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A matter of quantum voltages

Voltages inside matter are relevant to crystallization, materials science, biology, catalysis, and aqueous chemistry. The variation of voltages in matter can be measured by experiment, however, modern supercomputers allow the calculation of accurate quantum voltages with spatial resolutions of bulk systems well beyond what can currently be measured provided a sufficient level of theory is employed. Of particular interest is the Mean Inner Potential (V{sub o}) – the spatial average of these quantum voltages referenced to the vacuum. Here we establish a protocol to reliably evaluate V{sub o} from quantum calculations. Voltages are very sensitive to the distribution of electrons and provide metrics to understand interactions in condensed phases. In the present study, we find excellent agreement with measurements of V{sub o} for vitrified water and salt crystals and demonstrate the impact of covalent and ionic bonding as well as intermolecular/atomic interactions. Certain aspects in this regard are highlighted making use of simple model systems/approximations. Furthermore, we predict V{sub o} as well as the fluctuations of these voltages in aqueous NaCl electrolytes and characterize the changes in their behavior as the resolution increases below the size of atoms.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Engineered thermalization and cooling of quantum many-body systems

We develop a scheme for engineering genuine thermal states in analog quantum simulation platforms by coupling local degrees of freedom to driven, dissipative ancilla pseudospins. We demonstrate the scheme in a many-body quantum spin lattice simulation setting. A Born-Markov master equation describing the dynamics of the many-body system is developed, and we show that if the ancilla energies are periodically modulated, with a carefully chosen hierarchy of timescales, one can effectively thermalize the many-body system. Through analysis of the time-dependent dynamical generator, we determine the conditions under which the true thermal state is an approximate dynamical fixed point for general system Hamiltonians. Finally, we evaluate the thermalization protocol through numerical simulation and discuss prospects for implementation on current quantum simulation hardware.

36 MATERIALS SCIENCE↗

Sci—Sat AM: Stereo — 08: Stereotactic Ablative Radiotherapy (SABR) for low, intermediate and high risk prostate cancer using Volumetric Modulated Arc Therapy (VMAT) with a 10x Flattening Filter Free (FFF) beam

Purpose: To determine the feasibility of using Volumetric Modulated Arc Therapy (VMAT) with a 10x Flattening Filter Free (FFF) beam for Stereotactic Ablative Radiotherapy (SABR) for low, intermediate and high risk prostate cancer. Methods and Materials: Ten anonymized patient CT data sets were used in this planning study. For each patient CT data set, three sets of contours were generated: 1) low risk, 2) intermediate risk, and 3) high risk scenarios. For each scenario, a single-arc and a double-arc VMAT treatment plans were created. Plans were generated with the Varian Eclipse™ treatment planning system for a Varian TrueBeam™ linac equipped with Millenium 120 MLC. Plans were created using a 10x-FFF beam with a maximum dose rate of 2400 MU/min. Dose prescription was 36.25Gy/5 fractions with the planning objective of covering 99% of the Planning Target Volume with the 95% of the prescription dose. Normal tissue constraints were based on provincial prostate SABR planning guidelines, derived from national and international prostate SABR protocols. Plans were evaluated and compared in terms of: 1) dosimetric plan quality, and 2) treatment delivery efficiency. Results: Both single-arc and double-arc VMAT plans were able to meet the planning goals for low, intermediate and high risk scenarios. No significant dosimetric differences were observed between the plans. However, the treatment time was significantly lower for a single-arc VMAT plans. Conclusions: Prostate SABR treatments are feasible with 10x-FFF VMAT technique. A single-arc VMAT offers equivalent dosimetric plan quality and a superior treatment delivery efficiency, compared to a double-arc VMAT.

60 APPLIED LIFE SCIENCES↗

Sci-Fri AM: MRI and Diagnostic Imaging - 02: Quality Improvement: Diagnostic Reference Levels for Interior Health CT exams – L-Spine, Chest/Abdomen/pelvis, Abdomen/Pelvis, Head

Diagnostic Reference Levels are used to optimize patient dose and image quality in the clinical setting. It is assumed that the majority of exams are of diagnostic quality, or the radiologists would request protocol adjustments. By investigating the dose indicator distributions from all scanners, the upper DRL can be set to the 75th percentile of the distribution and a lower DRL can be set to the 10th percentile. Scanners using doses consistently outside the upper/lower DRL range can be adjusted accordingly. 11 CT scanners, all contributing to the American College of Radiology Dose Index Registry (ACR DIR) were used in this study. Dose indicator data were compiled from the ACR DIR data and local DRLs established. Scanners with median doses outside the upper/lower DRL were followed-up with. Using effective dose and exam volumes, collective dose was determined before and after protocol adjustments to evaluate the effect of this quality improvement effort. The quality initiative is complete for L-spine and Chest/Abdomen/Pelvis exams and only initial surveys were completed for Head and Abdomen/Pelvis examsg. Median Scanner Dose reductions were 8.8 and 4.9 % for L-spine and Chest/Abdomen/Pelvis exams, respectively, resulting with collective dose reductions of 0.7 and 3.2 person•Sv/yr. Follow-up is ongoing for Abdomen/Pelvis and Head exams.

60 APPLIED LIFE SCIENCES↗

Rethinking the Default Construction of Multimodel Climate Ensembles

Here, we discuss the current code of practice in the climate sciences to routinely create climate model ensembles as ensembles of opportunity from the newest phase of the Coupled Model Intercomparison Project (CMIP). We give a two-step argument to rethink this process. First, the differences between generations of ensembles corresponding to different CMIP phases in key climate quantities are not large enough to warrant an automatic separation into generational ensembles for CMIP3 and CMIP5. Second, we suggest that climate model ensembles cannot continue to be mere ensembles of opportunity but should always be based on a transparent scientific decision process. If ensembles can be constrained by observation, then they should be constructed as target ensembles that are specifically tailored to a physical question. If model ensembles cannot be constrained by observation, then they should be constructed as cross-generational ensembles, including all available model data to enhance structural model diversity and to better sample the underlying uncertainties. To facilitate this, CMIP should guide the necessarily ongoing process of updating experimental protocols for the evaluation and documentation of coupled models. Finally, with an emphasis on easy access to model data and facilitating the filtering of climate model data across all CMIP generations and experiments, our community could return to the underlying idea of using model data ensembles to improve uncertainty quantification, evaluation, and cross-institutional exchange.

54 ENVIRONMENTAL SCIENCES↗

Analysis, identification and confirmation of synthetic opioids using chloroformate chemistry: Retrospective detection of fentanyl and acetylfentanyl in urine and plasma samples by EI-GC-MS and HR-LC-MS

Electron Impact Gas Chromatography-Mass Spectrometry (EI-GC-MS) and High Resolution Liquid Chromatography-Mass Spectrometry (HR-LC-MS) have been used in the analysis of products arising from the trichloroethoxycarbonylation of fentanyl and acetylfentanyl in urine and plasma matrices. The method involves the initial extraction of both synthetic opioids separately from the matrices followed by detection of the unique products that arise from their reaction with 2,2,2-trichloroethoxycarbonyl chloride (Troc-Cl), namely Troc-norfentanyl and Troc-noracetylfentanyl. The optimized protocol was successfully evaluated for its efficacy at detecting these species formed from fentanyl and acetylfentanyl when present at low and high levels in urine (fentanyl: 5 and 10 ng/mL and acetylfentanyl: 20 and 100 ng/mL) and plasma (fentanyl: 10 and 20 ng/mL and acetylfentanyl: 50 and 200 ng/mL), values that reflect levels reported in overdose victims. The HR-LC-MS method’s LOQ (limit of quantitation) for the Troc-norfentanyl and Troc-noracetylfentanyl products was determined to be ~10 ng/mL for both species. Even though the superiority in the detection of these species by HR-LC-MS over EI-GC-MS, the latter method proved to be important in the detection of the second product from the reaction, namely 2-phenylethyl chloride that is crucial in the determination of the original opioid. This observation highlights the importance of using complimentary analytical techniques in the analysis of a sample, whether biological or environmental in nature. The method herein serves as a complementary, qualitative confirmation for the presence of a fentanyl in collected urine, plasma and by extension other biological samples amenable to the common extraction procedures described for opioid analysis. More importantly, the method’s main strength comes from its ability to react with unknown fentanyls to yield products that can be not only detected by EI-GC-MS and HR-LC-MS but can then be used to retrospectively identify an unknown fentanyl.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Preliminary study of PCBs in raccoons living on or near the Paducah Gaseous Diffusion Plant, Kentucky

The “Ecological Monitoring at the Paducah Gaseous Diffusion Plant: Historical Evaluation and Guidelines for Future Monitoring” report (Halbrook, et al. 2007) recommended the raccoon as a species for study at the Paducah Gaseous Diffusion Plant (PGDP). This species was selected to fill data gaps in ecological resources and provide resource managers with knowledge that will be valuable in making decisions and implementing specific actions to safeguard ecological resources and reduce human exposure. The current paper reports results of a preliminary evaluation to establish protocols for collection of tissues and initial screening of polychlorinated biphenyls (PCBs) in raccoons collected near the PGDP. These data are useful in developing future more comprehensive studies.

63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTAN↗

Hanford Double Shell Waste Tank Corrosion Studies (Final Report FY2023)

For fiscal year (FY) 2023, the Savannah River National Laboratory (SRNL) focused on two experimental tasks related to Hanford Double Shell Tank (DST) chemistry and integrity. The first task focused on understanding risk of corrosion due to formation of either continuous layers or discrete patches of solids on the tanks’ inner sidewalls and bottoms. Differences in the conductivity between various layers of the tank (e.g., solids, liquid, etc.) could result in differences in the electrochemical potential of the tank metal at various locations. The electrochemical potential difference may result in a corrosion current between the coupled surfaces. In FY23, SRNL investigated test configurations and protocols that could evaluate the presence of a galvanic couple between the tank bottom and the tank wall.

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

One Health Approach to Tackle Microbial Contamination on Poultries—A Systematic Review

This study reports the search of available data published regarding microbial occupational exposure assessment in poultries, following the PRISMA methodology. Air collection through filtration was the most frequently used. The most commonly used passive sampling method was material collection such as dust, cages, soils, sediment, and wastewater. Regarding assays applied, the majority of studies comprised culture-based methods, but molecular tools were also frequently used. Screening for antimicrobial susceptibility was performed only for bacteria; cytotoxicity, virological and serological assays were also performed. Most of the selected studies focused on bacteria, although fungi, endotoxins, and β-glucans were also assessed. The only study concerning fungi and mycotoxins reported the carcinogenic mycotoxin AFB1. This study gives a comprehensive overview of microbial contamination in the poultry industry, emphasizing this setting as a potential reservoir of microbial pathogens threatening human, animal, and environmental health. Additionally, this research helps to provide a sampling and analysis protocol proposal to evaluate the microbiological contamination in these facilities. Few articles were found reporting fungal contamination in poultry farms worldwide. In addition, information concerning fungal resistance profile and mycotoxin contamination remain scarce. Overall, a One Health approach should be incorporated in exposure assessments and the knowledge gaps identified in this paper should be addressed in further research.

Gomes, Bianca (ORCID:0000000262646072)↗

Towards optimized viral metagenomes for double-stranded and single-stranded DNA viruses from challenging soils

Soils impact global carbon cycling and their resident microbes are critical to their biogeochemical processing and ecosystem outputs. Based on studies in marine systems, viruses infecting soil microbes likely modulate host activities via mortality, horizontal gene transfer, and metabolic control. However, their roles remain largely unexplored due to technical challenges with separating, isolating, and extracting DNA from viruses in soils. Some of these challenges have been overcome by using whole genome amplification methods and while these have allowed insights into the identities of soil viruses and their genomes, their inherit biases have prevented meaningful ecological interpretations. Here we experimentally optimized steps for generating quantitatively-amplified viral metagenomes to better capture both ssDNA and dsDNA viruses across three distinct soil habitats along a permafrost thaw gradient. First, we assessed differing DNA extraction methods (PowerSoil, Wizard mini columns, and cetyl trimethylammonium bromide) for quantity and quality of viral DNA. This established PowerSoil as best for yield and quality of DNA from our samples, though ~1/3 of the viral populations captured by each extraction kit were unique, suggesting appreciable differential biases among DNA extraction kits. Second, we evaluated the impact of purifying viral particles after resuspension (by cesium chloride gradients; CsCl) and of viral lysis method (heat vs bead-beating) on the resultant viromes. DNA yields after CsCl particle-purification were largely non-detectable, while unpurified samples yielded 1–2-fold more DNA after lysis by heat than by bead-beating. Virome quality was assessed by the number and size of metagenome-assembled viral contigs, which showed no increase after CsCl-purification, but did from heat lysis relative to bead-beating. We also evaluated sample preparation protocols for ssDNA virus recovery. In both CsCl-purified and non-purified samples, ssDNA viruses were successfully recovered by using the Accel-NGS 1S Plus Library Kit. While ssDNA viruses were identified in all three soil types, none were identified in the samples that used bead-beating, suggesting this lysis method may impact recovery. Further, 13 ssDNA vOTUs were identified compared to 582 dsDNA vOTUs, and the ssDNA vOTUs only accounted for ~4% of the assembled reads, implying dsDNA viruses were dominant in these samples. This optimized approach was combined with the previously published viral resuspension protocol into a sample-to-virome protocol for soils now available at protocols.io, where community feedback creates ‘living’ protocols. This collective approach will be particularly valuable given the high physicochemical variability of soils, which will may require considerable soil type-specific optimization. This optimized protocol provides a starting place for developing quantitatively-amplified viromic datasets and will help enable viral ecogenomic studies on organic-rich soils.

Trubl, Gareth↗

Towards optimized viral metagenomes for double-stranded and single-stranded DNA viruses from challenging soils

Soils impact global carbon cycling and their resident microbes are critical to their biogeochemical processing and ecosystem outputs. Based on studies in marine systems, viruses infecting soil microbes likely modulate host activities via mortality, horizontal gene transfer, and metabolic control. However, their roles remain largely unexplored due to technical challenges with separating, isolating, and extracting DNA from viruses in soils. Some of these challenges have been overcome by using whole genome amplification methods and while these have allowed insights into the identities of soil viruses and their genomes, their inherit biases have prevented meaningful ecological interpretations. Here we experimentally optimized steps for generating quantitatively-amplified viral metagenomes to better capture both ssDNA and dsDNA viruses across three distinct soil habitats along a permafrost thaw gradient. First, we assessed differing DNA extraction methods (PowerSoil, Wizard mini columns, and cetyl trimethylammonium bromide) for quantity and quality of viral DNA. This established PowerSoil as best for yield and quality of DNA from our samples, though ~1/3 of the viral populations captured by each extraction kit were unique, suggesting appreciable differential biases among DNA extraction kits. Second, we evaluated the impact of purifying viral particles after resuspension (by cesium chloride gradients; CsCl) and of viral lysis method (heat vs bead-beating) on the resultant viromes. DNA yields after CsCl particle-purification were largely non-detectable, while unpurified samples yielded 1–2-fold more DNA after lysis by heat than by bead-beating. Virome quality was assessed by the number and size of metagenome-assembled viral contigs, which showed no increase after CsCl-purification, but did from heat lysis relative to bead-beating. We also evaluated sample preparation protocols for ssDNA virus recovery. In both CsCl-purified and non-purified samples, ssDNA viruses were successfully recovered by using the Accel-NGS 1S Plus Library Kit. While ssDNA viruses were identified in all three soil types, none were identified in the samples that used bead-beating, suggesting this lysis method may impact recovery. Further, 13 ssDNA vOTUs were identified compared to 582 dsDNA vOTUs, and the ssDNA vOTUs only accounted for ~4% of the assembled reads, implying dsDNA viruses were dominant in these samples. This optimized approach was combined with the previously published viral resuspension protocol into a sample-to-virome protocol for soils now available at protocols.io, where community feedback creates ‘living’ protocols. This collective approach will be particularly valuable given the high physicochemical variability of soils, which will may require considerable soil type-specific optimization. This optimized protocol provides a starting place for developing quantitatively-amplified viromic datasets and will help enable viral ecogenomic studies on organic-rich soils.

Trubl, Gareth↗

Towards optimized viral metagenomes for double-stranded and single-stranded DNA viruses from challenging soils

Soils impact global carbon cycling and their resident microbes are critical to their biogeochemical processing and ecosystem outputs. Based on studies in marine systems, viruses infecting soil microbes likely modulate host activities via mortality, horizontal gene transfer, and metabolic control. However, their roles remain largely unexplored due to technical challenges with separating, isolating, and extracting DNA from viruses in soils. Some of these challenges have been overcome by using whole genome amplification methods and while these have allowed insights into the identities of soil viruses and their genomes, their inherit biases have prevented meaningful ecological interpretations. Here we experimentally optimized steps for generating quantitatively-amplified viral metagenomes to better capture both ssDNA and dsDNA viruses across three distinct soil habitats along a permafrost thaw gradient. First, we assessed differing DNA extraction methods (PowerSoil, Wizard mini columns, and cetyl trimethylammonium bromide) for quantity and quality of viral DNA. This established PowerSoil as best for yield and quality of DNA from our samples, though ∼1/3 of the viral populations captured by each extraction kit were unique, suggesting appreciable differential biases among DNA extraction kits. Second, we evaluated the impact of purifying viral particles after resuspension (by cesium chloride gradients; CsCl) and of viral lysis method (heat vs bead-beating) on the resultant viromes. DNA yields after CsCl particle-purification were largely non-detectable, while unpurified samples yielded 1–2-fold more DNA after lysis by heat than by bead-beating. Virome quality was assessed by the number and size of metagenome-assembled viral contigs, which showed no increase after CsCl-purification, but did from heat lysis relative to bead-beating. We also evaluated sample preparation protocols for ssDNA virus recovery. In both CsCl-purified and non-purified samples, ssDNA viruses were successfully recovered by using the Accel-NGS 1S Plus Library Kit. While ssDNA viruses were identified in all three soil types, none were identified in the samples that used bead-beating, suggesting this lysis method may impact recovery. Further, 13 ssDNA vOTUs were identified compared to 582 dsDNA vOTUs, and the ssDNA vOTUs only accounted for ∼4% of the assembled reads, implying dsDNA viruses were dominant in these samples. This optimized approach was combined with the previously published viral resuspension protocol into a sample-to-virome protocol for soils now available at protocols.io , where community feedback creates ‘living’ protocols. This collective approach will be particularly valuable given the high physicochemical variability of soils, which will may require considerable soil type-specific optimization. This optimized protocol provides a starting place for developing quantitatively-amplified viromic datasets and will help enable viral ecogenomic studies on organic-rich soils.

dsDNA viruses↗

The effect of intermittent standing or walking during head down tilt bedrest on peak O2 consumption

The cardiovascular aspect of bedrest deconditioning is manifested by decreases in peak O2 uptake (VO(sub 2 peak)) during minimal exercise. The effect of intermittent standing (+G(z)) or walking (+G(z)W) during 4 days of 7 degree Head Down Tilt bedrest (HDT) on VO(sub 2 peak) was evaluated. Methods: Five protocols were performed by eight male subjects; control (C) consisting of complete bedrest, and 15 minute periods to total 2 or 4 hours daily of standing (+G(z)(exp 2) and +G(z)(exp 4) respectively) or walking at 3.0 MPH (+G(z)W2 and +G(z)W4 respectively). Subjects performed VO(sub 2 peak) tests prior to and on the final day of HDT. VO(sub 2 peak) was determined using open circuit indirect calorimetry during supine leg cycling ergometry. After a 5 minute warmup, three 2 minute incremental loads of 33 W previously determined to elicit VO(sub 2 peak) were given and the subject cycled to volitional fatigue. Results: The C protocol VO(sub 2 peak) decreased by 16 percent (2.71 plus or minus 0.16 to 2.27 plus or minus 0.14 L/min) and 11 percent in +G(z)(exp 4) (2.72 plus or minus 0.15 to 2.43 plus or minus 0.14 L/min). With +G(z)W2 VO(sub 2 peak) decreased by 9 percent (2.71 plus or minus 0.17 to 2.46 plus or minus 0.14 L/min) and with +G(z)W4, VO(sub 2 peak) decreased by 10 percent (2.71 plus or minus 0.14 to 2.43 plus or minus 0.14 L/min). VO(sub 2 peak) in all protocols decreased with HDT (P less than 0.05). The decrease in C VO(sub 2 peak) was significantly greater (P less than 0.05) than the decreases in either +G(z) or +G(z)W protocols. Conclusion: The deconditioning that occurs after only 4 days of HDT was demonstrated by decreases in VO(sub 2 peak). Intermittent +G(z) or +G(z)W attenuated, but did not prevent, the decrease in VO(sub 2 peak) with HDT.

Ertl, A. C.↗

Passive, Direct-Read Monitoring System for Selective Detection and Quantification of Hydrogen Chloride

Monitoring the exposure of an employee to hydrogen chloride or hydrochloric acid in the presence of other acids has been a challenge to the industrial hygiene community. The capability of a device to differentiate the levels of acid vapors would allow for more accurate determinations of exposure and therefore improved occupational health. In this work, a selective direct-read colorimetric badge system was validated for Short Term Exposure Limit (STEL) monitoring of hydrogen chloride. The passive colorimetric badge system consists of a direct reading badge and a color scale. The badge has a coated indicator layer with a diffusive resistance in the shape of an exclamation mark. An exclamation mark will appear if hydrogen chloride is present in the atmosphere at concentrations at or above 2.0 ppm. By using the color scale, the intensity of the color formed on the badge can be further quantified up to 25 ppm. The system was validated according to a protocol based on the NIOSH Protocol for the Evaluation of Passive Monitors. The badge was exposed to relative humidities ranging from 11% to 92%, temperatures ranging from 7 C to 400 C and air velocities ranging from 5 cm/sec to 170 cm/sec. All experiments were conducted in a laboratory vapor generation system. Hydrofluoric acid, nitric acid, sulfuric acid, chlorine, hydrogen sulfide and organic acids showed no effect on the performance of the hydrogen chloride monitoring system. The passive badge and color scale system exceeded the accuracy requirements as defined by NIOSH. At ambient conditions, the mean coefficient of variation was 10.86 and the mean bias was 1.3%. This data was presented previously at the American Industrial Hygiene Conference and Exposition in Toronto, Canada in June 1999.

Chapman, K. B.↗

NASA Space Technology Can Improve Soldier Health, Performance and Safety

One of the primary goals of NASA Life Sciences research is '... to enable a permanent human presence in space.' To meet this goal, NASA is creating alternative protocols designed to evaluate and test countermeasures that will account for and correct the environmental effects of space flight on crewmembers health, safety, and operational performance. NASA investigators have previously evaluated the effects of long-duration space flight on physiology and performance of cosmonauts aboard the MIR space station. They also initiated tests of a countermeasure, Autogenic-Feedback Training Exercise (AFTE) designed to prevent and/or correct adverse effects, i.e., facilitate adaptation to space and re-adaptation to Earth. AFTE is a six-hour physiological training program that has proven to be a highly efficient and effective method for enabling people to monitor and voluntarily control a range of their own physiological responses, thereby minimizing adverse reactions to environmental stress. However, because of limited opportunities to test this technology with space flight crews, it is essential to find operational or 'real world' environments in which to validate the efficacy of this approach.

Cowings, Patricia S.↗

Mutualism in a Reduced Gravity Environment (MuRGE)

Mutualism in a Reduced Gravity Environment (MuRGE) is a ground research study to determine the feasibility of assessing fungi-plant (Piriformospora indica-Arabidopsis thaliana) interactions in microgravity. Seeds from the plant Arabiddospsis thaliana (At) will be grown in the presence of Piriformospora indica (Pi) an endophytic Sebacinacae family fungus. Pi is capable of colonizing the roots of a wide variety of plant species, including non-mycorrhizal hosts like At, and promoting plant growth similarly to AMF (arbusuclar mychorrizal fungi) unlike most AMF, Pi is not an obligate plant symbiont and can be grown in the absence of a host. In the presence of a suitable plant host, Pi can attach to and colonize root tips. Interaction visualization is accomplished with strong autofluorescence in the roots, followed by root colonization via fungal hyphae, and chlamydospore production. Increased root growth can be observed even before root colonization is detectable. In addition, Pi chlamydospores generated from axenic culture in microgravity will be used to inoculate roots of At grown in 1g to determine the effect of microgravity upon the inherent virulence or beneficial effects. Based on recent reports of increased virulence of S. typhimurium, P. aeruginosa, and S. Pneumoniae in reduced gravity, differences in microbial pathogenic responses and host plant systemic acquired resistance are expected. The focus of this project within MuRGE involved the development P. indica culture media evaluation and microscopy protocol development. High, clean spore harvest yields for the detection of fungi-plant interactions microscopically was the immediate goal of this experiment.

Haire, Timothy C.↗

Analog Exercise Hardware to Implement a High Intensity Exercise Program During Bed Rest

Background: In order to evaluate novel countermeasure protocols in a space flight analog prior to validation on the International Space Station (ISS), NASA's Human Research Program (HRP) is sponsoring a multi-investigator bedrest campaign that utilizes a combination of commercial and custom-made exercise training hardware to conduct daily resistive and aerobic exercise protocols. This paper will describe these pieces of hardware and how they are used to support current bedrest studies at NASA's Flight Analog Research Unit in Galveston, TX. Discussion: To implement candidate exercise countermeasure studies during extended bed rest studies the following analog hardware are being utilized: Stand alone Zero-Gravity Locomotion Simulator (sZLS) -- a custom built device by NASA, the sZLS allows bedrest subjects to remain supine as they run on a vertically-oriented treadmill (0-15 miles/hour). The treadmill includes a pneumatic subject loading device to provide variable body loading (0-100%) and a harness to keep the subject in contact with the motorized treadmill to provide a ground reaction force at their feet that is quantified by a Kistler Force Plate. Supine Cycle Ergometer -- a commercially available supine cycle ergometer (Lode, Groningen, Netherlands) is used for all cycle ergometer sessions. The ergometer has adjustable shoulder supports and handgrips to help stabilize the subject during exercise. Horizontal Squat Device (HSD) -- a custom built device by Quantum Fitness Corp (Stafford, TX), the HSD allows for squat exercises to be performed while lying in a supine position. The HSD can provide 0 to 600 pounds of force in selectable 5 lb increments, and allows hip translation in both the vertical and horizontal planes. Prone Leg Curl -- a commercially available prone leg curl machine (Cybex International Inc., Medway, MA) is used to complete leg curl exercises. Horizontal Leg Press -- a commercially available horizontal leg press (Quantum Fitness Corporation) is used for leg press and heel raise exercises. Minor modifications were made to the device including adding 200 lbs to the weight stack, raising the frame by 12 inches, making the footplate adjustable, and providing removable handles. Conclusion: A combination of novel and commercial exercise hardware are used to mimic the exercise hardware capabilities aboard the ISS, allowing scientific investigation of new countermeasure protocols in a space flight analog prior to flight validation

Loerch, Linda↗

Ambulation Increases Decompression Sickness in Spacewalk Simulations

Musculoskeletal activity has the potential to both improve and compromise decompression safety. Exercise enhances inert gas elimination during oxygen breathing prior to decompression (prebreathe), but it may also promote bubble nuclei formation (nucleation), which can lead to gas phase separation and bubble growth and increase the risk of decompression sickness (DCS). The timing, pattern and intensity of musculoskeletal activity and the level of tissue supersaturation may be critical to the net effect. Understanding the relationships is important to evaluate exercise prebreathe protocols and quantify decompression risk in gravity and microgravity environments. Data gathered during NASA's Prebreathe Reduction Program (PRP) studies combined oxygen prebreathe and exercise followed by low pressure (4.3 psi; altitude equivalent of 30,300 ft [9,235 m]) microgravity simulation to produce two protocols used by astronauts preparing for extravehicular activity. Both the Phase II/CEVIS (cycle ergometer vibration isolation system) and ISLE (in-suit light exercise) trials eliminated ambulation to more closely simulate the microgravity environment. The CEVIS results (35 male, 10 female) serve as control data for this NASA/Duke study to investigate the influence of ambulation exercise on bubble formation and the subsequent risk of DCS. METHODS Four experiments will replicate the CEVIS exercise-enhanced oxygen prebreathe protocol, each with a different exception. The first of these is currently underway. Experiment 1 - Subjects complete controlled ambulation (walking in place with fixed cadence and step height) during both preflight and at 4.3 psi instead of remaining nonambulatory throughout. Experiment 2 - Subjects remain non-ambulatory during the preflight period and ambulatory at 4.3 psi. Experiment 3 - Subjects ambulate during the preflight period and remain non-ambulatory at 4.3 psi. Experiment 4 - The order of heavy and light exercise employed in the CEVIS protocol is reversed, with the light exercise occurring first (subjects remain non-ambulatory throughout). Decompression stress is assessed with non-invasive ultrasound during each of 14 epochs of a 4 hour simulated spacewalk at 4.3 psi; aural Doppler is used to monitor bubbles (Spencer grade 0-IV scale) passing through the pulmonary artery, and two-dimensional echocardiographic imaging is used to look for left ventricular gas emboli (LVGE; the presence of which is a test termination criterion). Venous blood is collected at baseline and twice following repressurization to determine if the decompression stress is correlated with microparticles (cell fragments) accumulation. The plan is to test 25-50 subjects in each experiment. Fisher Exact Tests (one-tailed) are used to compare test and control groups. Trials are suspended when the DCS or grade IV VGE observations reach 70% confidence of DCS risk >15% and grade IV VGE risk >20%. RESULTS Experiment 1 was concluded with 20 complete trials (15 male, 5 female) since the statistical outcome would not change with five additional trials. The observed DCS was significantly greater in Experiment 1 than in CEVIS trials (4/20 [20%] vs. 0/45 [0%], respectively, p=0.007), as was the frequency of peak grade IV VGE (6/21 [29%; including one additional subject that presented grade IV VGE but whose trial was ended before completion when LVGE were observed] vs. 3/45 [7%], respectively, p=0.024). Experiment 3 trials are now underway, with 11 trials completed (10 male, 1 female). Preliminary results indicate no difference in observed DCS between Experiment 3 and CEVIS trials (1/11 [9%] vs. 0/45 [0%], respectively, p=0.196), or between Experiment 3 and Experiment 1 trials (p=0.405). The frequency of peak grade IV VGE in Experiment 3 (2/11 [18%]) did not differ from CEVIS or Experiment 1 trials (p=0.251 and p=0.425, respectively). Microparticle patterns are widely variable and still under analysis. DISCUSSION The results of the Experiment 1 trials support the thesis that decompression stress is increased by ambulation exercise, given the higher incidence of DCS and grade IV VGE when compared to the non-ambulatory PRP CEVIS trials. Experiment 3 trials are incomplete, but suggest that the effect of ambulation during ground level preflight oxygen breathing alone, when subjects are undersaturated with inert gas, may not differ in risk from ambulation at both preflight and spacesuit pressures, the latter when subjects are supersaturated with inert gas. Further trials are needed to confirm the relative effects of ambulation in undersaturated vs. supersaturated states and to determine whether light exercise facilitates the removal of heavy exercise-induced nucleation (Experiment 4).

Pollock, N. W.↗