Search NASA⌕ Search

SEARCH · Search NASA

Results for “rodent”

Search indexed NASA NTRS and DOE OSTI research on propulsion, heat transfer, battery materials and energy systems. Follow report and document links to the original sources.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 235 records · Page 13
All sourcesNASA NTRSDOE OSTI
NASA NTRSDate not supplied

Somatic Mutation in Mice on the International Space Station (ISS): Guanine Substitution Suggests Link to Cancer Risk

We conducted comprehensive analysis of single nucleotide somatic mutations in mice exposed to microgravity and other factors aboard the International Space Station (ISS), using data archived in GeneLab. Animals in the experimental cohort consisted of mice that spent 37 days on the ISS within the Rodent Habitat. Ground control animals consisted of mice of identical age, sex, strain, in a terrestrial Rodent Habitat controlled for temperature, humidity and carbon dioxide levels, to match ISS conditions as closely as possible. RNA extracted from eye, liver, skeletal muscle, and kidney tissue specimens was subjected to next-generation sequencing to acquire primary data. Our analysis employed cutting-edge software developed at NASA Ames Research Center, executed on the NASA Ames Supercomputer and on another high-performance computer, for accurate variant calling of single point mutations. ISS-flown mice exhibited a notably heightened level of somatic mutation compared to control mice. The degree of somatic mutation correlated with the degree of gene expression across the four tissue types, i.e., the greatest rate of mutation accumulation was seen in highly expressed genes. We discovered that guanine substitutions were the most common type of somatic mutation. This observation is consistent with the hypothesis that DNA mutation events stem from reactive oxygen/nitrogen/chlorine species-mediated guanine oxidation induced by the spaceflight environment. Since guanine oxidation is a prominent feature of the DNA mutation landscape that accompanies malignant transformation, our findings suggest a possible link between the spaceflight environment and cancer risk that is independent of radiation carcinogenesis.

ISS↗
NASA NTRSDate not supplied

The NASA Biological Institutional Scientific Collection (NBISC): Tissue and Microbe Biospecimens to Advance Space Research

The NASA Biological Institutional Scientific Collection (NBISC) is a biorepository of non-human biospecimens from NASA-funded spaceflight investigations and correlative ground studies. The collection has its roots in the 1960s through collaborations by NASA Ames Research Center with other NASA centers, universities, and international space agencies by sharing non-human biospecimens from spaceflight and space-relevant ground experiments. These collaborations have advanced the field of space exploration by helping to maximize the data gained from spaceflight and other NASA-funded experiments. NBISC coordinates closely with the Biospecimen Sharing Program (BSP) which is responsible for organized sample collection from spaceflight and ground rodent experiments. In 2022 NBISC partnered with HRP’s Biospecimen and Tissue Sharing Collection (BTSC) Program to also archive samples from HRP’s Space Radiation Element-funded studies which primarily involve rodents exposed to various radiation protocols at the NASA Space Radiation Laboratory (NSRL) and other analog facilities. To date, more than fifty thousand biospecimens from BTSC have been transferred to NBISC for distribution to the community. In 2023, NBISC embarked on a new Space Biology funded endeavor to create the Space Microbial Culture Collection (SMCC) which will serve as a central repository for microbial isolates associated with space flight, gravitational, and space radiation research. This presentation will highlight several success stories of analyses carried out using archived NBISC samples received by researchers in recent years and will detail the process for proposing and receiving samples from NBISC, BTSC and SMCC. Making available these 150,000+ unique biospecimens to the scientific research community, NBISC not only serves as a repository for storing and distributing non-human biospecimens, but also acts as a resource to enable new discoveries that will benefit NASA and humankind.

NBISC↗
NASA NTRSDate not supplied

The NASA Biological Institutional Scientific Collection (NBISC): Tissue and Microbe Biospecimens to Advance Space Research

The NASA Biological Institutional Scientific Collection (NBISC) is a biorepository of non-human biospecimens from NASA-funded spaceflight investigations and correlative ground studies. The collection has its roots in the 1960s through collaborations by NASA Ames Research Center with other NASA centers, universities, and international space agencies by sharing non-human biospecimens from spaceflight and space-relevant ground experiments. These collaborations have advanced the field of space exploration by helping to maximize the data gained from spaceflight and other NASA-funded experiments. NBISC coordinates closely with the Biospecimen Sharing Program (BSP) which is responsible for organized sample collection from spaceflight and ground rodent experiments. In 2022 NBISC partnered with HRP’s Biospecimen and Tissue Sharing Collection (BTSC) Program to also archive samples from HRP’s Space Radiation Element-funded studies which primarily involve rodents exposed to various radiation protocols at the NASA Space Radiation Laboratory (NSRL) and other analog facilities. To date, more than fifty thousand biospecimens from BTSC have been transferred to NBISC for distribution to the community. In 2023, NBISC embarked on a new Space Biology funded endeavor to create the Space Microbial Culture Collection (SMCC) which will serve as a central repository for microbial isolates associated with space flight, gravitational, and space radiation research. This presentation will highlight several success stories of analyses carried out using archived NBISC samples received by researchers in recent years and will detail the process for proposing and receiving samples from NBISC, BTSC and SMCC. Making available these 150,000+ unique biospecimens to the scientific research community, NBISC not only serves as a repository for storing and distributing non-human biospecimens, but also acts as a resource to enable new discoveries that will benefit NASA and humankind.

Biospecimen↗
NASA NTRSDate not supplied

Biological Research and Space Health Enabled by Machine Learning to Support Deep Space Missions

A key science goal of the NASA “Moon to Mars” campaign is to understand how biology responds to the Lunar, Martian, and deep space environments in order to advance fundamental knowledge, reduce risk, and support safe, productive human space missions. Through the powerful emerging computer science approaches of artificial intelligence (AI) and machine learning (ML), a paradigm shift has begun in biomedical science and engineered astronaut health systems, to enable Earth-independence and autonomy of mission operations. We present a decadal view of AI/ML architecture to support deep space mission goals, developed in concert with leaders in the field. We describe current AI/ML methods to support 1) fundamental biology, 2) in situ analytics, 3) high performance computing hardware, 4) automated science, 5) self-driving labs, 6) remote data management, 7) integrated real-time mission biomonitoring, and 8) a Precision Space Health system. Cutting-edge AI/ML approaches that can be integrated to support these domains include active learning, explainable AI, adaptive learning, causal inference, knowledge graphs, federated learning, transfer learning, and large language models. Finally, we present results from several current ML projects that are underway in the field to address key challenges of small sample n, high feature count, heterogeneity, and sparse data. These include 1) connecting omics data to phenotypic data using an ensemble model to infer causality of spaceflight rodent liver health disruption, 2) usage of explainable ML to interrogate the muscular underpinnings of spaceflight muscle atrophy, 3) ML models analyzing and determining directed acyclic graphs of human space health risk leveraging rodent bone datasets, 4) usage of large pre-trained models connecting biomedical knowledgebases with small spaceflight datasets to understand gene-to-gene interaction networks, and 5) a suite of benchmarked open science datasets (spaceflight mouse liver; radiation DNA damage) enabling programmers to identify the best ML algorithms to answer space biological science questions.

space biology↗
DOE OSTI2020

Past and present disturbances generate spatial variation in seed predation

Seed survival is a key process for plant populations; variation in the activity and abundance of animals that consume seeds can lead to dramatic shifts in seed fate. Because granivores may respond to contemporary disturbance as well as to enduring changes in habitats caused by past disturbances, understanding seed fate requires studies capable of evaluating how past and present disturbances modify granivore communities, foraging activity, and ultimately, seed predation. Historic agricultural land use and contemporary canopy harvesting are widespread disturbances that could generate large-scale patterns of seed fate by modifying environmental characteristics that determine granivore identity and behavior. Here, to evaluate whether land-use history and canopy harvesting affect seed–animal interactions, we conducted an experiment distributed across 80,000 ha of longleaf pine woodlands that coupled large-scale canopy harvesting at seven 4-ha sites containing both post-agricultural land use and nonagricultural land use in South Carolina, United States. We deployed a total of 28,000 nail-tagged seeds and recovered the tags to quantify seed fate. Past agricultural land use and contemporary canopy harvesting interacted to affect the rate of seed predation. Seed predation rates in harvested sites depended on land-use history: Seed predation was 30% lower in post-agricultural plots than in nonagricultural plots. This interaction was driven by the differential effect of land-use history and canopy harvesting on rodent activity. Camera traps revealed that Sigmodon hispidus only foraged in harvested plots and was most active in nonagricultural plots. In harvested plots, seed removal increased with S. hispidus activity. In unharvested plots, seed removal increased with Sciurus niger activity, but S. niger was not affected by land-use history. In finding that land-use history and canopy harvesting determine the outcomes of seed–animal interactions, we show that understanding patterns of seed predation is contingent upon the interplay of disturbances in both the distant past and recent past. These results suggest that patterns of past land use and present land use may help reconcile the considerable variation in seed fate observed in ecological communities.

60 APPLIED LIFE SCIENCES↗
NASA NTRS2017

Advancing Translational Space Research Through Biospecimen Sharing: Amplifying the Impact of Ground-Based Studies

Biospecimen Sharing Programs (BSPs) have been organized by NASA Ames Research Center since the 1960s with the goal of maximizing utilization and scientific return from rare, complex and costly spaceflight experiments. BSPs involve acquiring otherwise unused biological specimens from primary space research experiments for distribution to secondary experiments. Here we describe a collaboration leveraging Ames expertise in biospecimen sharing to magnify the scientific impact of research informing astronaut health funded by the NASA Human Research Program (HRP) Human Health Countermeasures (HHC) Element. The concept expands biospecimen sharing to one-off ground-based studies utilizing analogue space platforms (e.g., Hind limb Unloading (HLU), Artificial Gravity) for rodent experiments, thereby significantly broadening the range of research opportunities with translational relevance for protecting human health in space and on Earth. In this presentation, we will report on biospecimens currently being acquired from HHC Award Head-Down Tilt as a Model for Intracranial and Intraocular Pressures, and Retinal Changes during Spaceflight, and their availability. The BSP add-on to the project described herein has already yielded for HHC-funded investigators more than 4,700 additional tissues that would otherwise have been discarded as waste, with additional tissues available for analysis. Young (3-mo old) male and female rats and Older (9-mo old) male rats are being exposed to HLU for either 7, 14, 28, or 90 days. Additional groups are exposed to 90 days of unloading followed by either 7, 14, 28 days or 90 days of recovery (normal loading). Comparisons are made with non-suspended controls. Unused tissues are: Skin, Lungs, Thymus, Adrenals, Kidneys, Spleen, Hindlimb Muscles (Soleus, Extensor Digitorum Longus, Tibialis Anterior, Plantaris Gastrocnemius), Fat Pads, Reproductive Organs, and Intestines. Tissues are harvested, weighed, preserved then archived (with metadata) using a sample tracking system (CryoTrack). Preservation techniques include snap-freezing and RNALatersnap-freezing. Specimens were weighed at the time of dissection, and organ mass: body mass ratios analyzed to determine unloading effects across conditions and durations. The results corroborate previously reported effects of short-term exposure to microgravity or unloading exposure on various organs, and provide new insights into adaptation to long-duration unloading relevant to sustained spaceflight exposures on ISS. Supported by the Human Research Program (HRP) Human Health Countermeasures (HHC) Element and NASA Grant NNX13AD94G (CAF).

rodent↗
NASA NTRS2017

Behavioral Adaptations of Female Mice on the International Space Station

Adult female mice were sent to the International Space Station (ISS) as part of an early life science mission utilizing NASA's Rodent Habitat. Its primary purpose was to provide further insight into the influence of a microgravity environment on various aspects of mammalian physiology and well-being as part of an ongoing program of research aimed ultimately at understanding and ameliorating the deleterious influences of space on the human body. The present study took advantage of video collected from fixed, in-flight cameras within the habitat itself, to assess behavioral adaptations observed among in-flight mice aboard the ISS and differences in behavior with respect to a control group on the ground. Data collection consisted of several behavioral measures recorded by a trained observer with the assistance of interactive behavior analysis software. Specific behavioral measures included frequencies of conspecific interactionsociability, time spent feeding and conducting hygienic behavior, and relative durations of thigmotactic behavior, which is commonly used as an index of anxiety. Data were used to test tentative hypotheses that such behaviors differ significantly across mice under microgravity versus 1g conditions, and the assumption that the novel experience of microgravity itself may represent an initially anxiogenic stimulus which an animal will eventually acclimate to, perhaps through habituation.

rodent behavior↗
NASA NTRS2017

Advancing Translational Space Research Through Biospecimen Sharing: Amplified Impact of Studies Utilizing Analogue Space Platforms

Biospecimen Sharing Programs (BSPs) have been organized by NASA Ames Research Center since the 1960s with the goal of maximizing utilization and scientific return from rare, complex and costly spaceflight experiments. BSPs involve acquiring otherwise unused biological specimens from primary space research experiments for distribution to secondary experiments. Here we describe a collaboration leveraging Ames expertise in biospecimen sharing to magnify the scientific impact of research informing astronaut health funded by the NASA Human Research Program (HRP) Human Health Countermeasures (HHC) Element. The concept expands biospecimen sharing to one-off ground-based studies utilizing analogue space platforms (e.g., Hindlimb Unloading (HLU), Artificial Gravity) for rodent experiments, thereby significantly broadening the range of research opportunities with translational relevance for protecting human health in space and on Earth. In this presentation, we will report on biospecimens currently being acquired from HHC Award Head-Down Tilt as a Model for Intracranial and Intraocular Pressures, and Retinal Changes during Spaceflight, and their availability. The BSP add-on to the project described herein has already yielded for HHC-funded investigators more than 4,700 additional tissues that would otherwise have been discarded as waste, with additional tissues available for analysis. Young (3-mo old) male and female rats and Older (9-mo old) male rats are being exposed to HLU for either 7, 14, 28, or 90 days. Additional groups are exposed to 90 days of unloading followed by either 7, 14, 28 days or 90 days of recovery (normal loading). Comparisons are made with non-suspended controls. Unused tissues are: Skin, Lungs, Thymus, Adrenals, Kidneys, Spleen, Hindlimb Muscles (Soleus, Extensor Digitorum Longus, Tibialis Anterior, Plantaris Gastrocnemius), Fat Pads, Reproductive Organs, and Intestines. Tissues are harvested, weighed, preserved then archived (with metadata) using a sample tracking system (CryoTrack). Preservation techniques include snap-freezing and RNALatersnap-freezing. Specimens were weighed at the time of dissection, and organ mass:body mass ratios analyzed to determine unloading effects across conditions and durations. The results corroborate previously reported effects of short-term exposure to microgravity or unloading exposure on various organs, and provide new insights into adaptation to long-duration unloading relevant to sustained spaceflight exposures on ISS.

biospecimen sharing↗
DOE OSTI2025

What's on the menu? Examining native apex‐ and invasive meso‐predator diets to understand impacts on ecosystems

Abstract Understanding how carnivores impact ecological communities is essential for guiding effective management actions and conserving biodiversity. Quantifying predators' diets, including prey selectivity, allows for the assessment of the relative effects native and invasive predators may have on prey populations. In Australia, populations of a native, terrestrial apex predator, the dingo Canis dingo/C. familiaris , and introduced and invasive subordinate mesopredators, the European red fox Vulpes vulpes and feral cat Felis catus , co‐occur, but there is limited understanding of their relative impacts on native and invasive prey in different ecosystems. To assess the possible effects of dingoes, foxes and cats on prey, we examined their diet and prey selectivity across a ~10,000 km 2 semi‐arid mallee ecosystem. Using macroscopic scat analysis, we identified strong dietary niche separation. Larger‐bodied dingoes primarily consumed large marsupial herbivores, whereas foxes and cats primarily consumed smaller prey, including introduced and native rodents and birds. Foxes had the broadest diet, and the greatest dietary overlap with cats ( = 0.81), compared with dingoes ( = 0.50) or between dingoes and cats ( = 0.36). Livestock were identified in 2% of dingo and 7% of fox scats. Cats and foxes consumed more than 15 times the volume of small native mammals compared with dingoes, including threatened species such as fat‐tailed dunnarts Sminthopsis crassicaudata . Cats and foxes also selectively consumed small mammals relative to their estimated availability and consumed fewer large mammals. In contrast, dingoes consumed fewer birds and more echidnas relative to their availability. Our results suggest limited intraguild competition within this semi‐arid ecosystem, as dingoes are primarily exerting top‐down pressure on large herbivores, whereas invasive mesopredators are disproportionately impacting smaller prey, including threatened native mammals. Practical implication . Our findings suggest that ongoing conservation management of dingoes, red foxes and feral cats must consider the variation in diets, impacts on prey and ecological roles of these different predator species, and avoid indiscriminate lethal control methods. Quantifying actual, rather than assumed, impacts of predators on threatened native species, large herbivores and livestock is essential to achieve effective and integrated ecosystem management.

Mason, Rachel T. [School of Life and Environmental↗
DOE OSTI2023

Treatment of cholinergic‐induced status epilepticus with polytherapy targeting GABA and glutamate receptors

Abstract Despite new antiseizure medications, the development of cholinergic‐induced refractory status epilepticus (RSE) continues to be a therapeutic challenge as pharmacoresistance to benzodiazepines and other antiseizure medications quickly develops. Studies conducted by Epilepsia . 2005;46:142 demonstrated that the initiation and maintenance of cholinergic‐induced RSE are associated with trafficking and inactivation of gamma‐aminobutyric acid A receptors (GABA A R) thought to contribute to the development of benzodiazepine pharmacoresistance. In addition, Dr. Wasterlain's laboratory reported that increased N‐methyl‐ d ‐aspartate receptors (NMDAR) and alpha‐amino‐3‐hydroxy‐5‐methyl‐4‐isoxazolepropionic acid receptors (AMPAR) contribute to enhanced glutamatergic excitation ( Neurobiol Dis . 2013;54:225; Epilepsia . 2013;54:78). Thus, Dr. Wasterlain postulated that targeting both maladaptive responses of reduced inhibition and increased excitation that is associated with cholinergic‐induced RSE should improve therapeutic outcome. We currently review studies in several animal models of cholinergic‐induced RSE that demonstrate that benzodiazepine monotherapy has reduced efficacy when treatment is delayed and that polytherapy with drugs that include a benzodiazepine (eg midazolam and diazepam) to counter loss of inhibition, concurrent with an NMDA antagonist (eg ketamine) to reduce excitation provide improved efficacy. Improved efficacy with polytherapy against cholinergic‐induced seizure is demonstrated by reduction in (1) seizure severity, (2) epileptogenesis, and (3) neurodegeneration compared with monotherapy. Animal models reviewed include pilocarpine‐induced seizure in rats, organophosphorus nerve agent (OPNA)‐induced seizure in rats, and OPNA‐induced seizure in two mouse models: (1) carboxylesterase knockout (Es1 −/− ) mice which, similarly to humans, lack plasma carboxylesterase and (2) human acetylcholinesterase knock‐in carboxylesterase knockout (KIKO) mice. We also review studies showing that supplementing midazolam and ketamine with a third antiseizure medication (valproate or phenobarbital) that targets a nonbenzodiazepine site rapidly terminates RSE and provides further protection against cholinergic‐induced SE. Finally, we review studies on the benefits of simultaneous compared with sequential drug treatments and the clinical implications that lead us to predict improved efficacy of early combination drug therapies. The data generated from seminal rodent studies of efficacious treatment of cholinergic‐induced RSE conducted under Dr. Wasterlain's guidance suggest that future clinical trials should treat the inadequate inhibition and temper the excess excitation that characterize RSE and that early combination therapies may provide improved outcome over benzodiazepine monotherapy.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2021

Neuroprotectin D1 Attenuates Blast Overpressure Induced Reactive Microglial Cells in the Cochlea

Objective/Hypothesis We examined a neuroinflammatory response associated with glial activation in the cochlea exposed to blast overpressure and evaluated the potential therapeutic efficacy of specialized pro‐resolving mediators such as neuroprotectin D1, NPD1; (10R, 17S‐dihydroxy‐4Z, 7Z, 11E, 13E, 15Z, 19Z‐docosahexaenoic acid) in a rodent blast‐induced auditory injury model. Study Design Animal Research. Methods A compressed‐air driven shock tube was used to expose anesthetized adult male Long‐Evan rats to shock waves simulating an open‐field blast exposure. Approximately 30 minutes after blast exposure, rats were treated with NPD1 (100 ng/kg body wt.) or vehicle delivered intravenously via tail vein injection. Rats were then euthanized 48 hours after blast exposure. Unexposed rats were included as controls. Tissue sections containing both middle and inner ear were prepared with hematoxylin–eosin staining to elucidate histopathological changes associated with blast exposure. Cochlear tissues were evaluated for relative expression of ionized calcium‐binding adaptor 1 (Iba1), as an indicator of microglial activation by immunohistochemistry and western blot analyses. Results Our animal model resulted in an acute injury mechanism manifested by damage to the tympanic membrane, hemorrhage, infiltration of inflammatory cells, and increased expression of Iba1 protein. Moreover, therapeutic intervention with NPD1 significantly reduced Iba1 expression in the cochlea, suggesting a reduction of a neuroinflammatory response caused by blast overpressure. Conclusions Blast overpressure resulted in an increased expression of proteins involved in gliosis within the auditory system, which were reduced by NPD1. Treatment of NPD1 suggests an effective strategy to reduce or halt auditory microglial cell activation due to primary blast exposure. Level of Evidence NA Laryngoscope , 131:E2018–E2025, 2021

Ríos, José David↗
DOE OSTI2022

Mouse models of Ebola virus tolerance and lethality: characterization of CD-1 mice infected with wild-type, guinea pig-adapted, or mouse-adapted virus

Development of lethal models of Ebola virus disease has been achieved by the serial passage of virus isolates from human cases in mice and guinea pigs. Use of mice infected with non-adapted virus has been limited due to the absence of overt clinical disease. In recent years, newly recognized sequelae identified in human cases has highlighted the importance of continued investigations of non-lethal infection both in humans and animal models. Here, we revisit the use of rodent-adapted and non-adapted Ebola virus (EBOV) in mice to investigate infection tolerance and future utility of these models in pathogenesis and therapeutic intervention studies. We found that like non-adapted wild-type EBOV, guinea pig-adapted EBOV resulted in widespread tissue infection, variably associated with tissue pathology, and alterations in clinical and immunological analytes in the absence of overt disease. Notably, infection with either non-lethal variant did not greatly differ from lethal mouse-adapted EBOV until near the time end-point criteria are reached in these mice. Furthermore, these data support future investigations of pathogenesis, convalescence, and sequelae in mouse models of virus tolerance.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2022

Asymmetric and non-stoichiometric glycoprotein recognition by two distinct antibodies results in broad protection against ebolaviruses

Several ebolaviruses cause outbreaks of severe disease. Vaccines and monoclonal antibody cocktails are available to treat Ebola virus (EBOV) infections, but not Sudan virus (SUDV) or other ebolaviruses. Current cocktails contain antibodies that cross-react with the secreted soluble glycoprotein (sGP) that absorbs virus-neutralizing antibodies. By sorting memory B cells from EBOV infection survivors, we isolated two broadly reactive anti-GP monoclonal antibodies, 1C3 and 1C11, that potently neutralize, protect rodents from disease, and lack sGP cross-reactivity. Both antibodies recognize quaternary epitopes in trimeric ebolavirus GP. 1C11 bridges adjacent protomers via the fusion loop. 1C3 has a tripartite epitope in the center of the trimer apex. One 1C3 antigen-binding fragment anchors simultaneously to the three receptor-binding sites in the GP trimer, and separate 1C3 paratope regions interact differently with identical residues on the three protomers. A cocktail of both antibodies completely protected nonhuman primates from EBOV and SUDV infections, indicating their potential clinical value.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2024

Crimean-Congo hemorrhagic fever survivors elicit protective non-neutralizing antibodies that target 11 overlapping regions on glycoprotein GP38

Crimean-Congo hemorrhagic fever virus can cause lethal disease in humans yet there are no approved medical countermeasures. Viral glycoprotein GP38, exclusive to Nairoviridae , is a target of protective antibodies and is a key antigen in preclinical vaccine candidates. Here, we isolate 188 GP38-specific antibodies from human survivors of infection. Competition experiments show that these antibodies bind across 5 distinct antigenic sites, encompassing 11 overlapping regions. Additionally, we show structures of GP38 bound with 9 of these antibodies targeting different antigenic sites. Although these GP38-specific antibodies are non-neutralizing, several display protective efficacy equal to or better than murine antibody 13G8 in two highly stringent rodent models of infection. Together, these data expand our understanding regarding this important viral protein and may inform the development of broadly effective CCHFV antibody therapeutics.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2021

Benzo[a]pyrene (BaP) metabolites predominant in human plasma following escalating oral micro-dosing with [ 14 C]-BaP

Benzo[a]pyrene (BaP) is formed by incomplete combustion of organic materials (petroleum, coal, tobacco, etc.). BaP is designated by the International Agency for Research on Cancer as a group 1 known human carcinogen; a classification supported by numerous studies in preclinical models and epidemiology studies of exposed populations. Risk assessment relies on toxicokinetic and cancer studies in rodents at doses 5–6 orders of magnitude greater than average human uptake. Using a dose–response design at environmentally relevant concentrations, this study follows uptake, metabolism, and elimination of [ 14 C]-BaP in human plasma by employing UPLC - accelerator mass spectrometry (UPLC-AMS). Volunteers were administered 25, 50, 100, and 250 ng (2.7–27 nCi) of [ 14 C]-BaP (with interceding minimum 3-week washout periods) with quantification of parent [ 14 C]-BaP and metabolites in plasma measured over 48 h. [ 14 C]-BaP median T max was 30 min with C max and area under the curve (AUC) approximating dose-dependency. Marked inter-individual variability in plasma pharmacokinetics following a 250 ng dose was seen with 7 volunteers as measured by the C max (8.99 ± 7.08 ng × mL -1 ) and AUC0-48hr (68.6 ± 64.0 fg × hr -1 × mL -1 ). Approximately 3–6% of the [ 14 C] recovered (AUC 0-48 hr ) was parent compound, demonstrating extensive metabolism following oral dosing. Metabolite profiles showed that, even at the earliest time-point (30 min), a substantial percentage of [ 14 C] in plasma was polar BaP metabolites. The best fit modeling approach identified non-compartmental apparent volume of distribution of BaP as significantly increasing as a function of dose (p = 0.004). Bay region tetrols and dihydrodiols predominated, suggesting not only was there extensive first pass metabolism but also potentially bioactivation. AMS enables the study of environmental carcinogens in humans with de minimus risk, allowing for important testing and validation of physiologically based pharmacokinetic models derived from animal data, risk assessment, and the interpretation of data from high-risk occupationally exposed populations.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2021

Radiofluorination of oxazole-carboxamides for preclinical PET neuroimaging of GSK-3

Glycogen synthase kinase 3 (GSK-3) is an enzyme that is dysregulated in oncology neurodegeneration, neuroinflammation and several mental health illnesses. As such, GSK-3 is a long-sought after target for positron emission tomography (PET) imaging and therapeutic intervention. Herein, we report on the development and radiofluorination of two oxazole-4- carboxamides, including one bearing a non-activated aromatic ring. Both compounds demonstrated excellent selectivity in a kinase screen and inhibit GSK-3 with high affinity. [ 18 F]OCM-49 was synthesized from [ 18 F]fluoride using a copper-mediated reaction of an aryl boronic acid precursor, while [ 18 F]OCM-50 used a trimethylammonium triflate precursor, and both radiotracers were translated for preclinical PET imaging in rodents. Due to superior radiochemical yields and brain uptake (peak standardized uptake value of ~2.0), [ 18 F]OCM-50 was further evaluated in non-human primate and also showed good brain uptake and rapid clearance. Finally, further studies to consider clinical translation of both radiotracers are underway.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗
DOE OSTI2024

Non-DNA radiosensitive targets that initiate persistent behavioral deficits in rats exposed to space radiation

Predicting future CNS risks for astronauts during deep-space missions will rely substantially on ground-based rodent data with space-relevant ions and behaviors. For rats, the accumulated evidence indicates that less densely ionizing radiation, such as 4 He and 12 C ions, induce behavior deficits at lower doses than densely ionizing ions, such as 48 Ti and 56 Fe. However, this observation conflicts with standard somatic radiobiology, in which densely ionizing ions are generally more effective than less densely ionizing ions, and where the DNA/nucleus is the accepted target for radiation-induced tumorigenesis, cytogenetic aberrations, genetic mutations, and reproductive cell death. To gain deeper insight into the subcellular nature of the radiation targets for behavior risks, we compared the effects of dose, fluence, and linear energy transfer (LET) of 4 He and 56 Fe particles using existing datasets for four distinct behavioral outcomes in rats: elevated plus maze (EPM-anxiety), novel object recognition (NOR-memory), operant responding (OR-response to environmental stimuli), and attentional set-shifting (ATSET-cognitive flexibility). We confirmed that less densely ionizing particles (except protons) showed ~100-fold lower threshold doses than densely ionizing particles for behavioral deficits (0.1–1 cGy for 4 He vs. 15–100 cGy for 56 Fe). However, when analyzed by fluence the behavioral responses converged, indicating that 4 He and 56 Fe were equally effective on a per-track basis. When analyzed by LET, there were ~100-fold differences in the LET for maximum effectiveness for behavioral deficits and DNA endpoints (~1 vs ~100 keV/μm, respectively). These unique features of radiation-induced behavioral deficits (high sensitivity to particles in the 1-keV/μm range, insensitivity to protons in the 0.2 keV/μm range, and isofluence dependence for particles with LET>1 keV/μm) provide evidence in support of a new hypothesis of sub-micron sized radiosensitive targets for behavioral effects consistent with the thickness of plasma membranes and/or small subcellular structures, smaller than a whole synapse. Like our behavior findings, mouse immature oocyte killing which is known to have a plasma membrane target was also better explained by fluence, rather than dose. In contrast, fluence analyses for DNA/nuclear endpoints in somatic cells (e.g., tumor induction, chromosome aberrations) showed opposite results, suggesting that behavior targets are not DNA. Our findings raise questions regarding the identity of subcellular targets and the multi-cellular functional unit for behavior risks, low-dose susceptibility, and generalizability from rat to other species and astronauts.

63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTAN↗
DOE OSTI2025

Human in vitro metabolism of an environmental mixture of polycyclic aromatic hydrocarbons (PAH) found at the Portland Harbor Superfund Site

Polycyclic aromatic hydrocarbons (PAHs) are widespread environmental contaminants that pose health risks to humans. Toxicity testing approaches of PAHs have evolved from traditional rodent models to New Approach Methodologies (NAMs), such as high-throughput screening in zebrafish, enabling rapid evaluation of chemical hazards. However, translating toxicity findings from laboratory systems to humans remains difficult due to complexity and species-specific differences. Chemical dosimetry modeling offers a quantitative framework to bridge this gap, but its accuracy depends on robust knowledge of PAH metabolism. The objective of this study was to measure human metabolism rates of Supermix-10, the ten most abundant PAHs found at the Portland Harbor Superfund Site, to support development of human pharmacokinetic models. We incubated individual PAHs from Supermix-10 in pooled human liver microsomes and quantified parent PAH disappearance using high-performance liquid chromatography (HPLC) with UV and florescent detection. To assess the potential of mixture interactions, we also measured metabolism of all 10 compounds in an equimolar mixture and compared rates of parent disappearance to those observed for individual PAHs. All Supermix-10 PAHs demonstrated rapid parent compound disappearance in human hepatic microsomes. PAHs grouped into three metabolism patterns: high metabolism rates and capacity (2-methylnaphthalene, acenaphthylene, fluorene, naphthalene), high affinity metabolism that rapidly achieves low-level saturation (benzo[a]anthracene, chrysene), and moderate metabolism rates and capacity (fluoranthene, pyrene, retene, phenanthrene). Smaller PAHs exhibited faster metabolism, and higher metabolism rates correlated inversely with molecular weight. When incubated in an equimolar mixture, Supermix-10 demonstrated significantly slower metabolism (47–89 %) compared to metabolism of individual PAHs at the same concentration. These findings enhance our understanding of PAH metabolism in humans and demonstrate significant mixture interactions under the conditions tested. Furthermore, our findings offer insights into the metabolic behavior of Supermix-10 and provide critical metabolism rate data to support the development of physiological based pharmacokinetic (PBPK) models. Dosimetry models can translate PAH chemical dosimetry from high-throughput testing platforms, like zebrafish and cellular system assays, to human exposures enhancing the accuracy and reliability of PAH risk assessments.

2-methylnaphthalene↗