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Tracking DO 2 with Compensatory Reserve during Whole Blood Resuscitation in Baboons

Background Hemorrhagic shock can be mitigated by timely and accurate resuscitation designed to restore adequate delivery of oxygen (DO 2 ) by increasing cardiac output (CO). However, standard care of using systolic blood pressure (SBP) as a guide for resuscitation has proven ineffective and can be associated with increased morbidity. We have developed a novel vital sign called the compensatory reserve (CRM) generated from analysis of arterial pulse waveform feature changes that has been validated in experimental and clinical models of hemorrhage. We tested the hypothesis that thresholds of DO 2 could be accurately defined by CRM while avoiding over resuscitation during whole blood resuscitation following a 25% hemorrhage in non‐human primates. Methods To accomplish this, adult male baboons (n=12) were exposed to a progressive controlled hemorrhage that resulted in an average (± SD) maximal reduction of 601 ± 18 ml of their estimated circulating blood volume of 1,262 ± 64 ml based on body weight. Values of DO 2 were calculated as the product of CO times O 2 carrying capacity of hemoglobin (i.e., Hb content × 1.34). Results Average (± SD) CRM increased from 6 ± 5 % at the end of hemorrhage to 70 ± 7 % at the end of resuscitation. DO 2 of 12.1, 10.3, and 7.0 ml O 2 ·kg −1 ·min −1 were calculated at 0% (baseline), 12.5% and 25% blood loss. By linear regression, CRM values of 6% (decompensation), 35%, and 68% corresponded to calculated DO 2 values of 6.0, 9.2, and 12.1 ml O 2 ·kg −1 ·min −1 during resuscitation. As such, return of CRM to baseline during resuscitation with only ~ 450 ml provided adequate DO 2 (12.5 ml O 2 ·kg −1 ·min −1 ) while total blood volume replacement (~600 ml) resulted in over resuscitation as indicated by an average increase in SBP (+17 mmHg) and CO (+0.1 L/min) over baseline values. Conclusion Consistent with our hypothesis, thresholds of DO 2 were associated with specific CRM values. A target resuscitation CRM value of 70% minimized the requirement for whole blood, while avoiding over resuscitation. Furthermore, CRM at 0% provided a noninvasive metric for determining critical DO 2 at approximately 5.4 ml O 2 ·kg −1 ·min −1 . Support or Funding Information This research was supported in part by appointment of Ms. Koons to the Postgraduate Research Fellowship Program at the US Army Institute of Surgical Research administered by the Oak Ridge Institute for Science and Education through an interagency agreement between the US Department of Energy and USAMRMC. Study funding was provided by a grant from the US Army Combat Casualty Care Research Program (D‐009‐2014‐USAISR). Linearregression of the relationship between DO 2 and compensatory reserve during whole blood resuscitation in baboons. Data are expressed as mean ± SE. image Linearregression of the relationship between DO 2 and compensatory reserve during whole blood resuscitation in baboons. Data are expressed as mean ± SE. This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .

Koons, Natalie Jane↗

An Explainable Machine-Learning Model for Compensatory Reserve Measurement: Methods for Feature Selection and the Effects of Subject Variability

Tracking vital signs accurately is critical for triaging a patient and ensuring timely therapeutic intervention. The patient’s status is often clouded by compensatory mechanisms that can mask injury severity. The compensatory reserve measurement (CRM) is a triaging tool derived from an arterial waveform that has been shown to allow for earlier detection of hemorrhagic shock. However, the deep-learning artificial neural networks developed for its estimation do not explain how specific arterial waveform elements lead to predicting CRM due to the large number of parameters needed to tune these models. Alternatively, we investigate how classical machine-learning models driven by specific features extracted from the arterial waveform can be used to estimate CRM. More than 50 features were extracted from human arterial blood pressure data sets collected during simulated hypovolemic shock resulting from exposure to progressive levels of lower body negative pressure. A bagged decision tree design using the ten most significant features was selected as optimal for CRM estimation. This resulted in an average root mean squared error in all test data of 0.171, similar to the error for a deep-learning CRM algorithm at 0.159. By separating the dataset into sub-groups based on the severity of simulated hypovolemic shock withstood, large subject variability was observed, and the key features identified for these sub-groups differed. This methodology could allow for the identification of unique features and machine-learning models to differentiate individuals with good compensatory mechanisms against hypovolemia from those that might be poor compensators, leading to improved triage of trauma patients and ultimately enhancing military and emergency medicine.

60 APPLIED LIFE SCIENCES↗

Superiority of compensatory reserve measurement compared with the Shock index for early and accurate detection of reduced central blood volume status

Shock index (SI) equals the ratio of heart rate (HR) to systolic blood pressure (SBP) with clinical evidence that it is more sensitive for trauma patient status assessment and prediction of outcome compared with either HR or SBP alone. We used lower body negative pressure (LBNP) as a human model of central hypovolemia and compensatory reserve measurement (CRM) validated for accurate tracking of reduced central blood volume to test the hypotheses that SI: (1) presents a late signal of central blood volume status; (2) displays poor sensitivity and specificity for predicting the onset of hemodynamic decompensation; and (3) cannot identify individuals at greatest risk for the onset of circulatory shock. We measured HR, SBP, and CRM in 172 human subjects (19–55 years) during progressive LBNP designed to determine tolerance to central hypovolemia as a model of hemorrhage. Subjects were subsequently divided into those with high tolerance (HT) (n = 118) and low tolerance (LT) (n = 54) based on completion of 60 mm Hg LBNP. The time course relationship between SI and CRM was determined and receiver operating characteristic (ROC) area under the curve (AUC) was calculated for sensitivity and specificity of CRM and SI to predict hemodynamic decompensation using clinically defined thresholds of 40% for CRM and 0.9 for SI. The time and level of LBNP required to reach a SI = 0.9 (~60 mm Hg LBNP) was significantly greater (p < 0.001) compared with CRM that reached 40% at ~40 mm Hg LBNP. Shock index did not differ between HT and LT subjects at 45 mm Hg LBNP levels. ROC AUC for CRM was 0.95 (95% CI = 0.94–0.97) compared with 0.91 (0.89–0.94) for SI (p = 0.0002). Despite high sensitivity and specificity, SI delays time to detect reductions in central blood volume with failure to distinguish individuals with varying tolerances to central hypovolemia.

60 APPLIED LIFE SCIENCES↗

Hanford Site Bee Nest Box Monitoring Report for Calendar Year 2021

This report summarizes the results of the third year of monitoring bee nest boxes on the Hanford Site. Twenty bee nest boxes were installed as a component of compensatory mitigation for the installation of the L-894 water line between the 200-East and 200-West Areas of the Hanford Site in July 2019. The goal of this compensatory mitigation is to replace lost nesting habitat for above-ground nesting bees in a mature sagebrush ecosystem. Annual monitoring will track the condition and occupation of the bee nest boxes to determine if the compensatory mitigation was successful and to identify best practices when replacing nesting habitat for native bees.

54 ENVIRONMENTAL SCIENCES↗

Hanford Site Bee Nest Box Monitoring Report for Calendar Year 2020

This report summarizes the results of the second year of monitoring bee nest boxes on the Hanford Site. Twenty bee nest boxes were installed as a component of compensatory mitigation for the installation of the L-894 water line between the 200-East and 200-West Areas of the Hanford Site in July 2019. The goal of this compensatory mitigation is to replace lost nesting habitat for above-ground nesting bees in a mature sagebrush ecosystem. Annual monitoring will track the condition and occupation of the bee nest boxes to determine if the compensatory mitigation was successful and to identify best practices when replacing nesting habitat for native bees.

54 ENVIRONMENTAL SCIENCES↗

Hanford Site Bee Nest Box Monitoring Report for Calendar Year 2019

This report summarizes the results of the first year of monitoring bee nest boxes on the Hanford Site. Twenty bee nest boxes were installed as a component of compensatory mitigation for the installation of the L-894 water line between the 200-East and 200-West Areas of the Hanford Site. The goal of this compensatory mitigation is to replace lost nesting habitat for above-ground nesting bees in a mature sagebrush ecosystem. Annual monitoring will track the condition and occupation of the bee nest boxes to determine if the compensatory mitigation was successful and to identify best practices when replacing nesting habitat for native bees.

54 ENVIRONMENTAL SCIENCES↗

Draft Prototype Microreactor Transportation Safety Program

Microreactors are compact reactors capable of producing less than 50 megawatts of electrical energy. Typically, these reactors are factory-fabricated and designed to be easily transportable by truck, rail, vessel, or air. Microreactor designs often assume that the unit can be transported containing either unirradiated or irradiated fuel. The interest in microreactors is driven by several factors, including the need to generate power on at remote locations, at military installations, at facilities such as data centers, and in areas recovering from natural disasters. The U.S. Department of Defense is actively pursuing the microreactor concept to meet the increasing energy demands of military operations that require portable and dense power sources. Commercial vendors are also exploring microreactor concepts. The report Microreactor Transportation Emergency Planning Challenges (Maheras et al. 2024) outlined the emergency planning challenges associated with the transportation of microreactors by road, rail, and by barge/ship. The successful commercial deployment and redeployment of microreactors will also require the development of microreactor transportation safety programs. The elements in these safety programs are not specific to microreactors; however, the transport of microreactors may pose unique challenges in these areas. This report builds on the report Microreactor Transportation Emergency Planning Challenges (Maheras et al. 2024) and develops the elements of a prototype microreactor transportation safety program that describes the elements that should be contained vendor-developed microreactor transportation safety programs, identifying the unique elements associated with microreactor transport. This will provide vendors and their transportation contractors a basis for their transportation planning and will accelerate the commercial deployment and redeployment of microreactors by identifying those issues unique to microreactor transport. The emphasis of this report is on highway transport of microreactors. This is based on a U.S. Nuclear Regulatory Commission transportation package approval strategy of crawl-walk-run, where transport by highway is evaluated first (Coles et al. 2021, 2024, Maheras et al. 2021), then other surface modes (rail and barge/ship), and finally air transport. Evaluation of maritime transport of microreactors was recently initiated (Rigato et al. 2024, Maheras et al. 2025). The report first discusses microreactors in general and microreactor transportation safety program planning assumptions. The report then provides a description of the transportation safety planning process and provides an extensive discussion of the elements of transportation safety programs. Specific elements examined included transportation roles and responsibilities, transportation planning, transportation mode and route selection, carrier selection, transportation packaging, advance notification of shipments, public information and communications, emergency response plans and procedures, inspections, security, safe parking, shipment tracking, weather and road conditions, medical preparedness, training and exercises, and program evaluation. The report then identifies the unique elements of a transportation safety program associated with microreactor transport. These unique elements were in the areas of: the unusual nature of microreactor designs, compensatory measures, increased radiation dose rates in the vicinity of microreactors, transportation package approval versus 10 CFR 50.59, and the use of a risk-informed transportation package approval process.

22 GENERAL STUDIES OF NUCLEAR REACTORS↗

Precursor-Dependent Routing of Aromatic Amino Acids Determines Lignin Structure in Grasses by Sensitivity-Enhanced Solid-State NMR

Lignin biosynthesis in grasses exhibits unique metabolic flexibility, yet the precursor-specific routing of carbon into lignin polymers remains poorly resolved in planta. Here, we combine 13 C-isotope labeling with solid-state NMR under sensitivity-enhancement by dynamic nuclear polarization (DNP), to directly track phenylalanine- and tyrosine-derived carbon incorporation into the lignin polymer in Brachypodium distachyon. Precursor-specific 13 C labeling reveals that phenylalanine is the dominant contributor to canonical guaiacyl and syringyl lignins, whereas tyrosine preferentially enriches hydroxyphenyl lignin and hydroxycinnamates, including ferulates characteristic of grass cell walls. Two-dimensional 13 C− 13 C correlation NMR resolves distinct lignin moieties arising from each precursor. Disruption of pcoumarate 3-hydroxylase (C3H) selectively impairs phenylalanine-derived lignification, while tyrosine-derived lignin remains comparatively unchanged, maintaining polymer assembly through alternative metabolic routes. These findings show precursordependent control of lignin composition and reveal tyrosine-mediated lignification as a compensatory pathway in grasses. This work also establishes precursor-resolved solid-state NMR and DNP as a powerful framework for dissecting lignin biosynthesis and metabolic plasticity in plant cell walls.

Biopolymers↗

Potentially adaptive SARS-CoV-2 mutations discovered with novel spatiotemporal and explainable AI models

Abstract Background A mechanistic understanding of the spread of SARS-CoV-2 and diligent tracking of ongoing mutagenesis are of key importance to plan robust strategies for confining its transmission. Large numbers of available sequences and their dates of transmission provide an unprecedented opportunity to analyze evolutionary adaptation in novel ways. Addition of high-resolution structural information can reveal the functional basis of these processes at the molecular level. Integrated systems biology-directed analyses of these data layers afford valuable insights to build a global understanding of the COVID-19 pandemic. Results Here we identify globally distributed haplotypes from 15,789 SARS-CoV-2 genomes and model their success based on their duration, dispersal, and frequency in the host population. Our models identify mutations that are likely compensatory adaptive changes that allowed for rapid expansion of the virus. Functional predictions from structural analyses indicate that, contrary to previous reports, the Asp 614 Gly mutation in the spike glycoprotein (S) likely reduced transmission and the subsequent Pro 323 Leu mutation in the RNA-dependent RNA polymerase led to the precipitous spread of the virus. Our model also suggests that two mutations in the nsp13 helicase allowed for the adaptation of the virus to the Pacific Northwest of the USA. Finally, our explainable artificial intelligence algorithm identified a mutational hotspot in the sequence of S that also displays a signature of positive selection and may have implications for tissue or cell-specific expression of the virus. Conclusions These results provide valuable insights for the development of drugs and surveillance strategies to combat the current and future pandemics.

59 BASIC BIOLOGICAL SCIENCES↗