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DOE OSTI2021

CBL aggravates Ang II-induced cardiac hypertrophy via the VHL/HIF-1α pathway

Highlights: • 1. The expression of CBL is increased in Ang II-induced cardiac hypertrophy in vitro and in vivo. • The pro-hypertrophic function of CBL on Ang II-induced myocardial hypertrophy was mediated by HIF-1α. • CBL increased HIF-1α expression via regulating VHL. • CBL decreased VHL through directly promote its ubiquitination modification. CBL (Casitas B cell lymphoma), an important ubiquitin protein ligase, is involved in protein folding, protein maturation, and proteasome-dependent protein catabolism in different cells. However, its role in cardiac hypertrophy is still unclear. In this study, we found that expression of CBL is increased in an Ang II-induced mouse cardiac hypertrophy animal model and in Ang II-treated H9C2 cells. Interference with CBL expression attenuates the degree of myocardial hypertrophy as well as the expression of hypertrophy-related genes in H9C2 cells. Further research found that CBL aggravates myocardial hypertrophy by activating HIF-1α, which is an aggravating factor for hypertrophy. The effect of CBL on promoting myocardial hypertrophy was reversed by interference with HIF-1α. Mechanistically, we found that CBL directly interacted with and degraded VHL by increasing its ubiquitination level, which is a widely accepted regulatory factor of HIF-1α. Finally, our results showed that CBL was partially dependent on degradation of VHL and that activation of HIF-1α promoted myocardial hypertrophy. Collectively, these findings suggest that strategies based on activation of the CBL/HIF-1α axis might be promising for the treatment of hypertrophic cardiomyopathy.

60 APPLIED LIFE SCIENCES↗

DOE OSTI2026

Biogas Utilization in Refuse Power Plants (BURP 2 )

The BURP 2 project investigated the technical and financial viability of co firing biogas with waste coal for power generation while using carbon capture and sequestration to achieve net negative emissions. A comparative assessment integrating geospatial mapping, technoeconomic analysis, and life cycle analysis was carried out to evaluate retrofitting an existing coal fired facility in West Virginia, versus developing a new greenfield power plant in Kentucky located near a low quality coal resource. The study found that CO 2 capture rates of 90% or higher, in combination with biogas feedstocks such as animal manure, could significantly reduce global warming potential compared to plants using neither biogas nor CO 2 capture systems. Economic feasibility depended heavily on federal tax credits and proximity to fuel sources. In both greenfield and retrofit scenarios, access to biogas played a key role. Because the retrofit site was located close to existing biogas resources, it represented a feasible option, whereas the greenfield site, being far from pipelines or biogas sources, would require prohibitively expensive biogas transport infrastructure. Overall, the research showed that repurposing waste or low-quality coal with renewable biogas and CO 2 capture systems could provide a viable approach for reducing carbon emissions in power production, if biogas resources are easily accessible and available in sufficient quantities.

01 COAL, LIGNITE, AND PEAT↗

DOE OSTI2023

Endemic Seroprevalence and Farmer Control Measures of Anthrax Among Ruminate Animals in Pakistan

Anthrax, caused by Bacillus anthracis (B. anthracis), is an important zoonotic disease of livestock that poses a threat to public safety. The present study aimed to determine the seroprevalence of Bacillus anthracis in sheep and cattle in endemic areas (Punjab and Balochistan provinces) of Pakistan by enzyme-linked immunosorbent assay (ELISA). Crosssectional survey was also done to assess the control measures of the farmers. Out of 401 serum samples from sheep and cattle, a total of 68 (16.9%) samples were seropositive for anti-PA antibodies; seroprevalence of B. anthracis in sheep and cattle was 22.11% (48/217) and 10.86% (20/184), respectively. Seroprevalence was significantly higher (p<0.05) in Baluchistan province (33.3%, 44/132) compared to Punjab province (8.92%, 24/269). Most farmers (65.90%, 29/44) were not disposing carcasses as recommended. Around 77.28%, 17/22 of the veterinary officials reported the nonavailability of vaccine against anthrax and 63.63% (14/22) stated the lack of cold chain for the vaccine storage and transportation. We concluded that seroprevalence of B. anthracis in unvaccinated sheep and cattle is evidence that the pathogen is circulating in Baluchistan and Punjab provinces. So must applied an active surveillance and, strengthened veterinary practices vaccination programs to control anthrax outbreaks in endemic areas.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2023

Proof of principle study: synchrotron X-ray fluorescence microscopy for identification of previously radioactive microparticles and elemental mapping of FFPE tissues

Biobanks containing formalin-fixed, paraffin-embedded (FFPE) tissues from animals and human atomic-bomb survivors exposed to radioactive particulates remain a vital resource for understanding the molecular effects of radiation exposure. These samples are often decades old and prepared using harsh fixation processes which limit sample imaging options. Optical imaging of hematoxylin and eosin (H&E) stained tissues may be the only feasible processing option, however, H&E images provide no information about radioactive microparticles or radioactive history. Synchrotron X-ray fluorescence microscopy (XFM) is a robust, non-destructive, semi-quantitative technique for elemental mapping and identifying candidate chemical element biomarkers in FFPE tissues. Still, XFM has never been used to uncover distribution of formerly radioactive micro-particulates in FFPE canine specimens collected more than 30 years ago. In this work, we demonstrate the first use of low-, medium-, and high-resolution XFM to generate 2D elemental maps of ~ 35-year-old, canine FFPE lung and lymph node specimens stored in the Northwestern University Radiobiology Archive documenting distribution of formerly radioactive micro-particulates. Additionally, we use XFM to identify individual microparticles and detect daughter products of radioactive decay. The results of this proof-of-principle study support the use of XFM to map chemical element composition in historic FFPE specimens and conduct radioactive micro-particulate forensics.

62 RADIOLOGY AND NUCLEAR MEDICINE↗

DOE OSTI2020

Laser speckle contrast imaging of blood microcirculation in pancreatic tissues during laparoscopic interventions

Laser speckle contrast imaging of the microcirculatory bed of the pancreas is performed, which allows its condition to be assessed and thereby is an additional valuable tool for making a diagnostic decision and dynamically monitoring the effectiveness of the treatment for pathology of the abdominal organs. Laparoscopic operations on the pancreas are low-traumatic and in most cases avoid open surgical interventions. For the first time an experimental system for recording speckle images, combined with a commercially available five-millimetre rigid laparoscope, is presented. The sensitivity of the system to the fluid motion in a capillary at different velocities is determined, and the possibility of finding areas of blood microcirculation disturbance in modelling pancreatic ischemia in an experiment on laboratory animals is revealed. The laparoscope illumination channel is verified by comparison with speckle dynamics under external illumination of the studied object. (paper)

60 APPLIED LIFE SCIENCES↗

DOE OSTI2020

Identification of Novel Genomic Associations and Gene Candidates for Grain Starch Content in Sorghum

Starch accumulated in the endosperm of cereal grains as reserve energy for germination serves as a staple in human and animal nutrition. Unraveling genetic control for starch metabolism is important for breeding grains with high starch content. In this study, we used a sorghum association panel with 389 individuals and 141,557 single nucleotide polymorphisms (SNPs) to fit linear mixed models (LMM) for identifying genomic regions and potential candidate genes associated with starch content. Three associated genomic regions, one in chromosome (chr) 1 and two novel associations in chr-8, were identified using combination of LMM and Bayesian sparse LMM. All significant SNPs were located within protein coding genes, with SNPs ∼ 52 Mb of chr-8 encoding a Casperian strip membrane protein (CASP)-like protein (Sobic.008G111500) and a heat shock protein (HSP) 90 (Sobic.008G111600) that were highly expressed in reproductive tissues including within the embryo and endosperm. The HSP90 is a potential hub gene with gene network of 75 high-confidence first interactors that is enriched for five biochemical pathways including protein processing. The first interactors of HSP90 also showed high transcript abundance in reproductive tissues. The candidates of this study are likely involved in intricate metabolic pathways and represent candidate gene targets for source-sink activities and drought and heat stress tolerance during grain filling.

genomics↗

DOE OSTI2022

Community stability is related to animal diversity change

Abstract Understanding the drivers of community stability in times of increasing anthropogenic pressure is an urgent issue. Biodiversity is known to promote community stability, but studies of the biodiversity–stability relationship rarely consider the full complexity of biodiversity change. Furthermore, finding generalities that hold across taxonomic groups and spatial and temporal scales remains challenging because most investigations have narrow taxonomic, spatial, and temporal scopes. We used organismal data collected through the National Ecological Observatory Network (NEON) at sites across the contiguous United States to evaluate linkages between community stability and biodiversity change for four taxonomic groups: small mammals, ground beetles, fish, and freshwater macroinvertebrates. We defined community stability as constancy of aggregate species' abundance. We quantified change in biodiversity as (1) dissimilarity in community taxonomic and functional composition and species replacement and richness change components of that dissimilarity and (2) change in species' abundance distributions as captured by change in species rank, richness, and evenness. We found that community stability increased with species replacement and with contribution of species replacement to overall dissimilarity for all taxonomic groups, but declined with increasing change in species richness and evenness. This is consistent with the notion that temporal fluctuations in species abundance can help stabilize community properties. We also found that community stability was highest when change in community functional composition was either lower or higher than expected given reshuffling of each community's taxonomic composition. This suggests that long‐term community stability can result from fluctuations of functionally similar species in assemblages with high taxonomic reshuffling. On the contrary, the functional uniqueness of fluctuating species compensates for lower taxonomic reshuffling to drive stabilization of community properties. Our study provides an initial assessment of the relationship between community stability and biodiversity change and illustrates the utility of fine temporal resolution data collected across ecosystems and biomes to understand the general mechanisms underlying biodiversity–stability relationships.

54 ENVIRONMENTAL SCIENCES↗

DOE OSTI2021

The Energizer Bunny: Dual-Use Photovoltaic and Pasture-Raised Rabbit Farms (Final Report)

This case study project lays the groundwork for the broader realization of agrivoltaics by considering the techno-economic, environmental, and social aspects of system development and assessing a novel rabbit-based concept. Research on the social and political barriers to agrivoltaic development is sparse, which emphasizes the importance of this project as it provides key insight into the consequential socio-political challenges that may hinder the diffusion of agrivoltaic technology. Additionally, the rabbit agrivoltaic concept is novel and has not been applied in experimental or commercial applications to date. This project advances our understanding of the viability of this co-location scheme through empirical assessment of integrating grazing rabbits with solar. A total of five manuscripts (3 published in peer-reviewed journals, 2 under review) have resulted from this project. The techno-economic component of the project included a) a pilot test study investigating the feasibility of the rabbit agrivoltaic concept and b) an economic analysis of the advantages available to solar operators who leverage grazing animals for lawn maintenance and a dual-revenue stream. Our calculations indicate that the co-location of solar and rabbit farms is a viable form of agrivoltaics, increasing overall site revenue by 2.5%-24.0% above projected electricity revenue. The environmental component of the project entailed an ISO 14040 and 14044 compliant life cycle assessment to quantify the impacts of the rabbit agrivoltaic concept. The results indicate pasture-based agrivoltaic systems provide a dual synergy that produces 69.3% less emissions and demands 82.9% less fossil energy per functional unit of cumulative MWh output of electricity and cumulative kg of meat compared to non-integrated production. The social component of the project included conducting two separate interview studies with agriculture and solar industry professionals and the distribution of a public survey. The interview findings identify the key socio-political opportunities for agrivoltaics include: the retention of agricultural land and rural interests, and increased local level acceptance of solar development. The key barriers to agrivoltaics include: ensuring long term agricultural productivity is not compromised, and subnational zoning strategies. The survey results indicate that support for local solar projects increase by 10% when an agricultural function is incorporated. Survey respondents prefer agrivoltaic projects that are a) designed to provide economic opportunities for farmers and the local community b) located on private property or existing agricultural land, and c) do not threaten local interests. Cumulatively, this project can inform agrivoltaic decision making, solar development practices, rabbit-based applications, land use management, and policy making in a way that supports the furtherance of the renewable energy transition, conserves arable land, and utilizes innovative solar photovoltaic technologies.

14 SOLAR ENERGY↗

DOE OSTI2021

Multiscale Airborne Infectious Disease Transmission

Airborne disease transmission is central to many scientific disciplines, including agriculture, veterinary biosafety, medicine, and public health. Legal and regulatory standards are in place to prevent agricultural, nosocomial, and community airborne disease transmission. However, the overall importance of the airborne pathway is underappreciated; e.g., the U.S. National Library of Medicine’s Medical Subjects Headings (MESH) thesaurus lacks an airborne disease transmission indexing term. This has practical consequences, as airborne precautions to control epidemic disease spread may not be taken when airborne transmission is important but unrecognized. Publishing clearer practical methodological guidelines for surveillance studies and disease outbreak evaluations could help address this situation. To inform future work, this paper highlights selected, well-established airborne transmission events, largely cases replicated in multiple, independently conducted scientific studies. Methodologies include field experiments, modeling, epidemiology studies, disease outbreak investigations, and mitigation studies. Collectively, this literature demonstrates that airborne viruses, bacteria, and fungal pathogens have the ability to cause disease in plants, animals, and humans over multiple distances, from near range (<5 m) to continental (>500 km) in scale. The plausibility and implications of undetected airborne disease transmission are discussed, including the notable underreporting of disease burdens for several airborne-transmitted diseases.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2025

Anatomy of a Turtle Confiscation: What to Expect When You're Not Expecting and Recommendations for Improving Outcomes

Turtles are being subjected to unprecedented collection for the illegal wildlife trade, but only a portion of the trade is detected. When confiscations do happen, wildlife agencies must decide what to do with those animals—euthanize them, place them in permanent captivity, or release them back into the wild. Here, we present a case study of a confiscation of .200 eastern box turtles (Terrapene carolina carolina) and our efforts to repatriate them back to the wild. Twentyfive percent of turtles died in captivity, and at least another 33% died during the soft-release phase. Approximately half of the confiscated turtles survived until they were released from their soft-release pen 9 months post-confiscation. For each phase, from the time turtles were seized until released, we describe our objectives, the challenges we encountered, and our recommendations for improving future turtle confiscations. Given the extended stressful conditions that confiscated turtles often experience before being seized, it is important to recalibrate our expectations regarding future confiscation outcomes.

60 APPLIED LIFE SCIENCES↗

DOE OSTI2023

Interrogating the role of the milk microbiome in mastitis in the multi-omics era

There is growing interest in a functional understanding of milk-associated microbiota as there is ample evidence that host-associated microbial communities play an active role in host health and phenotype. Mastitis, characterized by painful inflammation of the mammary gland, is prevalent among lactating humans and agricultural animals and is associated with significant clinical and economic consequences. The etiology of mastitis is complex and polymicrobial and correlative studies have indicated alterations in milk microbial community composition. Recent evidence is beginning to suggest that a causal relationship may exist between the milk microbiota and host phenotype in mastitis. Multi-omic approaches can be leveraged to gain a mechanistic, molecular level understanding of how the milk microbiome might modulate host physiology, thereby informing strategies to prevent and ameliorate mastitis. In this paper, we review existing studies that have utilized omics approaches to investigate the role of the milk microbiome in mastitis. We also summarize the strengths and challenges associated with the different omics techniques including metagenomics, metatranscriptomics, metaproteomics, metabolomics and lipidomics and provide perspective on the integration of multiple omics technologies for a better functional understanding of the milk microbiome.

metabolomics↗

DOE OSTI2021

Normal Tissue Injury Induced by Photon and Proton Therapies: Gaps and Opportunities

Despite technological advances in radiation therapy (RT) and cancer treatment, patients still experience adverse effects. Proton therapy (PT) has emerged as a valuable RT modality that can improve treatment outcomes. Normal tissue injury is an important determinant of the outcome; therefore, for this review, we analyzed 2 databases: (1) clinical trials registered with ClinicalTrials.gov and (2) the literature on PT in PubMed, which shows a steady increase in the number of publications. Most studies in PT registered with ClinicalTrials.gov with results available are nonrandomized early phase studies with a relatively small number of patients enrolled. From the larger database of nonrandomized trials, we listed adverse events in specific organs/sites among patients with cancer who are treated with photons and protons to identify critical issues. The present data demonstrate dosimetric advantages of PT with favorable toxicity profiles and form the basis for comparative randomized prospective trials. A comparative analysis of 3 recently completed randomized trials for normal tissue toxicities suggests that for early stage non-small cell lung cancer, no meaningful comparison could be made between stereotactic body RT and stereotactic body PT due to low accrual (NCT01511081). In addition, for locally advanced non-small cell lung cancer, a comparison of intensity modulated RT with passive scattering PT (now largely replaced by spot-scanned intensity modulated PT), PT did not provide any benefit in normal tissue toxicity or locoregional failure over photon therapy. Finally, for locally advanced esophageal cancer, proton beam therapy provided a lower total toxicity burden but did not improve progression-free survival and quality of life (NCT01512589). The purpose of this review is to inform the limitations of current trials looking at protons and photons, considering that advances in technology, physics, and biology are a continuum, and to advocate for future trials geared toward accurate precision RT that need to be viewed as an iterative process in a defined path toward delivering optimal radiation treatment. A foundational understanding of the radiobiologic differences between protons and photons in tumor and normal tissue responses is fundamental to, and necessary for, determining the suitability of a given type of biologically optimized RT to a patient or cohort.

62 RADIOLOGY AND NUCLEAR MEDICINE↗

DOE OSTI2020

2020 State of the Science Report, Chapter 14: Summary and Path Forward

The 2020 State of the Science report collates and presents the current understanding of interactions between marine renewable energy (MRE) systems and the marine environment, with an emphasis on their effects on marine animals, habitats, and oceanographic systems, using publicly available information. The report places this information in context through lessons learned from research studies in the laboratory and in the field, modeling simulations, and deployments; monitoring around demonstration, pilot, and small commercial MRE projects; and identifies gaps in knowledge and makes recommendations for filling those gaps. In addition, strategies for moving toward a consistent and effective consenting or permitting (hereafter consenting) process and management of the potential effects of MRE development are highlighted. The status and recommendations from each of the priority interactions between MRE devices and the environment are summarized here, and the management strategies for facilitating development are discussed. Finally, a path forward toward commercial MRE development is explored. https://tethys.pnnl.gov/publications/state-of-the-science-2020-chapter-14-summary-path-forward

16 TIDAL AND WAVE POWER↗

DOE OSTI2024

Reduced ribonucleotide reductase RRM2 subunit expression increases DNA damage and mitochondria dysfunction in woody breast chickens

Abstract OBJECTIVE The aim of this study was to investigate the roles of ribonucleotide reductase subunit M2 (RRM2; subunit of ribonucleotide reductase) in severe woody breast (WB) and normal breast muscles. ANIMALS 40 8-week-old male Ross-708 broiler chickens. METHODS Quantitative PCR was performed to determine gene expression, and commercial ELISA/assay kits were used to obtain several enzymatic activities. RESULTS Results showed that RRM2 activity (P= .0002) and RRM2 (P= .05) and hydroxymethylbilane synthase expression (impaired oxygen transport and metabolism,P= .002) were reduced in WB, while caveolin-3 (defected membrane integrity,P= .09), endoglin (increased fibrosis,P= .06), and secreted protein acidic rich in cysteine (metabolic dysregulation,P= .09) expression tended to increase in WB. WB tended to have increased levels of homocysteine (P= .06), aspartate aminotransferase mitochondria (P= .02), pyruvate kinase (P= .04), DNA damage (P= .06), creatine kinase (P= .05), and triglyceride (P= .002) but decreased ATPase activity (P= .01), all indicating mitochondria dysfunction and tissue damage. CLINICAL RELEVANCE In this study, differences in various enzyme activities and increased DNA damage suggest that RRM2-mediated mitochondrial abnormalities may play a role in WB myopathy.

Veterinary Sciences↗

DOE OSTI2020

Radiobiological Studies Using Cs-137 Gamma or 320 kV X Rays

There are numerous self-shielded research irradiators used in various facilities throughout the United States. The irradiators employ radioactive sources containing either 137 Cs or 60 Co and the irradiators are used for a variety of radiobiological investigations involving cellular and/or animal models. A report from the National Academy of Sciences described security issues associated with particular radiation sources and the desire for their replacement with suitable X-ray irradiators. One possible replacement would be a 320 kV X-ray irradiator. The participants in this research successfully performed in vivo radiobiological studies involving mice exposed to filtered (HVL ≈ 4 mm Cu) 320 kV X rays. Two publications (one published and one submitted at the publishing of this report) documenting key findings are provided in Appendices A and B of this report. The 320 kV X rays were found suitable for in vivo (in mice) cell survival studies and are expected to be suitable for bone marrow transplantation studies using mice but this needs to be experimentally validated.

62 RADIOLOGY AND NUCLEAR MEDICINE↗

DOE OSTI2022

World Whaling: Historical and Contemporary Studies

Whales are omnipresent in the consciousness of modern society, but their public and scientific perceptions have significantly shifted over the last several hundred years. Today, there is a broad attempt to protect and study whales and their habitats, even while sustainable commercial and Indigenous fisheries continue. For the past several millennia humans have interacted with and exploited whales – but it is the critical period since ca. ~1700 that dominates our public understanding of these animals primarily due to their extreme over-exploitation during this era. Further, the dichotomy between protecting whales versus exploiting whales is real, not always mutually exclusive, and palpably stressful both within society and within this text. That is one reason why this Senri Ethnological Studies volume edited by Nobuhiro Kishigami is such a significant contribution to the whale literature, especially for those interested in the state of modern humanwhale interaction and the historical processes that led to the current structure of whale science, protection, regulation and exploitative fisheries.

60 APPLIED LIFE SCIENCES↗

DOE OSTI2024

Tracking animal movements via collaborative acoustic telemetry networks: Multiscale habitat use, phenology, and management insights

Abstract Estuaries support diverse fish and invertebrate communities, including resident species that rely on estuarine habitats year‐round and transient migratory species. The unique movement patterns of these animals connect habitats within and far beyond the estuary and are integrally linked to fisheries management objectives. With a focus on Chesapeake Bay, this study leveraged data from collaborative acoustic telemetry networks in the northwest Atlantic to assess habitat use and phenology of movements for seven species of fish (cownose rays, dusky sharks, smooth dogfish, alewife, striped bass, common carp, and blue catfish) and one invertebrate (horseshoe crabs). A total of 288 acoustically tagged individuals were detected >3.2 million times (6,743 to 2,095,717 detections per species) on receivers across ~20.5 degrees of latitude spanning the North American Atlantic seaboard from Florida, USA, to New Brunswick, Canada. Common metrics of movement and phenology grouped these species as resident (common carp, blue catfish, horseshoe crabs), primarily resident in estuaries (juvenile striped bass), and coastal migrant (cownose rays, dusky sharks, smooth dogfish, alewife); maximum distance traveled varied by three orders of magnitude among these species. Further analysis of phenology for coastal migrants elucidated the timing and duration of these species' use of Chesapeake Bay. Collectively, movements linked habitats within Chesapeake Bay and connected the estuary to coastal ecosystems both to the north (e.g., alewife) and south (e.g., cownose rays), creating networks of fisheries management jurisdictions that varied in complexity and identified opportunities for enhancement to current management or co‐management of some species. Our results elucidate the importance of estuaries to species with diverse movement behaviors, identify scales and pathways of habitat connectivity via animal movements, and highlight the utility of collaborative acoustic telemetry networks for quantifying movements relevant to both ecological research and fisheries management.

Livernois, Mariah C.↗

DOE OSTI2022

Multiomic atlas with functional stratification and developmental dynamics of zebrafish cis -regulatory elements

Zebrafish, a popular organism for studying embryonic development and for modeling human diseases, has so far lacked a systematic functional annotation program akin to those in other animal models. To address this, we formed the international DANIO-CODE consortium and created a central repository to store and process zebrafish developmental functional genomic data. Our data coordination center (https://danio-code.zfin.org) combines a total of 1,802 sets of unpublished and re-analyzed published genomic data, which we used to improve existing annotations and show its utility in experimental design. We identified over 140,000 cis-regulatory elements throughout development, including classes with distinct features dependent on their activity in time and space. We delineated the distinct distance topology and chromatin features between regulatory elements active during zygotic genome activation and those active during organogenesis. Finally, we matched regulatory elements and epigenomic landscapes between zebrafish and mouse and predicted functional relationships between them beyond sequence similarity, thus extending the utility of zebrafish developmental genomics to mammals.

59 BASIC BIOLOGICAL SCIENCES↗