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At least 73 records · Page 4

Rapid biodegradation of microplastics generated from bio-based thermoplastic polyurethane

The accumulation of microplastics in various ecosystems has now been well documented and recent evidence suggests detrimental effects on various biological processes due to this pollution. Accumulation of microplastics in the natural environment is ultimately due to the chemical nature of widely used petroleum-based plastic polymers, which typically are inaccessible to biological processing. One way to mitigate this crisis is adoption of plastics that biodegrade if released into natural environments. In this work, we generated microplastic particles from a bio-based, biodegradable thermoplastic polyurethane (TPU-FC1) and demonstrated their rapid biodegradation via direct visualization and respirometry. Furthermore, we isolated multiple bacterial strains capable of using TPU-FC1 as a sole carbon source and characterized their depolymerization products. Finally, to visualize biodegradation of TPU materials as real-world products, we generated TPU-coated cotton fabric and an injection molded phone case and documented biodegradation by direct visualization and scanning electron microscopy (SEM), both of which indicated clear structural degradation of these materials and significant biofilm formation.

09 BIOMASS FUELS↗

Characterizing intracellular proteomes for microbes: An experimental approach using label-free protein quantitation

Genomic and transcriptomic studies generate a rich source of molecular information; however, neither a static genome nor the presence of a messenger RNA provides the most direct insight into the functional state of a microbial cell at any given time. Rather, it is the ensemble of proteins (i.e., proteome) that is the primary workhorse for most biological processes that are concurrent and coordinately active in a living cell. Currently, mass-spectrometry-based (MS) technologies provide unprecedented information into the composition of the proteome, shedding light on the numerous complex biological processes actively shaping observed phenotypes. Herein, we detail a protocol for the exploration of intracellular proteomes through the large-scale, unbiased identification of proteins and their relative abundances using liquid chromatography (LC) coupled to tandem mass spectrometry (MS/MS). In general, this protocol is applicable to both microbial and eukaryotic systems.

Abraham, Paul↗

Advances in microscale separations towards nanoproteomics applications

Microscale separations (e.g., liquid chromatography or capillary electrophoresis) coupled with mass spectrometry (MS) has become the primary tool for advanced proteomics, an indispensable technology for gaining understanding of complex biological processes. While significant advances have been achieved in MS-based proteomics, the current platforms still face a significant challenge in overall sensitivity towards nanoproteomics (i.e., with less than 1 g total amount of proteins available) applications such as cellular heterogeneity in tissue pathologies. Herein, we review recent advances in microscale separation techniques and integrated sample processing systems that improve the overall sensitivity and coverage of the proteomics workflow, and their contributions towards nanoproteomics applications.

Environmental Molecular Sciences Laboratory↗

Function and Evolution of the Plant MES Family of Methylesterases

Land plant evolution has been marked by numerous genetic innovations, including novel catalytic reactions. Plants produce various carboxyl methyl esters using carboxylic acids as substrates, both of which are involved in diverse biological processes. The biosynthesis of methyl esters is catalyzed by SABATH methyltransferases, and understanding of this family has broadened in recent years. Meanwhile, the enzymes catalyzing demethylation—known as methylesterases (MESs)—have received less attention. Here, we present a comprehensive review of the plant MES family, focusing on known biochemical and biological functions, and evolution in the plant kingdom. Thirty-two MES genes have been biochemically characterized, with substrates including methyl esters of plant hormones and several other specialized metabolites. One characterized member demonstrates non-esterase activity, indicating functional diversity in this family. MES genes regulate biological processes, including biotic and abiotic defense, as well as germination and root development. While MES genes are absent in green algae, they are ubiquitous among the land plants analyzed. Extant MES genes belong to three groups of deep origin, implying ancient gene duplication and functional divergence. Two of these groups have yet to have any characterized members. Much remains to be uncovered about the enzymatic functions, biological roles, and evolution of the MES family.

59 BASIC BIOLOGICAL SCIENCES↗

Geometry and dynamics of lipid membranes: The Scriven-Love number

The equations governing lipid membrane dynamics in planar, spherical, and cylindrical geometries are presented here. Unperturbed and first-order perturbed equations are determined and nondimensionalized. In membrane systems with a nonzero base flow, perturbed in-plane and out-of-plane quantities are found to vary over different length scales. Here, a new dimensionless number, named the Scriven-Love number, and the well-known Föppl–von Kármán number result from a scaling analysis. The Scriven-Love number compares out-of-plane forces arising from the in-plane, intramembrane viscous stresses to the familiar elastic bending forces, while the Föppl–von Kármán number compares tension to bending forces. Both numbers are calculated in past experimental works, and span a wide range of values in various biological processes across different geometries. In situations with large Scriven-Love and Föppl–von Kármán numbers, the dynamical response of a perturbed membrane is dominated by out-of-plane viscous and surface tension forces—with bending forces playing a negligible role. Calculations of non-negligible Scriven-Love numbers in various biological processes and in vitro experiments show in-plane intramembrane viscous flows cannot generally be ignored when analyzing lipid membrane behavior.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Regulation mechanism of long non-coding RNA in plant response to stress

Highlights: • LncRNAs can regulate gene expression at transcription and post-transcription, epigenetic level, and many biological processes. • In this review, we discuss the regulation mechanism of long non-coding RNA in plant response to stress. • This review will provide strong evidence for lncRNAs to explore new ideas and new targets for prevention and control. Long non-coding RNA (lncRNA) is a non-coding RNA greater than 200 nucleotides in length. LncRNAs can regulate gene expression at transcription and post-transcription, epigenetic level, and plays an important role in a wide range of biological processes such as genomic imprinting, chromatin remodeling, transcriptional activation, transcriptional interference and cell cycle. It becomes the current hot topics in the study of molecular biology and genetics. Emerging evidence proposed that lncRNAs play important roles in response to both abiotic and biotic stress. In this review, we discuss the role of lncRNAs in drought resistance, salt resistance, disease resistance, and immunity of plants, providing strong evidence for exploring the important role of lncRNAs in plant resistance, in order to explore new ideas and new targets for prevention and control.

60 APPLIED LIFE SCIENCES↗

Life Sciences Accomplishments 1994

The NASA Life and Biomedical Sciences and Applications Division (LBSAD) serves the Nation's life sciences community by managing all aspects of U.S. space-related life sciences research and technology development. The activities of the Division are integral components of the Nation's overall biological sciences and biomedical research efforts. However, NASA's life sciences activities are unique, in that space flight affords the opportunity to study and characterize basic biological mechanisms in ways not possible on Earth. By utilizing access to space as a research tool, NASA advances fundamental knowledge of the way in which weightlessness, radiation, and other aspects of the space-flight environment interact with biological processes. This knowledge is applied to procedures and technologies that enable humans to live and work in and explore space and contributes to the health and well-being of people on Earth. The activities of the Division are guided by the following three goals: Goal 1) Use microgravity and other unique aspects of the space environment to enhance our understanding of fundamental biological processes. Goal 2) Develop the scientific and technological foundations for supporting exploration by enabling productive human presence in space for extended periods. Goal 3) Apply our unique mission personnel, facilities, and technology to improve education, the quality of life on Earth, and U.S. competitiveness. The Division pursues these goals with integrated ground and flight programs involving the participation of NASA field centers, industry, and universities, as well as interactions with other national agencies and NASA's international partners. The published work of Division-sponsored researchers is a record of completed research in pursuit of these goals. During 1993, the LBSAD instituted significant changes in its experiment solicitation and peer review processes. For the first time, a NASA Research Announcement (NRA) was released requesting proposals for ground-based and flight research for all programs. Areas of particular interest to NASA were defined Proposals due April 29, 1994, will be peer reviewed - externally for scientific merit. This annual NRA process is now the mechanism for recruiting both extramural and intramural investigations. As an overview of LBSAD activities in 1993, this accomplishments document covers each of the major organizational components of the Division and the accomplishments of each. The second section is a review of the Space Life Sciences Research programs Space Biology, Space Physiology and Countermeasures, Radiation Health, Environmental Health, Space Human Factors, Advanced Life Support, and Global Monitoring and Disease Prediction, The third section, Research in Space Flight, describes the substantial contributions of the Spacelab Life Sciences 2 (SLS-2) mission to life sciences research and the significant contributions of the other missions flown in 1993, along with plans for future missions. The Division has greatly expanded and given high priority to its Education and Outreach Programs, which are presented in the fourth section. The fifth and final section, Partners for Space, shows the Divisions Cooperative efforts with other national and international agencies to achieve common goals, along with the accomplishments of joint research and analysis programs.

Burnell, Mary Lou↗

Nitrogen derived from Combined Algal Processing supports algae cultivation for biofuels

Algae biomass, a biofuel feedstock with potential to reduce global CO 2 emissions, is currently too expensive Nitrogen, a significant input and life cycle cost, requires research on recycling and additional sources to reduce costs. Integral to nitrogen recycling is the knowledge of actual nitrogen requirements in outdoor ponds, thus data from the Unified Field Studies of the Algae Testbed Public-Private Partnership was analyzed. The data showed that Nannochloropsis oceanica used 14–24 mg/L NH 4 + per harvest cycle for robust growth. Nitrogen recycled directly from the algae conversion process will likely be the primary source of nitrogen for algae cultivation. Harvested algae can be fractionated via Combined Algal Processing (CAP), where algae are pretreated, fermented, and the fuel feedstocks (e. g. ethanol, lipids) are removed leaving a nitrogen rich slurry. Chlorella vulgaris grew robustly in nitrogen-free media supplemented with CAP residues through 4 harvest cycles with no inhibition. The amount of nitrogen as free ammonium available for recycling directly without further processing was insufficient (~4% of total needed), thus the nitrogen fixing bacteria, Azotobacter vinelandii, was investigated as a means for increasing ammonium concentration in CAP residues. This organism was shown to be proficient at growth on sugars liberated from algae biomass during pretreatment and sustained algae growth using A. vinelandii supernatants was also demonstrated. Using Azotobacter to increase ammonium concentrations on-site using an inexpensive biological process to generate the necessary ammonium for outdoor algae cultivation improves sustainability of algae biofuels by avoiding the energy intensive steps of typical ammonium production and transport. Furthermore, these results show that byproducts of algae conversion begin to close nitrogen demand and, with further supplementation by diazotrophic bacteria or protein hydrolysis, may completely replace current energyintensive and costly industrial nitrogen sources offering a path to increased economic feasibility and improved life cycle analysis of algae-based biofuels.

09 BIOMASS FUELS↗

Sirtuin 7 is involved in the consolidation of fear memory in mice

Highlights: • SIRT7 is involved in many biological processes. • The functional role of SIRT7 in the brain is unknown. • We demonstrate that SIRT7 is expressed in the amygdala and hippocampus. • Fear memory consolidation was impaired in Sirt7 knockout mice. • SIRT7 is involved in the consolidation of fear memory. Sirtuin 7 (SIRT7) is an NAD{sup +}-dependent deacetylase/deacylase, and is involved in a variety of biological processes relevant to the transcription of rRNA, the DNA damage response, tumorigenesis, and metabolism. SIRT7 mRNA is expressed ubiquitously, including in the brain, but there is no detailed information about the anatomical distribution and functional role of SIRT7 in the brain. Here, we demonstrated that SIRT7 is widely expressed in the mouse brain, including in the cortex, striatum, thalamus, hippocampus, and amygdala. Behavioral examinations revealed that Sirt7 knockout (KO) and control mice showed similar levels of freezing behavior immediately after a fear response, but a significant decrease of freezing behavior at 24 h after fear conditioning was observed in Sirt7 KO mice. Histological analysis revealed that there is no apparent structural abnormality of the amygdala and hippocampus, which are regions involved in fear memory consolidation, in Sirt7 KO mice. Our results indicate that SIRT7 is involved in the consolidation of fear memory.

60 APPLIED LIFE SCIENCES↗

Resveratrol Ameliorates Lipopolysaccharide-Induced Sudden Sensorineural Hearing Loss in In Vitro Model through Multitarget Antiapoptotic Mechanism Based on Network Pharmacology and Molecular Docking

Objective. To explore the effects of resveratrol (RSV) on hair cell apoptosis caused by sudden sensorineural hearing loss (SSNHL) and its effect on lipopolysaccharide-induced apoptosis of HEI-OC1 cells. Methods. We used the network pharmacology method to screen molecules related to RSV for the treatment of SSNHL and analyzed these molecules and their enriched biological processes and signaling pathways through Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) analysis. We selected hub genes related to apoptosis using protein-protein interaction (PPI) analysis for in vitro and molecular docking verification. Results. Eighty overlapping genes were identified as potential targets for RSV treatment of SSNHL. Further GO analysis showed that the biological processes were mainly related to toxicity, cell proliferation, and lipopolysaccharide reactions. KEGG analysis showed that the AGE-RAGE signaling pathway in diabetic complications, Kaposi’s sarcoma-associated herpesvirus infection, FoxO signaling pathway, PI3K-Akt signaling pathway, and other inflammatory signaling pathways were concentrated. AKT1, STAT3, JUN, TNF, TP53, MAPK3, CASP3, and VEGFA were screened as HUB genes using PPI analysis. The apoptosis-related proteins TNF, CASP3, AKT1, and TP53 were selected for in vitro experiments, which showed that mRNA was significantly different before and after RSV intervention, confirming that the corresponding protein receptors could bind well with RSV. Conclusion. RSV mainly affects the prognosis of SSNHL through anti-inflammatory effects and may improve hair cell apoptosis caused by inflammatory factors through multitargeted interventions involving TNF, CASP3, AKT1, and TP53.

Ye, Shiming↗

Optogenetics and biosensors set the stage for metabolic cybergenetics

We report cybergenetic systems use computer interfaces to enable feed-back controls over biological processes in real time. The complex and dynamic nature of cellular metabolism makes cybergenetics attractive for controlling engineered metabolic pathways in microbial fermentations. Cybergenetics would not only create new avenues of research into cellular metabolism, it would also enable unprecedented strategies for pathway optimization and bioreactor operation and automation. Implementation of metabolic cybergenetics, however, will require new capabilities from actuators, biosensors, and control algorithms. The recent application of optogenetics in metabolic engineering, the expanding role of genetically encoded biosensors in strain development, and continued progress in control algorithms for biological processes suggest that this technology will become available in the not so distant future.

59 BASIC BIOLOGICAL SCIENCES↗

Quantitative proteomics reveals extensive lysine ubiquitination and transcription factor stability states in Arabidopsis

Abstract Protein activity, abundance, and stability can be regulated by post-translational modification including ubiquitination. Ubiquitination is conserved among eukaryotes and plays a central role in modulating cellular function; yet, we lack comprehensive catalogs of proteins that are modified by ubiquitin in plants. In this study, we describe an antibody-based approach to enrich ubiquitinated peptides coupled with isobaric labeling to enable quantification of up to 18-multiplexed samples. This approach identified 17,940 ubiquitinated lysine sites arising from 6,453 proteins from Arabidopsis (Arabidopsis thaliana) primary roots, seedlings, and rosette leaves. Gene ontology analysis indicated that ubiquitinated proteins are associated with numerous biological processes including hormone signaling, plant defense, protein homeostasis, and metabolism. We determined ubiquitinated lysine residues that directly regulate the stability of three transcription factors, CRYPTOCHROME-INTERACTING BASIC-HELIX-LOOP-HELIX 1 (CIB1), CIB1 LIKE PROTEIN 2 (CIL2), and SENSITIVE TO PROTON RHIZOTOXICITY1 (STOP1) using in vivo degradation assays. Furthermore, codon mutation of CIB1 to create a K166R conversion to prevent ubiquitination, via CRISPR/Cas9-derived adenosine base editing, led to an early flowering phenotype and increased expression of FLOWERING LOCUS T (FT). These comprehensive site-level ubiquitinome profiles provide a wealth of data for future functional studies related to modulation of biological processes mediated by this post-translational modification in plants.

Biochemistry & Molecular Biology↗

Electronic control of redox reactions inside Escherichia coli using a genetic module

Microorganisms regulate the redox state of different biomolecules to precisely control biological processes. These processes can be modulated by electrochemically coupling intracellular biomolecules to an external electrode, but current approaches afford only limited control and specificity. Here we describe specific electrochemical control of the reduction of intracellular biomolecules in Escherichia coli through introduction of a heterologous electron transfer pathway. E . coli expressing cymAmtrCAB from Shewanella oneidensis MR-1 consumed electrons directly from a cathode when fumarate or nitrate, both intracellular electron acceptors, were present. The fumarate-triggered current consumption occurred only when fumarate reductase was present, indicating all the electrons passed through this enzyme. Moreover, CymAMtrCAB-expressing E . coli used current to stoichiometrically reduce nitrate. Thus, our work introduces a modular genetic tool to reduce a specific intracellular redox molecule with an electrode, opening the possibility of electronically controlling biological processes such as biosynthesis and growth in any microorganism.

59 BASIC BIOLOGICAL SCIENCES↗

Nonaxisymmetric Shapes of Biological Membranes from Locally Induced Curvature

In various biological processes such as endocytosis and caveolae formation, the cell membrane is locally deformed into curved morphologies. Previous models to study membrane morphologies resulting from locally induced curvature often only consider the possibility of axisymmetric shapes-an indeed unphysical constraint. Past studies predict that the cell membrane buds at low resting tensions and stalls at a flat pit at high resting tensions. In this work, we lift the restriction to axisymmetry to study all possible membrane morphologies. Only if the resting tension of the membrane is low, we reproduce axisymmetric membrane morphologies. When the resting tension is moderate to high, we show that 1) axisymmetric membrane pits are unstable and 2) nonaxisymmetric ridge-shaped structures are energetically favorable. Furthermore, we find the interplay between intramembrane viscous flow and the rate of induced curvature affects the membrane's ability to transition into nonaxisymmetric ridges and axisymmetric buds. In particular, we show that axisymmetric buds are favored when the induced curvature is rapidly increased, whereas nonaxisymmetric ridges are favored when the curvature is slowly increased. Overall, our results hold relevant implications for biological processes such as endocytosis and physical phenomena like phase separation in lipid bilayers.

59 BASIC BIOLOGICAL SCIENCES↗

Stable carbon isotope values of syndepositional carbonate spherules and micrite record spatial and temporal changes in photosynthesis intensity

Marine and lacustrine carbonate minerals preserve carbon cycle information, and their stable carbon isotope values (δ 13 C) are frequently used to infer and reconstruct paleoenvironmental changes. However, multiple processes can influence the δ 13 C values of bulk carbonates, confounding the interpretation of these values in terms of conditions at the time of mineral precipitation. Co-existing carbonate forms may represent different environmental conditions, yet few studies have analyzed δ 13 C values of syndepositional carbonate grains of varying morphologies to investigate their origins. Here, we combine stable isotope analyses, metagenomics, and geochemical modeling to interpret δ 13 C values of syndepositional carbonate spherules (>500 μm) and fine-grained micrite (<63 μm) from a ~1600-year-long sediment record of a hypersaline lake located on the coral atoll of Kiritimati, Republic of Kiribati (1.9°N, 157.4°W). Petrographic, mineralogic, and stable isotope results suggest that both carbonate fractions precipitate in situ with minor diagenetic alterations. The δ 13 C values of spherules are high compared to the syndepositional micrite and cannot be explained by mineral differences or external perturbations, suggesting a role for local biological processes. We use geochemical modeling to test the hypothesis that the spherules form in the surface microbial mat during peak diurnal photosynthesis when the δ 13 C value of dissolved inorganic carbon is elevated. In contrast, we hypothesize that the micrite may precipitate more continuously in the water as well as in sub-surface, heterotrophic layers of the microbial mat. Both metagenome and geochemical model results support a critical role for photosynthesis in influencing carbonate δ 13 C values. The down-core spherule–micrite offset in δ 13 C values also aligns with total organic carbon values, suggesting that the difference in the δ13C values of spherules and micrite may be a more robust, inorganic indicator of variability in productivity and local biological processes through time than the δ 13 C values of individual carbonate forms.

59 BASIC BIOLOGICAL SCIENCES↗

Production of diverse PET probes with limited resources: 24 18 F-labeled compounds prepared with a single radiosynthesizer

Significance Molecular imaging with PET can provide a dynamic, whole-body picture of the rate of biological processes or distribution of biological targets by tracking the distribution of radiolabeled molecules or particles in the body over time. Continual efforts to develop new PET probes are expanding the variety of processes and targets that can be visualized, facilitating basic research, drug development, and patient care. However, access to these probes at all stages of their development is hindered by high costs arising, in large part, from the significant resources that are typically dedicated to production of a single probe. Emerging technologies with increased synthesis flexibility are allowing increased probe diversity with fewer resources and could significantly increase access to new molecular imaging agents.

62 RADIOLOGY AND NUCLEAR MEDICINE↗

Data for Spatial Analysis of Cell Patterning to Aid Genetic and Phenotypic Understanding of Grass Stomatal Density: A Case Study in Maize

Biological processes involve complex hierarchies where composite traits result from multiple component traits. However, holistically understanding of how sets of component traits interact to underpin genotype-to-phenotype relationships is generally lacking. Stomatal density (SD) is a tractable model system for exploring how high-throughput phenotyping (HTP) data could be exploited by a new spatial analysis approach to better understand a developmentally and functionally important trait. SD is a composite trait, resulting from various components related to cell identity and size, which are themselves governed by a series of spatio-developmental processes. Data from 192 recombinant inbred lines of maize [Zea mays (L.)] were analyzed by a new stomatal patterning phenotype (SPP) to (1) describe the average spatial probability distribution of the nearest neighboring stomata; (2) derive a core set of component traits related to cell size, cell packing, and positional probabilities; (3) build a structural equation model of component traits underlying SD; and (4) identify stomatal patterning quantitative trait loci (QTL). The core set of SPP-derived traits explained 74% of the variation in SD. Analyzing SPP component traits allowed some loci previously identified as generic SD QTL to be recognized as specific to lateral versus longitudinal elements of stomatal patterning. Therefore, this study highlights how novel insights can be gained by decomposing a composite trait (e.g., SD) into a set of component traits that were present in HTP data but not previously exploited.

AI/ML↗

Is Genetic Engineering a Route to Enhance Microalgae-Mediated Bioremediation of Heavy Metal-Containing Effluents?

Contamination of the biosphere by heavy metals has been rising, due to accelerated anthropogenic activities, and is nowadays, a matter of serious global concern. Removal of such inorganic pollutants from aquatic environments via biological processes has earned great popularity, for its cost-effectiveness and high efficiency, compared to conventional physicochemical methods. Among candidate organisms, microalgae offer several competitive advantages; phycoremediation has even been claimed as the next generation of wastewater treatment technologies. Furthermore, integration of microalgae-mediated wastewater treatment and bioenergy production adds favorably to the economic feasibility of the former process—with energy security coming along with environmental sustainability. However, poor biomass productivity under abiotic stress conditions has hindered the large-scale deployment of microalgae. Recent advances encompassing molecular tools for genome editing, together with the advent of multiomics technologies and computational approaches, have permitted the design of tailor-made microalgal cell factories, which encompass multiple beneficial traits, while circumventing those associated with the bioaccumulation of unfavorable chemicals. Previous studies unfolded several routes through which genetic engineering-mediated improvements appear feasible (encompassing sequestration/uptake capacity and specificity for heavy metals); they can be categorized as metal transportation, chelation, or biotransformation, with regulation of metal- and oxidative stress response, as well as cell surface engineering playing a crucial role therein. This review covers the state-of-the-art metal stress mitigation mechanisms prevalent in microalgae, and discusses putative and tested metabolic engineering approaches, aimed at further improvement of those biological processes. Finally, current research gaps and future prospects arising from use of transgenic microalgae for heavy metal phycoremediation are reviewed.

Ranjbar, Saeed (ORCID:0000000256518605)↗