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At least 145 records · Page 8

Calcium metabolism before, during, and after a 3-mo spaceflight: kinetic and biochemical changes

The loss of bone during spaceflight is considered a physiological obstacle for the exploration of other planets. This report of calcium metabolism before, during, and after long-duration spaceflight extends results from Skylab missions in the 1970s. Biochemical and endocrine indexes of calcium and bone metabolism were measured together with calcium absorption, excretion, and bone turnover using stable isotopes. Studies were conducted before, during, and after flight in three male subjects. Subjects varied in physical activity, yet all lost weight during flight. During flight, calcium intake and absorption decreased up to 50%, urinary calcium excretion increased up to 50%, and bone resorption (determined by kinetics or bone markers) increased by over 50%. Osteocalcin and bone-specific alkaline phosphatase, markers of bone formation, increased after flight. Subjects lost approximately 250 mg bone calcium per day during flight and regained bone calcium at a slower rate of approximately 100 mg/day for up to 3 mo after landing. Further studies are required to determine the time course of changes in calcium homeostasis during flight to develop and assess countermeasures against flight-induced bone loss.

NASA Discipline Regulatory Physiology↗

Effect of spaceflight on the functional, biochemical, and metabolic properties of skeletal muscle

This paper summarizes the effects of spaceflight on the functional, morphological, and biochemical properties of human and rodent skeletal muscle. The findings suggest that following as little as 5-6 in space there are deficits in both human and rodent motor capacity, strength, and endurance properties of skeletal muscle. The reduced strength is associated, in part, with a reduction in muscle mass as reflected in smaller cross-sectional areas of both fast- and slow-twitch fibers. Available evidence in animal models suggests that slow-twitch fibers are more sensitive to the atrophying process. Accompanying the atrophy is a transformation of slow to fast protein phenotype involving myosin heavy chain and sarcoplasmic reticulum protein isoforms. These transformations appear to be regulated, in part, by pretranslational processes. Data on the oxidative capacity of rodent skeletal muscle suggest a bias toward preferential utilization of carbohydrate as the primary substrate. These collective findings suggest that skeletal muscles comprised chiefly of slow fibers are highly dependent on gravity for the normal expression of protein mass and slow phenotype. Future studies need to focus on elucidating the mechanisms associated with the atrophy response, as well as identifying suitable exercise and other countermeasures capable of preserving the structural and functional integrity of skeletal muscle.

NASA Discipline Musculoskeletal↗

Amorphous material of the skin in amyotrophic lateral sclerosis: a morphologic and biochemical study

We performed morphologic studies on skin from seven patients with ALS and seven control subjects. By light microscopy, the wide spaces that separated collagen bundles reacted strongly with colloidal iron and alcian blue in ALS patients. Electron microscopy revealed markedly increased amorphous material that was positive for ruthenium red in the ground substance. These findings were not present in controls. Quantitative amino acid analysis showed that the amount of total amino acids (nmoles per mg dry weight) was significantly decreased (p < 0.01) in ALS patients compared with that of controls, and there was a significant negative correlation between skin amino acid content and duration of illness in ALS patients (r = -0.83, p < 0.001). These morphologic findings and biochemical data indicate that the amorphous material, which is markedly increased in ALS skin, includes glycosaminoglycans.

Non-NASA Center↗

Spaceflight effects on biomechanical and biochemical properties of rat vertebrae

The biomechanical and biochemical responses of lumbar vertebral bodies during a 12.5-day spaceflight (Cosmos 1887 biosatellite) were determined for rapidly growing rats (90-day-old, Czechoslovakian-Wistar). By use of age-matched vivarium controls (normal cage environment) and synchronous controls (simulated flight conditions), as well as a basal control group (killed before lift-off on the 1st day of flight), the combined influences of growth and space-flight could be examined. Centra of the sixth lumbar vertebrae (L6) were compressed to 50% strain at a fast strain rate while immersed in physiological buffer (37 degrees C). The body masses of vivarium and synchronous controls were significantly heavier than either the flight or basal controls. The flight group had an L6 vertebral body compressional stiffness that was 39% less than the vivarium controls, 47% less than the synchronous control, and 16% less than the basal controls. In addition, the average initial maximum load of the flight L6 was 22% less than vivarium controls and 18% less than the synchronous controls, whereas the linear compressional load of the flight group averaged 34% less than the vivarium and 25% less than the synchronous groups. The structural properties of the vertebrae from the 12.5-day-younger basal group closely resembled the flight vertebrae. Calcium, phosphorous, and hydroxyproline concentrations were not significantly different among the groups. Nevertheless, the lack of strength and stiffness development in spaceflight, coupled with a smaller proportion of mature hydroxypyridinoline cross-links, suggested that the 12.5 days of spaceflight slowed the maturation of trabecular bone in the vertebral bodies of rapidly growing rats.

Non-NASA Center↗

Blood biochemical and cellular changes during decompression and simulated extravehicular activity

Blood biochemical and cellular parameters were measured in human subjects before and after exposure to a decompression schedule involving 6 h of oxygen prebreathing. The exposure was designed to simulate extravehicular activity for 6 h (subjects performed exercise while exposed to 29.6 kPa). There were no significant differences between blood samples from subjects who were susceptible (n = 11) versus those who were resistant (n = 27) to formation of venous gas emboli. Although several statistically significant (P less than 0.05) changes in blood parameters were observed following the exposure (increases in white blood cell count, prothrombin time, and total bilirubin, and decreases in triglycerides, very-low-density lipoprotein cholesterol, and blood urea nitrogen), the changes were small in magnitude and blood factor levels remained within normal clinical ranges. Thus, the decompression schedule used in this study is not likely to result in blood changes that would pose a threat to astronauts during extravehicular activity.

Decompression Sickness/blood/etiology/prevention &↗

Biochemical Kinetics Model of DSB Repair and GammaH2AX FOCI by Non-homologous End Joining

We developed a biochemical kinetics approach to describe the repair of double strand breaks (DSB) produced by low LET radiation by modeling molecular events associated with the mechanisms of non-homologous end-joining (NHEJ). A system of coupled non-linear ordinary differential equations describes the induction of DSB and activation pathways for major NHEJ components including Ku(sub 70/80), DNA-PK(sub cs), and the Ligase IV-XRCC4 hetero-dimer. The autophosphorylation of DNA-PK(sub cs and subsequent induction of gamma-H2AX foci observed after ionizing radiation exposure were modeled. A two-step model of DNA-PK(sub cs) regulation of repair was developed with the initial step allowing access of other NHEJ components to breaks, and a second step limiting access to Ligase IV-XRCC4. Our model assumes that the transition from the first to second-step depends on DSB complexity, with a much slower-rate for complex DSB. The model faithfully reproduced several experimental data sets, including DSB rejoining as measured by pulsed-field electrophoresis (PFGE), quantification of the induction of gamma-H2AX foci, and live cell imaging of the induction of Ku(sub 70/80). Predictions are made for the behaviors of NHEJ components at low doses and dose-rates, where a steady-state is found at dose-rates of 0.1 Gy/hr or lower.

Cucinotta, Francis, A.↗

Thin membrane sensor with biochemical switch

A modular biosensor system for chemical or biological agent detection utilizes electrochemical measurement of an ion current across a gate membrane triggered by the reaction of the target agent with a recognition protein conjugated to a channel blocker. The sensor system includes a bioresponse simulator or biochemical switch module which contains the recognition protein-channel blocker conjugate, and in which the detection reactions occur, and a transducer module which contains a gate membrane and a measuring electrode, and in which the presence of agent is sensed electrically. In the poised state, ion channels in the gate membrane are blocked by the recognition protein-channel blocker conjugate. Detection reactions remove the recognition protein-channel blocker conjugate from the ion channels, thus eliciting an ion current surge in the gate membrane which subsequently triggers an output alarm. Sufficiently large currents are generated that simple direct current electronics are adequate for the measurements. The biosensor has applications for environmental, medical, and industrial use.

Case, George D.↗

Rapid Biochemical Analysis on the International Space Station (ISS): Preparing for Human Exploration of the Moon and Mars

The Lab-on-a-Chip Application Development - Portable Test System, known as LOCAD-PTS, was launched to the International Space Station (ISS) aboard Space Shuttle Discovery (STS-116) on December 9th,2006. Since that time, it has remained onboard ISS and has been operated by the crew on 10 separate occasions LOCAD-PTS is a handheld device for rapid biochemical analysis; it consists of a spectrophotometer, a series of interchangeable cartridges, a pipette and several clean/sterilized swabbing kits to obtain samples from ISS surfaces. Sampling, quantitative analysis and data retrieval is performed onboard, therefore reducing the need to return samples to Earth. Less than 20 minutes are required from sampling to data, significantly faster than existing culture-based methods on ISS, which require 3-5 days. Different cartridges are available for the detection of different target molecules (simply by changing the formulation within each cartridge), thereby maximizing the benefit and applications addressed by a single instrument. Initial tests on ISS have focused on the detection of the bact.erial macromolecule endotoxin, a component of bacterial cell walls. LOCAD-PTS detects endotoxin with a cartridge that contains a formulation known as Limulus Amebocyte Lysate (LAL) assay. LAL is derived from blood of the horseshoe crab, Limulus polyphemus, and detects enodotoxin with an enzyme cascade that triggers generation Of a yellow colored dye, p-nitroanaline. The more p-nitroanaline product, the more endotoxin is in the original sample. To enable quantitative analysis, the absorbance of this color is measured by LOCAD-PTS through a 395 nm filter and compared with an internal calibration curve, to provide a reading on the LED display that ranges from 0.05 Endotoxin Units (EU)/ml to 5 EU/ml. Several surface sites were analyzed within ISS between March 2007 and February 2008, including multiple locations in the US Laboratory Destiny, Node 1 Unity, AMock, and Service Module Zvezda. The goals of this initial study were to i) test the cleanliness of reagents/supplies on orbit, ii) test the crew's ability to collect and process a sample in microgravity without contamination, iii) demonstrate nominal function of the LOCAD-PTS, and iv) provide a general survey of endotoxin within the ISS. The surface sites varied greatly in terms of their frequency-of-use and material texture/composition; from relatively smooth aluminum, to fabric, to the room temperature vulcanizing (RTV) rubber of a Extravehicular Mobility Unit (EMU) spacesuit. Results showed that: i) the swabbing kits and reagents remained clean on orbit, ii) the crew could collect and process a sample without contamination, and iii) the LOCAD-PTS functioned nominally in > 99% of the 55 tests completed. We will present detailed results of the survey of endotoxin on ISS surfaces. These results and technology are important in the near-term - by providing an extra tool in the toolbox for ISS microbial monitoring. They are also important in the longer term as valuable preparation for human exploration of the Moon and Mars. One of the proposed science goals for the human exploration of Mars will be to detect and characterize any indigenous biological molecules that may exist on the Martian surface. To achieve that goal, the crew must have the technology available onboard to differentiate indigenous biology from any terrestrial biological material brought to Mars by the spacecraft and crew (termed 'forward contamination'). The LAL assay is already one of the official methods used by NASA's planetary protection program to certify cleanliness of interplanetary robotic spacecraft prior to launch; and therefore endotoxin is a good marker of forward contamination (as well as other microbial molecules detectable with LOCAD-PTS e.g. box-1, 3-glucan and lipoteichoic acid). Furthermore, the distribution and abundance of these molecules on the ISS provides a good indicator of what to expect on the Crew Exploratioehicle Orion, the lunar lander Antares, and future crewed spacecraft destined for Mars. In addition, technology such as LOCAD-PTS has been proposed to help evaluate forward contamination during lunar surface operations by the crew, as preparation for the human exploration of Mars.

Maule, J.↗

Alendronate and Resistive Exercise Countermeasures Against Bed Rest-Induced Bone Loss: Biochemical Markers of Bone and Calcium Metabolism

Weightlessness-induced bone loss must be counteracted to ensure crew health during extendedduration space missions. Studies were conducted to assess two bone loss countermeasures in a ground-based model: horizontal bed rest. Following a 3-wk ambulatory adaptation period, male and female subjects (aged 21-56 y) completed a 17-wk bed rest protocol. Subjects were assigned to one of three treatments: alendronate (ALEN; 10 mg/d, n=6), resistive exercise (RE; 1.5 h/d, 6 d/wk, n=8), or control (CN; no countermeasure, n=8). Dietary intake was adjusted to maintain body weight. Endocrine and biochemical indices were measured in blood and urine using standard laboratory methods. All data reported are expressed as percent change from individual pre-bedrest data. Serum calcium changed little during bed rest, and tended to decrease (4-8%) in ALEN subjects. In RE subjects, bone alkaline phosphatase and osteocalcin were increased >65 and >30%, respectively, during bed rest, while these were unchanged or decreased in ALEN and CN subjects. Urinary calcium was increased 50% in CN subjects, but was unchanged or decreased in both ALEN and RE groups. Urinary n-telopeptide excretion was increased 40-50% in CN and RE subjects, but decreased 20% in ALEN subjects. Pyridinium crosslink and deoxypyridinoline excretion were increased 20-50% during bed rest. These data suggest that RE countermeasures are effective at increasing markers of bone formation in an analog of weightlessness, while ALEN reduces markers of bone resorption. Counteracting the bone loss of space flight may require both pharmacologic and exercise countermeasures.

Smith, Scott M.↗

Biochemical Detection and Identification False Alarm Rate Dependence on Wavelength Using Laser Induced Fluorescence

Most organic and many inorganic materials absorb strongly in specific wavelength ranges in the deep UV between about 220nm and 300nm. Excitation within these absorption bands results in native fluorescence emission. Each compound or composite material, such as a bacterial spore, has a unique excitation-emission fingerprint that can be used to provide information about the material. The sensitivity and specificity with which these materials can be detected and identified depends on the excitation wavelength and the number and location of observation wavelengths.We will present data on our deep ultraviolet Targeted Ultraviolet Chemical Sensors that demonstrate the sensitivity and specificity of the sensors. In particular, we will demonstrate the ability to quantitatively differentiate a wide range of biochemical agent targets against a wide range of background materials. We will describe the relationship between spectral resolution and specificity in target identification, as well as simple, fast, algorithms to identify materials.Hand-held, battery operated instruments using a deep UV laser and multi-band detection have been developed and deployed on missions to the Antarctic, the Arctic, and the deep ocean with the capability of detecting a single bacterial spore and to differentiate a wide range of organic and biological compounds.

native flourescence↗

Relationships Between Seed Weight, Germination Potential and Biochemical Reserves of Maritime Pine in Morocco: Elements for Tree Seedlings Improvement

Selection of quality seeds in breeding programs can significantly improve seedling productivity. Germination and biochemical analyses on seeds from ten natural populations of maritime pine (Pinus pinaster Ait.) in Morocco reveals significant differences among populations in seed weight, germination characters and protein content in both dry seeds and megagametophytes. During germination, the mobilization of protein content in megagametophyte is significantly different among populations than sugar content. A strong positive correlation between the germination capacity and the protein content in both dry seeds and megagametophytes indicates that the best populations in term of germination capacity may also be the richest in protein content. The present study finds that seed weight is not a good indicator for quality seed selection, nor is it recommended to increase the degree of germinability. Our results suggest that the pine population in southern Morocco might have adapted to drought conditions as it is characterized by heavy seed weight and lower speed of protein content mobilization in megagametophyte compared to northern populations growing in temperate climate.

Wahid, Nadya↗

Characterization of Plastic Degrading Bacteria from Environmental Samples by Genetic and Biochemical Analysis

Plastic is the major waste-product during NASA space missions, recycling this waste-stream to produce other beneficial materials would decrease upmass. Bacterial called plastisomes have been demonstrated to metabolize non-biodegradable plastics such as polyethylene and polystyrene. Characterization and engineering of these bacteria, and their eventual incorporation as life support systems would enable space flight beyond lower earth orbit. We will utilize molecular techniques to identify and isolate the most productive plastisome. Environmental samples obtained from locations known to be rich in plastic will be cultured in a laboratory defined-media supplemented with plastic as the sole carbon source. Cultures will be monitored for growth over time. Ribosomal DNA will be amplified from cultures that exhibit growth using PCR. These amplified fragments will be sequenced to determine the identity of the consortia in the cultures. We will then perform bioinformatics analysis on the data to identify the plastisomes and generate phylogenetic trees. Morphological and physiological profile of the plastisomes will also be conducted by microscopy and biochemical tests. Our results would reveal a bacterial strain that can break down plastics efficiently. The implication for this project would not only benefit space exploration but also make a major impact towards sustainability development on Earth.

plastic conversion↗

Nanoscale biochemical sample preparation and analysis

Provided herein are methods and systems for biochemical analysis, including compositions and methods for processing and analysis of small cell populations and biological samples (e.g., a robotically controlled chip-based nanodroplet platform). In particular aspects, the methods described herein can reduce total processing volumes from conventional volumes to nanoliter volumes within a single reactor vessel (e.g., within a single droplet reactor) while minimizing losses, such as due to sample evaporation.

Kelly, Ryan T.↗

Mechanical and biochemical recovery of landfill waste in an underserved community

Historically in the United States, waste collected for recycling has been sold and shipped to processors in China. In 2013 and 2018, China introduced the Green Fence and National Sword policies which restricts the import of contaminated materials and banned the import of many recyclables. The cost of recycling in the United States has increased following these policy changes, which has led to many communities reducing their recycling programs or halting them altogether. Rural and underserved communities that don’t have resources to afford sophisticated recycling programs have been heavily impacted. Previous work at INL demonstrated that MSW is a potentially viable feedstock for both biochemical and thermochemical conversion. The goal of this project is to assess preprocessing tools that can produce consistent feedstocks that meet conversion specifications, remove problematic contaminants, and reduce the amount of waste that is landfilled. Municipal solid waste was collected from an underserved community in southeast Idaho, contaminants were characterized, and mechanically separated into two discrete fractions. The unit operations identified during mechanical separation trials will be mobilized to on-site with a goal of 50% recovery of paper and plastic waste.

09 - BIOMASS FUELS↗

Effect of biomass liquefaction on glucose and xylose prices predicted by National Renewable Energy Laboratory biochemical sugar model

Abstract The National Renewable Energy Laboratory (NREL) published a model in 2017 that enables the minimum selling price of lignocellulosic sugar to be calculated. The model can be modified to suit any biomass feedstock and operational design. In the present case, the model is used to understand the economics of a process configuration that incorporates liquefaction as a preprocessing step in corn‐stover‐fed biorefinery. This study demonstrates a quantitative approach utilizing an existing biorefinery setup to simulate the biomass liquefaction technique while estimating the price of the resulting sugar product. The objective is to understand whether the addition of liquefaction methodology – and the substitution of acid pretreatment – can reduce the cost of lignocellulosic sugars. The reason for setting up a liquefaction unit at the start is to reduce the yield stress of biomass slurry so that the flow remains unrestricted downstream. The liquefaction process is also unique because it uses no chemicals and saves the cost of pretreatment. The pretreatment step is bypassed because the output of liquefaction can be fed into enzyme hydrolysis just after simple cooking. The liquefaction process can be performed in three modes: enzyme, enzyme mimetic, and a combination of enzyme and enzyme mimetic. The results from the BC1707 model indicate the minimum cost for the enzyme liquefaction route. © 2022 The Authors. Biofuels, Bioproducts and Biorefining published by Society of Industrial Chemistry and John Wiley & Sons Ltd.

09 BIOMASS FUELS↗

Biochemical inhibition of acid phosphatase activity in two mountain spruce forest soils

The product inhibition of the 4-Methylumbelliferyl phosphate (MUB-P) decay rate, a measure of potential acid phosphatase activity, has not been considered in most of the published kinetic studies. The aim of this study was to determine the type and strength of the product inhibition in order to better define reaction conditions at which the acid phosphatase activity assay produces unbiased results. The MUB-P decay rate was measured in the forest floor and topsoil organic horizons of two spruce forest catchments at different initial MUB-P and P-PO4 3- concentrations. Furthermore, the type and strength of the inhibition was analyzed by non-linear regression. We found that MUB-P decay was competitively inhibited by the P-PO4 3- but also by dissolved organic P. By using estimated kinetic parameters, we calculated an underestimation of potential acid phosphatase activity by up to ~20% at concentrations of added MUB-P as high as 300 µmol g-1 dry weight and an incubation time one hour due to background inorganic and organic P concentrations in range of units of µmol per gram of dry soil. We discuss approaches that can be used to minimize the underestimation and show that the previously defined recommendations might not be always sufficient to avoid the bias. Therefore, we recommend to analyze progress curves at a wide range of initial MUB-P concentrations using non-linear methods when the enzyme assay is optimized for a given soil. When changes in inorganic and/or organic P concentrations are expected, we further recommend measuring background inorganic and organic P concentrations.

59 BASIC BIOLOGICAL SCIENCES↗