Search NASA⌕ Search

SEARCH · Search NASA

Results for “Biological Transport, Active”

Search indexed NASA NTRS and DOE OSTI research on propulsion, heat transfer, battery materials and energy systems. Follow report and document links to the original sources.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 271 records · Page 15

mTOR regulates aerobic glycolysis through NEAT1 and nuclear paraspeckle-mediated mechanism in hepatocellular carcinoma

Hepatocellular Carcinoma (HCC) is a major form of liver cancer and a leading cause of cancer-related death worldwide. New insights into HCC pathobiology and mechanism of drug actions are urgently needed to improve patient outcomes. HCC undergoes metabolic reprogramming of glucose metabolism from respiration to aerobic glycolysis, a phenomenon known as the ‘Warburg Effect’ that supports rapid cancer cell growth, survival, and invasion. mTOR is known to promote Warburg Effect, but the underlying mechanism(s) remains poorly defined. The aim of this study is to understand the mechanism(s) and significance of mTOR regulation of aerobic glycolysis in HCC. We profiled mTORC1-dependent long non-coding RNAs (lncRNAs) by RNA-seq of HCC cells treated with rapamycin. Chromatin immunoprecipitation (ChIP) and luciferase reporter assays were used to explore the transcriptional regulation of NEAT1 by mTORC1. [U- 13 C]-glucose labeling and metabolomic analysis, extracellular acidification Rate (ECAR) by Seahorse XF Analyzer, and glucose uptake assay were used to investigate the role of mTOR-NEAT1-NONO signaling in the regulation of aerobic glycolysis. RNA immunoprecipitation (RIP) and NONO-binding motif scanning were performed to identify the regulatory mechanism of pre-mRNA splicing by mTOR-NEAT1. Myristoylated AKT1 (mAKT1)/NRASV 12 -driven HCC model developed by hydrodynamic transfection (HDT) was employed to explore the significance of mTOR-NEAT1 signaling in HCC tumorigenesis and mTOR-targeted therapy. mTOR regulates lncRNA transcriptome in HCC and that NEAT1 is a major mTOR transcriptional target. Interestingly, although both NEAT1_1 and NEAT1_2 are down-regulated in HCC, only NEAT1_2 is significantly correlated with poor overall survival of HCC patients. NEAT1_2 is the organizer of nuclear paraspeckles that sequester the RNA-binding proteins NONO and SFPQ. We show that upon oncogenic activation, mTORC1 suppresses NEAT1_2 expression and paraspeckle biogenesis, liberating NONO/SFPQ, which in turn, binds to U5 within the spliceosome, stimulating mRNA splicing and expression of key glycolytic enzymes. This series of actions lead to enhanced glucose transport, aerobic glycolytic flux, lactate production, and HCC growth both in vitro and in vivo. Furthermore, the paraspeckle-mediated mechanism is important for the anticancer action of US FDA-approved drugs rapamycin/temsirolimus. These findings reveal a molecular mechanism by which mTOR promotes the ‘Warburg Effect’, which is important for the metabolism and development of HCC, and anticancer response of mTOR-targeted therapy.

59 BASIC BIOLOGICAL SCIENCES↗

Configuration and Projected Capabilities of the Common Habitat Medical Care Facility

The Common Habitat is a large, long-duration habitat being explored as part of a conceptual study (not an active NASA program) that uses an SLS core stage Liquid Oxygen (LOX) tank as its primary structure. It is intended for use on the Moon as part of a permanently occupied outpost, on Mars as part of an outpost that will be occupied for hundreds of days at a time, and in deep space as part of the Deep Space Exploration Vehicle where it will support crewed missions up to 1200 days in duration. A study of internal orientation and crew size resulted in a Common Habitat configuration sized for a crew of eight with a three-deck horizontal orientation. Additional work outside the scope of this paper is developing a vertical translation system, a crew mobility aids system based on wearable gecko-derived grippers, and a crew seating/restraint system. These systems are all assumed for use in conjunction with the Medical Care Facility, which is needed to maintain crew well-being during these missions, where distance from Earth precludes the possibility of evacuation to Earth. This paper describes recent improvements in the Common Habitat Medical Care Facility and associated benefits for crew survivability in long duration missions beyond Earth orbit. These improvements were made with the assistance of a NASA Pathways intern whose experience includes a tour of duty in Afghanistan as an Army combat medic with the 691st GHOST-T, attached to the 1st and 7th US Special Forces Groups as part of Operation Freedom’s Sentinel, where he helped provide far-forward surgical capabilities in austere combat environments. The initial baseline Medical Care Facility was developed working in conjunction with University of Houston Space Architecture graduate students. The facility was placed on the upper deck of the Common Habitat in a location that provided privacy, operational volume, and was close to the vertical translation pathway. The notional outfitting repurposed component CAD models from unrelated studies and notionally indicated a level of care roughly equivalent to that aboard the International Space Station. The CAD modeling provided notional stowage volumes, a deployable surface, some fixed equipment, an ultrasound, and a potentially reconfigurable treatment table. While this facility is clearly a competent arrangement, it was desired to leverage available expertise and upgrade the station given the vast distances from Earth to be experienced by the Common Habitat. Key driving requirements applied to the upgrade included to provide NASA’s Medical Level of Care V, offer enhanced telemedicine capabilities, provide patient physical accommodation, provide caregiver access to the patient from all sides, include sliding pocket doors for access to hygiene and to the Vertical Translation System, and to add any additional capability possible for the best achievable medical care. The first step in the facility upgrade was to quantify the current medical inventory on the International Space Station and ensure that sufficient stowage volume was present for this purpose. To that end, the ISS medical kits were reviewed, and eight full size mid deck lockers were placed in the facility. A number of additional devices were also added, based on the co-author’s combat medic experience. Also, two fixed shelves and one horizontal work surface were added to the Medical Care Facility, with the shelves providing storage space for the additional devices and the work surface providing a location for the caregiver to work or stage equipment. Four display monitors were added to the wall above the horizontal work surface, supporting data display, telemedicine, conferencing, or other needs. The existing treatment table was replaced with a mobile surgical stretcher-chair. Two additional doors were added to the Medical Care Facility. One leads directly to the hygiene compartment, allowing it to support medical operations in addition to providing galley/wardroom support. The other door leads directly into the Vertical Translation System. The wall adjacent to the subsystems bay was moved, adding additional volume to the Medical Care Facility. This improved caregiver access to the patient and allowed for a larger number of caregivers to be present. It also provided options for relocation of support equipment relative to the patient as needed. In the upgraded Medical Care Facility, the Surgical Stretcher-Chair and the Vertical Translation System can work together to provide incapacitated crew member transport from a site of injury on any deck of the Common Habitat to the Medical Care Facility. It can also support patient treatment in a variety of positions including a variety of sitting postures and a supine posture at a variety of pitch angles. The facility can also support caregiver office work for review of examination results, private consultation, inventory and maintenance, and a variety of other purposes. A forward activity will be to conduct evaluations of the Medical Care Facility with different medical scenarios. Additionally, ambient and task lighting selections remain as forward work. The eight middeck lockers can be augmented to use as portable equipment carts, similar to a manner in which maintenance facility stowage was used as portable carts during the NASA Desert Research and Technology Studies in the Constellation Program. Trash accommodation will also need forward work to assess, including provision for wet trash, dry trash, and biological waste. It will be important to assess a redesign of the surgical stretcher-chair. The commercial version used in the upgrade can only enable vertical translation in the seated configuration, requiring the patient to bend both hips and knees. A possible redesign of the chair will allow for vertical translation without requiring any bending at the hip or knees. Also, the commercial version is wheeled, making it mobile in gravity but unanchored in microgravity. Work will be needed to adapt the chair for gravity-independent performance. The hygiene compartment can be redesigned to serve both as a medical scrub facility and for galley hand washing. Pending sufficient volume, it may also be possible to place sanitation equipment in this location to clean medical tools. Finally, most space architectures have never allowed for more than one incapacitated crew member, but several scenarios could potentially injure two or more crew in the same incident. This facility could be assessed to determine its present ability to address two or more injured crew in parallel and determine the potential upper limit for number of treatable crew in a multi-crew injury scenario, or to treat polytrauma of a single patient.

Common Habitat↗

Cryo-EM structures of engineered active bc 1- cbb 3 type CIII 2 CIV super-complexes and electronic communication between the complexes

Respiratory electron transport complexes are organized as individual entities or combined as large supercomplexes (SC). Gram-negative bacteria deploy a mitochondrial-like cytochrome (cyt) bc 1 (Complex III, CIII 2 ), and may have specific cbb 3 -type cyt c oxidases (Complex IV, CIV) instead of the canonical aa 3 -type CIV. Electron transfer between these complexes is mediated by soluble ( c 2 ) and membrane-anchored ( c y ) cyts. Here, we report the structure of an engineered bc 1 - cbb 3 type SC (CIII 2 CIV, 5.2Å resolution) and three conformers of native CIII 2 (3.3 Å resolution). The SC is active in vivo and in vitro, contains all catalytic subunits and cofactors, and two extra transmembrane helices attributed to cyt c y and the assembly factor CcoH. The cyt c y is integral to SC, its cyt domain is mobile and it conveys electrons to CIV differently than cyt c 2 . The successful production of a native-like functional SC and determination of its structure illustrate the characteristics of membrane-confined and membrane-external respiratory electron transport pathways in Gram-negative bacteria.

42 ENGINEERING↗

A Rhodopseudomonas strain with a substantially smaller genome retains the core metabolic versatility of its genus

ABSTRACT Rhodopseudomonas are a group of phototrophic microbes with a marked metabolic versatility and flexibility that underpins their potential use in the production of value-added products, bioremediation, and plant growth promotion. Members of this group have an average genome size of about 5.5 Mb, but two closely related strains have genome sizes of about 4.0 Mb. To identify the types of genes missing in a reduced genome strain, we compared strain DSM127 with other Rhodopseudomonas isolates at the genomic and phenotypic levels. We found that DSM127 can grow as well as other members of the Rhodopseudomonas genus and retains most of their metabolic versatility, but it has many fewer genes associated with high-affinity transport of nutrients, iron uptake, nitrogen metabolism, and biodegradation of aromatic compounds. This analysis indicates genes that can be deleted in genome reduction campaigns and suggests that DSM127 could be a favorable choice for biotechnology applications using Rhodopseudomonas or as a strain that can be engineered further to reside in a specialized natural environment. IMPORTANCE Rhodopseudomonas are a cohort of phototrophic bacteria with broad metabolic versatility. Members of this group are present in diverse soil and water environments, and some strains are found associated with plants and have plant growth-promoting activity. Motivated by the idea that it may be possible to design bacteria with reduced genomes that can survive well only in a specific environment or that may be more metabolically efficient, we compared Rhodopseudomonas strains with typical genome sizes of about 5.5 Mb to a strain with a reduced genome size of 4.0 Mb. From this, we concluded that metabolic versatility is part of the identity of the Rhodopseudomonas group, but high-affinity transport genes and genes of apparent redundant function can be dispensed with.

59 BASIC BIOLOGICAL SCIENCES↗

Configuration and Projected Capabilities of the Common Habitat Medical Care Facility

The Common Habitat is a large, long-duration habitat being explored as part of a conceptual study (not an active NASA program) that uses an SLS core stage Liquid Oxygen (LOX) tank as its primary structure. It is intended for use on the Moon as part of a permanently occupied outpost, on Mars as part of an outpost that will be occupied for hundreds of days at a time, and in deep space as part of the Deep Space Exploration Vehicle where it will support crewed missions up to 1200 days in duration. A study of internal orientation and crew size resulted in a Common Habitat configuration sized for a crew of eight with a three-deck horizontal orientation. Additional work outside the scope of this paper is developing a vertical translation system, a crew mobility aids system based on wearable gecko-derived grippers, and a crew seating/restraint system. These systems are all assumed for use in conjunction with the Medical Care Facility, which is needed to maintain crew well-being during these missions, where distance from Earth precludes the possibility of evacuation to Earth. This paper describes recent improvements in the Common Habitat Medical Care Facility and associated benefits for crew survivability in long duration missions beyond Earth orbit. These improvements were made with the assistance of a NASA Pathways intern whose experience includes a tour of duty in Afghanistan as an Army combat medic with the 691st GHOST-T, attached to the 1st and 7th US Special Forces Groups as part of Operation Freedom’s Sentinel, where he helped provide far-forward surgical capabilities in austere combat environments. The initial baseline Medical Care Facility was developed working in conjunction with University of Houston Space Architecture graduate students. The facility was placed on the upper deck of the Common Habitat in a location that provided privacy, operational volume, and was close to the vertical translation pathway. The notional outfitting repurposed component CAD models from unrelated studies and notionally indicated a level of care roughly equivalent to that aboard the International Space Station. The CAD modeling provided notional stowage volumes, a deployable surface, some fixed equipment, an ultrasound, and a potentially reconfigurable treatment table. While this facility is clearly a competent arrangement, it was desired to leverage available expertise and upgrade the station given the vast distances from Earth to be experienced by the Common Habitat. Key driving requirements applied to the upgrade included to provide Medical Level of Care V, offer enhanced telemedicine capabilities, provide patient physical accommodation, provide caregiver access to the patient from all sides, include sliding pocket doors for access to hygiene and to the Vertical Translation System, and to add any additional capability possible for the best achievable medical care. The first step in the facility upgrade was to quantify the current medical inventory on the International Space Station and ensure that sufficient stowage volume was present for this purpose. To that end, the ISS medical kits were reviewed, and eight full size mid deck lockers were placed in the facility. A number of additional devices were also added, based on the intern’s combat medic experience. Also, two fixed shelves and one horizontal work surface were added to the Medical Care Facility, with the shelves providing storage space for the additional devices and the work surface providing a location for the caregiver to work or stage equipment. Four display monitors were added to the wall above the horizontal work surface, supporting data display, telemedicine, conferencing, or other needs. The existing treatment table was replaced with a mobile surgical stretcher-chair. Two additional doors were added to the Medical Care Facility. One leads directly to the hygiene compartment, allowing it to support medical operations in addition to providing galley/wardroom support. The other door leads directly into the Vertical Translation System. The wall adjacent to the subsystems bay was moved, adding additional volume to the Medical Care Facility. This improved caregiver access to the patient and allowed for a larger number of caregivers to be present. It also provided options for relocation of support equipment relative to the patient as needed. In the upgraded Medical Care Facility, the Surgical Stretcher-Chair and the Vertical Translation System can work together to provide incapacitated crew member transport from a site of injury on any deck of the Common Habitat to the Medical Care Facility. It can also support patient treatment in a variety of positions including a variety of sitting postures and a supine posture at a variety of pitch angles. The facility can also support caregiver office work for review of examination results, private consultation, inventory and maintenance, and a variety of other purposes. A forward activity will be to conduct evaluations of the Medical Care Facility with different medical scenarios. Additionally, ambient and task lighting selections remain as forward work. The eight mid deck lockers can be augmented to use as portable equipment carts, similar to a manner in which maintenance facility stowage was used as portable carts during the NASA Desert Research and Technology Studies in the Constellation Program. Trash accommodation will also need forward work to assess, including provision for wet trash, dry trash, and biological waste. It will be important to assess a redesign of the surgical stretcher-chair. The commercial version used in the upgrade can only enable vertical translation in the seated configuration, requiring the patient to bend both hips and knees. A possible redesign of the chair will allow for vertical translation without requiring any bending at the hip or knees. Also, the commercial version is wheeled, making it mobile in gravity but unanchored in microgravity. Work will be needed to adapt the chair for gravity-independent performance. The hygiene compartment can be redesigned for dual-use medical scrub and galley handwash facility. Pending sufficient volume, it may also be possible to place sanitation equipment in this location to clean medical tools. Finally, most space architectures have never allowed for more than one incapacitated crew member, but several scenarios could potentially injure two or more crew in the same incident. This facility could be assessed to determine its present ability to address two or more injured crew in parallel and determine the potential upper limit for number of treatable crew in a multi-crew injury scenario, or to treat polytrauma of a single patient.

Habitat↗

Light-Dependence of Formate (C1) and Acetate (C2) Transport and Oxidation in Poplar Trees

Although apparent light inhibition of leaf day respiration is a widespread reported phenomenon, the mechanisms involved, including utilization of alternate respiratory pathways and substrates and light inhibition of TCA cycle enzymes are under active investigation. Recently, acetate fermentation was highlighted as a key drought survival strategy mediated through protein acetylation and jasmonate signaling. Here, we evaluate the light-dependence of acetate transport and assimilation in Populus trichocarpa trees using the dynamic xylem solution injection (DXSI) method developed here for continuous studies of C1 and C2 organic acid transport and light-dependent metabolism. Over 7 days, 1.0 L of [13C]formate and [13C2]acetate solutions were delivered to the stem base of 2-year old potted poplar trees, while continuous diurnal observations were made in the canopy of CO2, H2O, and isoprene gas exchange together with δ13CO2. Stem base injection of 10 mM [13C2]acetate induced an overall pattern of canopy branch headspace 13CO2 enrichment (δ13CO2 +27‰) with a diurnal structure in δ13CO2 reaching a mid-day minimum followed by a maximum shortly after darkening where δ13CO2 values rapidly increased up to +12‰. In contrast, 50 mM injections of [13C]formate were required to reach similar δ13CO2 enrichment levels in the canopy with δ13CO2 following diurnal patterns of transpiration. Illuminated leaves of detached poplar branches pretreated with 10 mM [13C2]acetate showed lower δ13CO2 (+20‰) compared to leaves treated with 10 mM [13C]formate (+320‰), the opposite pattern observed at the whole plant scale. Following dark/light cycles at the leaf-scale, rapid, strong, and reversible enhancements in headspace δ13CO2 by up to +60‰ were observed in [13C2]acetate-treated leaves which showed enhanced dihydrojasmonic acid and TCA cycle intermediate concentrations. The results are consistent with acetate in the transpiration stream as an effective activator of the jasmonate signaling pathway and respiratory substrate. The shorter lifetime of formate relative to acetate in the transpiration stream suggests rapid formate oxidation to CO2 during transport to the canopy. In contrast, acetate is efficiently transported to the canopy where an increased allocation towards mitochondrial dark respiration occurs at night. The results highlight the potential for an effective integration of acetate into glyoxylate and TCA cycles and the light-inhibition of citrate synthase as a potential regulatory mechanism controlling the diurnal allocation of acetate between anabolic and catabolic processes.

59 BASIC BIOLOGICAL SCIENCES↗

Plant myo -inositol transport influences bacterial colonization phenotypes

Plant microbiomes are assembled and modified through a complex milieu of biotic and abiotic factors. Despite dynamic and fluctuating contributing variables, specific host metabolites are consistently identified as important mediators of microbial interactions. We combine information from a large-scale metatranscriptomic dataset from natural poplar trees and experimental genetic manipulation assays in seedlings of the model plant Arabidopsis thaliana to converge on a conserved role for transport of the plant metabolite myo-inositol in mediating host-microbe interactions. While microbial catabolism of this compound has been linked to increased host colonization, we identify bacterial phenotypes that occur in both catabolism-dependent and -independent manners, suggesting that myo-inositol may additionally serve as a eukaryotic-derived signaling molecule to modulate microbial activities. Our data suggest host control of this compound and resulting microbial behavior are important mechanisms at play surrounding the host metabolite myo-inositol.

59 BASIC BIOLOGICAL SCIENCES↗

The time course of acclimation to the stress of triose phosphate use limitation

Abstract Triose phosphate utilisation (TPU) limits the maximum rate at which plants can photosynthesise. However, TPU is almost never found to be limiting photosynthesis under ambient conditions for plants. This, along with previous results showing adaptability of TPU at low temperature, suggest that TPU capacity is regulated to be just above the photosynthetic rate achievable under the prevailing conditions. A set of experiments were performed to study the adaptability of TPU capacity when plants are acclimated to elevated CO 2 concentrations. Plants held at 1500 ppm CO 2 were initially TPU limited. After 30 h they no longer exhibited TPU limitations but they did not elevate their TPU capacity. Instead, the maximum rates of carboxylation and electron transport declined. A timecourse of regulatory responses was established. A step increase of CO 2 first caused PSI to be oxidised but after 40 s both PSI and PSII had excess electrons as a result of acceptor‐side limitations. Electron flow to PSI slowed and the proton motive force increased. Eventually, non‐photochemical quenching reduced electron flow sufficiently to balance the TPU limitation. Over several minutes rubisco deactivated contributing to regulation of metabolism to overcome the TPU limitation.

59 BASIC BIOLOGICAL SCIENCES↗

Applications of MODIS Fluorescent Line Height Measurements to Monitor Water Quality Trends and Algal Bloom Activity

Recent advances in satellite and airborne remote sensing, such as improvements in sensor and algorithm calibrations, processing techniques and atmospheric correction procedures have provided for increased coverage of remote-sensing, ocean-color products for coastal regions. In particular, for the Moderate Resolution Imaging Spectrometer (MODIS) sensor calibration updates, improved aerosol retrievals and new aerosol models has led to improved atmospheric correction algorithms for turbid waters and have improved the retrieval of ocean color in coastal waters. This has opened the way for studying ocean phenomena and processes at finer spatial scales, such as the interactions at the land-sea interface, trends in coastal water quality and algal blooms. Human population growth and changes in coastal management practices have brought about significant changes in the concentrations of organic and inorganic, particulate and dissolved substances entering the coastal ocean. There is increasing concern that these inputs have led to declines in water quality and have increase local concentrations of phytoplankton, which cause harmful algal blooms. In two case studies we present MODIS observations of fluorescence line height (FLH) to 1) assess trends in water quality for Tampa Bay, Florida and 2) illustrate seasonal and annual variability of algal bloom activity in Monterey Bay, California as well as document estuarine/riverine plume induced red tide events. In a comprehensive analysis of long term (2003-2011) in situ monitoring data and satellite imagery from Tampa Bay we assess the validity of the MODIS FLH product against chlorophyll-a and a suite of water quality parameters taken in a variety of conditions throughout a large optically complex estuarine system. A systematic analysis of sampling sites throughout the bay is undertaken to understand how the relationship between FLH and in situ chlorophyll-a responds to varying conditions and to develop a near decadal trend in water quality changes. In situ monitoring locations that correlated well with satellite imagery were in depths greater than seven meters and were located over five kilometers from shore. Water quality parameter of total nitrogen, phosphorous, turbidity and biological oxygen demand had high correlations with these sites, as well. Satellite FLH estimates show improving water quality from 2003-2007 with a slight decline up through 2011. Dinoflagellate blooms in Monterey Bay, California (USA) have recently increased in frequency and intensity. Nine years of MODIS FLH observations are used to describe the annual and seasonal variability of bloom activity within the Bay. Three classes of MODIS algorithms were correlated against in situ chlorophyll measurements. The FLH algorithm provided the most robust estimate of bloom activity. Elevated concentrations of phytoplankton were evident during the months of August-November, a period during which increased occurrences of dinoflagellate blooms have been observed in situ. Seasonal patterns of FLH show the on- and offshore movement of areas of high phytoplankton biomass between oceanographic seasons. Higher concentrations of phytoplankton are also evident in the vicinity of the land-based nutrient sources and outflows, and the cyclonic bay-wide circulation can transport these nutrients to the northern Bay bloom incubation region. Both of these case studies illustrate the utility MODIS FLH observations in supporting management decisions in coastal and estuarine waters.

Fischer, Andrew↗

Mechanistic Duality of Bacterial Efflux Substrates and Inhibitors: Example of Simple Substituted Cinnamoyl and Naphthyl Amides

Antibiotic resistance poses an immediate and growing threat to human health. Multidrug efflux pumps are promising targets for overcoming antibiotic resistance with small-molecule therapeutics. Previously, we identified a diaminoquinoline acrylamide, NSC-33353, as a potent inhibitor of the AcrAB–TolC efflux pump in Escherichia coli. This inhibitor potentiates the antibacterial activities of novobiocin and erythromycin upon binding to the membrane fusion protein AcrA. It is also a substrate for efflux and lacks appreciable intrinsic antibacterial activity of its own in wild-type cells. Here, we have modified the substituents of the cinnamoyl group of NSC-33353, giving rise to analogs that retain the ability to inhibit efflux, lost the features of the efflux substrates, and gained antibacterial activity in wild-type cells. The replacement of the cinnamoyl group with naphthyl isosteres generated compounds that lack antibacterial activity but are both excellent efflux pump inhibitors and substrates. Surprisingly, these inhibitors potentiate the antibacterial activity of novobiocin but not erythromycin. Surface plasmon resonance experiments and molecular docking suggest that the replacement of the cinnamoyl group with naphthyl shifts the affinity of the compounds away from AcrA to the AcrB transporter, making them better efflux substrates and changing their mechanism of inhibition. These results provide new insights into the duality of efflux substrate/inhibitor features in chemical scaffolds that will facilitate the development of new efflux pump inhibitors.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Exponential Escape Rate of Filamentary Incubation in Mott Spiking Neurons

Mott materials such as vanadium oxides, when subject to a strong applied voltage, present an inhomogeneous insulator-to-metal transition with formation of metallic filaments within the insulating bulk. This property is enabling the development of compact and power-efficient neuromorphic devices known as Mott neurons. However, the nature of the transition has not been fully understood yet, as it may be attributed to different effects, including Joule self-heating and hot-carrier injection. Moreover, the experimental determination of the threshold voltage needed to induce the transition has proven to be challenging, as the transition becomes increasingly unpredictable when the threshold is approached. The physical understanding of these issues would not only deepen our understanding of Mott insulators, but would also be an important step toward the realization of neuromorphic devices based on such materials. In this work we use numerical simulations based on the Mott resistor network model to study the nature of the filament incubation and formation process. Here, we show that both electronic and thermal effects, in the form of current density focusing and Joule self-heating, respectively, contribute to the filamentary incubation and growth. Remarkably, we find that the percolation of the metallic filaments near the threshold is intrinsically stochastic, qualitatively similar to the familiar Arrhenius activated behavior and to the stochastic firing of biological neurons. More precisely, we characterize the filament percolation as a Poisson point process, which has the same probability distribution as mathematical models of neuronal firing with an exponential escape rate. Finally, we support the numerical simulation results by performing experiments in VO 2 that are in agreement with the exponential escape rate behavior. Thus, we establish a functionality of Mott insulators that opens a path toward implementing neuromorphic hardware with quantum materials.

71 CLASSICAL AND QUANTUM MECHANICS, GENERAL PHYSIC↗

Electrochromic shift supports the membrane destabilization model of Tat-mediated transport and shows ion leakage during Sec transport

Significance Due to the transient nature of the active Tat complex, much of the knowledge available has been hard won. Electric-field-indicating absorbance transients allow investigation of transmembrane ion movement using native electric potential in thylakoids while circumventing challenges of reconstitution. Electrical behavior of transiently and/or constitutively assembled complexes suggests that Sec translocation occurs through a barrel-stave-type proteinaceous pore, while Tat translocation likely occurs through a toroidal pore. This would be the first instance of toroidal pores being exploited by a general protein translocation system. Our electrochemical analysis offers insight into how protein transport might be linked to light stimulation. Localized membrane destabilization presents a rational link between the proton motive force, membrane thinning, and Tat-mediated transport of substrates critical to photosynthesis.

59 BASIC BIOLOGICAL SCIENCES↗

Cerebellar dysfunction in a mouse model of childhood-onset manganese-induced dystonia parkinsonism

Humans with pathogenic variants of the manganese (Mn) transporter gene SLC39A14 exhibit highly elevated brain Mn concentrations and childhood-onset dystonia-parkinsonism. Here we show that Slc39a14-knockout (KO) mice, a preclinical model of the disease with elevated Mn concentrations in the CB, express deficits in physiological tremor implicating cerebellar (CB) dysfunction. Imaging of intracellular Mn in Purkinje cells (PCs) using synchrotron-based X-ray fluorescence microscopy confirmed highly elevated Mn concentrations in the PCs of Slc39a14-KO mice. To determine biological pathways altered in the CB of Slc39a14-KO mice relative to wildtype (WT), we performed RNA sequencing and discovered significant upregulation of pathways and genes regulating immune response and cell death. To substantiate these findings, we performed quantitative autoradiography of the neuroinflammation biomarker Translocator Protein 18 kDa (TSPO) which was significantly increased in the CB of Slc39a14-KO mice relative to WT. The latter findings were confirmed via immunostaining with the microglial marker Iba-1, revealing widespread microglia activation and clustering in the CB cortex. Immunostaining for cleaved caspase-3 (cCASP3), a marker of apoptosis, showed increased number of PCs with positive immunolabeling for cCASP3 in Slc39a14-KO mice relative to WT. Degeneration of PCs was confirmed by Hematoxylin and Eosin (H&E) staining. Lastly, functional electrophysiological assessment of CB neurocircuitry revealed a marked decrease in firing rates of cerebellar nuclei (CN) neurons and increased variability of PC simple spikes firing. Collectively, these findings show, for the first time, Mn-induced PC degeneration and dysfunctional CB circuitry in Slc39a14-KO mice providing additional evidence for the pathological underpinnings of the dystonia-like movements, balance, and gait abnormalities in SLC39A14 mutation carriers.

36 MATERIALS SCIENCE↗

Structural basis for human NKCC1 inhibition by loop diuretic drugs

Abstract Na + –K + –Cl − cotransporters functions as an anion importers, regulating trans-epithelial chloride secretion, cell volume, and renal salt reabsorption. Loop diuretics, including furosemide, bumetanide, and torsemide, antagonize both NKCC1 and NKCC2, and are first-line medicines for the treatment of edema and hypertension. NKCC1 activation by the molecular crowding sensing WNK kinases is critical if cells are to combat shrinkage during hypertonic stress; however, how phosphorylation accelerates NKCC1 ion transport remains unclear. Here, we present co-structures of phospho-activated NKCC1 bound with furosemide, bumetanide, or torsemide showing that furosemide and bumetanide utilize a carboxyl group to coordinate and co-occlude a K + , whereas torsemide encroaches and expels the K + from the site. We also found that an amino-terminal segment of NKCC1, once phosphorylated, interacts with the carboxyl-terminal domain, and together, they engage with intracellular ion exit and appear to be poised to facilitate rapid ion translocation. Together, these findings enhance our understanding of NKCC-mediated epithelial ion transport and the molecular mechanisms of its inhibition by loop diuretics.

Biochemistry & Molecular Biology↗

Refinement of Protocols for Measuring the Apparent Optical Properties of Seawater

Ocean color satellite missions, like the Sea-viewing Wide Field-of-view Sensor (SeaWiFS) or the Moderate Resolution Imaging Spectroradiometer (MODIS) projects, are tasked with acquiring a global ocean color data set, validating and monitoring the accuracy and quality of the data, processing the radiometric data into geophysical units using a set of atmospheric and bio-optical algorithms, and distributing the final products to the scientific community. The long-standing requirement of the SeaWiFS Project, for example, is to produce spectral water-leaving radiances, LW(lambda), to within 5% absolute (lambda denotes wavelength) and chlorophyll a concentrations to within 35% (Hooker and Esaias 1993), and most ocean color sensors have the same or similar requirements. Although a diverse set of activities are required to ensure the accuracy requirements are met (Hooker and McClain 2000), the perspective here is with field observations. The accurate determination of upper ocean apparent optical properties (AOPs) is essential for the vicarious calibration of ocean color data and the validation of the derived data products, because the sea-truth measurements are used to evaluate the satellite observations (Hooker and McClain 2000). The uncertainties with in situ AOP measurements have various sources: a) the sampling procedures used in the field, including the environmental conditions encountered; b) the absolute characterization of the radiometers in the laboratory; c) the conversion of the light signals to geophysical units in a processing scheme, and d) the stability of the radiometers in the harsh environment they are subjected to during transport and use. Assuming ideal environmental conditions, so this aspect can be neglected, the SeaWiFS ground-truth uncertainty budget can only be satisfied if each uncertainty is on the order of 1-2%, or what is generally referred to as 1% radiometry. In recent years, progress has been made in estimating the magnitude of some of these uncertainties and in defining procedures for minimizing them. For the SeaWiFS Project, the first step was to convene a workshop to draft the SeaWiFS Ocean Optics Protocols (hereafter referred to as the Protocols). The Protocols initially adhered to the Joint Global Ocean Flux Study (JGOFS) sampling procedures (JGOFS 1991) and defined the standards for optical measurements to be used in SeaWiFS calibration and validation activities (Mueller and Austin 1992). Over time, the Protocols were revised (Mueller and Austin 1995), and then recurringly updated on essentially an annual basis (Mueller 2000, 2002, and 2003) as part of the Sensor Inter-comparison and Merger for Biological and Interdisciplinary Oceanic Studies (SIMBIOS) project. 98

Hooker, Stanford B.↗

Overview of Mars Science Laboratory (MSL) Environmental Program

This viewgraph presentation is an overview of the Mars Science Laboratory (MSL) program. The engineering objectives of the program are to create a Mobile Science Laboratory capable of one Mars Year surface operational lifetime (670 Martian sols = 687 Earth days). It will be able to land and operation over wide range of latitudes, altitudes and seasons It must have controlled propulsive landing and demonstrate improved landing precision via guided entry The general science objectives are to perform science that will focus on Mars habitability, perform next generation analytical laboratory science investigations, perform remote sensing/contact investigations and carry a suite of environmental monitoring instruments. Specific scientific objectives of the MSL are: (1) Characterization of geological features, contributing to deciphering geological history and the processes that have modified rocks and regolith, including the role of water. (2) Determination of the mineralogy and chemical composition (including an inventory of elements such as C, H, N, O, P, S, etc. known to be building blocks for life) of surface and near-surface materials. (3) Determination of energy sources that could be used to sustain biological processes. (4) Characterization of organic compounds and potential biomarkers in representative regolith, rocks, and ices. (5) Determination the stable isotopic and noble gas composition of the present-day bulk atmosphere. (6) Identification potential bio-signatures (chemical, textural, isotopic) in rocks and regolith. (7) Characterization of the broad spectrum of surface radiation, including galactic cosmic radiation, solar proton events, and secondary neutrons. (8) Characterization of the local environment, including basic meteorology, the state and cycling of water and C02, and the near-surface distribution of hydrogen. Several views of the planned MSL and the rover are shown. The MSL environmental program is to: (1) Ensure the flight hardware design is capable of surviving all the environments throughout its mission life time, including ground, transportation, launch, cruise, entry decent and landing (EDL) and surface operation environments. (2) Verify environmental testing and analysis have adequately validated the flight hardware's ability to withstand all natural, self-induced, and mission-activity-induced environments. The planned tests to ascertain the capability of the MSL to perform as desired are reviewed.

spacecraft↗

Proteomic Analysis of Methanococcus voltae Grown in the Presence of Mineral and Nonmineral Sources of Iron and Sulfur

Iron sulfur (Fe-S) proteins are essential and ubiquitous across all domains of life, yet the mechanisms underpinning assimilation of iron (Fe) and sulfur (S) and biogenesis of Fe-S clusters are poorly understood. This is particularly true for anaerobic methanogenic archaea, which are known to employ more Fe-S proteins than other prokaryotes. Here, we utilized a deep proteomics analysis of Methanococcus voltae A3 cultured in the presence of either synthetic pyrite (FeS 2 ) or aqueous forms of ferrous iron and sulfide to elucidate physiological responses to growth on mineral or nonmineral sources of Fe and S. The liquid chromatography-mass spectrometry (LCMS) shotgun proteomics analysis included 77% of the predicted proteome. Through a comparative analysis of intra- and extracellular proteomes, candidate proteins associated with FeS 2 reductive dissolution, Fe and S acquisition, and the subsequent transport, trafficking, and storage of Fe and S were identified. The proteomic response shows a large and balanced change, suggesting that M. voltae makes physiological adjustments involving a range of biochemical processes based on the available nutrient source. Among the proteins differentially regulated were members of core methanogenesis, oxidoreductases, membrane proteins putatively involved in transport, Fe-S binding ferredoxin and radical S-adenosylmethionine proteins, ribosomal proteins, and intracellular proteins involved in Fe-S cluster assembly and storage. This work improves our understanding of ancient biogeochemical processes and can support efforts in biomining of minerals. Clusters of iron and sulfur are key components of the active sites of enzymes that facilitate microbial conversion of light or electrical energy into chemical bonds. The proteins responsible for transporting iron and sulfur into cells and assembling these elements into metal clusters are not well understood. Using a microorganism that has an unusually high demand for iron and sulfur, we conducted a global investigation of cellular proteins and how they change based on the mineral forms of iron and sulfur. Understanding this process will answer questions about life on early earth and has application in biomining and sustainable sources of energy.

59 BASIC BIOLOGICAL SCIENCES↗

Comparative effects of selenite and selenate on nitrate assimilation in barley seedlings

The effect of SeO3= and SeO4= on NO3- assimilation in 8-d-old barley (Hordeum vulgare L.) seedlings was studied over a 24-h period. Selenite at 0.1 mol m-3 in the uptake solutions severely inhibited the induction of NO3- uptake and active nitrate reductases. Selenate, at 1.0 mol m-3 in the nutrient solution, had little effect on induction of activities of these systems until after 12 h; however, when the seedlings were pretreated with 1.0 mol m-3 SeO4= for 24 h, subsequent NO3- uptake from SeO4(=) -free solutions was inhibited about 60%. Sulphate partially alleviated the inhibitory effect of SeO3= when supplied together in the ambient solutions, but had no effect in seedlings pretreated with SeO3=. By contrast, SO4= partially alleviated the inhibitory effect of SeO4= even in seedlings pretreated with SeO4=. Since uptake of NO3- by intact seedlings was also inhibited by SO3=, the percentage of the absorbed NO3- that was reduced was not affected. By contrast, SeO4=, which affected NO3- uptake much less, inhibited the percentage reduced of that absorbed. However, when supplied to detached leaves, both SeO3= and SeO4= inhibited the in vivo reduction of NO3- as well as induction of nitrate reductase and nitrite reductase activities. Selenite was more inhibitory than SeO4= ; approximately a five to 10 times higher concentration of SeO4= than SeO3= was required to achieve similar inhibition. In detached leaves, the inhibitory effect of both SeO3= and SeO4= on in vivo NO3- reduction as well as on the induction of nitrate reductase activity was partially alleviated by SO4=. The inhibitory effects of Se salts on the induction of the nitrite reductase were, however, completely alleviated by SO4=. The results show that in barley seedlings SeO3= is more toxic than SeO4=. The reduction of SeO4= to SeO3= may be a rate limiting step in causing Se toxicity.

NASA Discipline Life Support Systems↗