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1. NSAID-mediated cyclooxygenase inhibition disrupts ectodermal derivative formation in axolotl embryos

   https://doi.org/10.1016/j.diff.2025.100856  

   Marshall, Emma J; Ramarapu, Raneesh; Leathers, Tess A; Morrison-Welch, Nikolas; Sandberg, Kathryn; Kawashima, Maxim; Rogers, Crystal D (May 2025, Differentiation)

   Embryonic exposures to non-steroidal anti-inflammatory drugs (NSAIDs) have been linked to preterm birth, neural tube closure defects, abnormal enteric innervation, and craniofacial malformations, potentially due to disrupted neural tube or neural crest (NC) cell development. Naproxen (NPX), a common non-steroidal anti-inflammatory drug (NSAID) used to relieve pain and inflammation, exerts its effects through non-selective cyclooxygenase (COX) inhibition. Our lab has identified that the cyclooxygenase (COX-1 and COX-2) isoenzymes are expressed during the early stages of vertebrate embryonic development, and that global inhibition of COX-1 and COX-2 function disrupts NC cell migration and differentiation in Ambystoma mexicanum (axolotl) embryos. NC cells differentiate into various adult tissues including craniofacial cartilage, bone, and neurons in the peripheral and enteric nervous systems. To investigate the specific phenotypic and molecular effects of NPX exposure on NC development and differentiation, and to identify molecular links between COX inhibition and NC derivative anomalies, we exposed late neurula and early tailbud stage axolotl embryos to various concentrations of NPX and performed immunohistochemistry (IHC) for markers of migratory and differentiating NC cells. Our results reveal that NPX exposure impairs the migration of SOX9+ NC cells, leading to abnormal development of craniofacial cartilage structures, including Meckel's cartilage in the jaw. NPX exposure also alters the expression of markers associated with peripheral and central nervous system (PNS and CNS) development, suggesting concurrent neurodevelopmental changes. 

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2. Spatiotemporal characterization of cyclooxygenase pathway enzymes during vertebrate embryonic development

   https://doi.org/10.1016/j.ydbio.2024.11.009  

   Leathers, Tess A; Ramarapu, Raneesh; Rogers, Crystal D (November 2024, Developmental Biology)

   Vertebrate development is regulated by several complex well-characterized morphogen signaling pathways, transcription factors, and structural proteins, but less is known about the enzymatic pathways that regulate early development. We have identified that factors from the inflammation-mediating cyclooxygenase (COX) signaling pathway are expressed at early stages of development in avian embryos. Using Gallus gallus (chicken) as a research model, we characterized the spatiotemporal expression of a subset of genes and proteins in the COX pathway during early neural development stages. Specifically, here we show expression patterns of COX-1, COX-2, and microsomal prostaglandin E synthase-2 (mPGES-2) as well as the genes encoding these enzymes (PTGS1, PTGS2, and PTGES-2). Unique expression patterns of individual players within the COX pathway suggest that they may play non-canonical/non-traditional roles in the embryo compared to their roles in the adult. Future work should examine the function of the COX pathway in tissue specification and morphogenesis and determine if these expression patterns are conserved across species. 

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3. Quantification of naproxen in equine plasma for doping control in horse racing using strong anion exchange solid phase extraction followed by liquid chromatography with UV detection

   Ogunleye, T.; Chao, M.; Song, L. (April 2019, 111th Illinois State Academy of Science Annual Meeting)

   A method using strong anion exchange solid phase extraction (SAX-SPE) followed by liquid chromatography ultraviolet detection (LC-UV) for the analysis of flunixin in equine plasma for doping control in horse racing has been developed. By using SAX-SPE, commonly regulated non-steroidal anti-inflammatory drugs (NSAIDs) by the United States Equestrian Federation (USEF), i.e. PBZ, OPBZ, diclofenac, flunixin, ketoprofen, meclofenamic acid and naproxen, and an internal standard, i.e. flurbiprofen, were first selectively extracted. Then, baseline separation of flunixin from other NSAIDs, the internal standard, and residual components of equine plasma was achieved using LC-UV. Finally, flunixin in equine plasma was quantified after an internal calibration curve was created. 

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4. Quantification of phenylbutazone in equine plasma for doping control in horse racing using strong anion exchange solid phase extraction followed by liquid chromatography with UV detection

   Lin, F.; Heiser, N.; Song, L. (April 2019, 111th Illinois State Academy of Science Annual Meeting)

   A method using strong anion exchange solid phase extraction (SAX-SPE) followed by liquid chromatography ultraviolet detection (LC-UV) for the analysis of phenylbutazone (PBZ) and its metabolite oxyphenbutazone (OPBZ) in equine plasma for doping control in horse racing has been developed. By using SAX-SPE, commonly regulated non-steroidal anti-inflammatory drugs (NSAIDs) by the United States Equestrian Federation (USEF), i.e. PBZ, OPBZ, diclofenac, flunixin, ketoprofen, meclofenamic acid and naproxen, and an internal standard, i.e. tolfenamic acid, were first selectively extracted. Then, baseline separation of PBZ, OPBZ from other NSAIDs, internal standard, and residual components of equine plasma was achieved using LC-UV. Finally, PBZ and OPBZ in equine plasma were quantified after an internal calibration curve was created. 

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5. Single-cell sequencing provides clues about the developmental genetic basis of evolutionary adaptations in syngnathid fishes

   https://doi.org/10.7554/eLife.97764  

   Healey, HM; Penn, HB; Small, CM; Bassham, S; Goyal, V; Woods, MA; Cresko, WA (February 2025, eLife)

   Seahorses, pipefishes, and seadragons are fishes from the family Syngnathidae that have evolved extraordinary traits including male pregnancy, elongated snouts, loss of teeth, and dermal bony armor. The developmental genetic and cellular changes that led to the evolution of these traits are largely unknown. Recent syngnathid genome assemblies revealed suggestive gene content differences and provided the opportunity for detailed genetic analyses. We created a single-cell RNA sequencing atlas of Gulf pipefish embryos to understand the developmental basis of four traits: derived head shape, toothlessness, dermal armor, and male pregnancy. We completed marker gene analyses, built genetic networks, and examined the spatial expression of select genes. We identified osteochondrogenic mesenchymal cells in the elongating face that express regulatory genesbmp4, sfrp1a, andprdm16. We found no evidence for tooth primordia cells, and we observed re-deployment of osteoblast genetic networks in developing dermal armor. Finally, we found that epidermal cells expressed nutrient processing and environmental sensing genes, potentially relevant for the brooding environment. The examined pipefish evolutionary innovations are composed of recognizable cell types, suggesting that derived features originate from changes within existing gene networks. Future work addressing syngnathid gene networks across multiple stages and species is essential for understanding how the novelties of these fish evolved. 

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