The major objectives of this research were to examine the bioaccumulation parameters [depuration rates (kd), half life (t1/2) and biomagnification factor (BMF)] of individual isomers of hexabromocyclododecane (HBCD, C12H18Br6) in fish and to test the hypothesis of in vivo bioisomerization. This was done by exposing three groups of juvenile rainbow trout (Oncorhynchus Mykiss) to food fortified with known concentrations of an individual diastereoisomer (α, β, γ) for 56 days (uptake phase) followed by 112 days (depuration phase) of unfortified food. A fourth group of fish were exposed to unfortified food for the duration of the experiment. Fish (n=4) from all four aquaria were sacrificed on days 0, 7, 14, 56, 63, 70, 112 and 168 and muscle tissue was extracted and analyzed for diastereoisomer concentrations by high performance liquid chromatography tandem mass spectrometry (LC/MS/MS). Bioaccumulation of the γ diastereoisomer was linear during the uptake phase while the α and β diastereoisomers were found to increase exponentially with respective doubling times of 14.1 and 20.5 days. Both the β and γ diastereoisomers followed first order depuration kinetics with calculated t1/2’s of 94 ± 25 and 84 ± 51 (± 1 × standard error) days, respectively. The BMF for the α diastereoisomer (BMF = 4.1) was one and a half times greater than the β-diastereoisomer (BMF = 2.6) and about one fifth larger than the γ-diastereoisomer (BMF = 3.6). The large BMF for the α diastereoisomer is consistent with this diastereoisomer dominating higher trophic level organisms in wildlife. Although the BMF of the β diastereoisomer suggests that it will biomagnify, because it is present in small quantities in commercial mixtures it is rarely detected in environmental samples. Results from these studies also provide evidence of bioisomerization of the β and γ diastereoisomers. Most importantly, the α diastereoisomer which was recalcitrant to bioisomerization by juvenile rainbow trout in this study and known to be the dominant diastereosiomer in fish, was bioformed from both the β and γ diastereoisomers. To our knowledge, this is the first report of bioisomerization of a halogenated organic pollutant in biota.

The major objectives of this research were to examine the bioaccumulation parameters [depuration rates (kd), half life (t1/2) and biomagnification factor (BMF)] of individual isomers of hexabromocyclododecane (HBCD, C12H18Br6) in fish and to test the hypothesis of in vivo bioisomerization. This was done by exposing three groups of juvenile rainbow trout (Oncorhynchus Mykiss) to food fortified with known concentrations of an individual diastereoisomer (α, β, γ) for 56 days (uptake phase) followed by 112 days (depuration phase) of unfortified food. A fourth group of fish were exposed to unfortified food for the duration of the experiment. Fish (n=4) from all four aquaria were sacrificed on days 0, 7, 14, 56, 63, 70, 112 and 168 and muscle tissue was extracted and analyzed for diastereoisomer concentrations by high performance liquid chromatography tandem mass spectrometry (LC/MS/MS). Bioaccumulation of the γ diastereoisomer was linear during the uptake phase while the α and β diastereoisomers were found to increase exponentially with respective doubling times of 14.1 and 20.5 days. Both the β and γ diastereoisomers followed first order depuration kinetics with calculated t1/2’s of 94 ± 25 and 84 ± 51 (± 1 × standard error) days, respectively. The BMF for the α diastereoisomer (BMF = 4.1) was one and a half times greater than the β-diastereoisomer (BMF = 2.6) and about one fifth larger than the γ-diastereoisomer (BMF = 3.6). The large BMF for the α diastereoisomer is consistent with this diastereoisomer dominating higher trophic level organisms in wildlife. Although the BMF of the β diastereoisomer suggests that it will biomagnify, because it is present in small quantities in commercial mixtures it is rarely detected in environmental samples. Results from these studies also provide evidence of bioisomerization of the β and γ diastereoisomers. Most importantly, the α diastereoisomer which was recalcitrant to bioisomerization by juvenile rainbow trout in this study and known to be the dominant diastereosiomer in fish, was bioformed from both the β and γ diastereoisomers. To our knowledge, this is the first report of bioisomerization of a halogenated organic pollutant in biota.

To address my first objective, I opportunistically collected road-killed bobcats (n=44) throughout Illinois during 2013-2014, and analyzed their liver tissue for the presence of PCBs, PBDEs, and various other alternative halogenated flame retardants. Concentrations of ∑PCBs ranged from 76.4 ng/g lw to 3782 ng/g lw (median 562.97 ng/g lw). Male bobcats had significantly higher concentrations of PCBs than females (p = 0.04). Concentrations of ∑PBDEs (including all detectable PBDE congeners) ranged from 8.3 to 1920 ng/g lipid weight (median: 50.3 ng/g lw). Among the alternative flame retardants screened, Dechloranes (including anti- and syn-Dechlorane Plus and Dechlorane-602, 603, and 604), tetrabromo-o-chlorotoluene (TBCT), and hexabromocyclododecane (HBCD) were also detected frequently, with median concentrations of 28.7, 5.2, and 11.8 ng/g lw, respectively. Dechlorane analogue compositions in bobcats were different from that reported in other studies, suggesting species- or analogue-dependent bioaccumulation, biomagnification, or metabolism of Dechlorane chemicals in different food webs. My findings, along with previously reported food web models, suggest Dechloranes may possess substantial bioaccumulation and biomagnification potencies in terrestrial mammalian food webs. Thus, attention should be given to these highly bioavailable flame retardants in future environmental biomonitoring and risk assessments in a post-PBDE era. To address my second objective I collected raccoons (n=32) from various sites across Illinois and Missouri during 2013-2015. Liver tissues were analyzed for the presence of PBDEs and Dechloranes. ∑PBDE concentrations ranged from 19.1 ng/g lw to 2124 ng/g lw (median = 98.0 ng/g lw) and did not differ between gender or age of raccoon. Although nonsignificant (p=0.06), adult raccoons appeared to have greater PBDE concentration loads compared to juveniles. These nonsignificant differences are likely due to large variation in contaminant concentrations, possibly reflecting differences in individual raccoon diet and behavior. This is the first study reporting bioaccumulation of halogenated flame retardants in a wild felid in North America, and also the first report of PBDE accumulation in North American raccoons. The wide detection of Dechloranes, HBCD and TBCT in bobcats suggests a broad exposure of these alternative flame retardants in terrestrial apex predators in terrestrial ecosystems. The comparatively high levels of PBDEs in raccoons also suggest that biota in terrestrial habitats are still widely exposed to and susceptible to the bioaccumulation of these flame retardants.

No description available.

No description available.

Phenolic resins are important components of the composite industry because of their excellent flame retardance and cost effectiveness. However, the common procedure for curing phenolic novolac resins uses hexamethylenetetramine (HMTA) and releases volatiles during the cure, which produce networks with numerous voids. This results in materials that lack the toughness necessary for structural applications. An alternative to curing with HMTA is to crosslink the pendant phenolic groups in the novolac resin with epoxy reagents. This reaction proceeds by nucleophilic addition without the release of any volatiles, thereby creating a void-free network. Flame retardance can be achieved by using an excess of the phenolic component. Network densities can also be controlled to maximize both toughness and stiffness by tailoring the stoichiometry of the reagents. Structure-property relationships of phenolic/epoxy networks have been investigated. Glass transitions decreased, and toughness increased, as the phenolic content in the network was increased. Both results could be correlated to the decrease in network densities along this series, which was investigated by measuring the rubbery moduli well above T_g. Fracture toughness of phenolic/epoxy networks measured by K_1c reached 1.03 MPa-m^1/2, compared with an epoxy control with K^1c = 0.62 MPa-m^1/2 and phenolic control with K_1c = 0.16 MPa-m^1/2. In addition, an increase in novolac content improves flame retardance rather dramatically. The peak heat release rate (PHRR) dropped from 1230 kW/m²⁺ for the epoxy control to 260 kW/m²⁺ for the phenolic/epoxy networks, which approached that of a phenolic resol (PHRR = 116 kW/m²⁺). Phenolic/epoxy composite flame retardance also showed significant improvement when compared to epoxy composites. Melt processability of phenolic/epoxy composites has been achieved through the use of latent nucleophilic initiators. Kinetics of the phenolic/epoxy cure reactions with latent initiators demonstrated that monomeric phosphine initiators yielded faster cure reactions as compared to polymeric initiators. These latent initiators allow composite melt processing, such as prepregging or pultrusion, without premature curing. In addition, cure cycles can be reduced from 4 hours to less than 30 minutes. Composites prepared using these latent initiators had toughness exceeding that of epoxy composites and fatigue limits significantly higher than those of vinyl ester composites. <i>Vita removed, June 10, 2013, per author's request. GMc</i> / Ph. D.

Herbaceous peonies (Paeonia lactiflora Pall.) are common perennials used both in gardens and the landscape as well as for cut flowers. Peonies require a chilling period to break dormancy but not for flower bud differentiation. For all studies discussed in this dissertation, two peony cultivars, Sarah Bernhardt and Inspecteur Lavergne, small (3–5 eye) crowns from Holland were potted in 3.8-L pots in mid-November of 2017 and 2018. Our overall objective was to determine if we could manipulate chilling time, along with application of gibberellic acid (GA3) and growth retardants, to produce marketable containerized peonies from a small crown in a single season (November to May). We evaluated chilling, GA3 and a growth retardant (uniconazole; UNZ) under controlled chilling and greenhouse forcing conditions. All potted plants were held outdoors at Battlefield Farms (Rapidan, VA, 38˚ N) for 4 weeks [in 2017, 400 chilling units (CU) according to Fulton Chilling Model] or in a 10°C cooler for 5.5 weeks (in 2018, 400 CU) to root, then placed in a 5°C cooler for 3, 4 or 5 weeks (total 752, 869 or 986 CU). GA3 was applied as a 0 or 100 mg·L-1 drench at 250 ml/pot after the plants were moved into the Virginia Tech greenhouse (Blacksburg, VA, 37˚ N) for forcing. Uniconazole drenches were applied to each cultivar under each chilling treatment at 355 ml/pot at 0, 15, or 20 mg·L-1 at 7 days after the GA3 drench applications. Three weeks chilling at 5°C (752 CU total) provided sufficient chilling for 'Sarah Bernhardt' and 'Inspecteur Lavergne'. Application of GA3 reduced production time and resulted in a greater number of shoots, and, in three of the four studies, increased the number of flowering shoots in three of the four studies. Substrate drench application of 15 mg·L-1 UNZ prior to spring emergence reduced plant width moderately resulting in improved compactness of both cultivars. We evaluated the effects of plant growth retardants applied with different methods at different stages of production on the growth and development of containerized peony under nursery conditions. All potted plants were placed in an unheated coldframe at the Virginia Tech Urban Horticulture Center (Blacksburg, VA, 37˚ N) for one month after potting to promote rooting and then were moved outdoors to a gravel pad to receive natural chilling from November to February. In 2017–18, substrate drenches of UNZ at 0, 15, 30 or 45 mg·L-1 or paclobutrazol (PBZ) at 0, 30, 60 or 90 mg·L-1 at 237 mL/pot were applied about 4 weeks after potting for both cultivars in mid-December 2017. In 2018–19, fall drenches of uniconazole at 0, 15, 30 or 45 mg·L-1 at 237 mL/pot were applied about 4 weeks after potting in mid-December 2018, or spring sprenches of uniconazole were applied at 0, 15, 30 or 45 mg·L-1 at 840 mL·m-2 in March 2019 after 50% shoot emergence for each cultivar. Plant growth retardant applications had little effect on plant growth of either cultivar, but treated plants were of a darker green color compared to the control plants. In addition, higher rates of uniconazole applied as a fall drench increased the number of flowering shoots of both cultivars and the percentage of plants flowering for 'Sarah Bernhardt' in the second season of the study where plants were more protected from spring freezes. Fall paclobutrazol drenches or spring uniconazole sprenches had little effect on flowering. To determine the best timing for spring GA3 applications under nursery conditions, we applied three models based on natural chilling accumulation. The models were a modified Fulton Chilling Model (FCM) for herbaceous peonies, Blackberry Chilling Model 5 (BCM5) for blackberry, or a visual development model (VDM) which was 10% of plants showing shoot emergence in the spring. We choose 1,000 CU for the first two chilling models as the chilling required to break dormancy and promote normal plant growth and flowering. All plants were held in an unheated coldframe at the Virginia Tech Urban Horticulture Center for one month after potting to promote rooting, then were moved outdoors to a gravel pad to receive natural chilling over the winter months. Drenches of 0 or 100 mg·L-1 GA3 were applied at 250 mL/pot to each cultivar under each chilling model when the specific conditions were met. Due to greater winter injury in the 2017–18 season, results varied by year. In the 2017–18 season, GA3 applied according to BCM5 reduce days to emergence for both cultivars and reduce the plant width of 'Inspecteur Lavergne', and later application according to BCM5 and VDM reduced plant length and diameter of 'Sarah Bernhardt'. Reductions in plant size may have been due to greater winter injury due to the earlier emergence of GA3 treated plants. In the 2018–19 season, earlier GA3 drench applications tended to reduce days to emergence for both cultivars and the FCM application reduced days to bud for 'Inspecteur Lavergne', but GA3 drench applications had no effect on plant size. GA3 can be applied after chilling (1,000 CU) using a suitable chilling model such as FCM for peonies, or BCM5, or VDM, but GA3 had little effect on plant development under nursery conditions. We also evaluated GA3 effects on peony bud differentiation and development during controlled chilling and early forcing, as well as effects on growth and flowering. All potted plants were held in a 10°C cooler for 5.5 weeks (400 CU) to root, then placed in a 5°C cooler for 4 weeks (total 869 CU). GA3 was applied at 0 or 100 mg·L-1 pre-chilling or post-chilling as a 250 ml/pot drench. Bud differentiation and development of excised buds were evaluated using a stereomicroscope at potting, after rooting (before chilling), after 1, 2, 3 or 4 weeks of chilling, and at 5, 10 or 15 days after the beginning of forcing. All buds were removed from the sample plants, measured for bud length and diameter, and dissected under a stereomicroscope to assess differentiation stages. Root dry weights and crown dry weights were also determined after rooting, after chilling, and at 15 days of forcing. Ten plants of each treatment were grown in the Virginia Tech greenhouse after chilling until flowering. GA3 applications did not advance the bud development stage because most of buds were already in the reproductive stages before dormancy, but GA3 enhanced bud elongation during chilling and the early forcing period. Our findings suggest that GA3 applications can reduce the time to emergence and flowering, as well as increase the numbers of shoots and flowering shoots. GA3 applied right after rooting in, prior to the chilling period, or before greenhouse forcing, resulted in earlier emergence and flowering with higher quality plants. However, earlier applications, pre-chilling, tended to produce plants with more shoots. Overall, our experiments indicate that three weeks of chilling at 5°C (752 CU total) is a sufficient chilling regime for forcing 'Sarah Bernhardt' and 'Inspecteur Lavergne' peonies, and 1,000 CU of naturally accumulated chilling is sufficient for nursery production. GA3 applications can reduce the time to emergence and flowering, as well as increase the numbers of total shoots and flowering shoots. Timing of GA3 application is flexible; it can be applied right after rooting, before the chilling period, just before greenhouse forcing, or after shoots have begun to emerge. Plant growth retardant applications had a little effect on the growth of tested cultivars, but all plants treated with growth retardants are generally darker green in color. Additionally, growth retardant applications have some positive effects on flowering. / Doctor of Philosophy / Herbaceous peonies (Paeonia lactiflora Pall.) are common perennials used both in gardens and the landscape as well as for cut flowers. Peonies require a chilling period to break dormancy but not for flower bud differentiation. For all studies, two peony cultivars, Sarah Bernhardt and Inspecteur Lavergne, 3 to 5 eye small crowns from Holland were potted in 3.8-L pots in mid November of 2017 and 2018. Our overall objective was to determine if we could manipulate chilling time, along with application of gibberellic acid (GA3) and growth retardants, to produce marketable containerized peonies from a small crown in a single season (November to May). We evaluated chilling, GA3 and a growth retardant (uniconazole) under controlled chilling and greenhouse forcing conditions. We evaluated the effects of plant growth retardants (uniconazole or paclobutrazol) applied with different methods (fall drenches or spring sprenches) at different stages of production on the growth and development of containerized peony under nursery conditions. To determine the best timing for spring GA3 applications under nursery conditions, we applied three models based on natural chilling accumulation. We also evaluated GA3 effects on peony bud differentiation and development during controlled chilling and early forcing, as well as growth and flowering. Overall, 3 weeks chilling at 5°C [752 chilling units (CU) total] is a sufficient chilling regime for forcing 'Sarah Bernhardt' and 'Inspecteur Lavergne' peonies, and 1000 CU naturally accumulated chilling is sufficient for nursery production. GA3 applications can reduce the time to emergence and flowering, as well as increase the numbers of shoots and flowering shoots. Timing of GA3 application is flexible, it can be applied right after rooting, after the chilling period, or after shoots have begun to emerge. Plant growth retardant applications had little effect on plant growth of either cultivar, but all plants treated with growth retardants were darker green in color. Additionally, growth retardant applications had some positive effects on flowering.

The studies in this dissertation research were conducted to investigate the possible mode of action by which a brominated flame retardant, 2, 2-Bis (bromomethyl)-1, 3-propanediol (BMP) causes genotoxicity. Binding of BMP to DNA and BMP induced DNA strand breaks were investigated in SV-40 immortalized human uroepithelial cells (UROtsa) as an in vitro model for the bladder (a tissue that developed cancer after two year exposure to BMP in rodents). Results showed binding of [¹⁴C]-BMP equivalents to DNA increased with increased exposure time and concentration of [¹⁴C]-BMP. Comet analysis indicated BMP significantly increased the extent of DNA strand breaks at 1 and 3 h of incubation. However, strand breaks were repaired by 6 h of incubation. The DNA damaging effects of BMP at 1 h was concentration dependent. Compared with the parent compound, BMP-glucuronide (the predominant metabolite of BMP) bound less to DNA and produced less DNA strand breaks in UROtsa cells. Evidences that the BMP induced strand breaks were the result of an oxidative stress include: a concentration and time dependent increase in ROS generation; increased expression of Nrf2 and HSP70; complete attenuation of BMP induced DNA strand breaks by the antioxidant, NAC; and the presence of the oxidized base 8-OHguanine. UROtsa cells appear to be target cells for BMP because, as compared to rat hepatocytes (non-target cells), these cells lack the ability to detoxify BMP via glucuronidation and also because they are deficient in glutathione, a major intracellular antioxidant molecule. Both of these genotoxic events, DNA binding and oxidative DNA damage may, in part, contribute to BMP carcinogenicity observed in rodents. The relevance of current results to humans is remained to be established.

<p>Background: Organophosphate (OP) pesticides are well-known developmental neurotoxicants that have been linked to abnormal cognitive and behavioral endpoints through both epidemiological studies and animal models of behavioral teratology, and are implicated in the dysfunction of multiple neurotransmitters, including dopamine. Chemical similarities between OP pesticides and organophosphate flame retardants (OPFRs), a class of compounds growing in use and environmental relevance, have produced concern regarding whether developmental exposures to OPFRs and OP pesticides may share behavioral outcomes, impacts on dopaminergic systems, or both. Methods: Using the zebrafish animal model, we exposed developing fish to two OPFRs, TDCIPP and TPHP, as well as the OP pesticide chlorpyrifos, during the first 5 days following fertilization. From there, the exposed fish were assayed for behavioral abnormalities and effects on monoamine neurochemistry as both larvae and adults. An experiment conducted in parallel examined how antagonism of the dopamine system during an identical window of development could alter later life behavior in the same assays. Finally, we investigated the interaction between developmental exposure to an OPFR and acute dopamine antagonism in larval behavior. Results: Developmental exposure to all three OP compounds altered zebrafish behavior, with effects persisting into adulthood. Additionally, exposure to an OPFR decreased the behavioral response to acute D2 receptor antagonism in larvae. However, the pattern of behavioral effects diverged substantially from those seen following developmental dopamine antagonism, and the investigations into dopamine neurochemistry were too variable to be conclusive. Thus, although the results support the hypothesis that OPFRs, as with OP pesticides such as chlorpyrifos, may present a risk to normal behavioral development, we were unable to directly link these effects to any dopaminergic dysfunction.</p> / Dissertation

Numerous studies have focused on enhancing the flame retardant behavior of cotton and polyurethane foam. Some of the most commonly used treatments (e.g., brominated compounds) have raised concerns with regard to toxicity and environmental persistence. These concerns have led to significant research into the use of alternative approaches, including polymer nanocomposites prepared from more environmentally benign nanoparticles. These particles migrate to the surface from the bulk during fire exposure to form a barrier on the surface that protects the underlying polymer. This theory of fire suppression in bulk nanocomposites inspired the use of layer-by-layer (LbL) assembly to create nanocoatings in an effort to produce more effective and environmentally-benign flame retardant treatments. Negatively charged silica nanoparticles of two different sizes were paired with either positively charged silica or cationic polyethylenimine (PEI) to create thin film assemblies. When applying these films to cotton fabric, all coated fabrics retained their weave structure after being exposed to a vertical flame test, while uncoated cotton was completely destroyed. Micro combustion calorimetry confirmed that coated fabrics exhibited a reduced peak heat release rate, by as much as 20% relative to the uncoated control. Even so, this treatment would not pass the standard UL94 vertical flame test, necessitating a more effective treatment. Positively- charged chitosan (CH) was paired with montmorillonite (MMT) clay to create a renewable flame retardant nanocoating for polyurethane foam. This coating system completely stops the melting of a flexible polyurethane foam when exposed to direct flame from a butane torch, with just 10 bilayers (~ 30 nm thick). The same coated foam exhibited a reduced peak heat release rate, by as much as 52%, relative to the uncoated control. This same nanobrick wall coating is able to impart gas barrier to permeate plastic film. Multilayered thin films were assembled with "green" food contact approved materials (i.e., chitosan, polyacrylic acid (PAA) and montmorillonite clay). Only ten CH-PAA-CH-MMT quadlayers (~90 nm thick) cause polylactic acid (PLA) film to behave like PET in terms of oxygen barrier. A thirty bilayer CH-MMT assembly (~100 nm thick) on PLA exhibits an oxygen transmission rate (OTR) below the detection limit of commercial instrumentation (<= 0.005 cm^3/(m^2*day*atm)). This is the same recipe used to impart flame retardant behavior to foam, but it did not provide effective FR to cotton fabric, so a very different recipe was used. Thin films of fully renewable electrolytes, chitosan and phytic acid (PA), were deposited on cotton fabric in an effort to reduce flammability through an intumescent effect. Altering the pH of aqueous deposition solutions modifies the composition of the final nanocoating. Fabrics coated with highest PA content multilayers completely extinguished the flame and reduced peak heat release (pkHRR) and total heat release of 60% and 76%, respectively. This superior performance is believed to be due to high phosphorus content that enhances the intumescent behavior of these nanocoatings.