimming activity at 168 hpf. Additionally, BDE 47 significantly decreased the swimming rate at 168 hpf. Other endpoints included malformations and mortality. Congeners with fewer bromines (BDE 28, 47, 99, and 100) also induced a curved body axis starting around 120 hpf, which was followed by mortality. BDEs 153 and 183, however, did not elicit these adverse effects. A relationship was found between log K(OW) and median lethal concentration (LC50) and median effective concentration (EC50). Structure-activity relationships in this study suggest that PBDE acute toxicity results from a receptor-mediated effect and further studies are necessary to determine these pathways.

Hydroxylated PBDEs induce developmental arrest in zebrafish

The ubiquitous spread of polybrominated diphenyl ethers (PBDEs) has led to concerns regarding the metabolites of these congeners, in particular hydroxylated PBDEs. There are limited studies regarding the biological interactions of these chemicals, yet there is some concern they may be more toxic than their parent compounds. In this study three hydroxylated PBDEs were assessed for toxicity in embryonic zebrafish: 3-OH-BDE 47, 5-OH-BDE 47, and 6-OH-BDE 47. All three congeners induced developmental arrest in a concentration-dependent manner; however, 6-OH-BDE 47 induced adverse effects at lower concentrations than the other congeners. Furthermore, all three induced cell death; however apoptosis was not observed. In short-term exposures (24-28 hours post fertilization), all hydroxylated PBDEs generated oxidative stress in the region corresponding to the cell death at 5 and 10 ppm. To further investigate the short-term effects that may be responsible for the developmental arrest observed in this study, gene regulation was assessed for embryos exposed to 0.625 ppm 6-OH-BDE 47 from 24 to 28 hpf. Genes involved in stress response, thyroid hormone regulation, and neurodevelopment were significantly upregulated compared to controls; however, genes related to oxidative stress were either unaffected or downregulated. This study suggests that hydroxylated PBDEs disrupt development, and may induce oxidative stress and potentially disrupt the cholinergic system and thyroid hormone homeostasis.

UPTAKE AND METABOLISM OF INDIVIDUAL POLYBROMINATED DIPHENYL ETHER CONGENERS BY EMBRYONIC ZEBRAFISH

Embryonic zebrafish were used to compare the uptake and metabolism of six polybrominated diphenyl ether (PBDE) congeners (BDEs 28, 47, 99, 100, 153, and 183) and identified metabolites from static exposures at 24 and 120 h postfertilization (hpf). An inverse relationship was observed between uptake of PBDEs and their octanol-water partitioning coefficients (uptake of BDEs 28 and 47>99 and 100>153 and 183). Debromination metabolites were identified in all congeners (excluding BDE 28) tested in the 120-hpf tissue samples. Interestingly, BDE 153 underwent meta-debromination, forming BDEs 47 and 99. Gene transcription analysis was conducted at 120 hpf to identify potential metabolic pathways for the PBDEs examined in the present study (gstpi, deiodinases 1 and 2, cyp1a1, cyp1b1, and ugt5g). The greatest induction was of ugt5g for all congeners and deiodinase transcription was also upregulated by BDEs 28, 47, and 183. The cyp1a1 and cyp1b1 were upregulated by BDEs 28, 47, 99, and 183. The least alterations in gene transcription were in the BDE 153-exposed embryos. A clear primary pathway of debromination metabolism was not identified; however, upregulation of these different genes indicated that fish were responding to exposure of PBDEs. Furthermore, the present study demonstrated that the most bioavailable congeners are also those with the highest reported toxicity.

Particle uptake efficiency is significantly affected by type of capping agent and cell line

Surface‐functionalized silver nanoparticles (AgNPs) are the most deployed engineered nanomaterials in consumer products because of their optical, antibacterial and electrical properties. Almost all engineered nanoparticles are coated with application‐specific capping agents (i.e. organic/inorganic ligands on particle surface) to enhance their stability in suspension or increase their biocompatibility for biomedicine. The aim of this study was to investigate the contribution of the selected capping agents to their observed health impacts using realistic dose ranges. AgNPs capped with citrate, polyvinylpyrrolidone (PVP) and tannic acid were studied with human bronchoalveolar carcinoma (A549) and human colon adenocarcinoma (Caco‐2) cell lines and compared against exposures to Ag ions. Cellular uptake and cytotoxicity were evaluated up to 24 h. Tannic acid capped AgNPs induced higher cellular uptake and rate in both cell lines. Citrate‐capped and PVP‐capped AgNPs behaved similarly over 24 h. All three of the capped AgNPs penetrated more into the A549 cells than Caco‐2 cells. In contrast, the uptake rate of Ag ions in Caco‐2 cells (0.11 ± 0.0001 µg h–1) was higher than A549 cells (0.025 ± 0.00004 µg h–1). The exposure concentration of 3 mg l–1 is below the EC50 value for all of the AgNPs; therefore, little cytotoxicity was observed in any experiment conducted herein. Exposure of Ag ions, however, interrupted cell membrane integrity and cell proliferation (up to 70% lysed after 24 h). These findings indicate cellular uptake is dependent on capping agent, and when controlled to realistic exposure concentrations, cellular function is not significantly affected by AgNP exposure. Copyright

Evaluation of Common Use Brominated Flame Retardant (BFR) Toxicity Using a Zebrafish Embryo Model

Brominated flame retardants (BFRs) are used to reduce the flammability of plastics, textiles, and electronics. BFRs vary in their chemical properties and structures, and it is expected that these differences alter their biological interactions and toxicity. Zebrafish were used as the model organism for assessing the toxicity of nine structurally-diverse BFRs. In addition to monitoring for overt toxicity, the rate of spontaneous movement, and acetylcholinesterase and glutathione-S-transferase (GST) activities were assessed following exposure. The toxicities of BFRs tested can be ranked by LC50 as tetrabromobisphenol A (TBBPA) < 4,4′-isopropylidenebis[2-(2,6-dibromophenoxyl)ethanol] (TBBPA-OHEE) < Pentabromochlorocyclohexane (PBCH) < 2-ethylhexyl 2,3,4,5-tetrabromobenzoate (TBB) < hexabromocyclododecane (HBCD) < hexabromobenzene (HBB) < Tetrabromophthalic anhydride (PHT4). No adverse effect was observed in di(2-ethylhexyl) tetrabromophthalate (TBPH) or dibromoneopentyl glycol (DBNPG)-treated embryos. The rate of spontaneous movement was decreased in a concentration-dependent manner following exposure to four of the nine compounds. GST activity was elevated following treatment with PBCH, TBBPA, HBCD, and HBB. The results indicate that exposure to several BFRs may activate an antioxidant response and alter behavior during early development. Some of the BFRs, such as TBBPA and TBBPA-OHEE, induced adverse effects at concentrations lower than chemicals that are currently banned. These results suggest that zebrafish are sensitive to exposure to BFRs and can be used as a comparative screening model, as well as to determine alterations in behavior following exposure and probe mechanisms of action.

Evaluating a novel oxygenating therapeutic for its potential use in the advancement of wound healing

Non-gaseous oxygen therapeutics are emerging technologies in regenerative medicine that aim to sidestep the undesirable effects seen in traditional oxygen therapies, while enhancing tissue and wound regeneration. Using a novel oxygenating therapeutic (Ox66™) several in vitro models including fibroblast and keratinocyte monocultures were evaluated for potential drug toxicity, the ability of cells to recover after chemical injury, and cell migration after scratch assay. It was determined that in both cell lines, there was no significant cytotoxicity found after independent treatment with Ox66™. Similarly, after DMSO-induced chemical injury, the health parameters of cells treated with Ox66™ were improved when compared to their untreated counterparts. Particles were also characterized using scanning electron microscopy and electron dispersive spectroscopy both individually and in conjunction with fibroblast growth. The data in this study showed that the novel wound healing therapeutic has potential in advancing the treatment of various types of acute and chronic wounds.

Binary Mixtures of Polycyclic Aromatic Hydrocarbons Display Nonadditive Mixture Interactions in an In Vitro Liver Cell Model.

Polycyclic aromatic hydrocarbons (PAHs) have been labeled contaminants of concern due to their carcinogenic potential, insufficient toxicological data, environmental ubiquity, and inconsistencies in the composition of environmental mixtures. The Environmental Protection Agency is reevaluating current methods for assessing the toxicity of PAHs, including the assumption of toxic additivity in mixtures. This study was aimed at testing mixture interactions through in vitro cell culture experimentation, and modeling the toxicity using quantitative structure-activity relationships (QSAR). Clone-9 rat liver cells were used to analyze cellular proliferation, viability, and genotoxicity of 15 PAHs in single doses and binary mixtures. Tests revealed that many mixtures have nonadditive toxicity, but display varying mixture effects depending on the mixture composition. Many mixtures displayed antagonism, similar to other published studies. QSARs were then developed using the genetic function approximation algorithm to predict toxic activity both in single PAH congeners and in binary mixtures. Effective concentrations inhibiting 50% of the cell populations were modeled, with R(2) = 0.90, 0.99, and 0.84, respectively. The QSAR mixture algorithms were then adjusted to account for the observed mixture interactions as well as the mixture composition (ratios) to assess the feasibility of QSARs for mixtures. Based on these results, toxic addition is improbable and therefore environmental PAH mixtures are likely to see nonadditive responses when complex interactions occur between components. Furthermore, QSAR may be a useful tool to help bridge these data gaps surrounding the assessment of human health risks that are associated with PAH exposures.

Non-specific interactions between soluble and induce irreversible changes in the properties of bilayers.

Soluble in the extracellular matrix experience a crowded environment. However, most of the biophysical studies performed to date have focused on concentrations within the dilute regime (well below the mM range). Here, we systematically studied the interaction of model cell membrane systems (giant unilamellar vesicles and supported bilayers) with soluble globular , bovine serum albumin, and lysozyme at physiologically relevant concentrations. To mimic the extracellular environment more closely, we also used fetal bovine serum as a good representative of a biomimetic mixture. We found that regardless of the used (and thus of their biological function), the interactions between a model cell membrane and these are determined by their physico-chemical characteristics, mainly their dipolar character (or charged patches). In this paper we discuss the specificity and reversibility of these interactions and their potential implications on the living cells. In particular, we report initial evidence for an additional role of in cell membranes: that of reducing the effects of non-specific of soluble on the cell membrane.

Using in vitro to in vivo extrapolation (IVIVE) to develop toxicity metrics for human health risk assessment of polybrominated diphenyl ethers (PBDE)

Abstract Polybrominated diphenyl ethers (PBDE) are flame retardants found in many industrial components, such as furniture foam, consumer electronics, plastics, and textiles and levels have increased in human beings over the past few decades. PBDE (1) produce adverse neurotoxic effects in mice, (2) are lipophilic, and (3) bioaccumulate in fish. Consequently, PBDE biomagnification may occur, which may be transferred to human beings in amounts that may exert adverse health effects. This investigation uses in vitro models to calculate bioassay-based reference doses in order to develop a human health risk assessment based on the consumption of PBDE-contaminated fish, using in vitro to in vivo extrapolation. The toxicity effects of 10 PBDE congeners were examined and compared among those developed using traditional in vivo mouse studies and in vitro models in this investigation employing rat (Clone-9), HEPG2 and zebrafish liver cells to determine the feasibility of using alternative approaches to develop toxicity metrics to evaluate human health risk.

Emerging Associations of the ALDH2*2 Polymorphism with DiseaseSusceptibility

Ethanol is metabolized by Alcohol Dehydrogenase (ADH) to acetaldehyde and then irreversibly oxidized by Aldehyde Dehydrogenase (ALDH) to nontoxic acetate. In individuals expressing the ALDH2*2 variant enzyme, the rate of conversion from acetaldehyde to acetate is reduced and leads to flushing, nausea, and tachycardia due to increased blood levels of acetaldehyde. The ALDH2*2 variant has a lowered NAD+ coenzyme binding affinity, which results in a lowered clearance capacity toward acetaldehyde. This polymorphism is caused by the substitution of glutamate for lysine at position 487 within the catalytic active site of ALDH2, resulting in effects on subunit and quaternary complex activity. ALDH2*2 alleles are dominant over ALDH2*1 and therefore are expected to contribute to the formation of inactive heterotetramers decreased enzymatic activity in both homozygous and heterozygous individuals. Consequently, a higher susceptibility to various diseases such as Alzheimer’s, osteoporosis, and acute coronary syndrome has been associated with ALDH2*2 carriers. Additionally, the polymorphism seems to affect the efficacy of Glyceryl Trinitrate (GTN), a drug intended to treat coronary heart disease, in carriers of ALDH2*2 alleles. However, the polymorphism is believed to afford a protective effect against alcoholism as the side effects of acetaldehyde build-up are undesirable. Disulfiram, a drug historically used to treat alcohol dependency, induces the same undesirable physiological effects as the variant enzyme in non-carriers by inhibiting the normal functioning of ALDH2 enzyme.