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At least 667 records · Page 37

Waste to worth: a circular solution through lignin engineering

The inefficient management of industrial waste threatens environmental sustainability, requiring scalable resource recovery solutions. To address these challenges, we propose a novel and sustainable in-situ one-step strategy for the simultaneous recovery and functionalization of lignin from industrial black liquor, using a sustainable organic acid system assisted by choline chloride. The functionalized lignin adsorbent material (FLAM) exhibited superior adsorption performance, effectively treating animal feedlot wastewater to recover ammonia. The ammonia enriched FLAM was subsequently repurposed as a slow-release fertilizer, enhancing plant growth and demonstrating a closed-loop pathway for waste valorization and nutrient recycling. Methylene blue was used as a model cationic compound to simulate the adsorption behavior and interactions. FLAM achieved a high adsorption capacity of 725.98 mg/g across a broad pH range, significantly outperforming commercial kraft lignin. This sustainable approach advances circular economy principles by integrating remediation and resource recovery, offering a scalable, eco-friendly solution for environmental applications.

32 ENERGY CONSERVATION, CONSUMPTION, AND UTILIZATI↗

High-level expression, purification, crystallization and preliminary X-ray crystallographic studies of the receptor-binding domain of botulinum neurotoxin serotype D

Botulinum neurotoxins (BoNTs) are highly toxic proteins for humans and animals that are responsible for the deadly neuroparalytic disease botulism. Here, details of the expression and purification of the receptor-binding domain (HCR) of BoNT/D in Escherichia coli are presented. Using a codon-optimized cDNA, BoNT/D{_}HCR was expressed at a high level (150-200 mg per litre of culture) in the soluble fraction. Following a three-step purification protocol, very pure (>98%) BoNT/D{_}HCR was obtained. The recombinant BoNT/D{_}HCR was crystallized and the crystals diffracted to 1.65 {angstrom} resolution. The crystals belonged to space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 60.8, b = 89.7, c = 93.9 {angstrom}. Preliminary crystallographic data analysis revealed the presence of one molecule in the asymmetric unit.

59 BASIC BIOLOGICAL SCIENCES↗

Bayesian estimation of dose thresholds

An example is described of Bayesian estimation of radiation absorbed dose thresholds (subsequently simply referred to as dose thresholds) using a specific parametric model applied to a data set on mice exposed to 60Co gamma rays and fission neutrons. A Weibull based relative risk model with a dose threshold parameter was used to analyse, as an example, lung cancer mortality and determine the posterior density for the threshold dose after single exposures to 60Co gamma rays or fission neutrons from the JANUS reactor at Argonne National Laboratory. The data consisted of survival, censoring times and cause of death information for male B6CF1 unexposed and exposed mice. The 60Co gamma whole-body doses for the two exposed groups were 0.86 and 1.37 Gy. The neutron whole-body doses were 0.19 and 0.38 Gy. Marginal posterior densities for the dose thresholds for neutron and gamma radiation were calculated with numerical integration and found to have quite different shapes. The density of the threshold for 60Co is unimodal with a mode at about 0.50 Gy. The threshold density for fission neutrons declines monotonically from a maximum value at zero with increasing doses. The posterior densities for all other parameters were similar for the two radiation types.

NASA Discipline Radiation Health↗

Improved Method for Isolation of Microbial RNA from Biofuel Feedstock for Metatranscriptomics

Metatranscriptomics—gene express profiling via DNA sequencing—is a powerful tool to identify genes that are ac- tively expressed and might contribute to the phenotype of individual organisms or the phenome (the sum of several phenotypes) of a microbial community. Furthermore, metatranscriptome studies can result in extensive catalogues of genes that encode for enzymes of industrial relevance. In both cases, a major challenge for generating a high quality metatranscriptome is the extreme lability of RNA and its susceptibility to ubiquitous RNAses. The microbial commu- nity (the microbiome) of the cow rumen efficiently degrades lignocelullosic biomass, generates significant amounts of methane, a greenhouse gas twenty times more potent than carbon dioxide, and is of general importance for the physio- logical wellbeing of the host animal. Metatranscriptomes of the rumen microbiome from animals kept under different conditions and from various types of rumen-incubated biomass can be expected to provide new insights into these highly interesting phenotypes and subsequently provide the framework for an enhanced understanding of this socio- economically important ecosystem. The ability to isolate large amounts of intact RNA will significantly facilitate accu- rate transcript annotation and expression profiling. Here we report a method that combines mechanical disruption with chemical homogenization of the sample material and consistently yields 1 mg of intact RNA from 1 g of rumen-in- cubated biofuel feedstock. The yield of total RNA obtained with our method exceeds the RNA yield achieved with pre- viously reported isolation techniques, which renders RNA isolated with the method presented here as an ideal starting material for metatranscriptomic analyses and other molecular biology applications that require significant amounts of starting material.

Environmental Molecular Sciences Laboratory↗

Neuro-Vestibular Examination During and Following Spaceflight (Vestibular Health)

BACKGROUND Adaptation to microgravity during spaceflight causes neurological disturbances that are either directly or indirectly mediated by the vestibular system. These disturbances can include space motion sickness, spatial disorientation, and cognitive impairment, as well as changes in head-eye coordination, vestibulo-ocular reflexes, and control of posture and locomotion. Otolith-mediated reflex gains appear to adapt rapidly during spaceflight and after landing. However, animal studies have shown that structural modifications of the vestibular sensory apparatus develop during long-duration spaceflight. To date, no studies have characterized the severity of vestibular syndromes experienced by astronauts as a function of the duration of spaceflight or whether the effects are caused by changes at the peripheral end organs, midbrain, cerebellum, or vestibular cortex. OBJECTIVES We are investigating temporal vestibular changes in crewmembers of short, 6-month, and one-year missions to identify trends in adaptation of vestibular health and performance in orbit and after landing. We are also differentiating between peripheral and central vestibular forms of vertigo and oculomotor disorders. METHODS Recordings of eye, head, and body movements, as well as subjective reports of perception of motion, are being used to determine the presence of abnormal eye movements, dysmetria, motion sickness symptoms, and illusions of motion during head or body movements. This includes characterization of temporal trends in central compensation for vestibular (otolith) asymmetry. In-flight examinations are being performed early in the mission (Flight Days 1 and 30) and once every 2-3 months thereafter. Postflight examinations are performed after return (R) from the mission on R+0, R+4, R+9, and R+30. The inflight and postflight motion sickness questionnaires are customized to support data sharing across related studies. Ground-based control testing has been performed on healthy volunteers(18 females, 14 males;38.6 ± 9.2 years) in the laboratory to estimate mean normative responses, and on patients with bilateral vestibulopathy (BVP) (17 females, 13 males; 60.6 ± 13.0 years) at the University of Caen. RESULTS As of September 2024, two crewmembers have completed all preflight, inflight, and postflight testing. Additional crewmembers are currently enrolled and data collection is currently ongoing. For ground testing, BVP patients performed similarly to previous postflight astronauts on R+0 in various walking performance tasks. Three additional body movement perception tasks have been tested. For the Triangle Completion Task, BVP patients had a larger mean angle of deviation and longer mean distance than healthy controls. For the Self-Rotation Task and Distance Perception Task, BVP patients had larger errors than healthy controls. These data suggest that vestibular deficiencies impact all aspects of body movement perception tested; whereas previous studies suggest that vestibular deficiencies are only associated with directional errors, not with overall trajectories/path lengths. These data will be compared to those of crewmembers during early postflight readaptation. RELEVANCE If the observed symptoms in crewmembers are more deleterious after the year-long missions than those documented after 6-month missions, then relevant countermeasures will be required to maintain the health and operational performance of astronauts during longer missions. Depending on the etiology of the vestibular syndrome revealed by these tests, countermeasures will be proposed based on vestibular rehabilitation therapies currently used in patients with vestibular disorders, such as habituation, gaze stabilization, and/or balance training exercises. ACKNOWLEDGEMENT This work is supported by NASA’s Human Research Program Human Health Countermeasures Element.

T R Macaulay↗

Effect of track structure and radioprotectors on the induction of oncogenic transformation in murine fibroblasts by heavy ions

The oncogenic potential of high-energy 56Fe particles (1 GeV/nucleon) accelerated with the Alternating Gradient Synchrotron at the Brookhaven National Laboratory was examined utilizing the mouse C3H 10T1/2 cell model. The dose-averaged LET for high-energy 56Fe is estimated to be 143 keV/micrometer with the exposure conditions used in this study. For 56Fe ions, the maximum relative biological effectiveness (RBEmax) values for cell survival and oncogenic transformation were 7.71 and 16.5 respectively. Compared to 150 keV/micrometer 4He nuclei, high-energy 56Fe nuclei were significantly less effective in cell killing and oncogenic induction. The prostaglandin E1 analog misoprostol, an effective oncoprotector of C3H 10T1/2 cells exposed to X rays, was evaluated for its potential as a radioprotector of oncogenic transformation with high-energy 56Fe. Exposure of cells to misoprostol did not alter 56Fe cytotoxicity or the rate of 56Fe-induced oncogenic transformation.

NASA Discipline Radiation Health↗

TU-H-CAMPUS-TeP2-02: FLASH Irradiation Improves the Therapeutic Index Following GI Tract Irradiation

Purpose: To investigate and characterize the radiobiological effectiveness of very high dose rate radiotherapy (FLASH) compared to conventional irradiation in an in vivo model. Methods: The gastrointestinal (GI) tract of C57BL/6 mice were irradiated with doses ranging between 10 and 18 Gy using a custom stereotactic jig. A Varian Clinac 21EX was modified to allow dose rates ranging from 0.05 to 240 Gy/s at the position of the mirror. With the gantry at 180 degrees, the jig holding the individual animals was placed above the mirror to take advantage of the reduced source to target distance. Mice were irradiated with 20MeV electrons. Following irradiation, the mice were monitored twice daily for morbidity and daily for weight changes. Results: Mice irradiated with FLASH irradiation had lower weight loss compared to the mice receiving conventional irradiation. Following FLASH irradiation, a maximum weight loss of ∼20% was observed at day 6 with subsequent recovery, while following conventional irradiation, higher weight losses was observed with fewer instances of recovery. Concerning survival, all mice in the conventionally irradiated groups had a 100% mortality in the range of 15.5–18 Gy, while the mice irradiated with FLASH irradiation had a 100% survival in the same range. Conclusion: These results have demonstrated proof of principle that FLASH irradiations have a dramatic impact on the overall survival of mice following GI tract irradiations. If the increase in the therapeutic window can be validated and understood, this would revolutionize the field of radiation oncology and lead to increased cure rates with reduced side effects following treatment, resulting in increased quality of life for cancer survivors. Funding: DoD, Award#:W81XWH-14-1-0014, Weston Havens Foundation, Bio-X (Stanford University), the Office of the Dean of the Medical School, the Office of the Provost (Stanford University), and the Swedish Childhood Cancer Foundation; BL and PM are founders of TibaRay,Inc.; BL and PM have received research grants from Varian and RaySearch Laboratory.

60 APPLIED LIFE SCIENCES↗

Annual report of monitoring at Morrill, Kansas, in 2010.

Carbon tetrachloride contamination in groundwater at Morrill, Kansas, was initially identified in 1985 during statewide testing of public water supply wells for volatile organic compounds (VOCs). High levels of nitrate were also present in the wells. The city of Morrill is located in Brown County in the northeastern corner of the state, about 7 mi east of Sabetha. The population of Morrill as of the 2000 census was approximately 277. All residents of Morrill now obtain their drinking water from the Sabetha municipal water system via a pipeline constructed in 1991. Starting in 1922, eight different public wells formerly served the Morrill municipal system at some time. Because of poor water quality, including high nitrate levels attributed to numerous animal feeding operations in the vicinity and application of fertilizer on agricultural lands, use of the local groundwater from any public well for municipal supply purposes was terminated in 1991 in favor of obtaining water from the Sabetha municipal water system. Investigations of the carbon tetrachloride and nitrate contamination by the Kansas Department of Health and Environment (KDHE) in 1989, 1994, and 1996 (KDHE 1989; GeoCore 1994a-e, 1996) identified a localized plume of carbon tetrachloride in groundwater extending downgradient from a grain storage facility located in the northwestern section of Morrill. The facility was formerly operated by the Commodity Credit Corporation (CCC), an agency of the U.S. Department of Agriculture (USDA), from 1950 to 1971. Since termination of the CCC/USDA grain storage operations in 1971, the property and existing grain bins have been used for private grain storage up to the present time. Prior to 1986, commercial grain fumigants were commonly used by the CCC/USDA, as well as private and commercial grain storage operations, to preserve grain. Because the identified carbon tetrachloride contamination could in part be linked to historical use of carbon tetrachloride-based fumigants at its former facility, in 2003 the CCC/USDA assumed responsibility for the site investigation of the carbon tetrachloride contamination. The CCC/USDA involvement began with development and implementation of a work plan for a Phase I expedited site characterization (Argonne 2003). That investigation and subsequent investigations (Argonne 2004, 2005a) were performed by the Environmental Science Division of Argonne National Laboratory. Argonne is a nonprofit, multidisciplinary research center operated by UChicago Argonne, LLC, for the U.S. Department of Energy (DOE). The CCC/USDA has entered into an interagency agreement with DOE, under which Argonne continues to provide technical assistance to the CCC/USDA with environmental site characterization and remediation at its former grain storage facilities. The initial investigation by the CCC/USDA in 2003 determined that soils at the former facility have not been impacted by grain fumigation activities. Neither carbon tetrachloride nor chloroform was detected in near-surface soils or in subsurface soils collected to bedrock or to a depth of 15 ft below ground level (BGL). Therefore, no identifiable human health risk is associated with either carbon tetrachloride or chloroform in shallow soils, which additionally pose no further threat of contamination to groundwater. High carbon tetrachloride concentrations in groundwater (maximum 390 {micro}g/L in a sample collected from monitoring well MW3S - located on the former CCC/USDA property - in 1995) have declined significantly during long-term monitoring by the KDHE and currently by the CCC/USDA. Maximum levels within the plume of < 50 {micro}g/L at present confirm that no continuing soil source remains at the former CCC/USDA facility. Nevertheless, carbon tetrachloride concentrations exceeding the KDHE Tier 2 risk-based screening level of 5.0 {micro}g/L remain. In September 2005, the CCC/USDA initiated periodic sampling of groundwater at Morrill, in accord with a monitoring program approved by the state (KDHE 2005), to monitor carbon tetrachloride concentrations in the groundwater. Under the KDHE-approved monitoring plan (Argonne 2005b), groundwater was initially sampled twice yearly for a period of two years. The samples were analyzed for VOCs, as well as for selected geochemical parameters to aid in the evaluation of possible natural contaminant degradation (reductive dechlorination) processes in the subsurface environment. The data have been inconclusive overall, possibly because of the relatively low contaminant concentrations in the plume. Nevertheless, consistently low levels of dissolved oxygen (DO) and oxidation-reduction potential (ORP) at monitoring well MW1D (in the deepest portion of the contaminated aquifer) and the presence of chloroform (the primary degradation product of carbon tetrachloride) suggest that some degree of reductive dechlorination is occurring.

54 ENVIRONMENTAL SCIENCES↗

Oxidative Stress and Skeletal Health with Low-Dose, Low-LET (Linear Energy Transfer) Ionizing Radiation

We performed in vivo and in vitro experiments to accomplish the following specific aims of this project: 1) determine if low dose, low LET radiation affects skeletal remodeling at structural, cellular and molecular levels and 2) determine if low dose, low LET radiation modulates skeletal health during aging via oxidative mechanisms. A third aim is supported by NASA supplement to this DOE grant focusing on the influence of high LET radiation on bone. A series of experiments were conducted at the NASA Space Radiation Laboratory at Brookhaven, NSRL-BNL, using iron (56Fe) or a sequential exposure to protons / iron / protons, and separate experiments at NASA Ames Research Center (ARC) using 137Cs. The following provides a summary of key findings. (1) Exposure of nine-week old female mice to priming doses of gamma radiation (10cGy x 5) did not significantly affect bone volume/total volume (BV/TV) or microarchitecture as analyzed by 3D microcomputed tomography. As expected, exposure to the challenge dose of 2 Gy gamma irradiation resulted in significant decreases in BV/TV. The priming dose combined with the 2Gy challenge dose had no further effect on BV/TV compared to challenge dose alone, with the sole exception of the Structural Model Index (SMI). SMI reflects the ratio of rods-to-plates in cancellous bone tissue, such that higher SMI values indicate a tendency toward a weaker structure compared to lower SMI values. Mice treated with both priming and challenge dose had 25% higher SMI values compared to sham-irradiated controls and 7% higher values compared to mice treated with the challenge dose alone. Thus, although this priming regimen had relatively modest effects on cancellous tissue, the difference in SMI suggests this fractionated priming doses have adverse, rather than beneficial, effects on bone structure. (2) In 10-week old male mice, a single exposure to 100cGy of 137Cs reduces trabecular bone number and connectivity density by 20% and 36% respectively one month after irradiation (IR). At four months post-IR, these animals were comparable to sham-treated controls with regards to the abovementioned structural parameters. Irradation at 1 or 10 cGy did not result in any significant changes in bone structural parameters. (3) Irradiation of 16-wk old male mice with high doses of 56Fe or proton (50 or 200cGy), but not at low doses (5 or 10cGy), showed a similar loss of cancellous BV/TV and trabecular number at five weeks post-IR. (4) Age-related bone loss overtook acute radiation-induced decrements in bone structure within four months post-IR with 100 cGy gamma and 12 months post-IR with 200 cGy iron. Transgenic mice globally overexpressing human catalase gene in mitochondria did not exhibit cancellous bone loss as assessed at four month post-IR with 10 cGy proton, 50 cGy iron, or in combination. (5) The cellular and molecular mechanisms responsible for loss of bone with radiation are mediated primarily through increased osteoclastogenesis. Our data provide evidence that there are increases in gene expression of TNF alpha and MCP1 in the bone marrow cells 24 hours post-IR and of osteoclastogenic differentiation factor RANKL by day 3. These cytokines in the marrow may stimulate mature osteoclasts or drive osteoclastogenesis from precursors. (6) Osteoblastogenesis from marrow progenitors evaluated ex vivo decreased following whole body 56Fe irradiation at a dose threshold between 20 and 50 cGy whereas osteoclastogenesis ex vivo increased with doses as low as 10cGy two days post-IR of mice. However, the latter finding was not observed in more than a single experiment. (7) Gamma irradiation of cells in vitro requires relatively high doses (200cGy) to disturb normal osteoblastogenesis and osteoclastogenesis as evidenced by decrements in mineralized nodule formation, osteoclast counts, and expression of osteoblast related genes such as runx2, col1a1. (8) We also investigated the effect of antioxidants on osteoblastogenesis following low dose in vitro gamma irradiation (15cGy) on day four bone marrow stromal cell cultures. Superoxide dismutase (SOD) was added to the cell culture medium for 2 or 3 days post-irradiation and cell colonies were counted on days 7 and 10. SOD treatment increased cell growth as measured by DNA content and colony forming units (CFU) in both irradiated cells and 0 cGy control groups. However, low dose radiation of 15cGy abolished SOD stimulatory effects on cell growth and CFU number. These results suggest that exogenous SOD increases osteoblast cell growth and colony formation and that low-dose radiation (15cGy) can interfere with the antioxidant effects. In summary, our findings indicate that acute, whole body irradiation at high doses (50-200 cGy) results in prompt tissue degradation and bone loss. Lower doses (<50 cGy) do not cause bone structural deterioration but may deplete stem/progenitor cell pools in the bone marrow.

59 BASIC BIOLOGICAL SCIENCES↗

Neuro-Vestibular Examination During and Following Spaceflight (Vestibular Health)

BACKGROUND Adaptation to microgravity during spaceflight causes neurological disturbances that are either directly or indirectly mediated by the vestibular system. These disturbances could include space motion sickness, spatial disorientation, and cognitive impairment, as well as changes in head-eye coordination, vestibulo-ocular reflexes, and control of posture and locomotion. Otolith-mediated reflex gains appear to adapt rapidly during spaceflight and after landing. However, animal studies have shown that structural modifications of the vestibular sensory apparatus develop during long-duration spaceflight. To date, no studies have characterized the severity of vestibular syndromes experienced by astronauts as a function of the duration of spaceflight or whether the effects are caused by changes at the peripheral end organs, midbrain, cerebellum, or vestibular cortex. OBJECTIVES We will investigate temporal vestibular changes in crewmembers of short, 6-month, and one-year missions to identify trends in adaptation of vestibular health and performance in orbit and after landing. We will also determine whether the vestibular organs and/or the central vestibular system undergo structural changes during long-duration exposure to microgravity, which could cause vestibular disorders when transitioning to a different gravitational environment. METHODS Recordings of eye, head, and body movements, as well as subjective reports of perception of motion, will be used to determine the presence of abnormal eye movements, dysmetria, motion sickness symptoms, and illusions of motion during head or body movements. This includes characterization of temporal trends in central compensation for vestibular (otolith) asymmetry. Pre-flight data will be collected 90 days before the flight. In-flight tests will be performed early in the mission and once every one or two months thereafter. Post-flight examinations will be performed on the following days after return (R) from the mission: R+0, R+4, R+9, and R+30. Ground-based control tests will be performed on healthy volunteers in the laboratory to estimate mean normative responses. CONTROL RESULTS Thirty-two healthy (non-astronaut) control subjects performed the same ground test procedures as planned for crewmembers. In addition to establishing a normative database, these data were used to calculate vestibular asymmetries from perceptual reports during unilateral centrifugation, oculomotor responses during visual-alignment tasks, vestibulo-ocular reflex gain during head-impulse tests, and body rotation during stepping tests. A significant correlation was observed between asymmetries of subjective visual vertical and verbal report during unilateral centrifugation. Another significant correlation was observed between the asymmetries of ocular alignment, vestibulo-ocular reflex gain, and body rotation. These findings in a healthy cohort may help us better understand changes in vestibular asymmetries in crewmembers during and following spaceflight. RELEVANCE If the observed symptoms in crewmembers are more deleterious after the year-long missions than those documented after 6-month missions, then relevant countermeasures will be required to maintain the health and operational performance of astronauts during longer missions. Depending on the etiology of the vestibular syndrome revealed by these tests, countermeasures will be proposed based on vestibular rehabilitation therapies currently used in patients with vestibular disorders, such as habituation, gaze stabilization, and/or balance training exercises. ACKNOWLEDGEMENT This work is supported by the NASA’s Human Research Program Human Health Countermeasures Element.

T R Macaulay↗

MALDI MS Imaging at Acquisition Rates Exceeding 100 Pixels per Second

The practicality of matrix-assisted laser desorption/ionization (MALDI) time-of-flight (TOF) mass spectrometry (MS) applied to molecular imaging of biological tissues is limited by the analysis speed. Typically, a relatively low speed of stop-and-go micromotion of XY stages is considered as a factor substantially reducing the rate with which fresh sample material can be supplied to the laser spot. The sample scan rate in our laboratory-built high-throughput imaging TOF mass spectrometer was significantly improved through the use of a galvanometer-based optical scanner performing fast laser spot repositioning on a target plate. The optical system incorporated into the ion source of our MALDI TOF mass spectrometer allowed focusing the laser beam via a modified grid into a 10-μm round spot. This permitted the acquisition of high-resolution MS images with a well-defined pixel size at acquisition rates exceeding 100 pixel/s. The influence of selected parameters on the total MS imaging time is discussed. The new scanning technique was employed to display the distribution of an antitumor agent in 3D colorectal adenocarcinoma cell aggregates; a single MS image comprising 100 × 100 pixels with 10-μm lateral resolution was recorded in approximately 70 s. .

46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND ↗

Microbiomes of frozen blood plasma samples reveal potential pathogens in wild birds and rodents

The lack of genomic data on pathogens from wildlife severely limits our ability to track transmission patterns and trace the origins of an outbreak. There are currently millions of wildlife samples in biobanks around the world, including blood samples. Blood has traditionally been viewed as a sterile environment in healthy individuals, but recent evidence suggests that this is not the case, especially for wild animals. Our goal was to determine whether frozen plasma samples can be surveyed using 16S sequencing to provide information about potential hosts for pathogens for a more complete understanding of disease systems. We sequenced blood plasma from wild North American deer mice ( Peromyscus maniculatus ) and American kestrels ( Falco sparverius ) that were cryogenically stored for 7 and 13 years, respectively, and compared two DNA extraction kits. The kestrel samples contained a very high number of reads that could not be identified to phylum compared to the mouse samples. The two kits differed in the phyla and genera that were detected, and the Zymo kit, which is optimized for plasma and serum, produced more high-quality reads for both kestrel and mouse samples. We identified several pathogenic genera, including Mycoplasma, Escherichia-Shigella , and Bartonella . Sequencing blood samples for pathogens could potentially have broad applications for identifying important reservoir hosts for pathogen transmission and provide a reduced set of species on which to follow up.

59 BASIC BIOLOGICAL SCIENCES↗

Monte Carlo Simulations Of The Dose Distributions From Carbon Microbeams Used In An Experimental Radiation Therapy Method

Recent upgrades of the MCNPX Monte Carlo code include transport of heavy ions. We employed the new code to simulate the energy and dose distributions produced by carbon beams in rabbit's head in and around a brain tumor. The work was within our experimental technique of interlaced carbon microbeams, which uses two 90 deg. arrays of parallel, thin planes of carbon beams (microbeams) interlacing to produce a solid beam at the target. A similar version of the method was earlier developed with synchrotron-generated x-ray microbeams. We first simulated the Bragg peak in high density polyethylene and other materials, where we could compare the calculated carbon energy deposition to the measured data produced at the NASA Space Radiation Laboratory (NSRL) at Brookhaven National Laboratory (BNL). The results showed that new MCNPX code gives a reasonable account of the carbon beam's dose up to {approx}200 MeV/nucleon beam energy. At higher energies, which were not relevant to our project, the model failed to reproduce the Bragg-peak's extent of increasing nuclear breakup tail. In our model calculations we determined the dose distribution along the beam path, including the angular straggling of the microbeams, and used the data for determining the optimal values of beam spacing in the array for producing adequate beam interlacing at the target. We also determined, for the purpose of Bragg-peak spreading at the target, the relative beam intensities of the consecutive exposures with stepwise lower beam energies, and simulated the resulting dose distribution in the spread out Bragg-peak. The details of the simulation methods used and the results obtained are presented.

62 RADIOLOGY AND NUCLEAR MEDICINE↗

Magnetic levitation-based Martian and Lunar gravity simulator

Missions to Mars will subject living specimens to a range of low gravity environments. Deleterious biological effects of prolonged exposure to Martian gravity (0.38 g), Lunar gravity (0.17 g), and microgravity are expected, but the mechanisms involved and potential for remedies are unknown. We are proposing the development of a facility that provides a simulated Martian and Lunar gravity environment for experiments on biological systems in a well controlled laboratory setting. The magnetic adjustable gravity simulator will employ intense, inhomogeneous magnetic fields to exert magnetic body forces on a specimen that oppose the body force of gravity. By adjusting the magnetic field, it is possible to continuously adjust the total body force acting on a specimen. The simulator system considered consists of a superconducting solenoid with a room temperature bore sufficiently large to accommodate small whole organisms, cell cultures, and gravity sensitive bio-molecular solutions. It will have good optical access so that the organisms can be viewed in situ. This facility will be valuable for experimental observations and public demonstrations of systems in simulated reduced gravity. c2005 Published by Elsevier Ltd on behalf of COSPAR.

Non-NASA Center↗

Neuro-Vestibular Examination During and Following Spaceflight (Vestibular Health)

BACKGROUND Adaptation to microgravity during spaceflight causes neurological disturbances that are either directly or indirectly mediated by the vestibular system. These disturbances can include space motion sickness, spatial disorientation, and cognitive impairment, as well as changes in head-eye coordination, vestibulo-ocular reflexes, and control of posture and locomotion. Otolith-mediated reflex gains appear to adapt rapidly during spaceflight and after landing. However, animal studies have shown that structural modifications of the vestibular sensory apparatus develop during long-duration spaceflight. To date, no studies have characterized the severity of vestibular syndromes experienced by astronauts as a function of the duration of spaceflight or whether the effects are caused by changes at the peripheral end organs, midbrain, cerebellum, or vestibular cortex. OBJECTIVES We will investigate temporal vestibular changes in crewmembers of short, 6-month, and one-year missions to identify trends in adaptation of vestibular health and performance in orbit and after landing. We will also determine whether the vestibular organs and/or the central vestibular system undergo structural changes during long-duration exposure to microgravity, which could cause vestibular disorders when transitioning to a different gravitational environment. METHODS Recordings of eye, head, and body movements, as well as subjective reports of perception of motion, will be used to determine the presence of abnormal eye movements, dysmetria, motion sickness symptoms, and illusions of motion during head or body movements. This includes characterization of temporal trends in central compensation for vestibular (otolith) asymmetry. Pre-flight data will be collected 90 days before launch. In-flight examinations will be performed early in the mission (Flight Days 1 and 30) and once every two or three months thereafter. Post-flight examinations will be performed on the following days after return (R) from the mission: R+0, R+4, R+9, and R+30. Ground-based control tests have been performed on healthy volunteers in the laboratory to estimate mean normative responses. RESULTS Data collection for this study is ongoing. Data processing techniques are being refined. Our eye tracking method for measuring three-dimensional eye movement takes advantage of modern computer vision software (OpenCV) and advances in the field of iris recognition to improve measurement of ocular-counterolling. RELEVANCE If the observed symptoms in crewmembers are more deleterious after the year-long missions than those documented after 6-month missions, then relevant countermeasures will be required to maintain the health and operational performance of astronauts during longer missions. Depending on the etiology of the vestibular syndrome revealed by these tests, countermeasures will be proposed based on vestibular rehabilitation therapies currently used in patients with vestibular disorders, such as habituation, gaze stabilization, and/or balance training exercises.

T R Macaulay↗

Maternal undernutrition modulates hepatic MicroRNAs expression in the early life of offspring

Emerging studies revealed that a poor intrauterine environment elicited by maternal nutrient restriction (MNR) is associated with an increased risk of metabolic diseases in adulthood. Previous research has shown that microRNAs (miRNAs) exert pivotal roles in modulating molecular pathways involved in disease pathogenesis and progression. In this respect, we herein examined miRNA profiles in samples of liver from offspring whose mothers were fed either with a 50% food-restricted diet or standard laboratory chow during pregnancy. Our findings enumerated that miR-181a, involved in lipid metabolism, was found to be downregulated in the liver of MNR offspring at 1 day of age when compared to that of control offspring. We also noted that overexpression of miR-181a reduced the lipid droplets after treatment with oleic acid for 48 h, which suppressed the expressions levels of SIRT1, FOXO1, KLF6 and PPARγ in BRL-3A cells, while the opposite results were observed with decreased expression of miR-181a. Furthermore, the luciferase reporter assay confirmed the direct interactions between miR-181a with KLF6 and SIRT1. In adults, the MNR offspring elucidated increased TG content, decreased expression of miR-181a, and increased expressions levels of SIRT1, FOXO1, KLF6, and PPARγ in liver tissues. Collectively, these findings provided novel evidence that MNR could regulate miRNAs expression, which might be related to lipid metabolism in MNR offspring.

60 APPLIED LIFE SCIENCES↗

ACBD3 is up-regulated in gastric cancer and promotes cell cycle G1-to-S transition in an AKT-dependent manner

Highlights: • ACBD3 is up-regulated in gastric cancer. • ACBD3 regulates cell-cycle of gastric cancer in an AKT-dependent manner. It has been reported that ACBD3 is closely related to the malignant process of cells, but its role in gastric cancer has not been elucidated. This study aims to investigate the expression and function of ACBD3 in human gastric cancer. The Cancer Genome Atlas (TCGA) database were selected to analyze mRNA levels of ACBD3 in gastric cancer tissues and normal gastric epithelial tissues. qPCR and Western blot were conducted to detect the expression of ACBD3 in two normal gastric epithelial cell lines and five gastric cancer cell lines which were cultured in our laboratory. To exclude differences in individual background between different patients, we further detected the expression of ACBD3 in 8 pairs of malignant/non-malignant clinical gastric tissues. Through the establishment of stable cells, in vitro cell experiments and in vivo xenotransplantation models in mice, the role of ACBD3 in the proliferation of gastric cancer cells has been further explored. AKT inhibitors were used to deeply explore the molecular regulation mechanism of ACBD3. The results showed that the elevated ACBD3 in gastric cancer tissue were positively correlated with the clinical grade and prognosis of gastric cancer. In terms of molecular function, we found that ACBD3 can enhance the production and growth of gastric cancer cells. At the same time, the activation of AKT kinase played an important role in ACBD3's promotion of G1-to-S transition. The experiments generally indicate that ACBD3 is expected to become a potential diagnostic molecule or therapeutic target for gastric cancer.

60 APPLIED LIFE SCIENCES↗

Generation of a Slc39a8 hypomorph mouse: Markedly decreased ZIP8 Zn{sup 2+}/(HCO{sub 3}{sup -}){sub 2} transporter expression

Highlights: {yields} The mouse Slc39a8 gene encodes the ZIP8 transporter. {yields} ZIP8 functions endogenously as a electroneutral Zn{sup 2+}/(HCO{sub 3}{sup -}){sub 2} symporter. {yields} A Slc39a8(neo/neo) hypomorph mouse, due to retention of the neo mini-gene, has been created. {yields} ZIP8 expression in utero is {approx}90% decreased in all tissues examined. {yields} This mouse model will be useful for studying developmental and in utero physiological functions of ZIP8. -- Abstract: Previously this laboratory has identified the mouse Slc39a8 gene encoding the ZIP8 transporter, important in cadmium uptake. ZIP8 functions endogenously as a electroneutral Zn{sup 2+}/(HCO{sub 3}{sup -}){sub 2} symporter, moving both ions into the cell. The overall physiological importance of ZIP8 remains unclear. Herein we describe generation of a mouse line carrying the Slc39a8(neo) allele, containing the Frt-flanked neomycin-resistance (neo) mini-cassette in intron 3 and loxP sites in introns 3 and 6. Cre recombinase functions correctly in Escherichia coli and in adeno-Cre-infected mouse fetal fibroblasts, but does not function in the intact mouse for reasons not clear. Slc39a8(neo) is a hypomorphic allele, because Slc39a8(neo/neo) homozygotes exhibit dramatically decreased ZIP8 expression in embryo, fetus, and visceral yolk sac - in comparison to their littermate wild-type controls. This ZIP8 hypomorph will be instrumental in studying developmental and in utero physiological functions of the ZIP8 transporter.

60 APPLIED LIFE SCIENCES↗