@article{phelps_palekar_conley_ferrero_driggers_linder_kullman_reif_sheats_dewitt_et al._2023, title={Legacy and emerging per- and polyfluoroalkyl substances suppress the neutrophil respiratory burst}, volume={20}, ISSN={["1547-6901"]}, url={https://doi.org/10.1080/1547691X.2023.2176953}, DOI={10.1080/1547691X.2023.2176953}, abstractNote={Abstract Per- and polyfluoroalkyl substances (PFASs) are used in a multitude of processes and products, including nonstick coatings, food wrappers, and fire-fighting foams. These chemicals are environmentally-persistent, ubiquitous, and can be detected in the serum of 98% of Americans. Despite evidence that PFASs alter adaptive immunity, few studies have investigated their effects on innate immunity. The report here presents results of studies that investigated the impact of nine environmentally-relevant PFASs [e.g. perfluorooctanoic acid (PFOA), perfluorooctane sulfonic acid potassium salt (PFOS-K), perfluorononanoic acid (PFNA), perfluorohexanoic acid (PFHxA), perfluorohexane sulfonic acid (PFHxS), perfluorobutane sulfonic acid (PFBS), ammonium perfluoro(2-methyl-3-oxahexanoate) (GenX), 7H-perfluoro-4-methyl-3,6-dioxa-octane sulfonic acid (Nafion byproduct 2), and perfluoromethoxyacetic acid sodium salt (PFMOAA-Na)] on one component of the innate immune response, the neutrophil respiratory burst. The respiratory burst is a key innate immune process by which microbicidal reactive oxygen species (ROS) are rapidly induced by neutrophils in response to pathogens; defects in the respiratory burst can increase susceptibility to infection. The study here utilized larval zebrafish, a human neutrophil-like cell line, and primary human neutrophils to ascertain whether PFAS exposure inhibits ROS production in the respiratory burst. It was observed that exposure to PFHxA and GenX suppresses the respiratory burst in zebrafish larvae and a human neutrophil-like cell line. GenX also suppressed the respiratory burst in primary human neutrophils. This report is the first to demonstrate that these PFASs suppress neutrophil function and support the utility of employing zebrafish larvae and a human cell line as screening tools to identify chemicals that may suppress human immune function.}, number={1}, journal={JOURNAL OF IMMUNOTOXICOLOGY}, author={Phelps, Drake W. and Palekar, Anika I. and Conley, Haleigh E. and Ferrero, Giuliano and Driggers, Jacob H. and Linder, Keith E. and Kullman, Seth W. and Reif, David M. and Sheats, M. Katie and DeWitt, Jamie C. and et al.}, year={2023}, month={Dec} }
 @article{weitekamp_kvasnicka_keely_brinkman_howey_gaballah_phelps_catron_zurlinden_wheaton_et al._2021, title={Monoassociation with bacterial isolates reveals the role of colonization, community complexity and abundance on locomotor behavior in larval zebrafish}, volume={3}, DOI={10.1186/s42523-020-00069-x}, abstractNote={Abstract Background Across taxa, animals with depleted intestinal microbiomes show disrupted behavioral phenotypes. Axenic (i.e., microbe-free) mice, zebrafish, and fruit flies exhibit increased locomotor behavior, or hyperactivity. The mechanism through which bacteria interact with host cells to trigger normal neurobehavioral development in larval zebrafish is not well understood. Here, we monoassociated zebrafish with either one of six different zebrafish-associated bacteria, mixtures of these host-associates, or with an environmental bacterial isolate. Results As predicted, the axenic cohort was hyperactive. Monoassociation with three different host-associated bacterial species, as well as with the mixtures, resulted in control-like locomotor behavior. Monoassociation with one host-associate and the environmental isolate resulted in the hyperactive phenotype characteristic of axenic larvae, while monoassociation with two other host-associated bacteria partially blocked this phenotype. Furthermore, we found an inverse relationship between the total concentration of bacteria per larvae and locomotor behavior. Lastly, in the axenic and associated cohorts, but not in the larvae with complex communities, we detected unexpected bacteria, some of which may be present as facultative predators. Conclusions These data support a growing body of evidence that individual species of bacteria can have different effects on host behavior, potentially related to their success at intestinal colonization. Specific to the zebrafish model, our results suggest that differences in the composition of microbes in fish facilities could affect the results of behavioral assays within pharmacological and toxicological studies.}, number={1}, journal={Animal Microbiome}, publisher={Springer Science and Business Media LLC}, author={Weitekamp, Chelsea A. and Kvasnicka, Allison and Keely, Scott P. and Brinkman, Nichole E. and Howey, Xia Meng and Gaballah, Shaza and Phelps, Drake and Catron, Tara and Zurlinden, Todd and Wheaton, Emily and et al.}, year={2021}, month={Jan} }
 @article{garbutt_konganti_konneker_hillhouse_phelps_jones_aylor_threadgill_2020, title={Derivation of stable embryonic stem cell-like, but transcriptionally heterogenous, induced pluripotent stem cells from non-permissive mouse strains}, volume={31}, ISSN={["1432-1777"]}, DOI={10.1007/s00335-020-09849-x}, abstractNote={Genetic background is known to play a role in the ability to derive pluripotent, embryonic stem cells (ESC), a trait referred to as permissiveness. Previously we demonstrated that induced pluripotent stem cells (iPSC) can be readily derived from non-permissive mouse strains by addition of serum-based media supplemented with GSK3B and MEK inhibitors, termed 2iS media, 3 days into reprogramming. Here, we describe the derivation of second type of iPSC colony from non-permissive mouse strains that can be stably maintained independently of 2iS media. The resulting cells display transcriptional heterogeneity similar to that observed in ESC from permissive genetic backgrounds derived in conventional serum containing media supplemented with leukemia inhibitor factor. However, unlike previous studies that report exclusive subpopulations, we observe both exclusive and simultaneous expression of naive and primed cell surface markers. Herein, we explore shifts in pluripotency in the presence of 2iS and characterize heterogenous subpopulations to determine their pluripotent state and role in heterogenous iPSCs derived from the non-permissive NOD/ShiLtJ strain. We conclude that heterogeneity is a naturally occurring, necessary quality of stem cells that allows for the maintenance of pluripotency. This study further demonstrates the efficacy of the 2iS reprogramming technique. It is also the first study to derive stable ESC-like stem cells from the non-permissive NOD/ShiLtJ and WSB/EiJ strains, enabling easier and broader research possibilities into pluripotency for these and similar non-permissive mouse strains and species.}, number={9-12}, journal={MAMMALIAN GENOME}, publisher={Springer Science and Business Media LLC}, author={Garbutt, Tiffany A. and Konganti, Kranti and Konneker, Thomas and Hillhouse, Andrew and Phelps, Drake and Jones, Alexis and Aylor, David and Threadgill, David W.}, year={2020}, month={Dec}, pages={263–286} }
 @article{phelps_fletcher_rodriguez-nunez_balik-meisner_tokarz_reif_germolec_yoder_2020, title={In vivo assessment of respiratory burst inhibition by xenobiotic exposure using larval zebrafish}, volume={17}, url={https://doi.org/10.1080/1547691X.2020.1748772}, DOI={10.1080/1547691X.2020.1748772}, abstractNote={Abstract Currently, assessment of the potential immunotoxicity of a given agent involves a tiered approach for hazard identification and mechanistic studies, including observational studies, evaluation of immune function, and measurement of susceptibility to infectious and neoplastic diseases. These studies generally use costly low-throughput mammalian models. Zebrafish, however, offer an excellent alternative due to their rapid development, ease of maintenance, and homology to mammalian immune system function and development. Larval zebrafish also are a convenient model to study the innate immune system with no interference from the adaptive immune system. In this study, a respiratory burst assay (RBA) was utilized to measure reactive oxygen species (ROS) production after developmental xenobiotic exposure. Embryos were exposed to non-teratogenic doses of chemicals and at 96 h post-fertilization, the ability to produce ROS was measured. Using the RBA, 12 compounds with varying immune-suppressive properties were screened. Seven compounds neither suppressed nor enhanced the respiratory burst; five reproducibly suppressed global ROS production, but with varying potencies: benzo[a]pyrene, 17β-estradiol, lead acetate, methoxychlor, and phenanthrene. These five compounds have all previously been reported as immunosuppressive in mammalian innate immunity assays. To evaluate whether the suppression of ROS by these compounds was a result of decreased immune cell numbers, flow cytometry with transgenic zebrafish larvae was used to count the numbers of neutrophils and macrophages after chemical exposure. With this assay, benzo[a]pyrene was found to be the only chemical that induced a change in the number of immune cells by increasing macrophage but not neutrophil numbers. Taken together, this work demonstrates the utility of zebrafish larvae as a vertebrate model for identifying compounds that impact innate immune function at non-teratogenic levels and validates measuring ROS production and phagocyte numbers as metrics for monitoring how xenobiotic exposure alters the innate immune system.}, number={1}, journal={Journal of Immunotoxicology}, publisher={Informa UK Limited}, author={Phelps, Drake W. and Fletcher, Ashley A. and Rodriguez-Nunez, Ivan and Balik-Meisner, Michele R. and Tokarz, Debra A. and Reif, David M. and Germolec, Dori R. and Yoder, Jeffrey A.}, year={2020}, month={Jan}, pages={94–104} }
 @article{catron_swank_wehmas_phelps_keely_brinkman_mccord_singh_sobus_wood_et al._2019, title={Microbiota alter metabolism and mediate neurodevelopmental toxicity of 17β-estradiol}, volume={9}, ISSN={2045-2322}, url={http://dx.doi.org/10.1038/s41598-019-43346-9}, DOI={10.1038/s41598-019-43346-9}, abstractNote={Estrogenic chemicals are widespread environmental contaminants associated with diverse health and ecological effects. During early vertebrate development, estrogen receptor signaling is critical for many different physiologic responses, including nervous system function. Recently, host-associated microbiota have been shown to influence neurodevelopment. Here, we hypothesized that microbiota may biotransform exogenous 17-βestradiol (E2) and modify E2 effects on swimming behavior. Colonized zebrafish were continuously exposed to non-teratogenic E2 concentrations from 1 to 10 days post-fertilization (dpf). Changes in microbial composition and predicted metagenomic function were evaluated. Locomotor activity was assessed in colonized and axenic (microbe-free) zebrafish exposed to E2 using a standard light/dark behavioral assay. Zebrafish tissue was collected for chemistry analyses. While E2 exposure did not alter microbial composition or putative function, colonized E2-exposed larvae showed reduced locomotor activity in the light, in contrast to axenic E2-exposed larvae, which exhibited normal behavior. Measured E2 concentrations were significantly higher in axenic relative to colonized zebrafish. Integrated peak area for putative sulfonated and glucuronidated E2 metabolites showed a similar trend. These data demonstrate that E2 locomotor effects in the light phase are dependent on the presence of microbiota and suggest that microbiota influence chemical E2 toxicokinetics. More broadly, this work supports the concept that microbial colonization status may influence chemical toxicity.}, number={1}, journal={Scientific Reports}, publisher={Springer Science and Business Media LLC}, author={Catron, Tara R. and Swank, Adam and Wehmas, Leah C. and Phelps, Drake and Keely, Scott P. and Brinkman, Nichole E. and McCord, James and Singh, Randolph and Sobus, Jon and Wood, Charles E. and et al.}, year={2019}, month={May} }
 @article{triclosan-selected host-associated microbiota perform xenobiotic biotransformations in larval zebrafish_2019, url={http://dx.doi.org/10.1093/toxsci/kfz166}, DOI={10.1093/toxsci/kfz166}, abstractNote={Microbiota regulate important physiologic processes during early host development. They also biotransform xenobiotics and serve as key intermediaries for chemical exposure. Antimicrobial agents in the environment may disrupt these complex interactions and alter key metabolic functions provided by host-associated microbiota. To examine the role of microbiota in xenobiotic metabolism, we exposed zebrafish larvae to the antimicrobial agent triclosan. Conventionally colonized (CC), microbe-free axenic (AX), or axenic colonized on day 1 (AC1) zebrafish were exposed to 0.16-0.30 uM triclosan or vehicle on days 1, 6, 7, 8, and 9 days post fertilization (dpf). After 6 dpf and 10 dpf, host-associated microbial community structure and putative function were assessed by 16S rRNA gene sequencing. At 10 dpf, triclosan exposure selected for bacterial taxa, including Rheinheimera. Triclosan-selected microbes were predicted to be enriched in pathways related to mechanisms of antibiotic resistance, sulfonation, oxidative stress, and drug metabolism. Furthermore, at 10 dpf, colonized zebrafish contained 2.5-3 times more triclosan relative to AX larvae. Non-targeted chemical analysis revealed that, relative to AX larvae, both cohorts of colonized larvae showed elevations in 23 chemical features, including parent triclosan and putative triclosan sulfate. Taken together, these data suggest that triclosan exposure selects for microbes that harbor the capacity to biotransform triclosan into chemical metabolites with unknown toxicity profiles. More broadly, these data support the concept that microbiota modify the toxicokinetics of xenobiotic exposure.}, journal={Toxicological Sciences}, year={2019}, month={Sep} }
 @article{catron_keely_brinkman_zurlinden_wood_wright_phelps_wheaton_kvasnicka_gaballah_et al._2018, title={Host developmental toxicity of BPA and BPA alternatives is inversely related to microbiota disruption in zebrafish}, volume={10}, DOI={10.1093/toxsci/kfy261}, abstractNote={Host-associated microbiota can biotransform xenobiotics, mediate health effects of chemical exposure, and play important roles in early development. Bisphenol A (BPA) is a widespread environmental chemical that has been associated with adverse endocrine and neurodevelopmental effects, some of which may be mediated by microbiota. Growing public concern over the safety of BPA has resulted in its replacement with structurally similar alternatives. In this study, we evaluated whether BPA and BPA alternatives alter microbiota and modulate secondary adverse behavioral effects in zebrafish. Zebrafish were developmentally exposed to BPA, Bisphenol AF (BPAF), Bisphenol B (BPB), Bisphenol F (BPF), or Bisphenol S (BPS). At 10 days post fertilization (dpf), toxicity assessments were completed and 16S rRNA gene sequencing was performed to evaluate potential chemical-dependent shifts in microbial community structure and predicted function. A standard light/dark behavioral assay was used to assess locomotor activity. Based on developmental toxicity assessments at 10 dpf, a range of potencies was observed: BPAF > BPB > BPF ∼ BPA > BPS. Analysis of 16S rRNA gene sequencing data showed significant concentration-dependent disruption of microbial community structure and enrichment of putative microbial functions with exposure to BPS, BPA, or BPF, but not BPB or BPAF. Interestingly, microbial disruption was inversely related to host developmental toxicity and estrogenicity. Exposure to BP analogs did not cause behavioral effects at 10 dpf. Our findings indicate that some BP analogs disrupt host microbiota early in life and demonstrate novel chemical-microbiota interactions that may add important context to current hazard identification strategies.}, journal={Toxicological Sciences}, publisher={Oxford University Press (OUP)}, author={Catron, Tara R and Keely, Scott P and Brinkman, Nichole E and Zurlinden, Todd J and Wood, Charles E and Wright, Justin R and Phelps, Drake and Wheaton, Emily Anneken and Kvasnicka, Allison and Gaballah, Shaza and et al.}, year={2018}, month={Oct} }
 @article{garbutt_konneker_konganti_hillhouse_swift-haire_jones_phelps_aylor_threadgill_2018, title={Permissiveness to form pluripotent stem cells may be an evolutionarily derived characteristic in Mus muscuius}, volume={8}, ISSN={["2045-2322"]}, url={https://doi.org/10.1038/s41598-018-32116-8}, DOI={10.1038/s41598-018-32116-8}, abstractNote={Abstract Mus musculus is the only known species from which embryonic stem cells (ESC) can be isolated under conditions requiring only leukemia inhibitory factor (LIF). Other species are non-permissive in LIF media, and form developmentally primed epiblast stem cells (EpiSC) similar to cells derived from post-implantation, egg cylinders. To evaluate whether non-permissiveness extends to induced pluripotent stem cells (iPSC), we derived iPSC from the eight founder strains of the mouse Collaborative Cross. Two strains, NOD/ShiLtJ and the WSB/EiJ, were non-permissive, consistent with the previous classification of NOD/ShiLtJ as non-permissive to ESC derivation. We determined non-permissiveness is recessive, and that non-permissive genomes do not compliment. We overcame iPSC non-permissiveness by using GSK3B and MEK inhibitors with serum, a technique we termed 2iS reprogramming. Although used for ESC derivation, GSK3B and MEK inhibitors have not been used during iPSC reprogramming because they inhibit survival of progenitor differentiated cells. iPSC derived in 2iS are more transcriptionally similar to ESC than EpiSC, indicating that 2iS reprogramming acts to overcome genetic background constraints. Finally, of species tested for ESC or iPSC derivation, only some M. musculus strains are permissive under LIF culture conditions suggesting that this is an evolutionarily derived characteristic in the M. musculus lineage.}, journal={SCIENTIFIC REPORTS}, author={Garbutt, Tiffany A. and Konneker, Thomas I and Konganti, Kranti and Hillhouse, Andrew E. and Swift-Haire, Francis and Jones, Alexis and Phelps, Drake and Aylor, David L. and Threadgill, David W.}, year={2018}, month={Oct} }
 @article{dan_yousef_campbell_phelps_burnett_kekkonen_shockley_lila_2017, title={Development, and genetic and metabolic characterization of new tomato mutants with enhanced and deficient carotenoid content}, volume={92}, ISSN={["2380-4084"]}, DOI={10.1080/14620316.2017.1301223}, abstractNote={ABSTRACT Tomato (Solanum lycopersicum) is a valuable vegetable crop rich in health-protective carotenoids, but breeding improvements are limited by its narrow genetic diversity. New mutants with enhanced and deficient carotenoid content in a single genetic background of tomato cv. MicroTom were developed via chemical mutagenesis. Genetic and metabolic analyses showed that mutant DC260, which exhibited fruit color alteration from red to deep red interlaced with orange color, had significant (P < 0.05) increases of lycopene (up to 42.8%) and ß-carotene (up to 61.5%) compared with control plants. Pearson correlation analysis of M1 and M2 generations in DC260 revealed that fruit color alteration was significantly (P < 0.05) correlated with lycopene (coefficient = 0.55) and ß-carotene content (coefficient = 0.63). The fruit color alteration of DC260 was controlled by a single gene at a heterozygous locus. In contrast, mutant DC107 and DC624, which exhibited fruit color alteration from red to orange-yellow, was significantly (P < 0.05) carotenoid-deficient with up to 346.3-, 10.8-, and 185.2- fold reductions of lycopene, ß-carotene, and total carotenoids, respectively, compared with the control plants. Carotenoid deficiency in DC170 and DC624 was responsible for the fruit color alteration and was controlled by a dominant gene at a homozygous locus.}, number={5}, journal={JOURNAL OF HORTICULTURAL SCIENCE & BIOTECHNOLOGY}, publisher={Informa UK Limited}, author={Dan, Yinghui and Yousef, Gad and Campbell, Faith N. and Phelps, Drake W. and Burnett, Callie and Kekkonen, Anni and Shockley, Alexa and Lila, Mary Ann}, year={2017}, pages={475–483} }
 @article{phelps_brinkman_keely_anneken_catron_betancourt_wood_espenschied_rawls_tal_2017, title={Microbial colonization is required for normal neurobehavioral development in zebrafish}, url={https://doi.org/10.1038/s41598-017-10517-5}, DOI={10.1038/s41598-017-10517-5}, abstractNote={Abstract Changes in resident microbiota may have wide-ranging effects on human health. We investigated whether early life microbial disruption alters neurodevelopment and behavior in larval zebrafish. Conventionally colonized, axenic, and axenic larvae colonized at 1 day post fertilization (dpf) were evaluated using a standard locomotor assay. At 10 dpf, axenic zebrafish exhibited hyperactivity compared to conventionalized and conventionally colonized controls. Impairment of host colonization using antibiotics also caused hyperactivity in conventionally colonized larvae. To determine whether there is a developmental requirement for microbial colonization, axenic embryos were serially colonized on 1, 3, 6, or 9 dpf and evaluated on 10 dpf. Normal activity levels were observed in axenic larvae colonized on 1–6 dpf, but not on 9 dpf. Colonization of axenic embryos at 1 dpf with individual bacterial species Aeromonas veronii or Vibrio cholerae was sufficient to block locomotor hyperactivity at 10 dpf. Exposure to heat-killed bacteria or microbe-associated molecular patterns pam3CSK4 or Poly(I:C) was not sufficient to block hyperactivity in axenic larvae. These data show that microbial colonization during early life is required for normal neurobehavioral development and support the concept that antibiotics and other environmental chemicals may exert neurobehavioral effects via disruption of host-associated microbial communities.}, journal={Scientific Reports}, author={Phelps, Drake and Brinkman, Nichole E. and Keely, Scott P. and Anneken, Emily M. and Catron, Tara R. and Betancourt, Doris and Wood, Charles E. and Espenschied, Scott T. and Rawls, John F. and Tal, Tamara}, year={2017}, month={Sep} }