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At least 505 records · Page 28

Nonclassical autophagy activation pathways are essential for production of infectious Influenza A virus in vitro

Autophagy is a critical mechanism deployed by eukaryotic cells in response to stress, including viral infection, to boost the innate antimicrobial responses. However, an increasing number of pathogens hijack the autophagic machinery to facilitate their own replication. Influenza A virus (IAV), responsible for several global pandemics, has an intricate dependence on autophagy for successful replication in mammalian cells. To elucidate key chokepoints in the host stress responses facilitating IAV replication, we constructed a meta-transcriptome of IAV and host gene expression dynamics during early (1–3 hpi), mid (4–6 hpi), and late (8–12 hpi) stages of the viral replication cycle at two multiplicities of infection (MOI): 1 and 5. We supplemented the global transcriptome study with phosphoproteomic analysis of stress-activated protein kinase (SAPK/JNK) signaling in lung carcinoma (predominantly used as an in vitro model of IAV replication) and normal human bronchial epithelial cells. We report significant differences in the activation profiles of autophagy regulating genes upon IAV infection at the two MOI as well as divergent dependence on ULK1 signaling within the normal and cancer cells. Regardless of the cell model, JNK-Thr187 signaling was crucial for the production of infectious viral particles.

59 BASIC BIOLOGICAL SCIENCES↗

Oxidative Stress and Autophagy Responses of Osteocytes Exposed to Spaceflight-like Radiation.

Weightlessness and radiation, two of the unique elements of the space environment, causes a profound decrement in bone mass that mimics aging. This bone loss is thought to result from increased activity of bone-resorbing osteoclasts and functional changes in bone-forming osteoblasts, cells that give rise to mature osteocytes. Our current understanding of the signaling factors and mechanisms underlying bone loss is incomplete. However, it is known that oxidative stress, characterized by the excess production of free radicals, is elevated during radiation exposure. The goals of this study is to examine the response of osteocytes to spaceflight-like radiation and to identify signaling processes that may be targeted to mitigate bone loss in scenarios of space exploration, earth-based radiotherapy and accidental radiation exposure. We hypothesize that (1) oxidative stress, as induced by radiation, decreases osteocyte survival and increases pro-osteoclastogenic signals and that (2) autophagy is one of the key cellular defenses against oxidative stress. Autophagy is the process by which cellular components including organelles and proteins are broken down and recycled. To test our hypothesis, we exposed the osteocyte-like cell line, MLO-Y4, to 0.5, 1, and 2 Gy of simulated space radiation (Iron-56 radiation at 600 MeV/n) and assessed cell numbers, cell growth-associated molecules as well as markers of autophagy and oxidative stress at various time points post-irradiation. We observed a reduction in cell numbers in the groups exposed to 1 and 2 Gy of Iron-56 radiation. Collectively, flow cytometry and gene expression analysis revealed that radiation caused a shift in cell cycle distribution consistent with growth arrest. Compared to sham-treatment, 2 Gy of Iron-56 increased FoxO3, SOD1, and RANKL gene expression yet unexpectedly decreased LC3B-II protein levels at 4 and 24 hours post-IR. Taken together, these findings suggest that simulated space radiation invoke antioxidant, pro-osteoclastogenic, and growth arrest responses in osteocytes. The implications of reduced autophagy flux at the time points examined remain to be elucidated.

Biological Sciences↗

Multi-omics profiling of the cold tolerant Monoraphidium minutum 26B-AM in response to abiotic stress

Microalgae that are of interest for biofuel production must be able to tolerate environmental changes that occur in outdoor cultivation systems. While algal cultures may experience daily temperature fluctuations and seasonal environmental changes, the underlying mechanisms that control and regulate physiological responses and adaptation to environmental pressures are largely unknown. Systems-level characterization enabled by functional genomics can help identify biochemical pathways that promote stability and productivity of algae in various environmental conditions. Monoraphidium minutum 26B-AM, a freshwater green microalga, was identified as a top performer in biomass production in winter season screens. We sequenced the genome of M. minutum 26B-AM and applied our multi-omics pipeline to profile this high potential strain under high salt and cold temperature perturbations. Through comparative analysis, including other green algae in the class Chlorophyceae, we identified gene families unique to the genus Monoraphidium, including a desaturase that has been linked to cold tolerance in plants. We observed that osmolytes, such as trehalose, proline and betaine, accumulate under salt stress, coinciding with upregulation of genes involved in biosynthesis of these metabolites. From the genome annotation, we reconstructed a metabolic model to provide a detailed map of the metabolic pathways and can be used to simulate growth and reaction fluxes. This multi-omics analysis provides a foundation to explore algal strain potential for biofuel applications, guides strain engineering, and expands our understanding of metabolic and regulatory mechanisms of algae in applied systems.

59 BASIC BIOLOGICAL SCIENCES↗

Effect of fluid countermeasures of varying osmolarity on cardiovascular responses to orthostatic stress

Current operational procedures for shuttle crewmembers include the ingestion of a fluid countermeasure approximately 2 hours before reentry into the earth's gravitational field. The ingestion of the fluid countermeasure is thought to restore plasma volume and improve orthostatic responses upon reentry. The present countermeasure consists of ingesting salt tablets and water to achieve an isotonic solution. It has yet to be determined whether this is the optimal drink to restore orthostatic tolerance. It is also not known whether the drink solution is effective in increasing plasma volume. The purpose here is to evaluate the effectiveness of drink solutions of different osmolarity on restoring plasma volume and orthostatic responses. A hypertonic drink solution was more effective in restoring plasma volume after dehydration than an isotonic solution. However, there were no differences in their effects on an orthostatic challenge. These data suggest that the plasma volume differences produced in this study were not sufficient to produce differences in the cardiovascular responses to an orthostatic challenge, or there are other changes that occur during space flight that are more important in determining orthostatic intolerance.

Davis, John E.↗

Global drivers of local water stresses and global responses to local water policies in the United States

Abstract The rapid depletion of US groundwater resources and rising number of dying wells in the Western US brings attention to the significance of groundwater governance and sustainability restrictions. However, such restrictions on groundwater withdrawals are likely to generate spillover effects causing further environmental stresses in other locations and adding to the complexity of sustainability challenges. The goal of this paper is to improve our understanding of the implications of growing global food demand for local sustainability stresses and the implications of local sustainability policies for local, regional, and global food production, land use, and prices. We employ SIMPLE-G-US (Simplified International Model of agricultural Prices, Land use, and the Environment—Gridded version for the United States) to distangle the significance or remote changes in population and income for irrigation and water resources in the US. Then we examine the local-to-global impacts of potential US groundwater sustainability policies. We find that developments in international markets are significant, as more than half of US sustainability stresses by 2050 are caused by increased commodity demand from abroad. Furthermore, a US sustainable groundwater policy can cause overseas spillovers of US production, thereby potentially contributing to environmental stresses elsewhere, even as groundwater stress in the US is alleviated. These unintended consequences could include deforestation due to cropland expansion, as well as degradation in water quality due to intensification of production in non-targeted areas.

54 ENVIRONMENTAL SCIENCES↗

An update of the total-strain version of SRP

An updated procedure for characterizing an alloy and predicting cyclic life by using the total strain range version of strainrange partitioning (TS-SRP) has been developed. The principal feature of this update is a new procedure for determining the intercept of time dependent elastic strain range versus cyclic life lines. The procedure is based on an established relation between failure and the cyclic stress-strain response of an alloy. The stress-strain response is characterized by empirical equations presented in this report. These equations were determined with the aid of a cyclic constitutive model. The procedures presented herein reduce the testing required to characterize an alloy. Failure testing is done only in the high strain, low life regime; cyclic stress-strain response is determined from tests conducted in both the high and low strain regimes. These tests are carried out to stability of the stress-strain hysteresis loop but not to failure. Thus both the time and costs required to characterize an alloy are greatly reduced. This approach was evaluated and verified for two nickel base superalloys, AF2-1DA and Inconel 718.

Saltsman, J. F.↗

An update of the total-strain version of SRP

An updated procedure for characterizing an alloy and predicting cyclic life by using the total strain range version of strainrange partitioning (TS-SRP) has been developed. The principal feature of this update is a new procedure for determining the intercept of time dependent elastic strain range versus cyclic life lines. The procedure is based on an established relation between failure and the cyclic stress-strain response of an alloy. The stress-strain response is characterized by empirical equations presented in this report. These equations were determined with the aid of a cyclic constitutive model. The procedures presented herein reduce the testing required to characterize an alloy. Failure testing is done only in the high strain, low life regime; cyclic stress-strain response is determined from tests conducted in both the high and low strain regimes. These tests are carried out to stability of the stress-strain hysteresis loop but not to failure. Thus both the time and costs required to characterize an alloy are greatly reduced. This approach was evaluated and verified for two nickel base superalloys, AF2-1DA and Inconel 718.

Saltsman, James F.↗

Microstructure characterization and elastic-plastic self-consistent simulation studies of anisotropic deformation of β -tin

Uniaxial compression tests of 99.9% pure polycrystalline $β$-tin (Sn) were conducted at various strain rates (10 -3 /s, 10 -1 /s and 1/s) and temperatures (294 K and 193 K) to understand the effects of these variables on the stress–strain response, microstructure evolution, and stress relaxation behavior. Multiple Sn specimens were subjected to complex compressive loading/unloading/reloading paths at different strain rates. Specimens initially compressed at higher strain rates showed strain rate-dependent texture evolution and more pronounced relaxation upon reloading, as compared to those pre-strained at lower rates at room temperature (RT). Compression tests conducted at low temperature (193 K) revealed increased strength, similar to the enhanced strength observed when the strain rate was increased at RT. Neutron diffraction was employed to characterize the initial and final bulk textures of the RT specimens. Electron backscatter diffraction was utilized to examine the crystallographic grain orientation and morphology, thereby identifying the signatures of dislocation-mediated deformation, grain refinement, recrystallization, and twinning behaviors. Elastic-plastic self-consistent simulations were performed to investigate the deformation modes responsible for the strain rate-dependent macroscopic stress–strain response and texture evolution. Effects of crystallographic orientation on stress relaxation behavior was also examined. In conclusion, the model predictions are in reasonable agreement with experimental observations.

36 MATERIALS SCIENCE↗

Sustained strain applied at high rates drives dynamic tensioning in epithelial cells

Epithelial cells experience long lasting loads of different magnitudes and rates. How they adapt to these loads strongly impacts tissue health. Yet, much remains unknown about the evolution of cellular stress in response to sustained strain. Here, by subjecting cell pairs to sustained strain, we report a bimodal stress response, where in addition to the typically observed stress relaxation, a subset of cells exhibits a dynamic tensioning process with significant elevation in stress within 100 s, resembling active pulling-back in muscle fibers. Strikingly, the fraction of cells exhibiting tensioning increases with increasing strain rate. The tensioning response is accompanied by actin remodeling, and perturbation to actin abrogates it, supporting cell contractility’s role in the response. Collectively, our data show that epithelial cells adjust their tensional states over short timescales in a strain-rate dependent manner to adapt to sustained strains, demonstrating that the active pulling-back behavior could be a common protective mechanism against environmental stress.

bioinformatics↗

Cellular Responses to Mechanical Stress Selected Contribution: A Three-Dimensional Model for Assessment of in Vitro Toxicity in Balaena Mysticetus Renal Tissue

This study established two- and three-dimensional renal proximal tubular cell cultures of the endangered species bowhead whale (Balaena mysticetus), developed SV40-transfected cultures, and cloned the 61-amino acid open reading frame for the metallothionein protein, the primary binding site for heavy metal contamination in mammals. Microgravity research, modulations in mechanical culture conditions (modeled microgravity), and shear stress have spawned innovative approaches to understanding the dynamics of cellular interactions, gene expression, and differentiation in several cellular systems. These investigations have led to the creation of ex vivo tissue models capable of serving as physiological research analogs for three-dimensional cellular interactions. These models are enabling studies in immune function, tissue modeling for basic research, and neoplasia. Three-dimensional cellular models emulate aspects of in vivo cellular architecture and physiology and may facilitate environmental toxicological studies aimed at elucidating biological functions and responses at the cellular level. Marine mammals occupy a significant ecological niche (72% of the Earth's surface is water) in terms of the potential for information on bioaccumulation and transport of terrestrial and marine environmental toxins in high-order vertebrates. Few ex vivo models of marine mammal physiology exist in vitro to accomplish the aforementioned studies. Techniques developed in this investigation, based on previous tissue modeling successes, may serve to facilitate similar research in other marine mammals.

Goodwin, T. J.↗

Stress Counter-Response Training Via Physiological Self-Regulation During Flight Simulation

This study provided the first evaluation of a new training concept and technology aimed at training pilots to maintain physiological equilibrium during circumstances in an airplane cockpit. Thirty healthy subjects (16 males and 14 females) between the ages of 18 and 35 were randomized into two study groups, A and B. Subjects participated individually in a sequence of four study sessions. In the first visit, subjects were taught to operate a desktop fighter jet flight simulation program. In the three sessions that followed, subjects in group A were trained to minimize their autonomic deviation from baseline values while operating the desktop flight simulation. This was done by making their skin conductance and hand temperature deviations from baseline impair the functionality of the aircraft controls. Subjects also received auditory and visual cues about their autonomic deviation, and were instructed to keep these within pre-set limits to retain full control of the aircraft. Subjects in group B were subjected to periods of impaired aircraft functionality independent of their physiologic activity, and thus served as a control group. No statistically significant group differences were found in the flight performance scores from the three training sessions, and post-training flight performance scores of the two groups were not different. We conclude that this study did not provide clear support for this training methodology in optimizing pilot performance. However, a number of shortcomings in the current status of this training methodology may account for the lack of demonstrable training benefit to the experimental group. Suggested future modifications for research on this training methodology include: Limiting the amount of instrument impairment resulting from physiological deviations; conducting a greater number of physiological training sessions per subject; using pre-post training performance tests which invoke a greater amount of stress in subjects; and developing a more detailed performance scoring system.

Palsson, Olafur S.↗

A view of the pan‐genome of domesticated Cowpea ( Vigna unguiculata [L.] Walp.)

Abstract Cowpea, Vigna unguiculata L . Walp., is a diploid warm‐season legume of critical importance as both food and fodder in sub‐Saharan Africa. This species is also grown in Northern Africa, Europe, Latin America, North America, and East to Southeast Asia. To capture the genomic diversity of domesticates of this important legume, de novo genome assemblies were produced for representatives of six subpopulations of cultivated cowpea identified previously from genotyping of several hundred diverse accessions. In the most complete assembly (IT97K‐499‐35), 26,026 core and 4963 noncore genes were identified, with 35,436 pan genes when considering all seven accessions. GO terms associated with response to stress and defense response were highly enriched among the noncore genes, while core genes were enriched in terms related to transcription factor activity, and transport and metabolic processes. Over 5 million single nucleotide polymorphisms (SNPs) relative to each assembly and over 40 structural variants >1 Mb in size were identified by comparing genomes. Vu10 was the chromosome with the highest frequency of SNPs, and Vu04 had the most structural variants. Noncore genes harbor a larger proportion of potentially disruptive variants than core genes, including missense, stop gain, and frameshift mutations; this suggests that noncore genes substantially contribute to diversity within domesticated cowpea.

59 BASIC BIOLOGICAL SCIENCES↗

Tracking Seasonal and Interannual Variability in Photosynthetic Downregulation in Response to Water Stress at a Temperate Deciduous Forest

Abstract The understanding and modeling of photosynthetic dynamics affected by climate variability can be highly uncertain. In this paper, we examined a well‐characterized eddy covariance site in a drought‐prone temperate deciduous broadleaf forest combining tower measurements and satellite observations. We find that an increase in spring temperature usually leads to enhanced spring gross primary production (GPP), but a GPP reduction in late growing season due to water limitation. We evaluated how well a coupled fluorescence‐photosynthesis model (SCOPE) and satellite data sets track the interannual and seasonal variations of tower GPP from 2007 to 2016. In SCOPE, a simple stress factor scaling of Vcmax as a linear function of observed predawn leaf water potential ( ψ pd ) shows a good agreement between modeled and measured interannual variations in both GPP and solar‐induced chlorophyll fluorescence (SIF) from the Global Ozone Monitoring Experiment‐2 (GOME‐2). The modeled and satellite‐observed changes in SIF yield are ~30% smaller than corresponding changes in light use efficiency (LUE) under severe stress, for which a common linear SIF to GPP scaling would underestimate the stress reduction in GPP. Overall, GOME‐2 SIF tracks interannual tower GPP variations better than satellite vegetations indices (VIs) representing canopy “greenness.” However, it is still challenging to attribute observed SIF variations unequivocally to greenness or physiological changes due to large GOME‐2 footprint. Higher‐resolution SIF data sets (e.g., TROPOMI) already show the potential to well capture the downregulation of late‐season GPP and could pave the way to better disentangle canopy structural and physiological changes in the future.

He, Liyin↗

The Ancient Salicoid Genome Duplication Event: A Platform for Reconstruction of De Novo Gene Evolution in Populus trichocarpa

Orphan genes are characteristic genomic features that have no detectable homology to genes in any other species and represent an important attribute of genome evolution as sources of novel genetic functions. Here, we identified 445 genes specific to Populus trichocarpa. Of these, we performed deeper reconstruction of 13 orphan genes to provide evidence of de novo gene evolution. Populus and its sister genera Salix are particularly well suited for the study of orphan gene evolution because of the Salicoid whole-genome duplication event which resulted in highly syntenic sister chromosomal segments across the Salicaceae. We leveraged this genomic feature to reconstruct de novo gene evolution from intergenera, interspecies, and intragenomic perspectives by comparing the syntenic regions within the P. trichocarpa reference, then P. deltoides, and finally Salix purpurea. Furthermore, we demonstrated that 86.5% of the putative orphan genes had evidence of transcription. Additionally, we also utilized the Populus genome-wide association mapping panel, a collection of 1,084 undomesticated P. trichocarpa genotypes to further determine putative regulatory networks of orphan genes using expression quantitative trait loci (eQTL) mapping. Functional enrichment of these eQTL subnetworks identified common biological themes associated with orphan genes such as response to stress and defense response. We also identify a putative cis-element for a de novo gene and leverage conserved synteny to describe evolution of a putative transcription factor binding site. Overall, 45% of orphan genes were captured in trans-eQTL networks.

59 BASIC BIOLOGICAL SCIENCES↗

Gravitropism in leafy dicot stems

In an attempt to separate plant responses to mechanical stresses from responses to gravity compensation, six treatments were automated: (1) upright stationary controls; (2) horizontal clinostat; (3) intermittent clinostat (plants upright 3.3 minutes out of every 4 minutes, horizontal and rotated once in the remaining time); (4) inversion every ten minutes (plants upside down half the time); (5) inversion and immediate return to the vertical; and (6) vertical rotation. Epinasty appeared only on clinostated and on inverted plants, both subjected to gravity compensation. The mechanics of gravitropic stem bending and the effects of a unilateral application of ethephon of gravitropic bending were also investigated.

Salisbury, F. B.↗

Real time acousto-ultrasonic NDE technique for monitoring damage in ceramic composites under dynamic loads

Research effort was directed towards developing a near real-time, acousto-ultrasonic (AU), nondestructive evaluation (NDE) tool to study the failure mechanisms of ceramic composites. Progression of damage is monitored in real-time by observing the changes in the received AU signal during the actual test. During the real-time AU test, the AU signals are generated and received by the AU transducers attached to the specimen while it is being subjected to increasing quasi-static loads or cyclic loads (10 Hz, R = 1.0). The received AU signals for 64 successive pulses were gated in the time domain (T = 40.96 micro sec) and then averaged every second over ten load cycles and stored in a computer file during fatigue tests. These averaged gated signals are representative of the damage state of the specimen at that point of its fatigue life. This is also the first major attempt in the development and application of real-time AU for continuously monitoring damage accumulation during fatigue without interrupting the test. The present work has verified the capability of the AU technique to assess the damage state in silicon carbide/calcium aluminosilicate (SiC/CAS) and silicon carbide/ magnesium aluminosilicate (SiC/MAS) ceramic composites. Continuous monitoring of damage initiation and progression under quasi-static ramp loading in tension to failure of unidirectional and cross-ply SiC/CAS and quasi-isotropic SiC/MAS ceramic composite specimens at room temperature was accomplished using near real-time AU parameters. The AU technique was shown to be able to detect the stress levels for the onset and saturation of matrix cracks, respectively. The critical cracking stress level is used as a design stress for brittle matrix composites operating at elevated temperatures. The AU technique has found that the critical cracking stress level is 10-15% below the level presently obtained for design purposes from analytical models. An acousto-ultrasonic stress-strain response (AUSSR) model for unidirectional and cross-ply ceramic composites was formulated. The AUSSR model predicts the strain response to increasing stress levels using real-time AU data and classical laminated plate theory. The Weibull parameters of the AUSSR model are used to calculate the design stress for thermo-structural applications. Real-time AU together with the AUSSR model was used to study the failure mechanisms of SiC/CAS ceramic composites under static and fatigue loading. An S-N curve was generated for a cross-ply SiC/CAS ceramic composite material. The AU results are corroborated and complemented by other NDE techniques, namely, in-situ optical microscope video recordings and edge replication.

Tiwari, Anil↗