Exposure to and Biomarker Responses From Legacy and Emerging Contaminants Along Three Drainages in the Milwaukee Estuary, Wisconsin, USA.

Legacy contaminants and contaminants of emerging concern (CECs) were assessed in tree swallow (Tachycineta bicolor) tissue and diet samples from three drainages in the Milwaukee estuary, Wisconsin, USA, to understand exposures and possible biomarker responses. Two remote Wisconsin lakes were assessed for comparative purposes. Bioaccumulative classes of contaminants, such as polybrominated diphenyl ethers and per- and polyfluoroalkyl substances, while at higher concentrations than the reference lakes, did not vary significantly among sites or among the three drainages. Polycyclic aromatic hydrocarbons were assessed in diet and sediment and were from primarily pyrogenic sources. Ten biomarkers were assessed relative to contaminant exposure. Polychlorinated biphenyls (PCBs) were elevated above reference conditions at all Milwaukee sites but did not correlate with any measured biomarker responses. Only one site, Cedarburg, just downstream from a Superfund site, had elevated PCBs compared to other sites in the Milwaukee estuary. Few non-organochlorine insecticides or herbicides were detected in tree swallow liver tissue, except for the atrazine metabolite desethylatrazine. Few pharmaceuticals and personal care products were detected in liver tissue except for N,N-diethyl-meta-toluamide, iopamidol, and two antibiotics. The present study is one of the most comprehensive assessments to date, along with the previously published Maumee River data, on the exposure and effects of a wide variety of CECs in birds. Environ Toxicol Chem 2024;43:856-877. © 2024 SETAC. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.

Free Ethynylarsinidene and Ethynylstibinidene: Heavier Analogues of Nitrenes and Phosphinidenes.

Until now, there has been very little experimental evidence for the existence of free arsinidenes and stibinidenes, apart from the hydrides, AsH and SbH. Here, we report on photogeneration of triplet ethynylarsinidene, HCCAs, and triplet ethynylstibinidene, HCCSb, from ethynylarsine and ethynylstibine, respectively, in solid argon matrices. The products were identified using infrared spectroscopy and the associated UV absorption spectra are interpreted with the aid of theoretical predictions.

Multi-omics responses in tree swallow (Tachycineta bicolor) nestlings from the Maumee Area of Concern, Maumee River, Ohio.

A multi-omics approach was utilized to identify altered biological responses and functions, and to prioritize contaminants to assess the risks of chemical mixtures in the Maumee Area of Concern (AOC), Maumee River, OH, USA. The Maumee AOC is designated by the United States Environmental Protection Agency as having significant beneficial use impairments, including degradation of fish and wildlife populations, bird or animal deformities or reproduction problems, and loss of fish and wildlife habitat. Tree swallow (Tachycineta bicolor) nestlings were collected at five sites along the Maumee River, which included wastewater treatment plants (WWTPs) and industrial land-use sites. Polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs), polycyclic aromatic hydrocarbons (PAHs), polychlorinated dibenzo p dioxins and furans (PCDD/Fs), and chlorinated pesticide concentrations were elevated in Maumee tree swallows, relative to a remote reference site, Star Lake, WI, USA. Liver tissue was utilized for non-targeted transcriptome and targeted metabolome evaluation. A significantly differentially expressed gene cluster related to a downregulation in cell growth and cell cycle regulation was identified when comparing all Maumee River sites with the reference site. There was an upregulation of lipogenesis genes, such as PPAR signaling (HMGCS2, SLC22A5), biosynthesis of unsaturated fatty acids (FASN, SCD, ELOVL2, and FADS2), and higher lipogenesis related metabolites, such as docosapentaenoic acid (DPA), docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), and arachidonic acid (AA) at two industrial land-use sites, Ironhead and Maumee, relative to WWTP sites (Perrysburg and SideCut), and the reference site. Toledo Water, in the vicinity of the other two industrial sites and also adjacent to a WWTP, showed a mix of signals between industrial land-use and WWTP land-use. PAHs, oxychlordane, and PBDEs were determined to be the most likely causes of the differentiation in biological responses, including de novo lipogenesis and biosynthesis of unsaturated fatty acids.

Unusual Quartet-Doublet Phosphorescence from the Phosphaethynyl Radical, CP.

We report the first detection of phosphorescence from the phosphaethynyl radical. This rare instance of quartet-doublet emission, studied here in solid argon, is presumably promoted by efficient intersystem crossing from the originally photoexcited doublet (B2 Σ+ ) to the adjacent second quartet state, 14 Δ. Vibronic progressions were traced for the a4 Σ+ -X2 Σ+ and a4 Σ+ -A2 Пi systems from their origins up to (v'=0)→(v''=5) and (v'=0)→(v''=2) bands, respectively. The measured phosphorescence lifetime is 108 ± 3 ms.

Using tree swallows to assess reductions in PCB exposure as a result of dredging at Great Lakes Restoration Initiative (GLRI) sites in the Upper Midwest, USA.

Tree swallows (Tachycineta bicolor) were used to assess the effectiveness of reducing polychlorinated biphenyl (PCB) exposure to wildlife as a result of contaminated sediment removal at locations across the Great Lakes under two dredging scenarios, full or spot dredging. For comparative purposes, other locations where no dredging occurred were also assessed. Calculating accumulation rate, from the mass of a contaminant in tree swallow eggs and nestling carcasses, is a useful tool to assess the effectiveness of sediment removal. It has the advantage over more commonly used metrics such as cubic yards of sediment removed or kg of a contaminant removed, because it assesses a biotic endpoint that has more societal understanding. Egg and nestling concentrations of total PCBs and accumulation rate (µg of total PCBs accumulated per day) were compared pre- and post-dredge. At the most contaminated site, Waukegan Harbor, Illinois, the accumulation rate decreased by 95% because of dredging. At less contaminated locations in Wisconsin and Ohio, the accumulation rate was reduced by dredging as well, but not to such a large extent (~50%). Even at reference locations, there was a very small amount (0.01-0.06 µg/day) of PCBs accumulated each day because of the prevalence of this contaminant in the environment. The profile of individual PCB congeners also differed pre-and post-dredge and demonstrated significant changes as a result of dredging activities.

An Efficient Photochemical Route Towards Triplet Ethynylphosphinidene, HCCP.

While the archetypal free phosphinidene, H-P, has been studied for over a century, reports on uncomplexed, univalent phosphorus compounds are very sparse. Here we demonstrate production of HCCP in solid argon through the UV-induced rearrangement and subsequent dehydrogenation of phosphapropyne, CH3 CP. Migration of H atoms along the CCP backbone of CH3 CP resulted in production of the previously unobserved species 1-phosphapropadiene, CH2 =C=PH, followed by ethynylphosphine, HCCPH2 .

Legacy and Contaminants of Emerging Concern in Tree Swallows Along an Agricultural to Industrial Gradient: Maumee River, Ohio.

Exposure to multiple classes of contaminants, both legacy and contaminants of emerging concern (CECs), were assessed in tree swallow (Tachycineta bicolor) tissue and diet samples from 6 sites along the Maumee River, Ohio, USA, to understand both exposure and possible effects of exposure to those CECs for which there are little avian data. The 6 sites represented a gradient from intensive agriculture upstream to highly urbanized and industrial landscapes downstream; 1 or 2 remote Wisconsin lakes were assessed for comparative purposes. Cytochrome P450 induction, DNA damage, and thyroid function were also assessed relative to contaminant exposure. Bioaccumulative CECs, such as polybrominated diphenyl ethers (PBDEs) and perfluorinated substances, did not follow any upstream to downstream gradient; but both had significantly greater concentrations along the Maumee River than at the remote lake sites. Greater exposure to PBDEs was apparent in swallows at or near wastewater-treatment facilities than at other sites. Total polychlorinated biphenyl and total polycyclic aromatic hydrocarbon concentrations were greater in swallows at downstream locations compared to upstream sites and were associated with higher ethoxyresorufin-O-dealkylase activity. Few herbicides or nonorganochlorine insecticides were detected in swallow tissues or their food, except for atrazine and its metabolite desethylatrazine. Few pharmaceuticals and personal care products were detected except for DEET and iopamidol. Both were detected in most liver samples but not in eggs, as well as detected at the remote lake sites. This is one of the most comprehensive assessments to date of exposure and effects of a wide variety of CECs in birds. Environ Toxicol Chem 2020;39:1936-1952. © 2020 SETAC. This article has been contributed to by US Government employees and their work is in the public domain in the USA.

Isomerization of cyanopropyne in solid argon.

Cyanopropyne, CH3-C[triple bond, length as m-dash]C-CN, is a simple molecule whose photochemistry is still unexplored. Here we investigate the UV photolysis of this astrophysically significant nitrile trapped in solid argon. The FTIR study was assisted with 15N-isotopic substitution data and with DFT-level computations including the analyses of ground- and excited-state potential energy surfaces. Cyanopropyne was found to decay mainly via a two-step isomerization process. Infrared absorption spectra evolved to show signals from allenyl cyanide, CH2[double bond, length as m-dash]C[double bond, length as m-dash]CH-CN, which then further convert into propargyl cyanide, H-C[triple bond, length as m-dash]C-CH2-CN. Some evidence for the presence of allenyl isocyanide, propargyl isocyanide, 3-cyanocyclopropene, and 1,2,3-butatrien-1-imine under particular experimental conditions was also observed. Although cyano/isocyano interconversion has been observed during photolysis of other closely related species in solid argon matrices, including H-C[triple bond, length as m-dash]C-CN, no evidence could be found for production of 1-isocyano-1-propyne, CH3-C[triple bond, length as m-dash]C-NC for these experiments.

Photochemistry of XCH2CN (X = -Cl, -SH) in Argon Matrices.

We report infrared spectra and photochemical behavior of the potentially astrochemically significant species, mercaptoacetonitrile (HS-CH2C≡N) and, for comparison purposes, chloroacetonitrile (Cl-CH2C≡N), both suspended in an argon matrix at 6 K. Photolytic formation of the isocyano products HS-CH2-NC and Cl-CH2-NC were observed as well as CH3NSC and CH3SCN (in HS-CH2CN photolysis). While no dissociation products were observed for Cl-CH2-CN, photolysis of HS-CH2-CN produced compounds necessitating the loss of the CN group to form CH2═S, the SH group to form H2C-CN and HC-CN, or both CN and SH to form CH3 and CH4. Observation of emission spectra upon annealing indicates the presence of free sulfur atom in matrices of photolyzed HS-CH2-CN.

Perfluoroalkyl Contaminant Exposure and Effects in Tree Swallows Nesting at Clarks Marsh, Oscoda, Michigan, USA.

A site in northeastern Michigan, Oscoda Township, has some of the highest recorded exposure in birds to perfluorinated substances (PFASs) in the United States. Some egg and plasma concentrations at that location exceeded the lowest reproductive effect threshold established for two avian laboratory species. The objectives of this study were to determine whether there were reproductive effects or physiological responses in a model bird species, the tree swallow (Tachycineta bicolor), associated with this extremely high exposure to PFASs. The lack of exposure above background to other contaminants at this site allowed for an assessment of PFAS effects without the complication that responses may be caused by other contaminants. A secondary objective was to determine the distribution of PFASs in multiple tissue types to better understand and interpret residues in different tissues. This can best be done at highly exposed locations where tissue concentrations would be expected to be above detectable levels if they are present in that tissue. There were no demonstrable effects of PFAS exposure on reproduction nor on most physiological responses.

Element Concentrations in Tree Swallows (Tachycineta bicolor) from the U.S. and Binational Great Lakes Areas of Concern.

Selected elements were targeted in state Remedial Action Plans as one group of chemicals affecting the Beneficial Use Impairments of Great Lakes Areas of Concern (AOCs). Livers of nestling tree swallows, Tachycineta bicolor, were harvested from 76 sites in the Great Lakes, which included multiple sites at 27 AOCs and 12 reference sites from 2010 to 2015, and were analyzed for 21 elements. Mercury concentrations were at background levels at all sites. Elevated cadmium (Cd) concentrations were associated with industry. The highest Cd values were from the Black River, OH AOC and were associated with historic coke production but were not at toxic levels. Lead (Pb) concentrations were highest on the Rouge River, MI AOC-the oldest and most heavily populated and industrialized area in southeast Michigan. Individual Pb concentrations were elevated to a level associated with delta-aminolaevulinic acid dehydratase inhibition but not to a level considered toxic. In contrast, livers harvested from sites on the southwest shore of Lake Michigan had selenium (Se) concentrations elevated to levels associated with reduced avian reproduction. One likely source of the high Se concentrations was pollution from a local coal-fired power plant. Concentrations of the remaining elements were at background levels.

Reproductive success and contaminant associations in tree swallows (Tachycineta bicolor) used to assess a Beneficial Use Impairment in U.S. and Binational Great Lakes' Areas of Concern.

During 2010-2014, tree swallow (Tachycineta bicolor) reproductive success was monitored at 68 sites across all 5 Great Lakes, including 58 sites located within Great Lakes Areas of Concern (AOCs) and 10 non-AOCs. Sample eggs were collected from tree swallow clutches and analyzed for contaminants including polychlorinated biphenyls (PCBs), dioxins and furans, polybrominated diphenyl ethers, and 34 other organic compounds. Contaminant data were available for 360 of the clutches monitored. Markov chain multistate modeling was used to assess the importance of 5 ecological variables and 11 of the dominant contaminants in explaining the pattern of egg and nestling failure rates. Four of 5 ecological variables (Female Age, Date within season, Year, and Site) were important explanatory variables. Of the 11 contaminants, only total dioxin and furan toxic equivalents (TEQs) explained a significant amount of the egg failure probabilities. Neither total PCBs nor PCB TEQs explained the variation in egg failure rates. In a separate analysis, polycyclic aromatic hydrocarbon exposure in nestling diet, used as a proxy for female diet during egg laying, was significantly correlated with the daily probability of egg failure. The 8 sites within AOCs which had poorer reproduction when compared to 10 non-AOC sites, the measure of impaired reproduction as defined by the Great Lakes Restoration Initiative, were associated with exposure to dioxins and furan TEQs, PAHs, or depredation. Only 2 sites had poorer reproduction than the poorest performing non-AOC. Using a classic (non-modeling) approach to estimating reproductive success, 82% of nests hatched at least 1 egg, and 75% of eggs laid, excluding those collected for contaminant analyses, hatched.

Annual variation in polychlorinated biphenyl (PCB) exposure in tree swallow (Tachycineta bicolor) eggs and nestlings at Great Lakes Restoration Initiative (GLRI) study sites.

Tree swallow (Tachycineta bicolor) eggs and nestlings were collected from 16 sites across the Great Lakes to quantify normal annual variation in total polychlorinated biphenyl (PCB) exposure and to validate the sample size choice in earlier work. A sample size of five eggs or five nestlings per site was adequate to quantify exposure to PCBs in tree swallows given the current exposure levels and variation. There was no difference in PCB exposure in two randomly selected sets of five eggs collected in the same year, but analyzed in different years. Additionally, there was only modest annual variation in exposure, with between 69% (nestlings) and 73% (eggs) of sites having no differences between years. There was a tendency, both statistically and qualitatively, for there to be less exposure in the second year compared to the first year.

EROD activity, chromosomal damage, and oxidative stress in response to contaminants exposure in tree swallow (Tachycineta bicolor) nestlings from Great Lakes Areas of Concern.

Tree swallow, Tachycineta bicolor, nestlings were collected from 60 sites in the Great Lakes, which included multiple sites within 27 Areas of Concern (AOCs) and six sites not listed as AOCs from 2010 to 2014. Nestlings, approximately 12 days-of-age, were evaluated for ethoxyresorufin-O-dealkylase (EROD) activity, chromosomal damage, and six measures of oxidative stress. Data on each of these biomarkers were divided into four equal numbered groups from the highest to lowest values and the groups were compared to contaminant concentrations using multivariate analysis. Contaminant concentrations, from the same nestlings, included polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs), perfluorinated compounds (PFCs), and 17 elements. Alkylated polycyclic aromatic hydrocarbons (aPAHs) and parent PAHs (pPAHs) were measured in pooled nestling dietary samples. Polychlorinated dibenzo-p-dioxins, polychlorinated dibenzofurans, and pesticides were measured in sibling eggs. Concentrations of aPAHs, pPAHs, chlordane, dieldrin, heptachlor, and PCBs, in that order, were the major contributors to the significant differences between the lowest and highest EROD activities; PFCs, PBDEs, the remaining pesticides, and all elements were of secondary importance. The four categories of chromosomal damage did not separate out well based on the contaminants measured. Concentrations of aPAHs, pPAHs, heptachlor, PCBs, chlordane, and dieldrin were the major contributors to the significant differences between the lowest and highest activities of two oxidative stress measures, total sulfhydryl (TSH) activity and protein bound sulfhydryl (PBSH) activity. The four categories of thiobarbituric acid reacting substances (TBARS), oxidized glutathione (GSSG), reduced glutathione (GSH), and the ratio of GSSG/GSH did not separate well based on the contaminants measured.

Cryogenic Photochemical Synthesis and Electronic Spectroscopy of Cyanotetracetylene.

HC9N is a molecule of astrochemical interest. In this study, it was produced in cryogenic Ar and Kr matrices from UV-photolyzed diacetylene/cyanodiacetylene mixtures. Its strong phosphorescence was discovered and served for the identification of the compound. Vibrationally resolved phosphorescence excitation spectra gave insight into excited singlet electronic states. Two electronic systems were observed around 26 000-34 000 cm-1 and 35 000-50 000 cm-1. Energies of the second excited singlet and the lowest triplet state were derived from analysis of these systems. Vibrational and electronic spectroscopic features were assigned with the assistance of density functional theory calculations. Some trends concerning the electronic spectroscopy of HC2n+1N family molecules are presented.

Structure and Spectroscopy of C2HNO Isomers.

Formyl cyanide has been detected toward Sgr B2 making this chemical and its potential isomers of astrophysical interest. We calculated the precise energies of the five most stable structural isomers of formyl cyanide and found that formyl isocyanide and 2H-azirin-2-one are less stable than formyl cyanide by 55 and 125 kJ/mol, respectively. We present our ab initio coupled cluster predictions of the spectroscopic parameters relevant to their gas-phase rotational and vibrational spectroscopy. Parameters include ground vibrational state rotational constants, centrifugal distortion parameters, equilibrium electric dipole moments, and nuclear quadrupole coupling constants. Anharmonic vibrational frequencies together with infrared intensities of fundamental and nonfundamental modes were also calculated. The broader potential energy surface, including transition states and the minima they connect, were explored for these isomers at the B3LYP/aug-cc-pVTZ level of theory. Formyl cyanide can dissociate in an exothermic reaction to form HCN and CO or in an endothermic reaction to form HNC and CO. The activation energies for both processes are close to 260 kJ/mol. The activation barrier for conversion of formyl cyanide into formyl isocyanide is ∼180 kJ/mol. The energetic barrier for the exothermic decomposition of formyl isocyanide into HNC and CO is ∼210 kJ/mol. The barrier for transformation of formyl isocyanide into formyl cyanide is ∼130 kJ/mol. Formyl isocyanide seems to be a stable chemical and could potentially survive in a dense molecular cloud if formed. These data are useful for future identification of members of this family of molecules in a laboratory or in space.

In vitro labeling strategies for in cellulo fluorescence microscopy of single ribonucleoprotein machines.

RNA plays a fundamental, ubiquitous role as either substrate or functional component of many large cellular complexes-"molecular machines"-used to maintain and control the readout of genetic information, a functional landscape that we are only beginning to understand. The cellular mechanisms for the spatiotemporal organization of the plethora of RNAs involved in gene expression are particularly poorly understood. Intracellular single-molecule fluorescence microscopy provides a powerful emerging tool for probing the pertinent mechanistic parameters that govern cellular RNA functions, including those of protein coding messenger RNAs (mRNAs). Progress has been hampered, however, by the scarcity of efficient high-yield methods to fluorescently label RNA molecules without the need to drastically increase their molecular weight through artificial appendages that may result in altered behavior. Herein, we employ T7 RNA polymerase to body label an RNA with a cyanine dye, as well as yeast poly(A) polymerase to strategically place multiple 2'-azido-modifications for subsequent fluorophore labeling either between the body and tail or randomly throughout the tail. Using a combination of biochemical and single-molecule fluorescence microscopy approaches, we demonstrate that both yeast poly(A) polymerase labeling strategies result in fully functional mRNA, whereas protein coding is severely diminished in the case of body labeling.

Organic contamination in tree swallow (Tachycineta bicolor) nestlings at United States and binational Great Lakes Areas of Concern.

Contaminant exposure of tree swallows, Tachycineta bicolor, nesting in 27 Areas of Concern (AOCs) in the Great Lakes basin was assessed from 2010 to 2014 to assist managers and regulators in their assessments of Great Lakes AOCs. Contaminant concentrations in nestlings from AOCs were compared with those in nestlings from nearby non-AOC sites. Polychlorinated biphenyl (PCB) and polybrominated diphenyl ether concentrations in tree swallow nestling carcasses at 30% and 33% of AOCs, respectively, were below the mean concentration for non-AOCs. Polycyclic aromatic hydrocarbon (PAH) concentrations in nestling stomach contents and perfluorinated compound concentrations in nestling plasma at 67% and 64% of AOCs, respectively, were below the mean concentration for non-AOCs. Concentrations of PCBs in nestling carcasses were elevated at some AOCs but modest compared with highly PCB-contaminated sites where reproductive effects have been documented. Concentrations of PAHs in diet were sufficiently elevated at some AOCs to elicit a measurable physiological response. Among AOCs, concentrations of the perfluorinated compound perfluorooctane sulfonate in plasma were the highest on the River Raisin (MI, USA; geometric mean 330 ng/mL) but well below an estimated toxicity reference value (1700 ng/mL). Both PAH and PCB concentrations in nestling stomach contents and PCBs in carcasses were significantly correlated with concentrations in sediment previously reported, thereby reinforcing the utility of tree swallows to assess bioavailability of sediment contamination. Environ Toxicol Chem 2017;36:735-748. Published 2016 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.

Electronic Spectroscopy of Methylcyanodiacetylene (CH3 C5 N).

The results of a study devoted to the electronic spectroscopy of gaseous, solid, and cryogenic matrix-isolated methylcyanodiacetylene (CH3 C5 N) are reported. UV absorption and optical phosphorescence spectra of the compound are described here for the first time, and the corresponding vibronic assignments are proposed. UV absorption, studied directly or through the excitation of phosphorescence, revealed the B˜ 1 E--X˜ 1 A1 system, very weak A˜ 1 A2 -X˜ 1 A1 bands, and a strong, broad absorption feature, tentatively identified as D˜ 1 E-X˜ 1 A1 . Spectral measurements were assisted by quantum chemical calculations at the DFT and ab initio (coupled cluster) levels of theory.

A novel method to accurately locate and count large numbers of steps by photobleaching.

Photobleaching event counting is a single-molecule fluorescence technique that is increasingly being used to determine the stoichiometry of protein and RNA complexes composed of many subunits in vivo as well as in vitro. By tagging protein or RNA subunits with fluorophores, activating them, and subsequently observing as the fluorophores photobleach, one obtains information on the number of subunits in a complex. The noise properties in a photobleaching time trace depend on the number of active fluorescent subunits. Thus, as fluorophores stochastically photobleach, noise properties of the time trace change stochastically, and these varying noise properties have created a challenge in identifying photobleaching steps in a time trace. Although photobleaching steps are often detected by eye, this method only works for high individual fluorophore emission signal-to-noise ratios and small numbers of fluorophores. With filtering methods or currently available algorithms, it is possible to reliably identify photobleaching steps for up to 20-30 fluorophores and signal-to-noise ratios down to ∼1. Here we present a new Bayesian method of counting steps in photobleaching time traces that takes into account stochastic noise variation in addition to complications such as overlapping photobleaching events that may arise from fluorophore interactions, as well as on-off blinking. Our method is capable of detecting ≥50 photobleaching steps even for signal-to-noise ratios as low as 0.1, can find up to ≥500 steps for more favorable noise profiles, and is computationally inexpensive.